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7 - 9 JANUARY 2008

Knowledge l Innovation l Experience

National Chemical Laboratory (NCL), ( Pune was established in 1950 as a

constituent laboratory of Council of Scientific and Industrial Research (CSIR). NCL is a science and
knowledge based research, development and consulting organization. It is internationally known for its
excellence in scientific research in chemistry and chemical engineering as well as for its outstanding track
record of industrial research involving partnerships with industry from concept to commercialization.

NCL's R&D functions span diverse areas of scientific and industrial research covering Catalysis
(Heterogeneous and Homogeneous), Biochemical Sciences (Enzymology and microbiology, Plant
biochemistry and molecular biology, and Plant tissue culture), Organic Chemistry (Chiral synthesis, New
Synthetic methods, Process chemistry for active pharmaceutical intermediates, Multistep organic
synthesis of complex organic molecules, Chemical biology and Bio-organic chemistry, Molecular diversity
based chemical genetics), Polymer Science and Engineering (Polymer chemistry, Polymer physics,
Complex fluids and polymer engineering, Polymer and materials modeling), Physical and Materials
Chemistry (Nanomaterials science and technology, Materials chemistry, Theory and computational
science), and Chemical Engineering Science (Reaction Engineering, Process simulation and modeling,
Biochemical engineering, Industrial flow modeling, Process design and development of processes for fine
chemicals and polymers). The laboratory interacts with its external clients through Business Development
Division which takes care of business development, marketing, technology management, and intellectual
property management. Information Division organizes the vital management information system (MIS)
and networking of information and communication across NCL and also acts as major digital information
resource center. NCL has well equipped laboratories with state-of-the-art facilities for synthesis, analysis,
computation and scale-up.

The total staff strength of NCL is about 1500 out of which 300 are scientific staff with Ph.Ds with post
doctoral and industrial R&D experience and 425 are research students, 300 are project assistants mostly
with post-graduate degrees (M.Sc./ B.E. / B.Tech.) working on contract R&D projects. NCL publishes the
second largest number of papers in chemical sciences (~440) and produces the largest number of Ph.Ds
(~50 every year) in chemical sciences in India.

Apart from well-equipped research laboratories that cater to the disciplines of interest, NCL has several
special R&D support facilities. These include Catalyst Pilot Plant, Center for Material Characterization,
Central NMR facility, Combi Chem-Bio Resource Center, Digital Information Resources Center,
Multipurpose Pilot Plants, National Collection of Industrial Microorganisms, Tissue Culture Pilot Plant, etc.

NCL has, over the years, forged strong industrial and academic collaborations in pursuit of its objectives.
NCL works closely with about 20 international companies and over 50 Indian companies. Some of the
largest companies, within and outside India, have benefited by such collaborations. Examples are
Reliance Industries Limited, Indian Oil Corporation, Oil & Natural Gas Corporation, pharma majors such
as Ranbaxy, Cipla, Lupin and Cadilla and global companies such as GE, DuPont, Dow Chemicals and
Unilever. NCL has close academic links with several Universities, both Indian and foreign as well as with
several international R&D organizations and has signed MoU with many of them for continuous
cooperation in R&D. On an average, NCL files about 30 foreign and 50 Indian patents per year.

Dr. Homi Bhabha Road, Pune 411 008 (INDIA).

Tel. : +91-20-2590 2600, 2590 2000, Fax : +91-20-2590 2601, 2590 2639
BSD - 2008
International Conference on
(green chemicals and materials, bioenergy, biorefinery and clean water)

7-9 January 2008



National Chemical Laboratory,

Pune, India
Lund University, Sweden


National Chemical Laboratory

Dr. Homi Bhabha Road, Pune 411 008, India.

Organising Committee

l Convener l

Mr. Sanjay Nene

l International Organizing Commitee l

Prof. Rajni Hatti-Kaul

Prof. Bo Mattiasson

Dr. Sandra D’Souza

l Local Organizing commitee l

Dr. Anjani Varma

Dr. Ramchandra Gadre

Dr. Digambar Gokhale

Dr. P. K. Ingle

Mr. V. V. Jogdand

Mr. S. B. Katte

Dr. U. K. Kharul

Dr. H. V. Thulasiram

Mr. B. K. Vaidya (Student Representative)




Passionate about Green Fuels


International Conference on
(green chemicals and materials, bioenergy, biorefinery and clean water)
7-9 January 2008

7 January 2008
Venue : NCL Auditorium

15.00-17.00 Registration (NCL Main building Foyer)


17.00 hrs. Welcome remarks by Dr. S. Sivaram, Director, NCL

17.20 hrs. Inaugural address : Mr. Pramod Chaudhari, Chairman, PRAJ Industries Ltd., Pune

17.50 hrs. The MISTRA Programme : Prof. Rajni Hatti-Kaul, Lund University, Sweden

18.10 hrs. Vote of Thanks : Mr. Sanjay Nene

18.15 hrs. Plenary Lecture : 'Is the promotion of biofuels contributing towards the sustainable use of
Wilfrid Legg, Head of Agricultural Policies and Environment Division Trade and Agriculture
Directorate OECD, Paris

19.30 hrs. Dinner at the NCL Guest House lawns

8 January 2008


09.00-09.45 Dr. Colja Laane, DSM, Netherlands

White (Industrial) Biotechnology: Clean, Clever & Competitive
09.45-10.30 Dr. Per Henning Nielsen, Novozymes, Denmark
Enzymatic solutions in industry - a contribution to sustainable development
10.30-11.00 COFFEE BREAK
11.00-11.45 Prof. Rajni Hatti-Kaul, Lund University, Sweden
Evaluating the potential of biotechnology for production of speciality chemicals and polymers
from renewable resources
11.45-12.30 Dr. Martin Patel, Utrecht University, Netherlands
Green polymers - how sustainable are they?
12.30-13.00 Questions and General Discussions
13.00-14.00 LUNCH
14.00-15.00 POSTER SESSION


15.00-15.45 Dr. Gopal Chotani, Danisco, USA

Dislocating technologies for biorefineries development
15.45-16.30 Prof. Irini Angelidaki, Danish Technical University, Denmark
Biofuel Production : A new Biorefinery for Sustainable Energy from Crops; Conversion of
Lignocellulose to Bioethanol, Biohydrogen and Biomethane
16.30-17.00 COFFEE BREAK
17.00-17.45 Mr.Tomas Hagström, Perstorp, Sweden
Perstorp BioProducts AB, taking bio-chemicals and fuels to the market
17.45-18.30 Mr. Ghanshyam Deshpande, Executive Vice-President, Praj Industries Ltd.
Lignocellulosic biomass materials Sustainable Feedstock for Ethanol Production.
18.30-19.00 Questions and General Discussions

9 January 2008

09.00-09.45 Dr. Malek Alkasrawi, Syngenta, Sweden

Bioenergy from Energy crops : Technical possibilities and economical feasibilty
09.45-10.30 Pätz Reinhard R., University of Applied Sciences, Köthen/Anhalt, Germany
Anaerobic Sequencing-Batch-Reactor (ASBR) Technology for High Performance Biogas

10.30-11.00 COFFEE BREAK

11.00-11.45 Prof. Bo Mattiasson, Lund University, Sweden
Biogas from surplus biomass the optimal alternative/(ultimate solution)?
11.45-12.30 Dr. Pål Börjesson, Lund University, Sweden
Environmental system analysis concerning bioenergy from crops
12.30-13.00 Questions and General Discussions
13.00-14.00 LUNCH
14.00-15.30 An Indian Industry Perspective
(Invited presentations from leading Indian Industries)
15.30-16.00 COFFEE BREAK


16.00-16.45 Prof. Galip Akay, University of Newcastle, UK

Bioprocess-Chemical process intensifications using nano-structured macroporous materials
and their application in wastewater treatment
16.45-17.30 Prof. N.H. Ravindranath, IISc, India
Modern bioenergy technologies
17.30-18.15 Prof. Ajit Annachhatre, AIT, Thailand
Sulfate reduction process for removal of heavy metals
18.15-18.45 Questions and General Discussions
18.45-19.30 Valedictory Ceremony
19.30-20.30 HIGH TEA


BSD-2008 Plenary Lecture

Is the Promotion of Biofuels Contributing towards the

Sustainable Use of Resources ?
Wilfrid Legg

Head of Agricultural Policies and Environment Division, Trade and Agriculture Directorate, OECD

The rapid growth in biofuels is largely due to government policies that subsidise the production of cereal
and oilseed feedstock, investment in biorefineries, impose import restrictions on cheaper ethanol from other
countries and require mandatory blending shares with petrol. Estimates of subsidies to promote biofuel
production and consumption may be in the order US 15 billion according to research presented to the OECD.
Production-linked support to agriculture in OECD countries, while high, is gradually decreasing as a share of
overall support, and this influences the incentives to produce agricultural commodities and the sustainability of
the sector.

Biofuel production and consumption for transport is not economically viable in OECD countries under
current market conditions. The economic viability of biofuel production depends on a number of factors
including the relative prices of biofuel and fossil fuels which crops are used, as well as where and how they are
grown. In some cases, biomass used for heating and electricity generation is economically viable.

Governments support biofuel production and consumption for a variety of reasons, but the reasons most
frequently cited are: to reduce greenhouse gas emissions (GHG); enhance energy security through oil
substitution; boost incomes for farmers arising from increased demand for feedstock; and, through higher farm
incomes, stimulate rural economies.

The benefits achieved appear so far to be smaller than many anticipated. In considering environmental
benefits, the impact on the use of water, energy, fertiliser and pesticides need to be taken into account as well as
the effects on biodiversity through land use change. Moreover, while the evidence suggests that greenhouse gas
emissions from biofuel production and consumption, measured on a life-cycle analysis basis, are lower than
from fossil fuel sources of transport fuels, there are big differences depending on the feedstock used.In the case
of India, ethanol from molasses, as currently produced, might be more sustainable than from sugar cane.
Moreover, there are important knock-on effects where the production of feedstocks for biofuels has diverted
crops and land from food, which has been one factor in the current rise in food prices, and there have been
differing impacts on crop and crop-using livestock farmers.

The technology associated with the production of biofuels (for example, the move to ligno-cellulosic
bioreactors) is evolving at an increasing pace. Much research effort and capital is being devoted by countries to
this “second generation” of biofuels, which will impact on the economics of biofuels and the environmental
impacts. Policies that can achieve goals more effectively and at lower costs would thus need to focus on
investment in energy efficiency, information dissemination, and facilitation of research and technology
development to help move to the commercialisation of second generation biofuels and other sources of
renewable energy.

The preliminary conclusion at this stage is that, when taking a holistic view of the economic,
environmental and social dimensions, the promotion of biofuels from current agricultural feedstocks in many
countries and regions is not a sustainable use of scarce resources. However, the calculation may well be different
with a wider source of feedstocks in second generation bioenergy production.

BSD-2008 IL-01

White (industrial) biotechnology: Clean, clever & competitive

Colja Laane

Corporate Science manager Life Science, DSM Corporate Communications,

PO Box 6500, NL 6401 JH Heerlen, Netherlands

White (industrial) biotechnology is rapidly gaining momentum as clean, clever & competitive
technology to produce bio-based fuels, chemicals, materials and specialties in a safe and sustainable way from
renewable resources using nature's toolset. Renewed interest in bio-based processes is triggered by high oil
prices, unprecedented advances in screening and omics technologies, and a growing interest in sustainable
solutions for today's problems and future opportunities. Recent estimates from McKinsey & Company indicate
that the white biotech market will grow from $ 77 b in 2006 to $ 125 b in 2010, including biofuels. Currently
strategic partnerships are being realized covering (parts) of the white biotech demand and supply chain to create
best value. Feedstock- and technology providers are teaming up with companies which have access to the
st nd
market. In this arena partners already moving from the 1 generation of sugar and starch-based biorefineries to 2
and 3 generation biorefineries, in which complex (ligno) cellulosic feedstocks will be converted into fuels and a
multitude of specialties, chemicals and materials. In this presentation the strategic choices DSM made will be
highlighted and the direction our company is moving with our partners to shape a clean, clever & competitive

BSD-2008 IL-02

Enzymatic solutions in industry - a contribution to sustainable development

Per H. Nielsen & Karen M. Oxenbøll

Novozymes, Krogshøjvej 36, DK-2880 Bagsværd, Denmark (Eco-Efficiency Assessment departm.)

Enzymes are biological catalysts with an enormous capacity to speed up biochemical reactions. They
are developed during millions of years of evolution and are essential to any type of life. Human beings have taken
advantage of enzymes since early days, for instance by using animal dungs for softening animal hides in leather
making. Today, enzymes are produced in industrial scale by fermentation and used in a broad range of industries
because they reduce raw material, energy and water expenditures, processing time, and/or improve quality of the
final product.
Novozymes is a major producer of industrial enzymes and the company has used life cycle assessment
(LCA) systematically for some years now to analyse 1) the environmental impacts associated with enzyme
production and 2) the environmental achievements when the enzymes are applied in private homes, in
agriculture and in various industries.
So far, nine different fields of enzyme applications have been analysed (detergent additive, animal feed
supplementation, vegetable oil production, leather production, bread production, fatty acid ester production,
textile production, cheese production and fuel ethanol production) and the results show that enzymatic solutions
generally add considerably to the customers' environmental profiles in terms of reduced energy consumption,
reduced contribution to global warming, acidification, nutrient enrichment, photochemical smog formation and
sometimes also consumption of limited resources.

The explanation is the catalytic properties, the high reaction rates, and specificity of the enzymes and
that small amounts of enzyme do the same or a better job in production than large amounts of chemicals and

Enzymatic solutions are widely used today, but there is still a great potential for further penetration in
existing fields as well as expansion into completely new. The total potential savings of for instance greenhouse
gasses by enzyme application are measured in millions of tonnes of CO2 equivalents. Such saving potentials are
interesting even at a national/global scale and it is interesting to note that most of them are free of charge, because
use of enzymes for the most is driven by cost saving and/or quality improvements.

All environmental assessments of Novozymes' enzymatic solution are based on market oriented LCA
principles, and the results are backed up with sensitivity assessment of the most uncertain and variable
assumptions to evaluate and document robustness of the results.

The observed environmental performance of enzymes is used in Novozymes' marketing after external
review of LCA documentation according to ISO 14040 standards.
All environmental assessments of Novozymes' enzymatic solution are based on market oriented LCA principles,
and the results are backed up with sensitivity assessment of the most uncertain and variable assumptions to
evaluate and document robustness of the results.
LCA is a holistic environmental assessment tool which addresses raw material uses and emissions in all
processes in the product chain from raw material extraction through production to use and final disposal

BSD-2008 IL-03

Evaluating the Potential of Biotechnology for Production of Speciality

Chemicals and Polymers from Renewable Resources
Rajni Hatti-Kaul

Department of Biotechnology, Lund University, Box 124, SE-221 00 Lund, Sweden

Shifting the resource base of chemical industry from fossil feedstocks to renewable raw materials
provides interesting opportunities for biotechnology based process tools. Industrial biotechnology, based on the
use of microorganisms and their enzymes that are naturally adapted to processing renewable resources, can offer
a potentially cleaner and more energy-efficient means of production than chemical processes, with additional
advantages of biodegradable products having a minimal environmental impact. Microbial diversity of extreme
environments provides interesting tools for industrial biotechnology. The lecture will present some of our efforts
in production of speciality chemicals and polymers using biocatalysis and fermentation, respectively, and will
further highlight the challenges that need to be addressed for implementation of biotechnology in chemical

In the Greenchem research programme at Lund University, biocatalysis is being used for the production
of a variety of speciality chemicals from renewable resources under solvent-free conditions with an aim to
provide environmentally benign processes and products. Among the chemicals being produced are epoxidized
oleochemicals for use in surface coatings, plasticizers and stabilizers, and biosurfactants such as alkyl
glycosides and alkanolamides for use in detergent, cosmetic and therapeutic applications. The enzyme lipase
from Candida antarctica is used for the production of epoxides and alkanolamides with high yields. Epoxides
have been produced from different raw materials e.g. rapeseed methyl ester, tall oil methyl ester, etc., and further
modification of the epoxides is done for improving their reactivity. The enzymatic production of alkanolamides
from fatty acids and ethanolamine is facilitated by running the process under conditions avoiding ion pair
formation and by removal of water generated. Novel glycosidase enzymes from hyperthermophilic Themotoga
neapolitana are used for synthesis of alkyl glycosides from sugars and alcohol.

As an example of the polymer will be described the production of poly (3-hydroxybutyrate) (PHB) from
sugars by a moderate halophile, Halomonas boliviensis isolated from Bolivian highlands. PHB has properties
similar to polyethylene and polypropylene but is biodegradable. A significant increase in the productivity and
yield of the polymer has been achieved by optimization of fermentation conditions.

Acknowledgement: The author is grateful to Mistra for financial support to the Greenchem programme, and to
Sida and Swedish Research Link for supporting the work on bioplastics.

BSD-2008 IL-04

Green Polymers - How sustainable are they ?

M. K. Patel, B. G. Hermann, L. Shen

Utrecht University, Department of Science, Technology and Society (STS) / Copernicus Institute,
Heidelberglaan 2, NL-3584 CS Utrecht, Netherlands. Tel.: 0031 30 253 7634, Fax: 0031 30 253 7601,,

There are several pathways to produce bio-based chemicals and polymers. The pathways that are most
important in terms of production volumes today and in the medium-term future are White Biotechnology
(industrial biotechnology) and the use of natural polymers (in particular cellulose and starch). This presentation
presents environmental assessments for these two categories of bio-based chemicals.

In the BREW project (compare we conducted detailed environmental

and economic assessments for the 21 White Biotechnology products. We found that White Biotechnology offers
very substantial opportunities for reducing non-renewable energy use, greenhouse gas emissions and the related
environmental impacts. The scenario analysis shows that under unfavourable conditions the share of chemicals
produced via White Biotechnology in Europe remains marginal (up to the year 2050), while the opposite is true
under favourable conditions. In the latter case, up to 39%-67% of the non-renewable energy demand for the
(conventional) production of the selected chemicals can be saved in the EU-25 by 2050 (lower value for starch,
higher value for lignocellulosics). Concerning economic aspects we find that, in 2050, White Biotechnology can
offer substantial macroeconomic savings.

The presentation further elaborates on the use of bio-based polymers in final applications. Both White
Biotechnology polymers and natural polymers are studied. The applications analysed are a housing for
electronic goods, an automotive panel and high-barrier food packaging films. The most important conclusions
are that the environmental attractiveness depends on application area and on waste management and that bio-
based polymers are in their infancy (production and conversion) and that big step change

BSD-2008 IL-05

Dislocating Technologies for Biorefineries

Gopal Chotani, Ph.D.

Research Fellow, Genencor®, a Danisco Division

Palo Alto, CA, USA

There are huge opportunities in industrial biotechnology that range from production of cocktails of
enzymes to biomaterials, biochemicals, and biofuels that are accompanied with challenges like cost-price
fluctuations, infrastructure development, energy usage, regulatory requirements, and sustainable research
funds. Just as petroleum refineries make more than one product that serves as feedstock for other industries,
biorefineries too will be able to deliver a multitude of products, including industrial chemicals and biofuels
independent of non-renewable feedstocks. To enable a rapid expansion of the biotechnology industry, however,
there is critical need for scientists and engineers skilled in process development to integrate protein expression
and biochemical/metabolic pathway designs with process engineering.

Biological cell factories are capable of replacing traditional chemical factories. By extracting,
regulating and assembling materials from the natural environment, microbial cells have the potential to provide
breakthroughs for the chemical industry. Biological systems are very complex much like chemical
manufacturing systems, comprised of large numbers of machines, information systems, and warehouses, yet
very versatile

This paper will illustrate some of Genencor's approaches for process and product development The
pathway engineering examples will show the benefits of a “rational approach” where knowledge regarding
microbial cell physiology is accumulated to improve processes, obtain new products, and solve unexpected
problems on a continuous basis. The result is the building of nano-cell factories and a new paradigm for the
manufacture of chemicals and materials.

BSD-2008 IL-06

Biofuel Production: A new Biorefinery for Sustainable Energy from Crops;

Conversion of Lignocellulose to Bioethanol, Biohydrogen and Biomethane
Irini Angelidaki and Prawit Kongjan

Institute of Environment & Resources DTU, Technical University of Denmark, Building 113, DK-2800,
Kgs. Lyngby, Denmark

Sustainable production and utilization of biofuels has an enormous potential for the world's energy
supply. Most research studies usually focus on individual conversion processes and specific end-products. To
gain full benefits however, it is important to investigate production and utilization cycles in an integrated way
and to consider all important aspects involved: e.g. crop production, residues, supply chain, flexibility of end-
products, and environmental aspects. Agricultural residues are excellent examples of crops with a large potential
for flexible production of several end-products, such as: bioethanol, biohydrogen, biogas as well as
biopolymers. In this multidisciplinary study we tried to integrated innovative approaches to produce a multi-
product biofuel from these important crops based on a number of novel and mutually synergistic production
methods, and at the same time include an assessment on the environmental benefits and drawbacks related to the
concept. The combined production methods will significantly simplify the overall production process and
thereby substantially reduce the product costs and the environmental impacts. The scheme for multi-product
biorefinery process that is under investigation is including the following steps: pretreatment of lignocellulosic
biomass for release of carbohydrates; conversion of hexoses to bioethanol; conversion of pentoses to
biohydrogen and finally waste streams treatment in anaerobic biogas reactors for production of biogas. In the
present paper we show that pretreated lignocellulosic material could successfully be converted to bioethanol (for
the solids hexoses part) with a yield of 90-100% and to biohydrogen for the hydrolysate mainly containing the
pentoses with a specific yield of 2.95 mmoleH2/gVSadded. The hydrolysate was converted to hydrogen in a
thermophilic process using continuous stirred tank (CSTR). A productivity of 113 ml H2/(d·l) was achieved from
the CSTR reactor experiments. The effluents of the process could be converted to methane in a CSTR reactor.
The present study shows that it is possible to extract different biofuels from lignocellulosic material in order to
optimize for the highest value products in a chain different production steps, where all available organic material
is optimally utilized.

BSD-2008 IL-07

Perstorp Bio Products AB, taking Bio-chemicals and Fuels to the Market
Mr.Tomas Hagstrom

Commercial Manager, Perstorp BioProducts AB

SE-284 80 Perstorp, Sweden

Perstorp is today a USD 1500 million company focused entirely on Specialty Chemicals and active on
the global arena with operations on 4 continents. The company has a 127 year history today holding unique
experience and knowledge within the field of Aldehyde chemistry. It is a technology driven company with a
proven track record of innovative growth.

Since the early 50-ies the company has based its production entirely on oil and natural gas as its upstream
raw materials. Going back to the pre WWII period the company did though utilise renewable raw materials for
substantial parts of its production. With increased costs for both oil and natural gas as well as for energy the
company is evaluating alternative production methods and new products to secure its competitiveness. In doing
so the matter of sustainability also plays a central role.

Not only is biomaterial based raw materials on the agenda, but also new process methods based upon
white biotechnology. Extensive R&D efforts are spent on developing and evaluating new process solutions and
products based upon Glycerol, Ethanol and Butanol among others.

In order to accelerate the speed of implementation and in order to reach critical size within renewable
chemistry the company has entered into the field of biofuels. Today Perstorp operates Scandinavia's largest and
most advanced biodiesel plant and is active also within other biofuels.

Perstorp is at present focusing its biochemicals activities to Europe but evaluating new establishments in
South America, Africa and Asia. The company being a specialist in chemical processes and products is seeking
suitable upstream partners for further ventures.

BSD-2008 IL-08

Lignocellulosic Biomass Materials Sustainable Feedstock for Ethanol Production

Ghanshyam Deshpande

Executive Vice President, Praj Industries Limited, Pune, India

The ever increasing crude prices and governmental initiatives world over has significantly enhanced
our interest in biofuels based on lignocellulosic materials. Fuel ethanol is being produced at present either from
molasses or from starch based feedstocks like corn or tubers. It is important to utilize abundantly available
biomass materials to meet future demand. Various agricultural residues like corn stover, wheat straw, rice straw,
agricultural processing by-products such as rice hulls, sugar cane bagasse and energy crops such as grasses and
tubers are considered to be attractive alternative sources for biofuel production.

Though several process configurations for fuel ethanol production from lignocellulosic biomass were
studied and tested fairly on large scale, cost effectiveness of processing technologies is still under evaluation.
Predominant technological challenges are feed stock collection, material handing issues, pretreatment
strategies, cellulose conversion costs and pentose sugar conversion to alcohol.

On the technological front several major players are in the arena. Various pretreatment methodologies,
biomass fractionation and their hydrolysis into pentose and hexose sugars have been reported. Attempts are
being made to develop specific microbial species which can utilize mixed sugars effectively. Emphasis is also
being given to develop new varieties of energy crops with very high yields per hectare. In addition to biological
routes, technologies are being attempted to convert biomass based syngas to ethanol either by chemical methods
are by biological methods.

The presentation will give a broad over view of the biomass based bio-refineries which includes the first
generation to third generation technologies.

BSD-2008 IL-09

Bioenergy from Energy Crops : Technical Possibilities

and Economical Feasibility
Malek Alkasrawi PhD.

Syngenta Seeds AB, Box 302, SE-261 23 Landskrona, Sweden

The energy crops are ideal feedstock for different types of biofuel production due to high sugar content
& low structural complexity. The energy specific breeding program is focusing on the maximum yield of
fermentable and other substances that might improve the conversion process. This would result in specific crop,
customised for biofuel production. Different hybrids were evaluated for biofuel production in a small scale, and
the results showed that the energy crops are an attractive feedstock alternative. Technical data for the complete
process integration including innovative preparation technique will be presented.

BSD-2008 IL-10

Anaerobic Sequencing-Batch-Reactor (ASBR) Technology for High

Performance Biogas Production
Pätz Reinhard R.

Laboratory of Energetic Biotechnology, University of Applied Sciences, Köthen/Anhalt, Germany

Sequencing-Batch Reactor-Technology (SBR-Technology) is a widely used modern technology for

wastewater treatment. First investigations were done by PASVEER, IRVINE and WILDERER. The advantage
is the use of a discontinuous process with a high turnover of the substrate as a result of biomass retention with a
quasi-continuous process regime. A process cycle consists of four phases: filling, reaction, sedimentation and
reduction of reaction volume. In terms of biotechnology it is a classic repeated bed-batch with a sedimentation
step for biomass sedimentation. Anaerobically it is more difficult because of floating effect as a result of biogas
production. Thus there are the following phases during a cycle:

 Filling
 Intensive biogas production
 Separation
 Volume reduction

ASBR-Technology is described by ZAIAT 2001. Besides ASBR a biofilm process is possible by using
suitable carriers. Then it is termed as ASBBR- Technology (Anaerobic Sequencing- Batch Biofilm Reactor) In
this presentation two simple reactor concepts are described that can be used for anaerobic treatment of sieved pig
manure in labscale (6 l, 20 L).

Reactors for ASBR-Technology

1. Solid-Retention by simple tube construction
2. Biofilm reactor concept with FLORAKLAER carrier

During investigation there is an increase in biogas productivity as a result of training of active biomass
and of biomass retention. LINKE defined a Methane Capacity Coefficient pCH4ODM as product of Methane yield (of
organic Dry Matter DM of substrate) and Methane production.

BSD-2008 IL-11

Biogas from Surplus Biomass the Optimal Alternative/ (Ultimate Solution)?

Bo Mattiasson

Department of Biotechnology, Lund University;

Conversion of biomass into bioenergy is today a dominating topic among industrial biotechnologists.
The market pull has led to a range of investments that seem profitable in a short run, but in a longer perspective
they may be more questionable.

Conversion of biomass into bioenergy is connected with questions like: How energy efficient is the
process? How much of the biomass is utilized? Etc.

Biogas is produced using a consortium of strict anaerobic microorganisms. An attractive feature is that
the consortium members are able to convert almost all types of biomass into smaller entities that eventually are
transferred into biogas and carbon dioxide. The technology to produce biogas has been around for a long time,
but still the process suffers from some technological/biological drawbacks: the low productivity and the
sensitivity of the consortium to overloading. Due to these features, very little new process technology has been
presented and there is still today a lack of suitable high producing biogas reactors.

This presentation will address both these issues and indicate a way ahead.

 Process monitoring and control. By using proper monitoring signals it is possible to get good control
over the biogas process, and then it is realistic to increase the load over the levels often used today.
Monitoring signals and control strategies make it possible not only to increase the loading, but also to
improve the process stability.

 A limiting factor during the initial phase of an anaerobic process is the easily degradable material that
will cause the pH to drop and then negatively influence the methanogenic bacteria. However, after this
initial phase, another step in the process is limiting, and that is mobilization of the biomass via
hydrolysis. To handle these challenges a two-stage strategy would be advantageous. In a methanogenic
stage soluble compounds are converted into biogas. By cultivating the methanogens as biofilm on solid
carriers high catalytic capacity will be obtained in the reactor. The easily degradable material can be fed
into the methanogenic filter under optimal conditions and later on the leachate from the hydrolysis step
can be fed. This gives a possibility to both make a high producing methanogenic step and also to design
the hydrolytic step such that it will efficiently supply the methanogenic step with substrate.

There are several strategies to improve the hydrolytic step and that is one of the hot spots in biogas
research today.

BSD-2008 IL-12

Environmental Systems Analysis Concerning Bioenergy from Crops

Pål Börjesson

Environmental and Energy Systems Studies, Department of Technology and Society, Lund University,
Box 118, SE-221 00 Lund, Sweden

Today, there are strong incentives promoting an increased use of renewable fuels in the worldwide
transport sector. Bioethanol from crops rich in starch and sugar and biodiesel from oil crops are fuels produced
by commercial technologies and used in conventional vehicles. However, some bioethanol and biodiesel
production routes have limitations concerning resource efficiency and reduction of greenhouse gases. More
efficient biofuel systems are those based on lignocelluloses and novel conversion technologies, such as thermal
gasification, which are as yet not commercially available. A complementary option is to increase the production
of biogas from the digestion of organic residues and energy crops, or from the by-products of ethanol and
biodiesel production, and use this biogas as a vehicle fuel. Another opportunity is to utilise ethanol, vegetable oil
and methane from biomass as feedstock in the chemical industry.

From the perspective of resource efficiency, the various production routes of biofuels differ. For
example, the gross energy output per hectare of arable land of upgraded gaseous vehicle fuel is often higher than
for ethanol produced from the same crops, or biodiesel from rape seed. The energy input in biogas production is
also calculated to be lower than in today's ethanol production, leading to a higher energy output/input ratio for
biogas systems. Biofuel based on woody energy crops (short-rotation energy forest), such as willow, and
produced by thermal gasification often have the highest energy output/input ratio. However, the energy input in
biofuel production systems may be calculated in different ways, e.g. depending on how potential by-products are
included and which allocation methods that are used to divide the energy input between biofuels and by-
products. Other parameters of importance influencing the energy balance are local production conditions, crop
yield, cultivation methods, process technologies, systems boundaries etc. Thus, there is no “right” or “wrong”
method to use to calculate energy balances of biofuel production as different methods may be relevant to used
under specific local conditions.

Substituting biofuels for fossil fuels for vehicles will lead to a reduction of greenhouse gas emissions.
However, biofuels produced from energy crops are not greenhouse gas neutral, since fossil fuels are used in the
cultivation of energy crops, causing carbon dioxide emissions, e.g. diesel fuel used by tractors and natural gas
used in the production of chemical fertilisers. Fossil fuels may also be used in the biofuel conversion processes.
Energy crop production often utilises large amounts of fertilisers. The production of chemical fertilisers also
causes emissions of nitrous gas (N2O), which is a 300-times more potent greenhouse gas than carbon dioxide.
Furthermore, depending on soil conditions, the spreading of nitrogen fertilizers may lead to biogenic nitrous gas
emissions. From a greenhouse-gas perspective, the emissions of nitrous oxide from the production and use of
nitrogen fertilisers will often exceed the emissions of carbon dioxide from the use of fossil fuels, directly or
indirectly, in the cultivation of energy crops.

To conclude, when comparing different production routes of biofuels (and feedstock for the chemical
industry) from crops, from a resource and an environmental

BSD-2008 IL-13

Bioprocess Intensification in Sustainable Development

G. Akay
Process Intensification and Miniaturization (PIM) Centre,
School of Chemical Engineering and Advanced Materials
The Institute of Stem Cell Biology and Regenerative Medicine
Institute of Nanoscale Science and Technology
Newcastle University, Newcastle upon Tyne NE1 7RU (UK)

Biomass based integrated energy and chemicals technologies are now emerging as important routes to combat
global warming and replace diminishing fossil fuels and chemicals feedstock1. Bio-refineries will replace oil
refineries and petrochemical plants in the simultaneous generation of chemicals, energy and power. However,
due to the distributed nature of the biomass and its wide variability, bio-refineries should have weak 'economies
of scale' dependency and should be responsive to the changes in supply and demand. These conditions are
necessary for the sustainability of this emerging technology. Sustainability condition also requires the
integration of agro-, bio- and chemical processes. Such integration will be seriously compromised due to the
mismatching of conversion rates achievable by chemical and biological processing. These constraints can be
circumvented if bioconversion rate is enhanced by 100-1000 fold without increasing the size of the bioreactors.
This in turn can be achieved through higher selectivity and enhanced productivity using genetic enhancement
and by using Bioprocess Intensification based on so called 'confinement phenomenon' in biological systems
according to which, the behaviour (differentiation, proliferation rate, viability, productivity) of microorganisms/
mammalian and plant cells is dependent on the size and micro-architecture of the three-dimensional
confinement/support environment2-9. It is postulated that controlled and sustained (in time and space)
physiological stress (as a result of confinement) result in the alteration of the metabolic pathway and gene
2-5 st
expression pattern. This phenomenon, first observed in tissue engineering , has provided the 1 example of
Bioprocess Intensification with 10-100 fold intensification . We have recently further illustrated the generic
nature of Bioprocess Intensification based on the 'confinement phenomenon' in the intensification of antibiotic
production as well as plant biomass and crop yields. The physical-chemical-biological nature of the support
environment for biocatalysis is important in obtaining bioprocess intensification due to the interactions between
the support, microorganisms and any additional intensification field. These interactions can be optimised by
using nano-structured macro-porous materials which do not hinder mass transfer or signalling. Due to these
restrictions, Bioprocess Intensification needs to be carried out in micro-bioreactors which can be formed from
polymeric, metallic or ceramic or composite nano-structured macro-porous monoliths as the catalytic reactor

1. 1. G. Akay, M. Dogru, OF Calkan and B Calkan, Biomass processing in biofuel applications. In: Biofuels for
Fuel Cells, Eds: P. Lens, P. Westermann, M. Haberbauer and A. Menero, IWA Publishing, London. Ch. 4.

2. G. Akay, S. Dawnes, VJ Price, Microcellular polymers as cell growth media and novel polymers, EP
1183328 A2 (2002); US 09, 856,182 (2002).

3. G. Akay, M.A. Birch, M. A. Bokhari, Microcellular Polyhipe polymer (PHP) supports osteoblastic growth
and bone formation in vitro, Biomaterials, 25 (2004) 3991-4000.

4. M.A. Bokhari, G. Akay, S Zhang and MA Birch, A hybrid biomaterial combining the peptide hydrogel RAD
16-1 with PolyHIPE Polymer (PHP) enhances osteoblast growth and differentiation in vitro. Biomaterials,
26 (2005) 5198-5208.

BSD-2008 IL-13
5. G. Akay, MA Bokhari, VJ Byron and M. Dogru, Development of nano-structured materials and their
application in bioprocess-chemical process intensification and tissue engineering. In: Chemical
Engineering Trends and Developments, Ed: MA Galan and E.M. Del Valle, Wiley, London, 2005. Ch. pp.
171-196. (2005).

6. G. Akay, Micro-porous materials, PCT WO 2004/005355 (2004)

7. E. Erhan, E. Yer, G. Akay, B. Keskinler, and D. Keskinler, Phenol degradation in a fixed-bed bioreactor using
micro-cellular polymer immobilized Pseudononas syringae, J. Chem. Technology and Biotechnology, 79
(2004) 195-206.

8. G. Akay, E. Erhan, B Keskinler, Bioprocess intensification in flow through micro-reactors with immobilized
bacteria, Bioengineering Biotechnology, 90 (2005) 180-190.

9. G. Akay, Bioprocess and chemical process intensification. In: Encyclopedia of Chemical Processing, pp.
185 198, Ed: S Lee, Marcel Dekker, NY. (2006).

10. G. Akay, M. Dogru, B. Calkan and O.F. Calkan, Flow induced phase inversion phenomenon in process
intensification and micro-reactor technology. In: Microreactor Technology and Process Intensification,
Eds: Y Wang and J Holladay, Oxford University Press, Oxford, Ch. 18. (2005).

BSD-2008 IL-14

Modern Bioenergy Technologies for India

Prof. N. H. Ravindranath

Centre for Sustainable Technologies, Indian Institute of Science

The global and Indian economies have been experiencing unprecedented growth accompanied by
growth in energy use. Now there is a rethink on the energy path adopted by India and all the countries globally,
due to factors such as firstly, GHG emissions resulting from fossil fuel use leading to climate change, secondly,
an unprecedented rise in oil prices as well as the uncertainty of supply and thirdly, and finally energy security in
the face of risk of access to sustainable supplies. Thus India and other countries are exploring sustainable
alternatives. In the recent past among all the renewable energy technologies, bioenergy seems to provide the
largest opportunity to meet the energy demands, increase energy security, substitute fossil fuels and reduce GHG
emissions. Further, bioenergy technologies provide opportunity to meet liquid (ethanol and bio-diesel), gaseous
(biogas and municipal solid waste gas), power and process heat requirements. India is in the forefront of
promoting bioenergy technologies. The dominant modern bioenergy technological option for India is biomass
combustion and gasification for power generation. Bio-diesel is being increasingly pursued for producing liquid
fuel. Biogas has the potential to provide cooking gas for rural households.

Modern bioenergy technological options are relatively cost-effective for decentralized applications.
Rural India, with a population of nearly 700 million, is currently dependent on low quality energy services such
as biomass fuels for cooking, high cost diesel and unreliable grid power supply. Rural India provides a great
opportunity for deployment of bioenergy technologies not only to provide quality energy, but also increase
energy security and provide local environmental as well as socio-economic benefits. Despite multiple benefits
and cost-effectiveness, the rate of spread of bioenergy technologies is limited compared to the potential.
Biotechnology also has the potential to enhance efficiency of biomass production as well as biomass conversion
processes. Bioenergy is likely to play an even greater role for its potential to substitute fossil fuels leading to
reduction of GHG emissions, to address climate change. Mitigation potential of bioenergy technologies is
attracting large investment for research and development of bioenergy technologies as well participation of
industry. Bioenergy technologies are already dominating the Clean Development Mechanism.

This presentation attempts to provide an overview of bioenergy technologies, an assessment of

opportunities for meeting energy needs as well as providing local and global environmental benefits. India has
formulated and implemented some of the most innovative financial incentives and policies. Barriers and policies
are presented briefly to promote bioenergy technologies in India. Sustainable biomass production and supply for
power generation and degraded status of lands for growing non-edible oil seeds as feedstock for bio-diesel
production are critical barriers for large-scale spread of biomass power and bio-diesel production options,
respectively. Research and development to generate advanced and cost-effective bioconversion technologies is
the first step in large-scale promotion of bioenergy technologies in India and globally.

BSD-2008 IL-15

Sulfate Reduction Process for Removal of Heavy Metals

Warounsak Liamleam, Zaw Ko Woo, Phan Thong Thai, Ajit P. Annachhatre

Environmental Engineering and Management, Asian Institute of Technology, PO Box 4, Klongluang,

Pathumthani 12120, Thailand Fax: 662-524-5644

Biological sulfate reduction is currently one of the widely practiced processes for treatment of sulfate-
containing wastewaters commonly generated from industries such as mining, tannery, pulp and paper, rayon,
and textile industry. In the biological reduction, sulfate is converted to hydrogen sulfide as the end product, the
process is, therefore, ideal for treating metal-containing wastewater from which heavy metals are removed
through the formation of metal sulfides.

Biological sulfate reduction process with molasses as an electron donor was used in this research for the
removal of zinc and sulfate from rayon industry wastewater. Sulfate was reduced to sulfide under anaerobic
conditions and sulfide rich effluent was then used to remove zinc as zinc sulfide precipitate. Investigations were
conducted at pilot scale with real wastewater from rayon industry as feed. Effects of sulfate loading rate and
temperature of feeding wastewater were evaluated. The experimental results showed that there was no
significantly different sulfide production when the reactor was operated at the temperature 50 2 C and 65 2 C.
Sulfide production was in the range of 500-515 mg l-1. Zinc sulfide precipitation at pH 7.0 could yield more than
96% of zinc removal.

BSD-2008 GC-01

Production and degradation of β-(1→3) glucan

Sri Lakshmi P., Sathyanarayana N Gummadi, Mukesh Doble*

Dept of Biotechnology, IIT Madras, Chennai 600036

Curdlan is a high molecular weight water insoluble extracellular polysaccharide composed of

exclusively β-(1→3) linked glucose residues, produced by fermentation using Alcaligenes faecalis var.
myxogenes under nitrogen limiting conditions. This biopolymer has drawn considerable interest
because of its unique rheological and thermal gelling properties .It is used as a formulation aid,
processing aid, stabilizer and thickener in food products. It is approved by US Food and Drug
Administration (FDA).

The production of curdlan using sucrose as the carbon source consisted of two steps, where the
cells were initially grown to substantial biomass, followed by conditions for the production of curdlan.
The polymer produced was filtered from the biomass and recovered by dissolving it in alkali, followed
by precipitation with acid. Effect of various parameters such as pH, concentrations of sucrose and
nitrogen on curdlan production was studied. The kinetics of the process was also determined.

Since curdlan is widely used both in food industry and medicine, it is important to know its
degradation pattern. Degradation of curdlan by Bacillus subtilis was carried out and the amount of
degradation by this strain was quantified by fluorescence micro assay. The polymer recovered after
degradation was structurally characterized by FTIR and HPLC analyses.

BSD-2008 GC-02

Novel approach of controlling microbial induced corrosion : potential of

natural biocides
M Lavania, P M Sarma, A K Mandal and B Lal*

TERI, Darbari Seth Block, India Habitat Center, Lodhi Road, New Delhi 110003 Fax: 24682144/2468 2145,

The efficacy of natural biocide has been compared with some of the chemical and biological agents,
which are known to control microbial influenced corrosion (MIC). The study demonstrated that cow urine has
the most potent effect among the tested biocides against Desulfotomaculum kuznetsovii, Clostridium sporogenes
and Thermotoga neapolitana (TERI M1 consortium) grown in the laboratory. Sulfide production was found to
be minimum with cow urine followed by metabolic extract from Pseudomonas fragi and chemical biocide
bromonitropropanediol. Similarly, as compared with the control, 2 mM of Fe (III) reduction was avoided with
cow urine followed 6.2 mM of metabolic extract from Pseudomonas fragi and 7.9 mM of
bromonitropropanediol. The scanning electron micrographs described the biofilm structure of metal coupons in
presence of biocides against TERI M1 consortium. Low exopolysachharide production was observed against
-2 -2
TERI M1 consortium in presence of cow urine (42.6 µg cm ), followed by bromonitropropanediol (73.0 µg cm )
-2 -2
and biological agents (74.8 µg cm ) as compared to control (583.2 µg cm ). The result suggested that cow urine
is an effective agent for controlling the microbial induced corrosion in oil industry by the virtue of its
antimicrobial properties.

BSD-2008 GC-03

Polyhydroxyalkanoic acid (PHA) production by bacterial strains isolated from

hydrocarbon-contaminated soils
J Dalal*, P M Sarma**, A K Mandal** and B Lal**

*TERI University, India Habitat Centre, Lodhi Road, New Delhi

**The Energy and Resources Institute (TERI), India Habitat Centre, Lodhi Road, New Delhi
*, Fax: 24682144/2468 2145

Two hundred and sixty two bacterial strains isolated from 14 petroleum hydrocarbon contaminated sites
located in different geoclimatic regions of India were screened by a phenotypic (Nile blue A colony staining) and
a genotypic (PCR based gene specific screening) technique for PHA production. Amongst the 262 bacterial
strains examined, 97 were positive with the genotypic method and 81 were positive with phenotypic method.
Forty-four bacterial strains were found positive with both the screening methods and were selected for further
studies. The identification of the selected bacterial strains indicated that six bacterial strains were earlier not
reported for PHA production. The quantification of PHA produced by bacterial strains was confirmed by Gas
Chromatographic analysis. The nature of the monomeric units was analyzed by Gas Chromatography-Mass
Spectroscopy and found to contain medium-chain-length hydroxyalkanoic acids. This study indicates that
stressed environments like petroleum-contaminated sites can be potential sources of medium-chain-length PHA

BSD-2008 GC-04

Identification of Novel Halotolerance Genes by Metagenomic Approach

Raj Kishor Kapardar 1,2, Ravi Ranjan 1, Munish Puri 2 and Rakesh Sharma1

1. Environmental Biotechnology, Institute of Genomics and Integrative Biology, Mall Road, Delhi-11000
2. Department of Biotechnology, Punjabi University, Patiala.

Molecular microbial diversity and phylogenetic analysis have revealed that more than 99% of the
bacteria present in environment cannot be cultivated in laboratory. Traditionally,
microbiologists relied on culture to study the microorganism and their exploitation for
biotechnological application. This allowed access to only less than 1% of the microbial
diversity. By the molecular analysis, remaining 'uncultivable' bacteria represent unrecognized classes and
divisions in bacterial and archaeal domain and are expected to possess novel physiologies and genes.
Metagenomic (Culture-independent methods) give access to the vast gene pool of the 'unculturable' bacteria.

Microorganisms in environment face different abiotic stress like high salt and toxic heavy metals. They
possess different stress tolerance machineries to overcome these stresses. 'Unculturable' bacteria present in
different environments are expected to possess unique stress tolerance mechanisms. Identification of stress
tolerance genes from 'unculturable' bacteria and their characterization will help in elucidation of these
mechanisms. These genes can be utilized for various bioremediation applications and also in plant
biotechnology for development of abiotic stress tolerant plants.

We prepared metagenomic libraries from different environmental samples and isolated salt tolerant
clones. Three salt tolerant clones were chosen for further characterization. These clones were able to grow at
inhibitory concentration of NaCl, KCl and LiCl. The inserts were completely sequenced and genes
responsible for salt tolerance were identified by insertional mutagenesis and subcloning. The identified genes
showed low homology match with the known sequences in the databases indicating their origin from yet
uncharacterized organisms. Their possible function and role in stress tolerance mechanisms was predicted using
in-silico approach.

BSD-2008 GC-05

Optimization of Renewable bioresource Orange juice using Membrane

Distillation Technology.
Mr. S. K. Deshmukh Dr. V. S. Sapkal Dr. R. S. Sapkal

Department of Chemical Technology, Sant Gadge Baba Amravati University, Amravati.

The concentration of orange juice, a renewable health product, provides a reduction of transport,
packaging, and storage cost. During the concentration process, the water should be removed selectively in order
to obtain a product with an appearance and taste as close as possible to the original juice. In that case, the
concentrated juice could be reconstituted by just adding the appropriate amount of water. The production of high
quality concentrated orange juice is obtained using new integrated membrane distillation processes, alternative
to thermal evaporation and is evaluated in terms of preservation of the total antioxidant activity and of bioactive
anti oxidant components of the juice (ascorbic acid, anthocyanins, hydroxyl cinnamic acids, flavaroids).
Traditionally orange juice concentration has been carried out by multistage vacuum evaporation, which results
in a loss of fresh juice flavours, color degradation and cooked taste.

The principal of Membrane Distillation is explored and compared to other membrane processes. The
flux of permeate in the Direct Contact Membrane Distillation process is found to be a function of membrane
characteristics and operating conditions. The correlation exists between vapor transfer through a hydrophobic
membrane, the temperature at the surface of the membrane, changes of temperature and concentration solutions
along the module. The operating parameters in this process are feed and permeate bulk temperature and their
difference; flow rate and feed concentration and the performing parameters are permeate flux and retention
efficiency which can be developed for each fluid and used for designing the processes.

Membrane Distillation has very important advantages; practically a complete rejection of dissolved,
non volatile species, lower operating pressure than pressure driven processes, reduce vapor space compared to
conventional distillation, low operating temperature of a feed that enables the utilization of waste heat as a
preferable energy source. The possibility of utilizing of alternative energy source such as solar, wave or
geothermal energy is particularly attractive.

BSD-2008 GC-06

Kinetic and thermodynamic analysis in microwave assisted lipase-catalyzed

Indrakant V. Borkar and Ganapati D. Yadav

Department of Chemical Engineering, University Institute of Chemical Technology, University of Mumbai,

Matunga, Mumbai 400 019,
Telefax: ++91-22-2410-2121, Fax: ++91-22-2414-5614,

Microwaves are powerful and reliable form of electromagnetic energy source that can be adapted to
many applications. Microwave-assisted lipase coupled activity (MALCA) of various commercially available
lipases, such as Candida antartica lipase B, Mucor miehei lipase and Thermomyces lanuginosus lipase were
tested and among them Candida antartica lipase was found to be more sensitive to microwave. A network of
weakly polar interactions between the flaps (lid) was proposed to be responsible for stabilizing the semi-open
flap conformation because Candida antartica lipase B has no such a flapping lid. It is hypothesized that such
interactions could be responsible for making flap opening a highly sensitive gating mechanism which control
access to the active site. The microwave-assisted lipase coupled activity (MALCA) was influenced by several
factors such as temperature, solvent and substrates. At each tested temperature, the initial reaction rate under
MALCA was higher than that under conventional heating. It may be due to the modulated configuration of
enzyme molecules by accelerating the molecular rotation and electron spin oscillation of the polar parts of
enzyme, which can provide more chance to make the substrates fit to the enzyme in unit of time. In case of
conventional heating, the initial reaction rate changed with different substrates (acyl donors). The reaction rate
altering with the different acyl donors are explained in respect of two important parameters-polarity and stearic
hindrance effects.

We have summarized the influence of organic solvents on the enzymatic reactions and concluded that
the lipase activity is higher in the environment surrounded by non-polar and mid-polar solvents. Various kinetic
parameters affecting the conversion and initial rates including mass transfer, mechanism, kinetic modeling, and
deactivation were studied. Microwave irradiation leads to an increase in the frequency factor as a result of
enhanced collision of molecules and an increase in the entropy of the system. The values of activation energy for
the conventional and microwave heating are almost the same. From detailed kinetic modeling studies it was
observed that the turnover number of lipase was increased, while Michaelis constants and inhibition constants
were reduced by the microwaves. The kinetics of deactivation was systematically studied. Further investigation
revealed that in the presence of microwaves, the deactivation of lipase was substantially reduced. In MALCA,
the polar molecules collide with each other because of thermal effect and microwave effect. Therefore, the
molecule collision under MALCA has extra driving force compared to that under conventional heating, which
results higher rate under MALCA as long as the enzyme is not deactivated by microwave.

BSD-2008 GC-07

Optimization of biosurfactant production from Acinetobacter calcoaceticus,

isolated from Iran oil wells
F. Nemati1, D. Arabian1, Prof. R. Roostaazad2

Department of Biotechnology, University of Pune, Pune, India

Department of Chemical & Petroleum Engineering, Sharif University of Technology, Teheran, Iran

Biosurfactants have become recently an important product of biotechnology for industrial and medical
applications. The unique properties of biosurfactants allow their use and possible replacement of chemically
synthesized surfactants in a great number of industrial operations.

In the present work, we have investigated the ability of some of bacteria, isolated from Iran oil wells, for
producing biosurfactant. We have isolated a lipo-polysaccharide biosurfactant producer and by taxonomic tests
identified it to be a Gram-negative cocobacili, Acinetobacter calcoaceticus. This was found to decrease the
surface tension by 42 mN/m value (i.e from 72 to 30). Biosurfactant production was optimized by use of various
carbons (Ethanol, Soybean oil) and nitrogen sources (yeast extract, (NH4)2SO4 and urea), pH, temperature,
rotation per minute and time for biosurfactant production by use of the Tagochi program. It was observed that the
best conditions for production of biosurfactant were using 0.5 % Ethanol, (NH4)2SO4, and incubation conditions
of 35ºC, and 72 h.

BSD-2008 GC-08

Conservation and sustainable utilization of the Hornstedtia fenzlii (Kurz)

K.Schum. - An endemic endangered honey bee repellent species of the Great
Nicobar Island through in vitro multiplication and formulation of an insect
repellent stuff from essential oil components
Radha RK, Sam P Mathew and Seeni S

Division of Plant Biotechnology and Bioinformatics, Tropical Botanic Garden and Research Institute,
Palode, Thiruvananthapuram 695 562, Kerala, India.

Andaman and Nicobar Islands, the densely wooded tropical archipelago over 700 nautical miles away
from the Coramandel coast of the Peninsular India in the Bay of Bengal, is the abode of a few primitive dwindling
aboriginal human races and natural habitat of several lesser known endemic plant species. The favorite food of
Shompens is the fruits of screw pines (Pandanus andamanensis Kurz) and honey from wild bees. The tropical
rain forests in the Great Nicobar Island have several Malesian floristic elements of economic value. Hornstedtia
fenzlii (Kurz) K. Schum. (Ammomum fenzlii Kurz) of the Zingiberaceae family is one of such species exclusively
used by the Shompens as a bee repellant source for their honey collection. They chew the plant parts and spit out
the sap filled in the mouth as coarse spray on the bee hives which tranquilize the honey bees and protect
them from bee stings.

Hornstedtia fenzlii (Kurz) K. Schum was selected for conservation and bioprospecting by the Tropical
Botanic Garden and Research Institute (TBGRI). The in vitro propagation of the species is a pioneering attempt.
The shoot tip and auxillary bud from young shoots were cultured on Murashige & Skoog medium variously
supplemented with 6- benzyladenine (BA) and naphthaleneacetic acid (NAA), indole-3-acetic acid (IAA). A
1 1
combination of 2mglˉ BA and 0.05 mglˉ NAA induced high frequency (77%) formation of up to 5 shoots with
well developed roots from each node in 8 weeks. Repeated subculturing of the shoots at 6 week intervals in
combination with 1mglˉ1BA and 0.02 mglˉ1NAA enabled mass multiplication of shoots without any evidence of
decline. The oil extracted from the two different parts of the plant (rhizome and leaf) showed significant variation
in both quantitative and qualitative aspects. The rhizome oil (0.02%) is yellowish whereas leaf oil (0.4%) is
colourless; the oil from both parts exhibited a pungent odor. Qualitative analysis of the oil samples on Gas
Chromatography - out of the total 98% components in the rhizome, 4 constituents were identified (eugenol-
14.864%,geraniol-29.419%,inalool-18.673% and methyl chavicol- 41.094%). Twenty four essential oils
including p- cymene, linalool and eugenol have proved to be effective repellants against worker bees and
mosquitoes, therefore, Hornstedtia fenzlii possesses both repellent and tranquillising properties.

As far as Hornstedtia germplasm is concerned, in vitro or conventional propagation using either rhizome
buds or rhizome cuttings is lacking. Thus our major objectives are to establish a suitable protocol for
conservation and development of a natural oral product from prioritized components of essential oil possessing
insect (blood sucking flies) repellent properties for sustainable utilization.

BSD-2008 GC-09

Production of biosurfactant from industrial wastewater using biotechnological

approach and its environmental applications
Asha A. Juwarkar*, Ravi Sharma, T. Chakrabarti and S.K. Singh

*Environmental Biotechnology Division, National Environmental Engineering Research Institute (NEERI),

Nehru Marg, Nagpur-440020, India
Tel No. +91-0712-2249764 Fax No. +91-0712-2249900

Surfactants are chemically synthesized surface-active compounds and are widely used in the
pharmaceutical, cosmetic, petroleum, food industry and several environmental remediation processes. But these
are toxic and either slow or non-biodegradable and persist in the environment for long, causing harm to the
ecosystem. Biosurfactants are microbial biomolecules, derived from biological sources, and like synthetic
surfactants, exhibit characteristic physico-chemical properties and can be a substitute to chemical counterparts.
Biosurfactants have several advantages over chemical counterparts, that include biodegradability, non-toxicity
and environment-friendly nature which favours their commercial production as specialty chemical.

NEERI isolated Pseudomonas azotoformans (BS2) (MTCC 7176) from an oily sludge of Mathura
Refinery, UP, India for biosurfactant production. The surface and interfacial tensions of aqueous solutions of the
biosurfactant compared well with the known synthetic surfactants and detergents. The reduction in surface and
interfacial tension of water and water:hexadecane mixture by the biosurfactant was 27 dynes/cm and less than 1
dyne/cm, respectively, which were at par with surface and interfacial tensions of the commercial surfactants and
detergents tested. The biosurfactant emulsified the crude oil and pesticides similarly to synthetic surfactants. The
detergency test performed with the biosurfactants showed high cleansing action of the biosurfactant comparable
to alpha-olefin sulphonate (chemical surfactant). The said microbial strain utilized the industrial wastes, such as
diluted distillery waste, whey wastes, food processing wastes and soybean oil as substrates, in batch
fermentation system and successfully produced the biosurfactant of good yield. Production of biosurfactant
from industrial wastes can be a cost effective process at industrial scale.

Biosurfacants are further reported to act as soil washing agents and mobilize metals off the soil and thus
decontaminate the soil. Biosurfactant, which is identified as rhamnolipid, has been demonstrated for
decontamination of metals like cadmium, chromium, arsenic and lead from contaminated sites. Promising
results have been obtained for lead, arsenic, cadmium and chromium removal from soil after four to five
sequential washings using biosurfactant at laboratory level.

The results have shown that biosurfactant can be produced economically by using industrial wastes thus
minimizing their cost of production. The biosurfactant are potential substitute to hazardous chemical surfactants
as demonstrated by emulsification and detergency tests.

BSD-2008 GC-10

Studies on phytochemicals from Abutilon pannosum (A. pannosum) (Kasali)

Eajaj Pathan, D. P. Nerkar and R. K. Bhadekar

Dept. Microbial Biotechnology, Rajiv Gandhi Institute of IT and Biotechnology, Bharati Vidyapeeth
University, Katraj, Pune 411046, India

The genus Abutilon belongs to the family Malvaceae. The plants belonging to this genus are herbaceous
or shrubby and abundantly found as a weed in different parts of India. Different species of Abutilon have been
studied viz. A. hirtum, A. indicum, A. grandifolium etc. The extracts of different parts of the plant (seeds, roots,
leaves, flower petals etc.) have shown anticancer, diuretic, antipyretic, anti-inflammatory activities. Here we
report antimicrobial activity of the roots of another species of Abutilon i.e. A. pannosum (common name: Kasali)
found in Maharashtra. Acetone extract of dried and powdered roots was obtained using soxhlet apparatus. It was
concentrated using rotavapour and used at a concentration of 8.00% (w/v) in acetone. It was tested for
antibacterial activity against various Gram negative and Gram positive bacteria including Escherichia coli,
Salmonella typhimurium, Klebsiella, Pseudomonas aeruginosa, Staphylococcus aureus by disc diffusion
method. The concentration of 0.8 mg showed in vitro antibacterial activity.

In another study we have used water extract of A. pannosum dried and powdered roots. The results of
antibacterial activity of acetone extract and water extract were found to be comparable (except for that of
Pseudomonas) which is very advantageous in further cytotoxic studies. The water extract was used at a
concentration of 12.00% (w/v) in water by well diffusion method. In both the studies increase in the diameter of
inhibition zone was obtained with increase in concentration of extract.

In case of A. persicum and A. striatum, 50% ethanolic extract of plant (excluding roots) were devoid of
antibacterial activity. The alcoholic extracts of roots of A. hirtum were also found to be devoid of antibacterial
activity. However ethanolic, methanolic and hexane extracts of roots of A. indicum have been reported to be
active against different fungi. Preliminary chemical studies on the roots of A. indicum have revealed presence of
tannins, saponins, flavonoids, steroids, terpenes, alkaloids and phenolic compounds. Our results of antibacterial
activity using acetone extract or water extract of A. pannosum roots seem to be very promising in comparison to
other species of Abutilon. Further work on the pathogenic organisms from hospital acquired infections will
definitely help to develop a suitable drug. The studies on chemical profiling of different extracts of A. panosum
and their cytotoxic effects are in progress.

BSD-2008 GC-11

Synthesis and characterization of esters for biolubricant applications

Cecilia Orellana-Coca, Dietlind Adlercreutz and Rajni Hatti-Kaul

Department of Biotechnology, Lund University, Box 124, SE-221 00 Lund, Sweden

Tel: int+46-46-222 3426;

Lubricants constitute a big market, about 127000 t being consumed in Europe in 2006. They are used
primarily for decreased friction and wear, the main consumption being in mechanical industry as hydraulic oils
and chain saw oils, and in automotive industry. Mineral oils are the most commonly used today. They consist
predominantly of hydrocarbons along with some sulfur and nitrogen containing compounds and traces of a
number of metals. There is an increasing demand for environment-friendly lubricants, particularly in areas
where they come into contact with water, food or people. Biolubricants based on vegetable oils are available but
have a drawback of poor cold-flow properties and poor oxidation stability at high temperatures. Modification of
fatty acids is being done within Greenchem research programme to provide good cold properties for use at sub-
zero temperatures in Nordic countries, while maintaining the biodegradability of oils and the compounds.

Esters from fatty acids (with chain length of C5-C18) and different alcohols (especially polyols) have
been synthesized chemically, purified and characterized with respect to lubricant properties. Product analysis
has been done by GPC-HPLC and LC-MS. The melting point of the product components is determined by
differential scanning calorimetry (DSC). A correlation between the melting points of individual product
components and the melting point and pour point of the total product mixture is under study.

Acknowledgement: The study is supported by funding from MISTRA (The Foundation for Strategic
Environmental Research).

BSD-2008 GC-12

Antimicrobial properties of Fresh and stored Banarasi aonla extracts

M.Talib *, V.S.Sapkal**, R.S.Sapkal**

*Shri Shivaji Science College Amravati-444601

**Deptt of Chemical Technology, Sant Gadge Baba Amravati University, Amravati-444602

The aonla extracts were prepared from unblanched and blanched Banarasi aonla cultivar. Antimicrobial
properties of fresh and 3.5 months old extracts were studied and tested against bacteria viz. Escherichia coli,
Klebsialla pneumoniae, Bacillus cereus, Salmonella paratyphi A., Enterobactter aerogens, Proteus vulgaris,
Proteus mirabilus, Bacillus thuringensis, Streptococcus pyrogens, Bacillus subtilis, pathogenic yeasts viz.
Candida albicans, C. blanki, C. glabrata , C. crusei , C. tropicalis and molds viz. Cladospora, Trichoderma
viridie and Aspergillus niger using cup plate method. Antimicrobial properties of blanched aonla extract were
found to be lower than that of unblanched aonla extract. Antimicrobial activity of unblanched and blanched
Banarasi aonla extract was found to be reduced on storage. The loss of antimicrobial properties against bacteria,
yeast and mold was found to vary with microbial species. More than 40% activity loss was observed in
unblanched aonla extract against Enterobacter aerogenes, Escherichia coli NCIM 2066, Proteus mirabilis and
Bacillus subtilis than in blanched aonla extract against Enterobacter aerogens , Streptococcus pyrogens, and
Bacillus subtilis. Unblanched aonla extract showed <10% activity loss against Candida crusei , while blanched
aonla extract showed very high activity loss of 35-37% against Candida albicans and Candida crusei .

Neither aonla extracts showed any significant loss in activity against Trichoderma viride. Greater than
50% activity loss against Bacillus subtilis. Fusarium solami was inhibited by unblanched aonla extract without
any loss in activity. Greater loss of antibacterial activity was observed than antifungal activity on storage of both
the aonla extracts.

BSD-2008 GC-13

Purification and characterization of glycerol stimulated extracellular lipase

from novel Trichosporon sp. MSR 17
Sonal Tonger*, S.Kumar and Rani Gupta

Department of Microbiology, University of Delhi, South Campus , Benito Juarez Marg , New Delhi- 400 021

A glycerol stimulated extracellular lipase (EC from novel dimorphic yeast Trichosporon
sp.MSR17 isolated from petroleum sludge has been purified and characterized. The strain showed 99%
homology with Trichosporon loubieri based on D1 D2 sequence. The enzyme was purified stepwise with ultra
filtration and hydrophobic column chromatography (octyl-sepharose). The molecular weight of purified
enzyme was determined by SDS PAGE. Further, the purified enzyme was characterized on the basis of substrate
specificity (triglycerides, methyl esters and pNP esters), pH and temperature optima, effect of metal ions and
solvent stability. Enzyme showed maximum hydrolytic activity towards mid and short chain fatty acids, pNP
myristate and methyl myristate. Enzyme showed a maximum activity at 40º C and pH 9.0 with 50% residual
activity at 50º C and 60º C. Enzyme activity was found to be enhanced to150% in glycerol after incubation of 30
min at room temperature whereas It showed poor solvent stability towards acetone, methanol, ethanol,
isopropanol and chloroform. Solvent stability was increased by lyophilization of enzyme with different
polymeric substances and dextrin was found to be most suitable. Using 1% dextrin, enzyme stability in different
organic solvents was tested and enzyme showed 50% stability in case of acetone, hexane and ethanol and 20% in
case of isopropanol.

BSD-2008 GC-14

Characterisation of Biosurfactant producing yeast isolates from petroleum

sludge and production of a thermostable glycolipid from Trichosporon asahii
MSR- 54
Richa Sharma, Kashika Paliwal, S.Suresh Kumar, Rani Gupta

Department of Microbiology, University of Delhi, South Campus , Benito Juarez Marg , New Delhi- 110 021

More than fifty yeast strains were isolated from petroleum sludge areas of Delhi, India using kerosene
enrichment method. Out of these, ten biosurfactant producing strains were selected on the basis of their ability to
form halo on hexadecane agar plates. Molecular identification of these revealed them to be Trichosporon sp.,
Yarrowia sp. and Candida sp. Comparative evaluation of their biosurfactant producing ability was done using
hemolytic assay, hydrophobic interaction chromatography (HIC), salt aggregation test (SAT), surface tension
measurements and emulsification index (EI). Overall results of these tests revealed Yarrowia lipolytica MSR 84
and Trichosporon asahii MSR 54 to be the best biosurfactant producers with a maximum surface tension
reduction of 32% and 28% respectively. Among them the biosurfactant produced by Trichosporon asahii MSR-
54 formed a highly stable xylene-water emulsion with an E-24 value of 100%. After solvent extraction this
extracellular water-soluble biosurfactant was identified as a glycolipid. Its production was favored by high
carbon (2%), low nitrogen (0.05%) and low phosphorous (0.1%) in the oil medium over a wide pH range of pH 2
to12 between 20oC to 45 oC and 50 to 200 rpm. This is a highly stable biosurfactant as it tolerated boiling
temperatures and ionic strength of 30% NaCl. This is the first report of production of a biosurfactant from
Trichosporon asahii.

BSD-2008 ENG-01

Renewable Energy as an Industrial Fuel

1 1 2 2
K.K.Meher D.R.Ranade , Sumedh Bapat and Deepak Shah

1. Agharkar Research Institute, Pune;

2. GreenLeaf Renewable Energy Pvt. Ltd.,Pune

Sustainable development certainly needs renewable energy technologies such as bio-diesel,

biomethanation and bio-fuels. India has undertaken a massive program on bio-diesel production. Currently the
focus is on the utilization of available non-edible oil seeds for the production of bio-diesel. This is going to
generate huge quantities of glycerol and de-oiled non-edible cakes as a byproduct. Most of these seed cakes are
used as organic manure. However, by doing this, carbon content in the cake gets converted to CO2 through
microbial degradation.

Green Leaf Renewable Energy Pvt. Ltd, a sister concern of Ashish Cans and Containers P.Ltd (ACCL),
Talegaon, under the technical guidance of Agharkar Research Institute (ARI) has developed a high rate
biomethanation process for de-oiled castor seed cake. The process uses a microbial consortium containing
appropriate concentrations of anaerobic bacteria and methanogenic archaea required for efficient degradation of
de-oiled cake. Castor (Ricinus commuinis) is an important cash crop and is grown widely in many states. India is
the second largest country in the world producing castor seed. After oil extraction, nearly 70% of the seed
remains as de-oiled seed cake. This cake is rich in organic matter but is non-edible due to the presence of an
alkaloid, ricin.

The ACCL-ARI process, after bench scale and pilot scale experimentation, is now being implemented in
the field. The process generates minimum 425 lit of biogas/kg cake/day with 77% methane. A year long study has
shown that the process is very stable, provided the microbial dynamics is well controlled. The digested sludge
being rich in NPK and low carbohydrate is useful as organic manure.

This technology is currently used by the company. The biogas produced is used as a fuel in furnace,
replacing ~ 50% of the daily requirement of diesel. The furnace is used in the manufacture of LDPE cans. The
process is an eco-friendly, green technology. Now, ACCL is commercializing this technology under the name of
Green Leaf Technologies Ltd.

BSD-2008 ENG-02

Biohydrogen production: the dark fermentation approach

Priyangshu M Sarma*, Anu Sudhakaran, Navarun Varma, Ajoy Mandal and Banwari Lal

Microbial Biotechnology, The Energy and Resources Institute (TERI), Lodhi Road,
New Delhi, 110003

Energy is vital to global prosperity, but dependence on fossil fuels as our primary source contributes to
global climate change, environmental degradation and other health problems. The awareness of environmental
concerns and energy insecurity has led to an increasing interest on research for alternative sources of energy. In
this context, hydrogen holds a promising option as an energy carrier of the future. The current industrial
methods, for producing hydrogen, like steam reformation of natural gas or splitting water with electricity
release carbon dioxide and other greenhouse gases as by-products and hence do not qualify as alternative
renewable energy sources.

In the scenario where there is a worldwide collaborative effort to curb down emissions, attention is
drawn towards microbial processes, and biotechnological interventions based on these microbial systems that
could lead to clean and renewable sources of hydrogen. Hydrogen production by using microbial strains
traditionally targeted the cyanobacterial cultures and through biophotolysis (hydrogen production through
photo-synthetically splitting water). However, a review of more recently published works led the focus on
fermentative approach of biohydrogen production, due to the dual benefits of energy production as well as waste
reduction. The important challenges faced in this approach are low hydrogen conversion efficiency and unstable
hydrogen production. Another limiting factor of biohydrogen production pertains to the enzymes
(hydrogenases) as they are sensitive to oxygen.

Microbial Biotechnology area of TERI in a recent corporate sponsored project attempted to explore and
isolate microbial cultures that have biocatalysts which has high turnover rate and higher degree of oxygen
tolerance and can efficiently produce hydrogen in a continuous manner. The approaches that we are currently
exploring are the dark fermentation mode. The sources of hydrogen producing microbes are targeted from river
bed sediments, sewage sludge, fruits and vegetable wastes and other industrial wastes. The initial screening of
strains indicated the capability of the selected strains to use various carbons sources and wastes to generate
hydrogen. The upscaling of the process is currently under process.

BSD-2008 ENG-03

Energy and Ethanol : Refueling India An Overview

Amit Shrivastava

Sagar Institute of Research & Technology, Bhopal (M.P)

Bioethanol is a fuel-grade ethanol made from trees, grasses, and waste materials. It represents a
sustainable substitute for gasoline in today's passenger cars. Modeling and design of processes for making
bioethanol are critical tools used in the US Department of Energy's bioethanol research and development
program. The authors have used such an analysis to discover new directions for research and to help them
evaluate the potential for bioethanol to achieve commercial success. This paper provides an update on their latest
estimates for current and projected costs of bioethanol. These estimates are the result of very sophisticated
modeling and costing efforts undertaken in the program over the past few years. Bioethanol could cost anywhere
from $1.16 to$1.44 per gallon, depending on the technology and the availability of low cost feedstocks for
conversion to ethanol. While this cost range opens the door to fuel blending opportunities, in which ethanol can
be used, for example, to improve the octane rating of gasoline, it is not currently competitive with gasoline as a
bulk fuel. Research strategies and goals described in this paper have been translated into cost savings for ethanol.
Their analysis of these goals shows that the cost of ethanol could drop by 40 cents per gallon over the next ten
years by taking advantage of exciting new tools in biotechnology that will improve yield and performance in the
conversion process.

The paper highlights:

 Ethanol as a future fuel,

 Effect of ethanol on economy,

 Types of ethanol fuel for eg: E85, E10 etc.

 Current status of ethanol in India- An Overview

BSD-2008 ENG-04

Fuel alcohol production from agricultural residues

Alemzadeh, R. Roosta Azad and M. Vossoughi

Chemical and Petroleum Engineering Department, Sharif University of Technology, 11365-6891, Tehran, Iran,

The bioconversion of abundant and renewable cellulosic biomass into ethanol as an alternative to petroleum is
gaining importance due to the realization of diminishing natural oil and gas resources. Agricultural and forest
plant residues are an abundant and renewable source of sugar substrates that could be fermented to ethanol. A
thermochemical treatment of biomass in which both cellulose and hemicellulose are hydrolyzed to soluble sugar
is necessary before yeast fermentation. After thermochemical treatment, cellulase enzymes must be introduced
in the system to hydrolyze any remaining cellulose. The simultaneous saccharification and fermentation (SSF),
is a method which converts agricultural residue to ethanol. In this investigation, pretreatmrnt of rice husk was
effected under acid, alkaline and mixed treatment, in which alkaline pretreatment showed higher biomass
degradation. Production of enzyme complex(cellulase) was studied by fungal strain cultivation on pretreated
biomass, using Trichoderma reesei on alkali pretreated rice husk. The biomass was then subjected to an SSF
process, using the cellulase enzyme complex extracted from fungal culture and Saccharomyces cerevisiae yeast
cells in batch scale fermentation system. Ethanol production was evaluated by changing various parameters
such as: substrate and enzyme concentration, corn steep liquor addition and inoculum percentage under SSF

BSD-2008 ENG-05

Analysis of Growth and Lipase Production of an Indigenous Candida sp. for

Biodiesel Production
Ritam Sarkara, Ankush Madaana, Anju Chadhaab*
a b
Department of Biotechnology & National Center for Catalysis Research ,
Indian Institute of Technology Madras, Chennai 600 036

Lipases are used for transesterification reactions and there is a constant search for robust lipases which
can be employed gainfully for making biodiesel from oil [triglycerides]. We have isolated from local soil
samples, a Candida sp. which is capable of producing an extracellular lipase that can transesterify the local, non-
edible Pongamia oil to pure biodiesel. This work presents optimization of i) growth of the organism ii) lipase
production and iii) protein. The growth of this organism in YMB [growth medium] was investigated with
varying amounts of the inoculum for a period of 24 hrs. The organism is in the lag phase for the first 4 hrs with 2-
6% inoculum and the maximum growth was achieved at 24 hrs for the 4% inoculum. As it is known that cell
th th
viability is best in the exponential growth phase [6 - 16 hour], investigations on protein, growth and lipase
production were carried out in production medium with inoculum grown from 6th -16th hrs. Production media
containing Tween-20 and Olive oil indicate that the growth was maximum at the 12th hr for the 6th hr inoculum
while the highest specific activity of enzyme (U*/mg protein) was found to be at 24 hrs for the 12th hr inoculum.
The growth and production of lipase was found to be the highest when the initial pH of the media was 8. The
correlation of enzyme activity as monitored by p-NPP assay, olive oil assay and transesterification of Pongamia
oil for the production of Biodiesel will be discussed.

*Unit Enzyme: One Unit of the enzyme activity is defined as the release of one μmole p-NP per minute under
standard assay conditions.

BSD-2008 ENG-06

Thermoethanologens: creating future fuel from agro waste

Sachin Kumar and D. K. Adhikari

Biotechnology Area/ PEACBD, Indian Institute of Petroleum, Dehradun (UK) INDIA

Due to growing population and increasing oil consumption, the government policies in India have
targeted at reducing petroleum dependence. With the rise in energy costs, India has also committed the utilization
of a large share of its lignocellulosic for bio-ethanol production. Bagasse, as lignocellulosic biomass, can be used
to produce bio-ethanol. Lignocellulosic materials consist of cellulose and hemicellulose which are formed by
polymerization of sugars. These can be liberated by acid or enzymatic hydrolysis of the polymers. The recent
development in acid and enzymatic hydrolysis of cellulose and hemicellulose to fermentable sugars (glucose
and xylose) may possibly lead to commercially viable production of ethanol from the vast and renewable
quantities of cellulose available on the earth.

In the present paper we have studied the hydrolysis of bagasse by dilute and concentrated acid and
fermented the hydrolysate by thermophilic Klyuveromyces sp. IIPE453 at 50°C. The overall yield of fermentable
sugars in dilute acid treatment was 62% w/w of total cellulose and hemicellulose in bagasse. Bagasse was treated
with 2% v/v H2SO4 at 121°C and 15psi for 1½hr. The overall yield of fermentable sugars in concentrated acid
treatment was 77% w/w of total cellulose and hemicellulose in bagasse with 70% w/w H2SO4 at 100°C and
normal pressure. The hydrolysate obtained from dilute acid treatment contains 38% glucose whereas from
concentrated acid treatment contains 80% glucose. The sugars obtained from bagasse hydrolysis were used for
bio-ethanol production. Fermentation of hydrolysate containing glucose and xylose as reducing sugars was
conducted at 50°C using thermophilic yeast IIPE453 with overall 52% of theoretical yield and 0.6gl-1h-1
productivity in a batch process.

BSD-2008 ENG-07

Biodiesel production: exploration towards option for bioresource technology

Bhagyashri G. Raut, Arti S. Shanware and Sudhir U. Meshram

*P.G. Department of Microbiology, L.I.T. Campus, Rashtrasant Tukdoji Maharaj

Nagpur University, Nagpur-440033(M.S.) India.
Rajiv Gandhi Biotechnology Centre, L.I.T. Campus, Rashtrasant Tukdoji Maharaj
Nagpur University, Nagpur-440033(M.S.) India.

Every year India imports 67%of its petroleum requirements from other countries, which involves a huge
amount in foreign exchange. A replacement of even 5% of this amount could help India save crores of rupees in
foreign exchange annually. Development of a renewable energy source like biodiesel is need of the hour.
Biodiesel is the monoalkyl ester of long chain fatty acids derived from renewable lipid feedstock such as
vegetable oils or animal fats.

In the present investigation, five different types of seeds i.e .Karanja, Soybean, Mustard, Jatropha and
Castor were used as the feedstock for biodiesel production. Oil was extracted from these plant seeds using a
soxhlet extraction assembly. This oil was then trans-esterified using potassium hydroxide as catalyst & methanol
in mass proportion of 20 % of oil in a reactor equipped with condenser & thermometer. The reaction mixture was
then kept overnight. Two layers were then recovered from reaction mixture i.e. upper layer of biodiesel & lower
layer of glycerin.

Besides being a renewable resource, environmental concerns are a greater concern than economic
benefits. The eco-friendly nature of this fuel far outweighs the economic constraints associated with the
manufacture of biodiesel. The higher cost of biodiesel as compared to conventional diesel is an area for future
work. The role of government subsidy, on environmental grounds, is another avenue, decreasing the cost of this
promising renewable energy resource.

BSD-2008 ENG-08

Bio-processing of coir dust for the production of methane

V. A.Selvi, Banerjee, R, Shalini J.M , Ram.L.C, Masto R.E

Environmental Management Division, Central Institute of Mining & Fuel Research, Digwadih campus,
(CSIR), PO.FRI. Dhanbad 828 108, Jharkhand
Microbial Biotechnology and Down Stream Processing, Agricultural & Food Engineering Department,
Indian Institute of Technology, Kharagpur, West Bengal, India

The increasing expansion of agro-industrial activity has led to the accumulation of a huge amount of
lignocellulosic residues all over the world including India. Due to increase in demand of energy, attempts are
being made all over the world to use alternative feedstocks such as agroindustrial wastes for biogas generation.
With the idea of using these wastes for an economically viable purpose, the present work is aimed at the
biomethanation of coir dust in the laboratory digester. Pretreatment (physical, chemical and biological)
followed by bio-methanantion was carried out for the production of methane from agro-wastes in a batch
digestion process. Coir pith - dust from husk of Cocos nucifera (L) was used as the substrate. Coir dust with
different particle size like 2 mm, 1mm, 18 mesh, 60 mesh and 72 mesh were pretreated by physical (autoclaving
up to 15 psi, heating at 65ºC and 130ºC, boiling for 30 min and milling), chemical (HCL: 0.1-0.2% and NaOH
:0.1-0.2%) and biological method with white rot fungus, Pleurotus djamor. Anaerobic digestion was carried
out in a laboratory model digester of 10 l capacity using different compositions of cow dung and coir in
percentage as follows: T1-20:80, T2-40:60, T3-60:40, T4-80:20, T5-100. Biogas production was studied at 10,
20, 30 and 40 day hydraulic retention time. Methane content was analyzed using gas chrotomograph at 2-day
interval. The GC with Porapak -Q column, nitrogen as carrier gas (40 ml/min) with thermal conductivity at an
oven temperature of 60ºC, injector 100ºC and detector 100ºC were used. Among the different pre-treatments,
fungal treated coir (60 and 72 mesh particle size) gave the highest yield of methane. Gas production in untreated
(raw) coir pith was less due to high lignin content (30%). There was a gradual increase in the methane content of
biogas, which rose from 21 percent on the 6th day to 61 percent on 35th day of biogas generation.

BSD-2008 ENG-09

K. Singh

Central Salt and Marine Chemical Research Institute, Bhavnagar.

The chief source of energy is fossil fuel. The resources of this fuel are finite and will last for few more
decades only. Thus, there is an urgent need to find alternative source of energy, which could be renewable. There
are very few options available to substitute/replace fossil fuel, such as methanol, vegetable oils, esters of
vegetable oils & fats etc. Methyl esters of fatty acids (vegetable oils) commonly known as bio-diesel is one most
reliable alternative fuel to replace fossil fuel because bio-diesel is renewable and it's fuel properties such as
octane number, energy content, viscosity and phase changes are similar to those of petroleum-based diesel fuel.
Moreover, biodiesel is essentially sulfur free and the engines fueled by biodiesel emit significantly fewer
particulates, hydrocarbons, and less carbon monoxide than those operating on conventional diesel fuel. Bio-
diesel is obtained by trans-esterification of vegetable oils by conventional chemical methods and by bio-
conversion using enzymes. Transesterification of vegetable oils has been successfully achieved by alkali-
catalyzed process and by acid catalyzed process. Alkali catalyzed trans-esterification process requires vegetable
oil of very low FFA value and moisture free because naturally occurring FFA in oil lead to soap formation, which
lowers the yield of biodiesel and renders the separation of bio-diesel and glycerol difficult. Transesterification of
vegetable oil by acid catalyzed process does not require highly pure oil, however requires extreme reaction
conditions and takes longer time to complete the reaction. Modification of oils and fats using biocatalysts would
overcome several problems associated with chemical catalysis. Enzyme bio-catalysis process has several
advantages over conventional chemical catalysis process such as moderate reaction conditions, stereo-
specificity of substrate, no side reaction products etc. Enzyme bio-conversion of oils / fats using lipase can
handle high FFA feed stocks and no by-products would form thereby enhancing the overall yield.

Lipase from different sources was immobilized on polysulfone, and polyethersulfone membranes by
ultrafiltration and covalent bonding method. The amount of lipase immobilized was estimated by Lowry method
and effect of various process parameters was studied. Immobilizing enzyme on membrane by ultrafiltration is
remarkably better method in comparison to covalent bonding. Transesterification of vegetable oil was studied on
enzyme immobilized membranes and effect of various process parameters viz., pH, time, substrate
concentration and enzyme concentration was determined.

BSD-2008 ENV-01

Optimization of caffeine degradation by immobilized cells of Pseudomonas

putida NCIM 5235 using various polymer matrices
Devarai Santhosh Kumar and Gummadi Sathyanarayana Naidu

Department of Biotechnology, Indian Institute of Technology-Madras, Chennai- 600 036, India

Tel: 91-44-2257-4114; Fax: 91-44-2257-4102

Caffeine is one of the major toxic compounds generated by solid wastes in the coffee and tea industries. The
concentration of caffeine in effluents from these industries is high. The waste is ultimately dumped into the
surrounding water bodies and soil, thereby causing serious ultimatum to health problems. In order to solve this
problem, decaffeination becomes a necessary step in coffee and tea processing. Biodecaffeination using
microbes and microbial enzymes is much more convenient and cheaper than the conventional chemical
processes of decaffeination. In this context, immobilization of Pseudomonas sp entrapped with different
synthetic polymer matrices viz. calcium alginate, agarose, agar-agar, polyacrylamide and polyvinyl alcohol was
carried out to find the best matrix in terms of higher caffeine degradation rates, stability and reusability. It was
observed that the cells entrapped in agar-agar matrix showed the highest rate of caffeine degradation (0.08 g/l/h)
with initial caffeine concentration of 1.2 (g/l). The rates of degradation of caffeine for calcium alginate, agarose,
polyacrylamide and polyvinyl alcohol were (g/l/h) 0.056, 0.054 ,0.014 and 0.07 respectively. Further,
optimization of three parameters of the agar-agar matrix i.e. size of bead (mm), agar-agar % (w/v) and cell
concentration (g/l) was carried out using central composite design. From the mathematical model developed on
the results obtained, the optimum values for the above mentioned parameters were determined to be 2.4 (mm),
2.8 % (w/v) and 6.73 (g/l) enhancing the caffeine degradation rate to 0.17 g/l/h with initial caffeine
concentration of 5 (g/l) at pH-7.0 and 30 C. The findings from this study can be potentially beneficial in
development of an efficient bioprocess for decaffeination on an industrial scale.

BSD-2008 ENV-02

Biodissolution of low-grade Malanjkhand Ore by Bacillus stearothermophilus

S.Singh, L.B.Sukla and B.K.Mishra

Biomineral Department, Institute of Mineral and Material Technology (CSIR),

Bhubaneswar 751013, Orissa, India.

Thermopiles are actively prevalent in hot springs and have the potential to grow and sustain at temperatures
above 70-80oC. Use of moderately thermophilic bacteria and thermophilic archaea is gaining significant
attention for commercial applications due to their higher oxidation rates in comparison to mesophilic bacteria.
Large quantities of the metals are embodied in the low-grade ores and mining residues which can be recovered
efficiently using thermophilic bacterial strains. The present study was initiated to determine the feasibility of
extraction of copper using thermophilic heterotrophic bacterial strain isolated from hot water spring at Atri,
Orissa, India. The bacterial strain was identified as Bacillus stearothermophilus (MTCC No. 8051).
Malanjkhand low-grade copper ore contains 0.3% Cu, in which, the major copper bearing mineral is
chalcopyrite associated with other minerals present as minor phase. The thermophilic bacterial strain was
adapted to 50 C and the bioleaching experiments were conducted in 250ml conical flask containing 10% ore
(w/v) in 90ml mineral salt media (yeast extract) and 10% (v/v) of inoculum. The pH was maintained at 6.5 and
agitated continuously in a rotary shaker at 150 rpm and incubated for a period of 14 days at 500C. Simultaneously,
a control was established by addition of 10 ml of mercuric chloride as bactericide. The leaching ability of
thermophilic heterotrophic bacterial strain showed maximum leaching of 43% of copper solubilized at pH-6.5
with 10%(w/v) at 50 C after 14 days of incubation.

BSD-2008 ENV-03

Bioleaching of copper from chalcopyrite concentrate by moderately

thermophilic acidophilic mixed consortium
S.Mohapatra*, B.D.Nayak, L.B.Sukla and B.K.Mishra

Biomineral Department
Institute of Mineral and Material Technology, Bhubaneswar 751013, Orissa, India

In the present study, an attempt was made to leach copper from chalcopyrite concentrate using thermophilic
acidophilic chemolithotrophic mixed consortia. The thermophile was isolated from lagoon material collected
from a copper mine site at Khetri, India. The moderate thermophile was adapted at 50 C and the bioleaching
experiments were conducted in 250ml conical flask containing 2, 5 and 10% (w/v) in 90 ml of 9K+ media and
10% (v/v) inoculum of thermophile consortia. The pH was maintained at 2 and the slurry was agitated
continuously in a rotary shaker at 150 rpm for a period of 42 days, at 50 C. Simultaneously, control experiments
were conducted for all the sets of experiments by the addition of 10 ml of mercuric chloride as bactericide.
Representative samples were taken at regular interval for analysis of copper through Atomic Absorption
Spectrophotometer (AAS). The maximum extraction of copper was observed at 2% pulp density (94.12 %)
within 42 days at pH 2 and least extraction in case of 10% pulp density, (78.99%) copper, respectively.

BSD-2008 ENV-04

Beneficiation of Iron ore by Removal of Alumina and Silica using Bacillus,

Pseudomonas and Aspergillus Sp.
M. Mishra, N.Pradhan, B.D. Nayak, L.B. Sukla, B.K. Mishra

Department of Biominerals, Institute of Minerals and Materials Technology (CSIR)

Bhubaneswar-751013, Orissa, India

Bio-beneficiation is considered as one of the most eco-friendly, promising and revolutionary technique. It
involves removal of undesirable mineral components from an ore through interaction with microorganisms
which bring about their selective removal and thereby, enriching the desired mineral constituents in the solid ore
matrix, mediated by a number of surface chemical and physiochemical phenomena. High percentage of alumina
present in iron ore has an adverse effect on reducibility, coke rate, productivity and blast furnace operation. To
meet the demand of steel plants it is required to reduce alumina percentage to <2.5% in iron ore. Microorganisms
degrade the alumino-silicates by using the soluble low molecular weight metabolites like organic acids, amino
acids etc. These metabolites dissolve metals from minerals by displacement of metal ions from the ore matrix by
hydrogen ion and formation of soluble metal complexes and chelators. Studies were carried out on the removal
of alumina and silica from iron ore containing (%) Fe2O3 84.4, Al2O3 7.18, SiO2 7.53, and Fe 59.05, with the help
of four bacterial isolates and two fungal strains. The ore was analyzed by high resolution synchrotron based X-
ray diffractometer and optical microscopy. Bromfield medium was used for the beneficiation studies. Bacillus
circulans showed the maximum removal of alumina and silica i.e.15.95% and 18.6% respectively at 20% pulp
density, 35 C, pH 6.8 at 150 rpm within 10 days. Under same conditions, Pseudomonas aeuroginosa,
Pseudomonas putida, Bacillus polymyxa Aspergillus niger and Aspergillus flavus showed 4.45, 14.71, 10.86,
16.9, 15.4% alumina and 3.03, 11.5, 14.53 17.95, 17.82% silica removal respectively.

BSD-2008 ENV-05

Microbial Solubilisation of Phosphorus from insoluble Tricalcium phosphate

N.P. Marhual, N. Pradhan, N.C Mohanta, L.B.Sukla, B.K.Mishra

Biominerals Department, Institute of Minerals and Materials Technology

Bhubaneswar, 751013, Orissa, India

Microbial beneficiation of different industrial and mine waste are gaining importance these days due to non
availability of raw materials. This process is eco-friendly and economical. Efforts are being made to remove
phosphorus from Manganese ore, Iron ore and LD slag. In all these cases, bacteria capable of solubilising
phosphorus are used. In the present study, phosphorus solubilizing bacteria (PSB) are used to solubilize
phosphorus from an insoluble form of phosphate i.e. tri-calcium phosphate (TCP). Different bacterial species
including two Gram ve bacteria like Pseudomonas aeruginosa, Pseudomonas putida and one Gram + ve
bacteria Bacillus sphaericus isolated from agricultural soil were used to see the efficiency of phosphorus
solubilisation. P.aeruginosa, P putida and B sphericus solubilized 323 µg/ml, 197 µg/ml and 179 µg/ml of
phosphorus, respectively, after 5 days of incubation at 35 C and 150 rpm. Total theoretical solubilisation would
have been 997.5 µg/ml (0.5%) tri-calcium phosphates. Concentrations of the bacterial cell also affect the
solubilisation of phosphorus. Phosphorus solubilisation rate was also related to pH decrease caused by the
growth of the bacteria in medium containing glucose as carbon source. PSB convert insoluble phosphates into
soluble forms generally through the process of acidification, chelation and exchange reactions.

BSD-2008 ENV-06

Oxidation of metal sulphides from mine by the novel bacterial strain

Daryush Arabian, Fahimeh Nemati, Rajendra Patil and W. N. Gade

Department of Biotechnology, University of Pune, Pune, India

In general, bioleaching is a process described as "The dissolution of metals from their mineral source by
certain naturally occurring microorganisms" or "the use of microorganisms to transform elements so that the
elements can be extracted from a material when water is filtered through it".

In this study, we have isolated six extreme acidophilic obligate chemolitotrophic bacteria and fungi from
complex Zn and Pb sulphide mine, Yazd, central of Iran that has showed the bioleaching properties. Two of the
isolates, MY1 and MY2, oxidized 65-80 % of ferrous to ferric which ultimately leached Zinc and Lead to about
60 %. The bioleaching process was optimized by varying different parameters viz, pH,
temperature, initial inoculum and concentration of ferrous iron and nitrogen source.
Optimization process showed that the best condition for growth and activity is: pH: 1.4-1.6,
º o
Temperature:30 C-30 C, initial inoculums:5% - 8%, ferrous concentration:300 g/l -20g/l, Nitrogen
concentration 4g/l -3g/l respectively. The optimization condition has increased the leaching capacity of these
bacteria to around 96%. This is a new bacterial strain isolated by us. It has been characterized.

BSD-2008 ENV-07

Bioremediation of hydrocarbon contaminated soil

Gheyrati Arani Leyla, and V.R.Gunale

Department of Environmental Science, University of Pune,

Pune-411007 (INDIA).

It is known that bacteria and fungi are the principal petroleum degrading microorganisms. Therefore, the
study of the petroleum hydrocarbon degradation by filamentous was accomplished fungi to evaluate the
potential of strain, isolated from soil, contaminated with engine and diesel oil. The results of this evaluation
ascertained the utility of the microorganism to degrade petroleum, added as the only carbon and energy source to
a mineral medium. The selected strain was identified as an Aspergillus sp. Degradation tests were subsequently
performed at 30 ºC, in the contaminated soil, working under the following process conditions: 60% w/w of water
holding capacity, periodic aeration, natural pH, with and without additional nutrient supply.

BSD-2008 ENV-08

Limnological Study to Assess the Tropic Status of Drinking

Water Reservoir near Mumbai
Sanyogita Verma, P.R. Chaudhari and S.R. Wate

National Environmental Engineering Research Institute, Nehru Marg, Nagpur 440020

A multipurpose upper Vaitarna Dam serves the dual purpose for power generation and water supply to
Mumbai. All the lakes undergo natural eutrophication process, which results in deterioration of water quality.
With this view present investigation has been undertaken to carry out detailed limnological study to evaluate the
tropic status of lake, the present impacts and possible mitigation measures.

The lake water was found to be clean with pH 7-8.1, low turbidity and low nutrient status. The study
indicated that nutrient loading of the lake was very low due to absence of agricultural activities and dwelling in
the catchment area. Physico-chemical observations were complemented by biological observations. Lake water
quality was observed to be oligotrophic in nature as indicated by low plankton count, dominance of clean water
indicator planktons, Palmer's Pollution Index between 1-4. Shannon Wiener Index was low indicating nutrient

Considering all the above factors it is concluded that there is no indication of occurrence of
eutrophication process and that the lake is in an oligotrophic conditions. With these conditions the lake water
quality will remain unchanged for a long time in near future. However, proper catchment treatment for soil
conservation would be useful to conserve the lake water quality.

BSD-2008 ENV-09

Sisal decortication fibre residues: A novel biofilm carrier in

anaerobic digestion of plant biomass leachate.
1 2 1 1 2
A.M. Mshandete , L. Björnsson , A.K. Kivaisi , M.S.T. Rubindamayugi and B. Mattiasson
Department of Molecular Biology and Biotechnology, University of Dar es Salaam, P.O.
Box 35179 Dar es Salaam, Tanzania. Tel:+255- 22- 4110223; Fax:+255-22- 2410 078.
Department of Biotechnology, Centre for Chemistry and Chemical Engineering, Lund
University, P.O. Box 124, SE-22100, Lund, Sweden. Tel: + 46 -46- 222 -8324;
Fax:+46-46- 222- 4713.

Sisal fibre decortication residue was evaluated as a biofilm carrier in anaerobic digestion
of plant biomass leachate. Three methanogenic biofilm bioreactors configurations were
studied; a sisal fibre decortications residue packed-bed bioreactor, a pumice stones
packed-bed bioreactor and porous glass beads packed-bed bioreactor. The bioprocess
evaluated was the methanogenesis anaerobic digestion of sisal leaf decortications
residues leachate. Process performance was investigated by increasing the organic
loading rates (OLRs) step-wise. The best results were obtained from the bioreactor
packed with sisal fibre decortications residue. It had the highest chemical oxygen demand
-1 -1
(COD) removal efficiencies ranged from 93-80% at OLRs from 2.4-25 g COD L d . It
also had superior biodegradation pattern of individual volatile fatty acids (VFAs). The
degradation of propionate and i-valerate were limiting at higher OLRs. However,
degradation efficiency of propionate was over 50% for the sisal fibre residue bed, even
-1 -1
when the OLR was increased to 25 g COD L d . On the other hand, the degradation of
i-valerate was negative for packed-bed containing pumice stones and porous glass beads
-1 -1
when OLR was increased to 25 g COD L d . This demonstrated microbial inhibition.
The conclusion is that sisal fibre residue, a new biofilm carrier, has proven to be promising
in the intended application and could withstand the variations in load and VFAs
concentrations that can occur in a two-stage anaerobic process. However, it remains to be
tested at pilot scale trials in the context of Tanzania's low technology environment.

BSD-2008 ENV-10

Membrane bioreactor equipped with low cost mesh filter:

application for distillery wastewater
Yamini Satyawali, Malini Balakrishnan

TERI University, Darbari Seth Block, India Habitat Centre, Lodhi Road, New Delhi 110003, India
Tel.: +9111 2468 2100; fax: +9111 2468 2144.

Membrane bioreactors (MBR) are a promising option for wastewater treatment and reuse. The MBR
system integrates biological treatment with membrane filtration using MF/UF membranes that act as a solid-
liquid separator. Thus the MBRs extend the application of membrane filtration from tertiary to secondary
treatment stage. Molasses based alcohol distilleries are one of the most polluting industries generating large
volumes of high strength effluent (spentwash). Distillery spentwash contains melanoidins, which are a class of
high molecular weight polymers with molecular weight between 5000 to 40000 Da. Melanoidins have
antioxidant properties rendering them toxic to many microorganisms involved in wastewater treatment.
Furthermore, the BOD: COD ratio of this effluent is relatively low (0.14) thus making it relatively difficult to

This work presents the results for distillery wastewater treatment in a MBR equipped with mesh filter.
Distillery wastewater, after anaerobic digestion, was used as the feed and the bioreactor was inoculated with
acclimatized municipal sludge. The acclimatization of municipal sludge to 100% distillery wastewater was
carried out for almost 100 days. It was noticed that even after acclimatization, the organic loading had to be
stepped up gradually. The highest organic loading investigated was 5.71 kg COD day-1 m-3 and the system
resulted in a maximum of up to 50% COD removal. Gel permeation chromatography results revealed that the
low molecular weight compounds were mainly degraded. The mesh filter was capable of acting as a good solid-
liquid separator, resulting in almost complete retention of biomass.

BSD-2008 ENV-11

Development of a Partial Nitrification System for Ammoniacal Nitrogen

Removal under DO Limited Condition
Samik Bagchi1, Nisha M. Jhon1, Rima Biswas1, Kunal Roychoudhury2 and Tapas Nandy1

1National Environmental Engineering Research Institute, Nehru Marg, Nagpur 440 020, India
2 PG Dept. of Microbiology, S.K.Porwal College, Kamptee, Nagpur 441002, India
Tel: +91-712-2249763; Fax: +91-712-2249900

Establishment of partial nitrification system under oxygen limiting condition in an up-flow fixed film bioreactor,
as a pretreatment step to anammox process has been investigated in the present study. The first phase of the work
comprised of developing a complete nitrification system. During this phase, the reactor was fed with synthetic
wastewater with initial ammonium loading of 0.03 Kg NH4-N/m .day. Loading rate was increased from 0.03 Kg
3 3
NH4-N/m .day to 0.175 Kg NH4-N/m .day in a phase wise manner over a period of 90 days. DO uptake increased
from 1.70.2 mg/l to 3.00.2 mg/l as ammonia oxidation rate increased from 0.004 Kg NH4-N/m .day to 0.135 Kg
NH4-N/m .day. Nitrate concentration in the effluent was in stoichiometric proportion to the ammonium
oxidation rate, indicating complete nitrification. After the development of stable nitrification during 90 days of
operation (i.e. Phase I), limiting DO condition was created by decreasing the DO concentration to 0.6-0.7 mg/l
(in Phase II). Under DO limited condition, the ratio of NO2-N generated to total ammonia oxidized () slowly
reached more than 0.9 indicating partial nitrification. After 60 days of operation under DO-limited condition
(0.6-0.7 mg/l), a stable partial nitrification system was achieved with 90% of the ammonia getting oxidized into
nitrite. The effluent of this phase had ammonia to nitrite ratio of 1:1.42, which can be suitably used as a substrate
for anammox process.

BSD-2008 ENV-12

Understanding and Developing Flora for Waste Management

P. M. Dongre, Aashutosh Mule, Kartik Bane, Rohan Kusnur, Geetanjali Ranade

Dept of Biophysics, University of Mumbai, Vidyanagari, Santacruz (E), Mumbai 400 098.
Goldfield Fragrances, 1/17 Prabhadevi Indl Estate, Veer Savarkar Marg, Prabhadevi, Mumbai 400 025,

Environmental balance would be best achieved by constructive waste management plan. The most
common approach for the same has been composting the waste for the use as bio- fertilizer. Further, it is possible
to convert the same to utilize any type of organic waste for biogas formation.

Here we are developing temple waste management system. The temple waste (Nirmalya i.e. floral
waste) of many temples in Mumbai has been utilized for the preparation of bio-fertilizers (reported in the local

The aim of this project was to understand in detail the biological and chemical processes involved in the
conversions during compost formation. The process involves addition of inoculum (1 gm for 1 kg) to the waste,
and incubating it under natural conditions. Samples were drawn periodically at an interval of one week from the
above reaction mixture over a period of a month. The sample was analyzed for microbial flora, at each interval.
We tried to understand the organisms that survive the entire process of composting. Further, we would try to
continue the process for the formation of biogas, and if possible, to use available mixed flora, to manage any type
of organic waste.

The pragmatism involved in this project would allow us to develop microbial flora using indigenous and
added species, for waste management. This protocol would be cost efficient, free from obnoxious odours and
toxic gas production. Detailed studies are to be continued.

BSD-2008 ENV-13

Studies on Microbial Degradation of Chlorinated Organic Compounds by

Paenibacillus macerans
Ranjani C, M B Saidutta, G Srinikethan

Department of Chemical Engineering, NITK, Surathkal

Chlorinated organic compounds which are toxic to higher forms of life are also recalcitrant to microbial
degradation. Among the toxic chlorinated organic compounds, chlorinated phenols have been chosen for this
study because of their widespread prevalence in the environment, affecting both soil and groundwater.
Chlorophenols especially mono- and dichlorophenols are formed during the chlorination of water and waste
water, in the presence of some pollutants.
Several techniques are available for the removal of contaminants from waste water, although not all are
efficient enough to reduce the contaminants to acceptable limits. This includes physical methods, chemical
methods and biological method. In most cases, physical and chemical methods lead to secondary pollutants,
disposal being a major problem for most secondary pollutants. Biological treatment using micro-organisms is
especially attractive because it has the potential to almost completely degrade chlorophenol while producing
innocuous end products. In addition, it has the advantage of reduced capital and operating cost because of
operating at ambient conditions.
A pure culture of chlorophenol degrading bacteria used in this study was isolated from soil near waste
water treatment tank of an industry manufacturing compounds using chlorophenols. The isolate was identified
by genotype and phenotypic characterization as Paenibacillus macerans. Comparative analysis of the 16S
rDNA sequence in the GenBank database revealed that these bacteria are related to Paenibacillus macerans.
Growth of Paenibacillus macerans was optimized in different concentration of chlorophenols. Degradation of
chlorophenol was studied using this culture in defined salt media under various ambient conditions of pH and
temperatures. Paenibacillus macerans can effectively degrade chlorophenols and degradation rates are
influenced strongly by temperature and pH.

BSD-2008 ENV-14

Use of Activated Sludge Process (ASP) for domestic wastewater treatment in

small communities
Shohreh Azizi, Dr. V. Kalyan Raman, Dr. V. S. Ghole

Department of Environmental Sciences, University of Pune, Pune -7

With increasing urbanization, the volume of domestic wastewater and urban run offs are steadily
growing. All these wastewaters must be assimilated into the environment without impairing the health and well-
being of man. Environmental awareness around the world has resulted in new waste management initiatives in
developed and developing countries. Assessing and controlling waste pollution now focuses on minimizing
waste, reuse and reduction of toxic chemicals. One of methods used for domestic wastewater treatment is
activated sludge process. The success of the activated-sludge process is dependent upon establishing a mixed
community of microorganisms that will remove and consume organic waste material. The process entails the
aggregation of suspended solids by the process of bio-flocculation, which causes settlement of solids to a
concentrated sludge, suitable for recycling. Any of several types of activated sludge solids separations problems
indicate an imbalance in the biological component of this process. This study on using an ASP surface aerator
optimized the optimum Hydraulic Retention Time (HRT) which can be used for domestic wastewater treatment
in small communities successfully. This information will help reduce the release of domestic wastewater into

BSD-2008 ENV-15

Use of water hyacinth (Eichhornia crassipes (Mart.) Solms) for domestic

wastewater treatment
Alireza Valipour i, Dr. V. Kalyan Raman, Dr. V. S. Ghole

Department of Environmental Sciences, University of Pune, Pune -7

The increasing scarcity of water in the world along with rapid population increase in urban areas gives
reason for concern and the need for appropriate water management practices. However, due to the trends in urban
development, domestic wastewater treatment deserves much greater emphasis. Safe, economic and effective
treatment of domestic wastewater is one of the most challenging problems being faced not only by India but
many other developing countries. During six months studies on water hyacinth, Eichhornia crassipes Mart.
Solms, a highly obnoxious weed, has shown great promising tool in domestic wastewater treatment. A small
pilot-scale continuuous flow system with water hyacinth was set up in the premises of Department of
Environmental Sciences, University of Pune, India, to find out the feasibility of using water hyacinth for sewage
degradation and characterizing the bacterial population during this treatment. The optimum hydraulic retention
time (HRT) for domestic wastewater treatment by water hyacinth was also investigated. In this study it was
observed that the optimum hydraulic retention time (HRT) for domestic wastewater treatment by water hyacinth
was much better than recent studies in the literature. Use of water hyacinths (Echhornia crassipes) for domestic
wastewater treatment is an emerging technology. The use of water hyacinth for nutrient absorption from waste
water assumes greater significance and promise.

BSD-2008 ENV-16

Dibenzofuran Mediated Improved Performance of Activated Sludge for

Treatment of Distillery Spentwash
Anshuman A. Khardenavis, Hemant J. Purohit

Environmental Genomics Unit, National Environmental Engineering Research Institute (NEERI)

Nehru Marg, Nagpur-440 020

Activated sludge from a wastewater treatment plant consists of consortium of microbes that utilize
various organic molecules including persistent organic pollutants for their survival. Phenolic compounds and
their derivatives such as dibenzofuran are found as dominating pollutants in distillery wastewater and are formed
as result of partial enzymatic degradation and depolymerization of lignin polymer. Primary carbon source for
microbial growth expected in distillery wastewater are the fermentable and non fermentable sugars. However,
the partial oxidation of lignin by peroxidases and lacasses leads to accumulation of heterocyclic hydrocarbon
molecules. Therefore, acclimatization, a process that leads to selective enrichment in microbial community has
been employed. In this study, we report the acclimatizing effect of dibenzofuran on improving the treatment
efficiency of distillery sludge. The adaptation dependent performance of activated biomass was studied by
monitoring colony-forming units (CFU) on mineral media and utilization pattern for phenol and dibenzofuran.
Study showed that the acclimatization process remarkably improved the treatment efficiency of distillery
sludge. 19% COD removal from distillery spentwash was observed in case of unacclimatized sludge which
improved to 31% in case of acclimatized sludge. The COD removal efficiency further increased to 80% and 85%
when 10 times and 50 times diluted distillery spentwash was used. Study proposes that such a step could be
included as a part of treatment plant where intermittently the activated biomass could be metabolically charged
by exposing to selected molecules for increasing the treatment efficiency.

BSD-2008 ENV-17

Study with Arthrobacter sp. strain HPC 891 showing flexible catabolic
capacity for degradation of nitro/amino aromatic compounds
Asifa Qureshi and Hemant J. Purohit

Environmental Genomics Unit, National Environmental Engineering Research Institute,

Nehru Marg, Nagpur 440 020 (MS) India

Arthrobacter sp. strain HPC 891 was isolated from a nitroaromatic contaminated waste soil. The strain
was identified by 16S rRNA gene sequencing (Gendata Bank Accession No. AY 948222). The initial degradative
studies were carried out using 2,4,6-trinitrophenol (2,4,6-TNP) as the sole source of carbon, nitrogen and energy.
Aerobic biodegradation process was performed at 30oC with agitation (150 rpm) and was evaluated by
spectrophotometry and high performance liquid chromatography (HPLC). It showed 70% degradation of 2,4,6-
TNP (1 mM) after 96 h with release of nitrite. Oxygen uptake studies demonstrated that Arthrobacter sp. strain
HPC 891 could consume various nitro and amino substituted compounds such as 4-nitrophenol, 2,4-
dinitrophenol, 2-aminophenol, 3-amino phenol and 4-aminophenol after growth on picric acid indicating the
diverse catabolic potential characteristics of the isolate. Since, the isolate has shown flexible degradative
capacity, PCR primer specific to the gene HDO loci, which shows the non-specific aromatic ring cleavage
activity has been used in the study. The studies are underway to further characterize possible genotype associated
for flexible catabolic capacity.

BSD-2008 ENV-18

Phytoremediation of phenol by Vetiveria zizanoides: Role of antioxidant enzymes

Sudhir Singh, J.S.Melo, Susan Eapen, S.F.D'Souza

Nuclear Agriculture & Biotechnology Division, Bhabha Atomic Research Centre,

Trombay, Mumbai 400085, India. Phone: 022-25592638.

The objective of the present study was to evaluate the potential of Vetiveria zizanoides plants for
remediation of phenol, a major contaminant of industrial effluents. When micropropagated plants were tested for
their potential for phenol remediation from Murashige and Skoog's basal medium under in vitro conditions,
phenol at 50 and 100 mgl were found to be completely removed, while 89%, 76% and 70% removal was seen at
200, 500 and 1000 mgL of phenol respectively at the end of 4 days. In the second phase of our study,
micropropagated vetiver plantlets were used under in vivo condition for phenol removal (200 mgL ) in relation
to plant growth and reusability. The growth of vetiver plantlets was affected in presence of phenol. However, on
repeated use, both uptake and growth remained similar to plants treated with phenol only once indicating that
they developed a mechanism for adaptation to phenol. Finally, fully grown vetiver plants were used for bench
scale study in hydroponics conditions and were also found to be efficient for phenol removal. Mechanism of
phenol removal by vetiver plants was found to be associated with inherent production of peroxidase and
hydrogen peroxide by the plant. Coupled with H2O2 formation, the levels of antioxidant enzymes like superoxide
dismutase and peroxidase showed an enhancement when plants were exposed to phenol, whereas catalase levels
initially showed a decline due to the utilization of H2O2 by peroxidase for phenol oxidation. However, when
peroxidase levels declined, there was an enhancement in catalase levels to minimize the presence of H2O2 in the
medium. The results of present study suggest the possibility of using V. zizanoides for phenol remediation as the
plant have evolved resistance to phenol induced H2O2 through a biochemical mechanism.

BSD-2008 ENV-19

Phytoremediation of xenobiotic pollutants with plants expressing fungal

Glutathione-S-transferase gene
Prachy Dixit, Prasun K. Mukherjee and Susan Eapen

Nuclear Agriculture & Biotechnology Division, Bhabha Atomic Research Centre, Mumbai 400085, India.

Glutathione-S-transferases (GSTs, E.C. are a multigene family of detoxification enzymes that

have a crucial role in degradation of a wide variety of exogenous substances including herbicides, pesticides,
insecticides and carcinogens. GSTs catalyze the nucleophilic attack of the S atom of glutathione (γ-Glu-Cys-Gly,
GSH) on electrophilic groups of hydrophobic substrates forming a less reactive and more polar glutathione-s-
conjugate. GSTs tag xenobiotics for vacuolar sequestration by ATP-binding cassette transporters that function as
glutathione-s-conjugate Mg ATPase pumps. GST gene was cloned from Trichoderma virens and introduced
into plant based vectors pCAMBIA1301 and pCAMBIA1302. These constructs were then electroporated into
Agrobacterium tumefaciens EHA 105 strain. Leaf discs of tobacco were co-cultivated with Agrobacterium
tumefaciens harbouring pCAMBIA1301 plasmid and putative transformed plants were selected on regeneration
medium containing hygromycin at 15mgl . The regenerated plants were further transferred for rooting. Putative
transgenic plants were confirmed using PCR with GST gene and selectable marker gene specific primers. RT-
PCR for GST gene has confirmed expression of this gene in transgenic tobacco lines. The results of response of
transgenic plants with GST gene to different xenobiotic compounds will be presented.

BSD-2008 ENV-20

Expression of the chitinase gene from Trichoderma virens in

Saccharomyces cerevisiae
Mamta R. Shah, George V. Sible, Prasun K. Mukherjee and Susan Eapen

Nuclear Agriculture and Biotechnology Division, Bhabha Atomic Research Centre, Mumbai 400085, India.
Phone: 022-25592638

Chitinases are a heterogeneous group of enzymes that catalyze the hydrolytic reaction of chitin, which is
the second most abundant polysaccharide (after cellulose) on the earth. Chitin is a long-chain polymer of N-
acetylglucosamine linked by ß-1, 4 glycosidic bonds. The important applications of chitinases are in the fields of
fungal disease control, pollution abatement and in basic and commercial biology. Chitinase can also be used for
the treatment of shellfish wastes. Biotechnological approaches can be used for the production of chitinases.
Trichoderma virens is a mycoparasitic fungus and possess strong chitinolytic system.

The ech42 gene encoding 42-kDa endochitinase enzyme from T. virens was cloned and expressed in
Saccharomyces cerevisiae. The ech42 gene was integrated into the genomic DNA of Yeast expression vector,
pYES2 by insertion into cloning site, yielding the recombinant pYES2-ech42. The S. cerevisiae BY4742 cells
were transformed with the expression vector pYES2-ech42. Chitinase expression was induced in the presence of
galactose and level of expression of endochitinase enzyme measured by fluorometric assay. Transformed cells of
S. cerevisiae showed high level of endochitinase expression and the activity ranged from 2 to 31 fold in the
transformants as compared to the parental strain. The present work has potential commercial applications and
can also be used in environmental bioremediation for the treatment of shellfish wastes.

BSD-2008 ENV-21

Endosulfan removal by Bacillus magaterium biomass

Deepika Upadhyay and A.K. Dikshit

Centre for Environmental Science and Engineering, IIT Bombay,

Powai, Mumbai 400 076, India,

Pesticides, are an integral part of modern agricultural practice. Conversely, extensive application of
these pesticides has caused the pollution of soil, ground and surface water, which involves a serious risk to the
environment and also to the human health. Some of these pesticides have been reported to be persistent, toxic,
mutagenic, carcinogenic, and tumorogenic. The organochlorine pesticides being lipophilic, hydrophobic and
with low chemical and biological degradation rates, have been found to accumulate in biological tissues and
subsequent magnification of concentrations in organisms, progressing up the food chain. Hence, the usage of
many organochlorine pesticides has been abandoned / banned in most of the developed countries. As per BIS
(Bureau of Indian Standards), the pesticide residue should be absent in drinking water and should not exceed
0.005 ppm in surface waters. Endosulfan, a highly toxic organochlorine insecticide and acaricide, is one of the
most widely used pesticides in developing countries as well as parts of Europe and Africa and has been found
contaminating surface waters, soils and ground water. The removal of pesticides by conventional methods is
found to be either uneconomical or insufficient.

In the present study, Bacillus magaterium is employed as model micro-organism for investigating
biosorption of endosulfan from aqueous phase. In the adsorption experiments carried out for duration of 3 days,
different doses of biosorbent prepared from bacterial biomass ranging from 2.5 to 50 mg were added in a
endosulfan solution of 1 mg/l. The adsorbed endosulfan on the bacterial biomass was extracted easily in n-
Hexane and the extract was analysed using high-resolution gas chromatograph. The biosorbent was found to
remove 49.6 % of endosulfan from water. The suitability of the Freundlich, Langmuir and BET adsorption
models to the equilibrium data was investigated for the endosulfan-biosorbent system. The results showed that
the equilibrium data for endosulfan-biosorbent system fitted the Freundlich model with a correlation coefficient
of 0.851.

BSD-2008 ENV-22

COD and color removal of distillery spent wash using fungal bioreactor
S. S. Singh, Prof. A. K. Dikshit

Centre for Environmental Science and Engineering, IIT Bombay, Powai, Mumbai 400 076, India,

Effluent originating from distilleries known as spent wash leads to extensive soil and water pollution.
Color of spent wash is largely due to melanoidin. Melanoidin is one of the biopolymers, which are difficult to
decompose by microorganisms. Several basidiomycetes and ascomycetes type fungi have been used in the
decolourization of melanoidins in connection with color and COD reduction of wastewaters from distilleries.
Several types of fungal bioreactors have been employed to study the degradation of these coloring compounds.
The common types of bioreactor used are fed batch reactor, bubble column bioreactor, fluidized bed reactor and
immobilized bioreactor. The dilution rate affects fungal biomass yield in the reactor. Different rates of aeration
are used in different types of bioreactors. The temperature range of 32 to 36 C is found optimum for fungal
activities. The hydraulic retention time of reactors ranges from 20 hrs to 16 days. The availability of carbon
source in form of simple sugars (glucose and sucrose) is required in all experiments. Requirement of nitrogen
and phosphorus source is also reported by some researchers. The efficiency of these reactors for decolorization
ranges from 37 to 80% and COD removal efficiency ranges from 34 to 77%.

This paper presents various types of fungal bioreactors for COD and color removal of distillery spent
wash; factors affecting decolorization of spent wash; problems associated with this type of treatment and
possible trouble shooting.

BSD-2008 ENV-23

Decolourisation of distillery wastewater : A biotechnological approach

M. S. Chauhan and A. K. Dikshit

Centre for Environmental Science and Engineering, IIT Bombay Powai, Mumbai 400 076, India,

Sugar is the largest industrial sector after textiles in India and India is among the top five sugar producing
countries in the world, with an annual production of 17.6 million tons. Sugar manufacturing produces molasses
as a by-product. Most of the distilleries in India use sugar cane molasses for alcohol production. Wastewater
coming out from distilleries, commonly known as spent wash, is characterized by its high percentage of
dissolved organic and inorganic matter, dark brown colour, high temperature, low pH and high ash content.
Indian spent wash contains very high amounts of potassium, calcium, chloride, sulfate and BOD as compared to
spent wash in other countries. Dark brown colour of distillery wastewater is mainly caused by high molecular
weight organic melanoidin compounds, produced from the non-enzymatic browning reaction between aldose
and amino compounds; and the caramels produced from thermal degradation and condensation reaction of
sugars. At present, various physico-chemical and biological treatment options are available for the removal of
colour from spent wash. However, these methods are either very costly or not very effective in removing colour
from distillery wastewater.

Present research work aims at integrated physico-chemical and fungal treatment approach for
decolourisation of anaerobically digested distillery wastewater. Efficacy of pretreatment was assessed using
various coagulants, oxidants and adsorbents. Following which, fungal treatment with locally isolated strain was
done. This technology is economically and technically viable and gives about 88% reduction in colour with
corresponding 75% reduction in COD value without any dilution and with minimal external nutrient addition.

BSD-2008 ENV-24

Opportunity, usefulness and efficiency of alumina ceramic membranes in

treatment of sewage water
S Mahesh Kumara, Pushpa Agrawal, Sukumar Royb
a, .
Dept of Biotechnology, R.V. College of Engineering, Bangalore 560 059, India.
Ceramic Technological Institute, Bharat Heavy Electricals Limited, Corporate Research & Development,
Bangalore 560 012, India.

This work highlights the recovery of water from sewage effluents using alumina ceramic membranes
with pore sizes of 0.2 and 0.45µm, in dead-end filtration mode. The work demonstrates the ability and
advantages of alumina-based microfiltration (MF) membranes in filtering microbes and other harmful pollutants
normally present in sewage effluents in dead-end filtration mode. The fouling behavior of the membranes in the
filtration cycle is identified, which, in turn, helped to regenerate the fouled membranes for subsequent usage.
Regeneration studies of fouled membranes also suggest that though chemical cleaning was effective in
recovering membrane performance, the fouling had still been progressed slowly and the membranes showed the
ability to perform at least five filtration cycles of highly-contaminated sewage effluents. As expected, the
filtration efficiency and flux characteristics at various trans-membrane pressure (TMP) of the membranes varies
with the pore size of the membrane and is explained in light of Darcy's and Poiseuille's laws of filtration. The
results show that alumina ceramic membrane with a pore size of 0.2µm is more effective in filtering the total
suspended solids, turbidity and microbes of the sewage effluents as compared to that of 0.45 µm membrane to a
level in which the permeate water appears to be benign for discharging into the surface or offering the possibility
of recycling or reusing for suitable purposes. The data could be useful in designing large-capacity ceramic
membrane based sewage filtration system.

BSD-2008 ENV-25

Biomonitoring of Industrial effluent and affected sites of Urban areas around

Jodhpur, Rajasthan
Sukha Ram Vishnoi, N.S Shekhawat, and P.N Srivastava

Biotech Unit, Department of Botany, JNV University, Jodhpur-342005

A huge volume (20-25 MLD) of industrial effluent is released from the industries at Jodhpur everyday.
The effluent is highly acidic /alkaline (pH 2.5-9.5), colored, turbid with elevated BOD, COD, TDS and heavy
metals. The untreated effluent runs over vast areas along Jodhpur-Pali railway track and is finally released in
Jojari river basin. As the flow is unchannalised the polluted water enters into agricultural fields. The
biomonitoring studies of the effluent and affected sites revealed that only blue-green algal patches survive in
toxic condition on the river banks. The algal flora observed from the river basin under such conditions were
Oscillatoria salina, O. subrevis, O. tenuis, O. jasorvansis, O. grunowiana, Phormidium tenuis,
Johonnesbaptistia pellucida and Scenedesmus dimorphus. We tried enrichment cultures of Nostoc calcicola,
Calothrix weberi, Ocillatoria tenuis, and Phormidium foveolarum from the contaminated soil. The
heterocystous forms were absent under the polluted conditions but these appeared in the enrichment cultures
indicating existence of viable spores in affected soils. The non-heterocystous forms show greater tolerance and
thrive well under stressed conditions. The ground vegetation of this site is adversely affected; certain tree species
of Prosopis juliflora, Eucalyptus camaldulensis, Withania sominifera, Chenopodium album, Salsola baryosma
and Heliotropium curassavicum can grow at these sites. Xerophytic plants namely Calotropis procera, Aerva
percica, Crotalaria burhia, Tecomella undulata and Prosopis cineraria cannot survive in this polluted site.

Agricultural residues for lipase production : Response surface optimization of
fermentation parameters
Aravindan Rajendran1 and Viruthagiri Thangavelu3
1 2
Lecturer, Professor and Head
Biochemical Engineering Laboratory, Department of Chemical Engineering,
Annamalai University, Annamalainagar-608 002, Tamil Nadu, India.
E.mail:; Phone: 04144-238150

Lipases [triacylglycerol acylhydrolases (EC] are ubiquitous enzymes found in animals, plants,
fungi and bacteria which are of considerable physiological significance, that catalyze the hydrolysis of
triacylglycerides to produce glycerides, monoglycerides, glycerol and fatty acids at the lipid water interface.
Candida rugosa, an industrial producer is the potential microbial source for lipase production. Among processes
used for enzyme production, industrially important enzymes have traditionally been obtained from submerged
culture because of ease of handling and greater control of environmental factors, such as temperature and pH.
Solid state fermentation (SSF) constitutes an interesting alternative, since metabolites so obtained are more
concentrated and purification procedures are less costly. SSF process offers more economical and abundant
sources of substrates such as agro-industrial residues. Usually SSF is a batch process that utilizes heterogeneous
natural residues, which provides essential carbon, nitrogen and energy sources to the microorganisms. Response
surface methodology using central composite design was adopted to design the experiments and to study the
independent and interaction effects of variables on lipase production.

Candida rugosa NCIM 3462 was used for the production of lipase in solid state fermentation. Agro
industrial wastes such as sesame oil cake, groundnut oil cake and coconut oil cake were used for the solid state
fermentation. Among the various oil cakes screened, maximum lipase activity of 12.47 U/g substrate was
obtained from sesame oilcake at the incubation time of 60 h. The response surface methodology was employed to
optimize the lipase production and to study the effect of temperature and substrate to moisture ratio. A 2 full
factorial central composite experimental design was employed for the response surface methodology. The
maximum lipase activity of 22.40 U/g substrate was obtained at the optimized temperature and substrate to
moisture ratio were found to be 32C and 1:3.2 g/ml respectively. A predictive model of the combined effects of
the independent variables using response surface methodology and artificial neural network was proposed.

Adhikari D. K (ENG-06) 37 Mohapatra. S (ENV-03) 43

Adlercreutz Dietlind (GC-11) 28 Mshandete1.A.M (ENV-09) 49
Agrawal Pushpa (ENV-24) 64 Mukherjee Prasun K (ENV-19,20) 59,60
Akay Galip (IL-13) 14 Mule Aashutosh (ENV-12) 52
Alemzadeh I (ENG-04) 35 Naidu Gummadi Sathyanarayana(ENV-01) 41
Alkasrawi Malek (IL-09) 10 Nandy Tapas 1 (ENV-11) 51
Angelidaki Irini (IL-06) 7 Nayak B.D (ENV-03,04) 43,44
Annachhatre Ajit (IL-15) 17 Nemati Fahimeh (GC-07,ENV-06) 24,37
Arabian Daryush (ENV-06, GC-07) 46, 24 Nerkar D. P (GC-10) 27
Azad R. Roosta (ENG-04) 35 Nielsen Per Henning (IL-02) 3
Azizi Shohreh (ENV-14) 54 Paliwal Kashika (GC-14) 31
Bagchi Samik (ENV-11) 51 Patel Martin K. (IL-04) 5
Balakrishnan Malini (ENV-10) 50 Pathan Eajaj (GC-10) 27
Bane Kartik (ENV-12) 52 Patil Rajendra (ENV-06) 46
Banerjee. R (ENG-08) 39 Pätz Reinhard (IL-14) 16
Bapat Sumedh (ENG-01) 32 Pradhan.N (ENV-04,05) 44,45
Bhadekar R. K (GC-10) 27 Puri Munish (GC-04) 21
Biswas Rima (ENV-11) 51 Purohit Hemant J (ENV-16,17) 56,57
Björnsson.L (ENV-09) 49 Qureshi Asifa (ENV-17) 57
Börjesson Pal (IL-12) 13 Radha RK (GC-08) 27
Borkar Indrakant V (GC-06) 23 Ram.L.C (ENG-08) 39
Chadha Anju (ENG-05) 36 Raman V. Kalyan (ENV-14,15) 54,55
Chakrabarti T (GC-09) 26 Ranade Geetanjali (ENV-12) 52
Chaudhari P.R (ENV-08) 48 Ranade.D.R (ENG-01) 32
Chauhan M. S) (ENV-23) 63 Ranjan Ravi (GC-04) 21
Chotani Gopal (IL-05) 6 Ranjani C (ENV-13) 53
Coca Cecilia Orellana (GC-11) 28 Raut Bhagyashri G (ENG-07) 38
D'Souza S.F (ENV-18) 58 Ravindranath N. H (IL-14) 16
Dalal J (GC-03) 20 Roostaazad.R (GC-07) 24
Deshpande Ghanshyam (IL-08) 9 Roy Sukumar (ENV-24) 64
Deshmukh S. K (GC-05) 22 Roychoudhury Kunal (ENV-11) 51
Dikshit A. K. (ENV-21,22,23) 61,62,63 Rubindamayugi M.S.T (ENV-09) 49
Dixit Prachy (ENV-19) 59 Saidutta M B (ENV-13) 53
Doble. Mukesh (GC-01) 18 Sam P Mathew (GC-08) 25
Dongre P. M (ENV-12) 52 Sapkal R.S (GC-05,12) 45,52
Eapen Susan (ENV-18,19,20) 58,59,60 Sapkal V.S. (GC-05,12) 45,52
Gade W. N (ENV-06) 46 Sarkar Ritam (ENG-05) 36
Ghole V. S (ENV-14,15) 54,55 Sarma Priyangshu M (GC-02,03,ENG-02) 19,20,33
Gummadi Sathyanarayana (GC-01) 18 Satyawali Yamini (ENV-10) 50
Gunale V.R (ENV-07) 47 Seeni S (GC-08) 25
Gupta Rani (GC-13,14) 30,31 Selvi V. A (ENG-08) 39
Hagström Tomas (IL-07) 8 Shah Deepak (ENG-01) 32
Hatti-Kaul Rajni (IL-03,GC-11) 4,28 Shah Mamta R (ENV-20) 60
Jhon Nisha M (ENV-11) 51 Shalini J.M (ENG-08) 39
Juwarkar Asha A (GC-09) 26 Shanware Arti S (ENG-07) 38
Kapardar Raj Kishor (GC-04) 21 Sharma Rakesh (GC-04) 21
Khardenavis Anshuman A (ENV-16) 56 Sharma Ravi (GC-09) 26
Kivaisi1.A.K (ENV-09) 49 Sharma Richa (GC-14) 31
Kumar Devarai Santhosh (ENV-01) 41 Shekhawat N.S (ENV-25) 65
Kumar S Mahesh (ENV-24) 64 Shrivastava Amit (ENG-03) 34
Kumar S (GC-13) 30 Sible George V (ENV-20) 60
Kumar S.Suresh (GC-14) 31 Singh K (ENG-09) 40
Kumar Sachin (ENG-06) 37 Singh S. S (ENV-22) 62
Kusnur Rohan (ENV-12) 52 Singh S.K (GC-09) 26
Laane Colja (IL-01) 2 Singh Sudhir (ENV-18) 58
Lal Banwari (GC-02,03,ENG-02) 19,20,33 Singh. S (ENV-02) 42
Lavania M (GC-02) 19 Sri Lakshmi P (GC-01) 18
Legg Wilfrid (Plenary Lecture) 1 Srinikethan G (ENV-13) 53
Leyla Gheyrati Arani (ENV-07) 47 Srivastava P.N (ENV-25) 65
Madaan Ankush (ENG-05) 36 Sudhakaran Anu (ENG-02) 42
Mandal A K (GC-02,03) 19,20 Sukla L.B (ENV-02,03,04,05) 42,43,44,45
Mandal Ajoy (ENG-02) 33 Talib M (GC-12) 29
Marhual N.P (ENV-05) 45 Tonger Sonal (GC-13) 30
Masto R.E (ENG-08) 39 Upadhyay Deepika (ENV-21) 61
Mattiasson Bo (IL-11,ENV-09) 12,49 Valipouri Alireza (ENV-15) 55
Meher. K.K (ENG-01) 32 Varma Navarun (ENG-02) 33
Melo J.S (ENV-18) 58 Verma Sanyogita (ENV-08) 48
Meshram Sudhir U (ENG-07) 38 Vishnoi Sukha Ram (ENV-25) 65
Mishra B.K (ENV-02,03,04,05) 42,43,44,45 Vossoughi M (ENG-04) 35
Mishra M (ENV-04) 44 Wate S.R (ENV-08) 48
Mohanta N.C (ENV-05) 45 Yadav Ganapati D (GC-06) 23



Passionate about Green Fuels

Praj began operations in 1984. Focused upon bioethanol & biodiesel technology, plant & equipment, Praj has
since become a world leader with a number of processes and systems for ethanol production to its credit. Today,
Praj has over 350 references in 40 countries across 5 continents. Praj has acquired an international repute and has
its own offices in in India and overseas. Praj also has a wholly owned subsidiary in USA called Praj Schnieder, an
engineering firm active in biofuels industry. Recently, Praj formed a JV in Europe with Aker Kvaerner, located
in Netherlands. Praj has also made an entry into Brazil and will be shortly setting up operations there.

Praj enjoys a unique position in the world of ethanol technology by virtue of its expertise which cuts across a
variety of feedstocks from sugar bearing ones like cane and beet to starch bearing ones like cassava, sorghum,
cane juice etc. In fact, Praj has developed a technology for multi-feed plants which can process, apart from
canemolasses, cane juice, tropical sugarbeet juice, sorghum, wheat and any other starch based raw materials.
This technology gives distillers the distinct advantage of operating a plant round the year and to even out the
price fluctuations in raw materials, so that profitability levels can be maintained.

In bio-diesel too, Praj has developed a robust design which offers high yields with high efficiency levels. Praj
extends its expertise in agri-based industrial processes to the field of biodiesel.

Praj works with Government and Private sector to promote biofuels. Recently, Praj has signed a Memorandum
of Association with the Department of Agriculture, Philippines to jointly develop the biofuels program. Praj also
represents the technology viewpoint on the National Planning Commission's Biofuels Sub-Committee (India)
and has assisted with the development of ethanol programme in Colombia. Pramod Chaudhari, Chairman, Praj
Industries is the President of CII-National Biofuels Committee.

With increasing awareness and application of fuel ethanol as an environment friendly additive to transport fuel in
many countries, there is a need to identify newer sources of ethanol production. Praj has already taken a lead in
this field. Praj has evaluated sweet sorghum as a viable alternate feedstock for ethanol production and has
introduced process innovations for which a patent has already been filed. The Company has received its first
commercial scale project based on sweet sorghum from Tata Chemicals, recently.

The backbone of Praj's technology development is Matrix, the Innovation Center. This Center has been
conceived and established with the objective of providing cutting edge technology and engineering solutions to
the distillery and brewery industry, backed up by research and pilot/commercial scale trials.

With a SEZ unit at Kandla (Gujarat, India) and four other manufacturing units, Praj is in a position to supply
fabricated equipment for all its projects. The units are accredited for ASME H & U stamp and are capable of
producing equipment to various standards.

Praj is now pursuing development of cellulose (biomass) to ethanol technology.

Praj also supplies brewery plant & equipment and process equipment for other process industries like chemical,
food, pharmaceutical, textiles, etc.

Praj employs more than 800 people in India and overseas, with over 80% being from engineering/technology

Praj is a listed Company (BSE/NSE) with market capitalization of around US$ 900 mln and revenues in excess of
US$ 145 million. Praj is a founder member of Global Growth Companies (WEF).


Established in 1939 at Mithapur, and part of the US$ 28.8 billion Tata Group, Tata Chemicals Limited (TCL)
today is the third largest producer of Soda Ash in the world with manufacturing facilities across three continents.
TCL is the pioneer and market leader in the branded, iodised salt segment. TCL is also India's leading
manufacturers of urea and phosphatic fertilisers. Its urea plant at Babrala is the country's most energy efficient
fertiliser unit and produces 12% of the country's urea output in the private sector. Phosphatic fertilisers are
manufactured at its plant in Haldia.

An ISO 9001/14001 certified company, TCL has a varied user industry base comprising glass, paper, textiles,
Consumer products, petroleum, refining, chemicals, dyes, pesticides, direct farm application etc. With an export
presence in Europe, Africa, Southeast Asia and the Middle East, TCL has set itself the objective of achieving
global cost competitiveness in soda ash. The acquisition of an equal partnership in Indo Maroc Phosphore S.A.
(IMACID) along with Chambal Fertilisers and the global phosphate major, OCP of Morocco in the year 2005
was the first step that TCL took towards internationalization. In early 2006, TCL completed the acquisition of the
UK based Brunner Mond Group (BM). The total soda ash production capacity of the combined entity is close to 3
MTPA. In the year 2007, TCL entered in to a 50:50 joint venture with Total Produce, Ireland, the third largest
fruits and vegetable distribution company in the world and Europe's largest and most accomplished fresh
produce provider, to set up a fruits and vegetables distribution business in India. TCL also plans to leverage its
expertise in chemistry & agriculture, together with its in-house research capabilities in biology and crop
genomics to develop a significant presence in Biofuels space. The TCL Innovation centre was created in mid
2004 to develop world-class R&D capability in the emerging areas of nanotechnology and biotechnology.

Domestic Products and Services

Chemicals : Its range of chemicals, produced at the company's integrated complex at Mithapur on the Gujarat
coast in Western India, includes apart from soda ash caustic soda, salt, cement, sodium bicarbonate, bromine
and bromine based compounds, and gypsum. The plant has an installed capacity of 875,000 tonnes of soda ash
per annum, nearly 35% of the country's capacity. The company manufactures bulk chemicals like sulphuric acid,
phosphoric acid and Sodium Tripoly Phosphate (STPP) at its plant in Haldia, West Bengal in eastern India. This
unit produces about 60,000 tonnes of STPP annually, about 65% of the country's capacity.
Fertilisers : The company manufactures nitrogenous fertilisers at its plant in Babrala in the state of Uttar Pradesh
in northern India. With a capacity of 864,000 tonnes of urea per annum, this plant matches global standards in
technology use, energy conservation, productivity and safety. The unit also houses an ammonia plant with a
capacity of 1,350 tonnes per day.
The company's phosphatic fertiliser plant is produced at its integrated complex at Haldia which, apart from bulk
chemicals, also makes phosphatic fertilisers like Di-ammonium Phosphate (DAP), NPK complexes and Single
Super Phosphate (SSP). Tata Chemicals is a market leader in phosphatic fertilisers in the eastern Indian states of
West Bengal, Bihar and Jharkhand.

Consumer products :
Tata Chemicals' consumer products business has two of the company's premium product groups under its wing :
branded salt and sodium bicarbonate.

Branded Salt: TCL manufactures four varieties of salt; iodised salt, crystalline salt, vacuum salt and pure salt.
With over 40% market share in the branded salt segment in India and a reach of close to 40 million households,
Tata Salt is the No.1 Food Brand in the list of India's Most Trusted Brands.
Sodium bicarbonate: TCL produces three varieties of sodium bicarbonate - technical, refined and granular at its
50,000 tonnes per annum plant and markets them in India and the West Asia.

The company also helps small farmers enhance the yield from their land by providing end-to-end solutions,
harnessing sophisticated modern technology such as satellite mapping and geographical information systems, at
its network of Tata Kisan Sansar's (Tata Farmer's Centres) in the northern Indian states of Uttar Pradesh, Punjab,
Haryana and Uttaranchal. The centres are one-stop resource centres; they stock seeds, pesticides and fertilisers;
lease out farm equipment and implements to farmers who cannot afford to buy expensive modern machinery;
provide agronomy services like soil testing and mapping and fertiliser testing; and extend credit finance, arrange
crop insurance and even provide buyback facilities.

Corporate Social Responsibility

The 'avoid, reduce and recycle' philosophy that the company follows is exemplified best by the company's
cement plant at Mithapur, originally set up to consume the solid waste generated at the plant. The company is also
a signatory to Responsible Care, a voluntary global initiative of the chemical industry that demonstrates
allegiance to safety, health and environmental issues. Its Mithapur and Babrala plants have won the highest
British Safety Council 5-star rating. The company has also developed the Mithapur salt works as a natural habitat
for thousands of migratory birds.
Tata Chemicals' Society for Rural Development (TCSRD) nurtures rural populations in and around the facilities,
and helps people achieve self-sufficiency in natural resource management, livelihood support through self-help
groups and the building of health and education infrastructure.

Contact : Head - Corporate Communications, Tata Chemicals, Bombay House, 24, Homi Mody Street, Mumbai
400 001, India.
Tel : +91-22-6665 8282 Fax : +91-22-6665 8144 Email :


Corporate Office
Chembond Centre,
EL-71, Mahape, Navi Mumbai 400 710

Chembond Chemicals Limited, established in 1974, manufactures metal treatment chemicals, water treatment
chemicals, construction chemicals, protective coatings, and enzymes. Chembond is located in Navi Mumbai,
has over 550 group employees across the country with group sales of over Rs. 150 crores.

The metal treatment and water treatment chemicals businesses are joint ventures with global leaders Henkel,
Germany and Ashland, USA respectively. Both businesses are Indian market leaders with sales to the
automotive, steel, petrochemical, power, appliance, engineering, fertilizer and pharmaceutical industries.

Chembond also manufactures construction chemicals for the building construction and infrastructure sectors
and high performance coatings for structural protection from corrosion, for floors and walls in clean rooms and
shop floors.

Chembond's newest project in biotechnology promises to improve industrial applications using the technology
of the future. The company's products are supplied for applications in animal feed, textile processing, and waste
water treatment. Development work is ongoing in the area of bio-refining.

Chembond's modern, well-equipped, ISO 9001:2000, TS 16949, and ISO 14000 certified plants are located at
Tarapur, Baddi, Balasore, Chennai, and Baroda (under construction). Additional pilot plants, warehouses and
branch offices are located at Pune, Ahmedabad, Delhi, Faridabad, Jamshedpur, and Chennai.

Chembond Group of Companies

 Chembond Chemicals Limited
 Chembond Drewtreat Limited
 Henkel Chembond Surface Technologies Limited
 Chembond Enzyme Company Limited
 CCL Building Systems Limited

Company growth over the years

1979: Construction of a manufacturing plant at Tarapur

1983: Diversification into manufacture of cooling water treatment
1992: Expansion of facilities at Tarapur
1994: Addition of anticorrosive coatings, construction chemicals,
maintenance products and water treatment plants
1995: Listed on the Bombay stock exchange
1997: Joint venture with Henkel KGaA for metal treatment chemicals
2001: Joint venture with Ashland for water treatment chemicals
2002: Launch of environmental bio-products, specialty chemicals,