Biomass and Bioenergy 119 (2018) 54–60

Contents lists available at ScienceDirect

Biomass and Bioenergy

journal homepage: www.elsevier.com/locate/biombioe

Research paper

Advances and opportunities in biomass conversion technologies and T

biorefineries for the development of a bio-based economy
Celina K. Yamakawa1, Fen Qin1, Solange I. Mussatto∗
Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Kemitorvet, Building 220, 2800, Kongens Lyngby, Denmark

A R T I C LE I N FO A B S T R A C T

Keywords: The desire to have a more sustainable future, with lower emissions of carbon and sulfur to the atmosphere, a
Biomass more appropriate reuse and valorization of wastes, and less dependency on oil has motivated the society to
Pretreatment develop processes where renewable biomass is used as a feedstock for the production of fuels, chemicals, energy
Fermentation and materials. In addition, a bio-based economy has also potential to generate new jobs and new opportunities
Biorefinery
for entrepreneurship, with further benefits to the society. In view of this, great efforts have been done in order to
Bio-based economy
develop efficient, sustainable and cost competitive bio-based processes able to be implemented in industrial
scale. Although important advances were achieved and some processes are already available in a large scale,
improvements are still needed to have a final product at a more competitive market price. In this sense, the
strategy of integrating biorefineries to produce a variety of products from biomass has been considered as an
important alternative to improve the financial performance. This paper highlights the most recent advances and
opportunities in biomass conversion technologies and biorefineries for the development of a bio-based economy.
Technological aspects including the hemicellulose integration and use of sugars for different products, lignin
valorization, development of efficient and low-cost pretreatment technologies and development of highly effi-
cient fermentation processes are also presented and discussed.

1. Introduction sustainable processes and a suitable waste management are used. In

A bio-based economy is defined as an economy predominantly
based on the use of biomass feedstock, including a full range of natural
and renewable biological resources and biological materials (rather
than fossil-fuel resources), for food, feed, energy, chemicals, materials
and other purposes. So, a bio-based product can be defined as a product
partly or fully derived from biomass.
The bio-based economy comprises different sectors including ma-
terials and chemicals (also pharma), bioenergy and biofuels. Currently,
the use of biomass as a raw material to produce fuels, chemicals and
materials is considered crucial to ensure a low-carbon economy for the
future, with potential economic, environmental and social benefits. In
the economic perspective, the benefits are related, for example, to the
possibility of supporting new investments in the industrial sector, en-
hancing regional growth and competitiveness, and reducing the de-
pendency on fossil fuels. In the environmental aspect, the benefits
comprise the reduction of wastes and their negative effects to the en-
vironment (soil, water and atmosphere, including greenhouse gas
(GHG) emissions), contributing to climate change mitigation when
addition, the use of biomass as a feedstock for processes induces a
sustainable use of carbon because plants, which absorb carbon for their
growth, are used to manufacture bio-based products by industries.
Then, the absorbed carbon is released at the end of the products life
cycle without increasing the atmospheric carbon dioxide concentration
[1]. Finally, in the social perspective, the benefits related to the de-
velopment of a bio-based economy reflect in a stimulation of regional
and rural development, and generation of more jobs both in farming
and in the industrial sector.
The renewable energy sector, for example, which includes solar
photovoltaic, bioenergy (liquid biofuels, solid biomass and biogas),
hydropower, and wind energy among others, has experienced a sig-
nificant growth in the last years, attracting more investors and creating
millions of new jobs. According to the International Renewable Energy
Agency [2], the global renewable energy sector employed 10.34 million
people in 2017, a number 45% higher than in 2012 (Fig. 1). Solar
photovoltaic was the largest employer in 2017, with 3.37 million jobs,
surpassing the bioenergy sector, which was in the second place with
3.06 million jobs. Liquid biofuels represent the major part of

∗
Corresponding author.
E-mail addresses: smussatto@biosustain.dtu.dk, solangemussatto@hotmail.com (S.I. Mussatto).
1
Both authors contributed equally to this work.

https://doi.org/10.1016/j.biombioe.2018.09.007
Received 17 February 2018; Received in revised form 12 August 2018; Accepted 11 September 2018
0961-9534/ © 2018 Elsevier Ltd. All rights reserved.
C.K. Yamakawa et al. Biomass and Bioenergy 119 (2018) 54–60

Fig. 1. Number of employments related to renewable en-

ergy technologies for the period 2012–2017 (data from
IRENA [2]). Other technologies include solar heating/
cooling, geothermal energy, small hydropower, con-
centrated solar power, municipal and industrial waste,
ocean energy and miscellaneous. Bioenergy includes liquid
biofuels, solid biomass and biogas.

employments in the bioenergy sector, corresponding to 1.93 million

jobs in 2017.
In fact, the production of liquid biofuels from biomass is a topic that
has received great attention for decades and nowadays the large-scale
production of second-generation ethanol, for example, is already a
reality in different countries. However, the current commercial-scale
facilities for second-generation biofuels production are still facing some
bottlenecks related to incomplete breakdown of biomass, incomplete
sugars fermentation, production of undesirable by-products, and re-
quirement of starter culture every new batch [3]. These factors re-
present a substantial challenge for a cost-competitive application of
second-generation technology and technical improvements are required Fig. 2. The basic process chain for conversion of lignocellulosic biomass into
to make the technology more efficient and commercially profitable. different valuable products in a biorefinery context.
Currently, there are six big companies around the world in a position to
be the first to make profitable the large-scale production of cellulosic
“sugars switch”) in order to obtain maximum economic turnover; 2)
ethanol, which include Raízen, Poet/DSM, GranBio, Beta Renewables,
using lignin not only for power/heat generation but also for the pro-
Dupont and Abengoa.
duction of valuable chemical compounds; 3) integrating nutrients and
In principle, the concept of integrating first- and second-generation
water recycling for a better use of these resources and to close the cycle
ethanol plants could be an alternative to reduce the process costs, due
from a sustainable perspective.
to the possibility of sharing facilities, equipments and labor. A good
A large variety of chemical products can be produced from biomass,
example is the integration of the first- and second-generation sugarcane
both via biological or chemical conversion. With innovative process
ethanol plants in Brazil, where the second-generation process shares
technologies, some high-value products with great commercial interest
many of the same unit operations used for first-generation process in-
can be obtained at a reasonable cost. In 2004, the U.S. Department of
cluding sugar extraction, juice preparation, fermentation, distillation,
Energy (DOE) identified the top 12 most promising building blocks that
and biomass combustion, resulting in a great opportunity for cost sav-
can be produced from biomass. In 2010, a revised and updated list was
ings by integrating the two processes [4]. Nevertheless, in many cases,
presented, which included succinic acid, lactic acid, 2,5-fur-
this integration strategy is still not enough to payback the investment.
andicarboxylicacid (FDCA), 3-hydroxypropionic acid, fumaric acid,
Therefore, policies and mandates actions have been adopted in many
glycerol, xylitol, sorbitol, levulinic acid, L-aspartic acid, L-glutamic acid,
countries to create a market demand for advanced biofuels from lig-
and itaconic acid [5].
nocellulosic feedstocks. This outlook also reinforces the necessity of
Integrating the production of bio-based chemicals with cellulosic
including the production of value-added chemicals in a lignocellulosic
biofuels in a biorefinery contributes to a more effective utilization of
biorefinery, since higher economic value can be achieved when biomass
feedstock fractions, provides a diversified value stream outputs, makes
is converted into multiple valued added chemicals, besides energy and
scale-up easier as more installations will be available, and results in an
fuels.
improvement of financial performance due to the highest market price
of chemicals compared to liquid biofuels. In general, chemicals have a
2. Development of biorefineries high market price because they either may be used in low concentra-
tions or can be obtained via sophisticated conversion routes [6]. When
Biorefinery refers to the conversion of biomass feedstock into a compared to biofuels, chemicals and materials from biomass present
spectrum of valuable products including food/feed ingredients, che- higher market price and low market volume, while liquid and gaseous
micals, materials, and bioenergy (biofuels, power and/or heat) [5]. biofuels present higher market volume but they are cheaper products
Depending on the main product, the biorefinery system can be named (Fig. 3). So, including chemicals and/or materials in an energy-driven
as energy-driven biorefinery or product-driven biorefinery. biorefinery can be an interesting strategy to improve the revenues.
A basic process chain for conversion of lignocellulosic biomass into In the environmental aspect, important benefits can also be ob-
fuels, chemicals, food/feed, and power/heat, in a biorefinery context, is tained as both, bio-based chemicals and cellulosic biofuels are able to
presented in Fig. 2. Biomass must be pretreated and hydrolyzed in order contribute to minimize GHG emissions. Second-generation biofuels
to release sugars for use on the production of fuels or chemicals. Lignin were estimated to be able to reduce GHG emissions in up to 90% de-
is also separated during this stage and, depending on the feedstock, pending on the technology and feedstock employed [8]. Bio-based
protein can also be recovered in significant amount and used for food/ chemicals are also able to significantly reduce the GHG emissions. For
feed applications. In order to have a sustainable process chain, three example, the GHG savings (t CO2/t of product) for succinic acid, bu-
important aspects must be considered: 1) determining the amount of tanol, adipic acid, and ethanol, were estimated in 5.0, 3.9, 3.3, and 2.7,
sugars to be used for the production of chemicals instead of fuels (the respectively, assuming a complete replacement of fossil-based chemical

55
C.K. Yamakawa et al.
Fig. 3. Market price and volume for different products that can be obtained
from lignocellulosic biomass (adapted from Jungmeier et al. [7]).
by bio-based chemical [9].
In brief, there are numerous opportunities for the development of
sustainable biorefineries integrating bio-based chemicals and biofuels.
However, the selection of the building block chemical is an important
factor to be considered when designing a biorefinery. This will depend,
among other factors, on the market size, price and opportunities for
such compound. In the technological aspect, several important aspects
must also be taken into account to develop efficient and sustainable
biorefineries, which include the hemicellulose integration and use of
sugars for different products, lignin valorization, development of effi-
cient and low-cost pretreatment technologies and development of
highly efficient fermentation processes. These aspects are discussed in
more details in the following sections.
3. Hemicellulose integration in a biorefinery and the “sugars
switch”
Hemicellulose is the second most abundant component (after cel-
lulose) of lignocellulosic biomass. It consists of a group of complex
heterogeneous polysaccharides composed of 5-carbon sugars (xylose
and arabinose), 6-carbon sugars (glucose, mannose and galactose), and
some acids (acetic acid, D-glucuronic acid, and D-galacturonic acid) [5].
Hemicellulose, together with lignin, contribute to the recalcitrance of
lignocellulosic materials to hydrolysis [10,11]. However, hemicellulose
is easier to break down with chemicals and/or heat when compared to
cellulose or lignin [12].
The recalcitrant nature of lignocellulosic biomass makes their ap-
plication a challenge. Pretreatment aims to convert native lig-
nocellulosic biomass into a form where hydrolysis could effectively be
achieved, providing either an increase in the accessible cellulose sur-
face area and a liquefaction of hemicellulose and lignin fractions [13].
However, promoting a selective separation of the main biomass frac-
tions is more convenient and indicated in order to maximize the ap-
plication/conversion opportunities. For example, in the pulp and paper
industry, chemicals and high temperature are used for biomass (e.g.
wood) pretreatment in order to produce insoluble cellulose fibers [14].
The by-product from pulping process, the black liquor, containing both
lignin and hemicellulose derived products, is usually burned to provide
energy [15]. However, hemicellulose has lower heating value compared
to lignin, and such application represents an uneconomical use of this
resource [10]. Separation of hemicellulose and lignin prior to com-
bustion would allow more efficient applications for both fractions.
Hemicellulose sugars, for example, can be used to produce ethanol
[16–18], acetone and butanol [19], and higher value chemicals such as
succinic acid [20], lactic acid [20,21], xylooligosaccharides [22], fur-
fural [23], and xylitol [24,25], among others.
56
Biomass and Bioenergy 119 (2018) 54–60
In fact, since hemicellulose is mainly composed of pentose sugars, it
offers numerous opportunities for biomass utilization. However, using
both cellulosic and hemicellulosic fractions for the production of fuels
(ethanol, for example) is a strategy that may not result in a positive
revenue depending on the technologies used for pretreatment and fer-
mentation. In a previous study on techno-economic assessment (un-
published results), two different scenarios were evaluated in a bior-
efinery for the production of fuels and chemicals. For analysis, raw
biomass (assuming two third of the composition being sugars (cellu-
lose + hemicellulose) and the remaining one third being lignin) was
considered at the gate in a biorefinery. In scenario I, all sugar fractions
were used to produce ethanol, while lignin was converted into power/
heat. In scenario II, 25% of the total sugars were used for the production
of chemicals (lactic acid), while the remaining 75% were converted into
ethanol, and lignin was converted into power/heat. According to the
results, the total revenue obtained for scenario II was almost twice the
revenue obtained for scenario I, confirming the importance of the
“sugar switch” for fuels and chemicals production in a lignocellulosic
biorefinery.
Another example was recently reported by Castro [26], who per-
formed a techno-economic analysis for comparison of different bior-
efinery scenarios using rice straw as a feedstock. In scenario I, the au-
thor considered all the sugars (from cellulose and hemicellulose
fractions) for ethanol production, and lignin for the production of
phenolic acids and power/heat. In this scenario, the total revenue was
US$ 295.1 million per year. However, the final net revenue was nega-
tive (US$ -37.9 million per year) (Table 1). In scenario II, it was con-
sidered using the hemicellulose sugars for the production of xylitol
instead of ethanol. All the other products were the same (cellulose to
ethanol and lignin to phenolic acids and power/heat). Using part of the
biomass sugars for the production of a more valuable compound (xy-
litol) resulted in higher total revenue (US$ 409.3 million per year), and
the final net revenue was also positive (US$ 73.5 million per year).
These results also confirm the importance of using part of the sugars
from biomass composition for the production of high-value products in
order to have a final positive balance.
Finally, integrating hemicellulose in a biorefinery, especially for the
production of high value compounds, is a strategy with great potential
to improve the revenues as well as to increase the diversity of products
that can be obtained. In a broader context, using all the different
components of lignocellulosic biomass for the production of multiple
commercially valuable products is a promising alternative to have a
‘zero-waste lignocellulosic biomass biorefinery’, to maximize the value
derived from biomass, as well as to created new bio-based industries
[5].
Table 1
Example of conversion of biomass to fuel (ethanol) and chemicals in two sce-
narios, predicted by simulation (adapted from Castro [26]).
Parameters Values predicted by simulation
Scenario I Scenario II
Production (kg per 100 kg rice straw)
Ethanol 17.42 13.96
Xylitol – 5.71
Lignin from alkali liquor 8.55 8.55
Unconverted material 21.82 21.82
Total revenues (Mil.US $ year−1) 295.1 409.3
Ethanol 65.8 52.8
Xylitol – 133.5
Phenolic acids 203.9 203.9
Others 25.4 19.1
Operation cost (Mil.US $ year−1) 361.0 367.0
Net operation cost (Mil.US $ year−1) 333.0 335.8
Net revenue (Mil.US $ year−1) −37.9 73.5
C.K. Yamakawa et al.
4. Lignin valorization
Lignin is a network polymer made up of phenylpropanoid units
(coniferyl alcohol, sinapyl alcohol, and p-coumaryl alcohol) with var-
ious carbon-to-carbon and ether linkages. Due to its molecular struc-
ture, lignins are extremely resistant to degradation and they are also
commonly insoluble in most solvents [27]. Lignin is usually separated
during the pretreatment of biomass, which has as objective promoting
at least a partial removal of this fraction in order to make the cellulose
fibers more accessible to enzymes [13].
Unlike cellulose and hemicellulose, lignin has been typically un-
derused as a feedstock for the production of fuels, chemicals and ma-
terials, being generally burned to provide power/heat. However, in
second-generation biorefineries, approx. 60% more lignin is generated
than is needed to meet plant energy needs through combustion [28]. In
addition, lignin is an important source of aromatic compounds and it
has a huge potential for the future chemicals and materials production
industries. With the development of integrated biorefineries and en-
couragement of a better use of natural resources, a growing interest in
the use of lignin has been observed, mainly for the production of more
valuable compounds instead of power/heat only.
In this sense, several valuable applications have been proposed for
lignin. Its use for the production of vanillin, for example, is a well-
established commercial process [29]. Aromatic hydrocarbons including
mixture of benzene, toluene and xylene (BTX) can be produced by
catalytic pyrolysis of lignin [30]. Phenol and BTX can also be produced
by photocatalysis of lignin [31]. Carbon fiber is another product of
interest that can be produced by catalytic pyrolysis of lignin [32] or via
processing by extruding filaments from a melt or solvent swollen gel
[33]. Dispersants, adhesives, resins, and bioactive agents can be pro-
duced by functionalization of lignin via various chemical reactions
[34]. Engineering plastics and thermoplastic elastomers, polymeric
foams and membranes can also be prepared from lignin [33].
Currently, the main lignin products in the market include vanillin,
cement additive and energy (Fig. 4). Among these products, energy
produced from lignin has the highest total market value, equivalent to
US$ 2.9 billion per year, followed by vanillin (US$ 192 million per
year) and cement additive (US$ 176 million per year). Other potential
markets for lignin products include phenol, carbon fiber, powered ac-
tivated carbon (PAC), and BTX. Phenol, in particular, has the highest
potential total market value (US$ 12 billion per year), followed by
Fig. 4. Total market volume (M$ per year) and market value ($ per ton) for
lignin products. Lignin products currently in the market (vanillin, cement ad-
ditive, and energy) and lignin products with potential markets (mixture of
benzene, toluene and xylene (BTX), powered activated carbon (PAC), carbon
fibers, and phenol) (adapted from Smith et al. [35]).
57
Biomass and Bioenergy 119 (2018) 54–60
carbon fiber (US$ 1.1 billion per year), PAC (US$ 677 million per year)
and BTX (US$ 123 million per year). Although phenol and carbon fiber
are poised to capture the largest market potential among the lignin
products, factors such as the production costs and viable chemical
conversion pathways still limit their commercial feasibility [33,35].
Despite all the potential applications and the significant increase in
the lignin product market observed in the last decades (Fig. 5) the main
current application for lignin is still for energy generation or landfill
(98%), while only 2% of the total lignin market is current related to the
production of higher valuable compounds. These numbers reveal that
there are still huge opportunities related to lignin conversion and ap-
plication to be explored. However, an important aspect to be considered
is that the chemical structure of lignin can be altered or degraded
during pretreatment, which can limit its application in some industrial
sectors. Such effect will depend on the pretreatment technology em-
ployed for biomass fractionation. Understanding the characteristics and
properties of the isolated lignin is therefore crucial to define the most
potential areas for its application.
5. Development of efficient and low-cost pretreatment
technologies
As mentioned before, pretreatment is a key step for biomass con-
version into biofuels or other bio-based products. Nevertheless, pre-
treatment is still one of the most expensive steps within the overall
biomass conversion chain. When considering the whole process for
biomass conversion into products, the major fraction of the costs
(nearly 40%) is associated with the steps for sugars recovery (including
pretreatment, enzyme production, and enzymatic hydrolysis), with
pretreatment being responsible for approx. 18% of the total costs [5].
A recent study on techno-economic assessment compared the costs
of different processes in a biorefinery for conversion of rice straw into
ethanol and other products [26]. The processes considered in this study
included a mild alkaline pretreatment, a dilute acid pretreatment,
glucose fermentation to ethanol, xylose fermentation to ethanol,
ethanol distillation, lignin recovery and also the utilities. According to
the results, the costs related to the dilute acid pretreatment were the
highest among all the considered options, mainly due to the high
temperature (120 °C) required to perform this stage. The cost to per-
form the fermentation steps was approx. half the value obtained for the
dilute acid pretreatment, but it was the second highest value when
compared to the other processes. These results confirm that pretreat-
ment is a very expensive step in a lignocellulosic biorefinery. In fact,
due to complexity of the lignocellulosic biomass matrix, usually a high
energy input is required to maximize the sugar release during pre-
treatment. Further studies are therefore needed in order to develop less
expensive and more sustainable pretreatment alternatives able to be
implemented in a large scale.
Another important point to highlight is regarding the formation of
toxic by-products during pretreatment, which should be avoided or at
least minimized in order to produce hydrolysates more suitable for use
as fermentation medium. The main toxic by-products include sugar
degradation products (furfural and hydroxymethylfurfural), lignin de-
gradation products, and acetic acid [36]. Acetic acid, in particular,
which is released from the acetyl groups present in the hemicelluloses,
is one of the most important fermentation inhibitors commonly found
in lignocellulosic hydrolysates. Due to the side effects for most of mi-
croorganisms during the fermentation, acetic acid has been denoted as
a potent inhibitor depending on its concentration. For ethanol pro-
duction, for example, the presence of this acid in the fermentation
medium in concentration of approx. 1 g/l may not be a problem for
fermentation, but when present in concentration higher than 5 g/l this
compound may cause a strong inhibition in the microbial metabolism
[37]. For the microbial production of xylitol, this acid was also de-
monstrated to be a potent inhibitor of the yeasts metabolism when
present in the medium in concentration higher than 3 g/l [38].
C.K. Yamakawa et al.
Fig. 5. Market value of lignin products from 1960 to 2014 (data from Smith et al. [35]).
Although numerous studies have been performed aiming to over-
come the toxicity of acetic acid to microorganisms, by reducing the
concentration of this acid in the fermentation medium through a de-
toxification procedure, or increasing the tolerance of the micro-
organism to this acid by means of adaptive laboratory evolution or
genetic modification, another interesting alternative could be pro-
moting a selective removal of this acid during the pretreatment step.
Acetic acid is a compound with numerous applications in the food in-
dustry and it can also be used in the production of vinyl acetate
monomers for further polymerization to produce polyvinyl acetate, or
as a green solvent in the production of terephthalic acid and ethyl
acetate [39]. If selectively recovered from biomass during pretreatment
and incorporated as a product in a biorefinery, acetic acid could be a
potential alternative to improve the revenues. Related to this idea, some
recent studies have evaluated the possibility of selectively recovering
acetic acid from biomass by applying a mild alkaline pretreatment [16]
or by pyrolysis [40], for example.
6. Development of highly efficient fermentation processes
The development of highly efficient fermentation processes is es-
sential to obtain technologies able to compete with the chemical con-
version routes currently available in large scale. In this sense, there are
some important points related to the fermentation of biomass hydro-
lysates that still require improvements. Currently, the production of a
bio-based product faces two main problems: the presence of undesirable
side-products (from sugars degradation or released during the biomass
fractionation) that act as inhibitors of the microbial metabolism, and
the absence of an efficient native microorganism able to consume
pentoses. To overcome these problems, some strategies have been
proposed.
Including an additional step for the hydrolysates detoxification be-
fore use as fermentation medium can be an alternative to reduce the
concentration of toxic compounds to levels that do not affect the mi-
crobial metabolism [36]. However, detoxification demands chemicals
or membranes resulting in more unit operations, more wastes forma-
tion, and increase in the overall cost and complexity of the process.
Regarding the sugars conversion, the well-known conventional yeast
Saccharomyces cerevisiae, widely applied in industrial ethanol produc-
tion from hexoses providing high yields and productivities, is unable to
metabolize pentoses (xylose and arabinose) due to the absence of a
pentose phosphate pathway. Nevertheless, a large number of natural
yeast strains are able to metabolize pentose sugars. Such strains, also
called as non-conventional yeasts, include Scheffersomyces stipitis (for-
merly known as Pichia stipitis), Yarrowia lipolytica, Kluyveromyces
marxianus, Debaromyces hansenii, Candida shehatea and Spathaspora
passalidarum strains. For the production of second-generation ethanol,
Scheffersomyces stipitis is the most widely studied yeast due to its great
ability to metabolize both, pentose and hexose sugars (glucose, xylose,
mannose, galactose, rhamnose, arabinose, and cellobiose) [41]. How-
ever, the conversion yield and resistance toward inhibitors are still
some points that should be improved when using non-conventional
58
Biomass and Bioenergy 119 (2018) 54–60
yeasts to obtain more efficient pentose conversion processes.
In this way, huge efforts have been done in metabolic engineering
and evolutionary engineering to create robust strains able to convert
pentoses and hexoses with high efficiency and with ability to tolerate
toxic compounds. A concrete success on the use of a genetically mod-
ified microorganism in large scale is the case of the second-generation
ethanol produced by Raízen Energia (Brazil), an industrial plant with
capacity to produce 40.1 Million liters of ethanol per year. In 2015, this
plant produced 91 L of ethanol per ton of biomass and in 2016 this
number was increased to 200 L per ton of biomass [42]. The process
currently used by Raízen is based on the use of sugarcane bagasse as
feedstock, acid pretreatment technology from Iogen, separate hydro-
lysis and fermentation, enzymes from the Danish company Novozymes,
and genetically modified yeasts for the conversion of pentoses and
hexoses to ethanol [43].
Evolutionary engineering, a strategy that consists in a strain de-
velopment by random mutation and direct selection on plates, also
encompassing recombination and continuous evolution of large popu-
lations over many generations [44], have also resulted in the devel-
opment of promising robust strains. Exposure of the Saccharomyces
cerevisiae strain TMB3500 to step-wise reduction in pH from 5 to 3.7
over 3600 h in a chemostat, for example, led to a successful growth and
ethanol production in the presence of inhibitory concentrations of
acetic acid, furfural, hydroxymethylfurfural and vanillin over the whole
pH range [45]. Evolutionary engineering was also successfully used to
develop an acetic acid tolerant Spathaspora passalidarum strain able to
produce ethanol even in the presence of 4.5 g/L of acetic acid in the
fermentation medium, with a productivity 7-fold higher and yield 2-
fold higher than the native strain [46]. This technique has also been
used in recombinant yeasts to generate evolved strains with better
tolerance to inhibitors and to improve productivity [47].
Apart from the second-generation ethanol, several bio-based che-
micals including succinic acid and aroma compounds have also been
produced in commercial scale using biomass as a feedstock and robust
microbial strains. Currently, bio-based succinic acid is commercially
produced by different companies including BioAmber, Reverdia,
Myriant and Succinity. The process used by BioAmber consists on the
use of corn as feedstock and a US DOE's E. coli as microorganism for
fermentation. Reverdia process is also based on the use of corn as
feedstock but a recombinant Saccharomyces cerevisiae for fermentation.
Myriant uses a genetically modified E. coli able to convert sugars from
lignocellulosic feedstocks and glucose from grain sorghum; while the
Succinity bioprocess uses a strain of Basfia succiniciproducens [48].
Among the aroma compounds, vanillin, for example, has been produced
in a commercial scale by Evolva using a recombinant Schizosacchar-
omyces pombe strain [49]. Amyris produces isoprenoids by using an
engineered Saccharomyces cerevisiae [50]. There are still various ali-
phatic and diamines compounds, promising candidates for the synthesis
of bio-based polyesters and polyamides, which can be produced from
biomass using metabolically engineered strains [51].
In summary, the use of robust microorganisms and optimized pro-
cess conditions is a promising strategy to achieve highly efficient
C.K. Yamakawa et al.
fermentation processes with potential to generate the bio-based product
in an accessible and commercially viable manner. This approach boosts
the construction of a bio-based economy, and enhances the valorization
of biomass feedstocks.
7. Conclusion
The development of sustainable and robust processes for biomass
conversion into fuels and chemicals in a biorefinery approach, at
competitive prices when compared to the fossil-based products, is the
ultimate goal to address societal needs around climate change and
energy security, resulting in more cost-competitive production pro-
cesses and products, reduced greenhouse gas emissions, and efficient
use of available resources (raw materials, minerals, water). Although
some bio-based processes are already available in industrial scale,
technological improvements, mainly in the pretreatment and fermen-
tation steps, are still necessary to obtain more efficient and economic-
ally viable processes. The lignin conversion to high-value products is
also an area with huge potential to be explored and incorporated in a
biorefinery perspective.
Acknowledgements
This work was supported by the Novo Nordisk Foundation,
Denmark (Grant number: NNF10CC1016517).
References
[1] M. Carus, Biobased economy and climate change - important links, pitfalls, and
opportunities, Ind. Biotechnol. 13 (2017) 41–51.
[2] IRENA. Renewable Energy and Jobs - Annual Review, International Renewable
Energy Agency, Abu Dhabi, 2018 (August 12, 2018) Available from: http://irena.
org/publications/2018/May/Renewable-Energy-and-Jobs-Annual-Review-2018.
[3] G. Bell, Second-generation biorefineries: optimization, opportunities, and implica-
tions for Australia, Ind. Biotechnol. 13 (2017) 76–84.
[4] Z. Losordo, J. McBride, J. van Rooyen, K. Wenger, D. Willies, A. Froehlich,
I. Macedo, L. Lynd, Cost competitive second-generation ethanol production from
hemicellulose in a Brazilian sugarcane biorefinery, Biofuels, Bioprod. Bioref. 10
(2016) 589–602.
[5] S.I. Mussatto, G.M. Dragone, Biomass pretreatment, biorefineries and potential
products for a bioeconomy development, Biomass Fractionation Technologies for a
Lignocellulosic Feedstock Based Biorefinery. Mussatto SI, Elsevier Inc., Waltham,
MA, 2016, pp. 1–22.
[6] W.M. Budzianowski, High-value low-volume bioproducts coupled to bioenergies
with potential to enhance business development of sustainable biorefineries,
Renew. Sustain. Energy Rev. 70 (2017) 793–804.
[7] G. Jungmeier, M. Hingsamer, R. van Ree, Biofuel-driven biorefineries, IEA
Bioenergy Report - Task 42 Biorefinery (2013) (August 03, 2017) Available from:
https://www.nachhaltigwirtschaften.at/resources/iea_pdf/iea_task_42_biofuel_
driven_biorefineries_lr.pdf.
[8] FAO, The State of Food and Agriculture - Biofuels: Prospects, Risks and
Opportunities, Food and Agriculture Organization of United Nations, 2008 (2008)
(August 03, 2017) Available from: http://ftp.fao.org/docrep/fao/011/i0100e/
i0100e.pdf.
[9] E. Jong, A. Higson, P. Walsh, M. Wellisch, Bio-based chemicals - value added
products from biorefineries, IEA bioenergy Report - Task 42 biorefinery (2011)
(August 03, 2017) Available from: http://www.qibebt.cas.cn/xscbw/yjbg/201202/
P020120223415452622293.pdf.
[10] M. FitzPatrick, P. Champagne, M.F. Cunningham, R.A. Whitney, A biorefinery
processing perspective: treatment of lignocellulosic materials for the production of
value-added products, Bioresour. Technol. 101 (2010) 8915–8922.
[11] S.I. Mussatto, M. Fernandes, A.M.F. Milagres, I.C. Roberto, Effect of hemicellulose
and lignin on enzymatic hydrolysis of cellulose from brewer's spent grain, Enzym.
Microb. Technol. 43 (2008) 124–129.
[12] F. Cherubini, The biorefinery concept: using biomass instead of oil for producing
energy and chemicals, Energy Convers. Manag. 51 (2010) 1412–1421.
[13] S.I. Mussatto (Ed.), Biomass Fractionation Technologies for a Lignocellulosic
Feedstock Based Biorefinery, Elsevier Inc., Waltham, MA, 2016.
[14] H. Krawczyk, Separation of Biomass Components by Membrane Filtration. Process
Development for Hemicellulose Recovery, PhD Thesis Lund University, Lund,
Sweden, 2013.
[15] A. Dieste, L. Clavijo, A.I. Torres, S. Barbe, I. Oyarbide, L. Bruno, F. Cassella, Lignin
from Eucalyptus spp. kraft black liquor as biofuel, Energy Fuels 30 (2016)
10494–10498.
[16] R.C.A. Castro, B.G. Fonseca, H.T.L. Santos, I.S. Ferreira, S.I. Mussatto, I.C. Roberto,
Alkaline deacetylation as a strategy to improve sugars recovery and ethanol pro-
duction from rice straw hemicellulose and cellulose, Ind. Crop. Prod. 106 (2017)
59
Biomass and Bioenergy 119 (2018) 54–60
65–73.
[17] F. Peng, P. Peng, F. Xu, R.-C. Sun, Fractional purification and bioconversion of
hemicelluloses, Biotechnol. Adv. 30 (2012) 879–903.
[18] J.P.A. Silva, S.I. Mussatto, I.C. Roberto, The influence of initial xylose concentra-
tion, agitation, and aeration on ethanol production by Pichia stipitis from rice straw
hemicellulosic hydrolysate, Appl. Biochem. Biotechnol. 162 (2010) 1306–1315.
[19] A. Sanchez, I. Valdez-Vazquez, A. Soto, S. Sánchez, D. Tavarez, Lignocellulosic n-
butanol co-production in an advanced biorefinery using mixed cultures, Biomass
Bioenergy 102 (2017) 1–12.
[20] M. Zhang, L. Xie, Z. Yin, S.K. Khanal, Q. Zhou, Biorefinery approach for cassava-
based industrial wastes: current status and opportunities, Bioresour. Technol. 215
(2016) 50–62.
[21] A.G. Daful, J.F. Görgens, Techno-economic analysis and environmental impact as-
sessment of lignocellulosic lactic acid production, Chem. Eng. Sci. 162 (2017)
53–65.
[22] C. Álvarez, A. González, M.J. Negro, I. Ballesteros, J.M. Oliva, F. Sáez, Optimized
use of hemicellulose within a biorefinery for processing high value-added xylooli-
gosaccharides, Ind. Crop. Prod. 99 (2017) 41–48.
[23] L. Mesa, M. Morales, E. González, C. Cara, I. Romero, E. Castro, S.I. Mussatto,
Restructuring the processes for furfural and xylose production from sugarcane ba-
gasse in a biorefinery concept for ethanol production, Chem. Eng. Process 85 (2014)
196–202.
[24] X. Jiang, P. He, X. Qi, Y. Lin, Y. Zhang, Q. Wang, High-efficient xylitol production
by evolved Candida maltosa adapted to corncob hemicellulosic hydrolysate, J.
Chem. Technol. Biotechnol. 91 (2016) 2994–2999.
[25] S.I. Mussatto, G. Dragone, I.C. Roberto, Kinetic behavior of Candida guilliermondii
yeast during xylitol production from brewer's spent grain hemicellulosic hydro-
lysate, Biotechnol. Prog. 21 (2005) 1352–1356.
[26] R.C.A. Castro, Produção de etanol a partir da palha de arroz por sacarificação e
fermentação simultânea empregando um reator agitado não convencional. PhD
thesis. Escola de Engenharia de Lorena, Universidade de São Paulo. Lorena, São
Paulo, Brazil, 2016.
[27] F. Lu, J. Ralph, Lignin, in: R.-C. Sun (Ed.), Cereal Straw as a Resource for
Sustainable Biomaterials and Biofuels, Elsevier, Oxford, 2010, pp. 169–207.
[28] P. Sannigrahi, Y. Pu, A. Ragauskas, Cellulosic biorefineries - unleashing lignin op-
portunities, Curr. Opin. Environ. Sustainability 2 (2010) 383–393.
[29] J.D.P. Araújo, C.A. Grande, A.E. Rodrigues, Vanillin production from lignin oxi-
dation in a batch reactor, Chem. Eng. Res. Des. 88 (2010) 1024–1032.
[30] D. Shen, J. Zhao, R. Xiao, S. Gu, Production of aromatic monomers from catalytic
pyrolysis of black-liquor lignin, J. Anal. Appl. Pyrolysis 111 (2015) 47–54.
[31] C.A. Lekelefac, N. Busse, M. Herrenbauer, P. Czermak, Photocatalytic based de-
gradation processes of lignin derivatives, Int. J. Photoenergy 2015 (2015)
13763418 pages https://doi.org/10.1155/2015/137634.
[32] B. Shrestha, Y. Brech, T. Ghislain, S. Leclerc, V. Carré, F. Aubriet, S. Hoppe,
P. Marchal, S. Pontvianne, N. Brosse, A. Dufour, A multi-technique characterisation
of lignin softening and pyrolysis, ACS Sustain. Chem. Eng. (2017), https://doi.org/
10.1021/acssuschemeng.7b01130.
[33] A.J. Ragauskas, G.T. Beckham, M.J. Biddy, R. Chandra, F. Chen, M.F. Davis,
B.H. Davison, R.A. Dixon, P. Gilna, M. Keller, P. Langan, A.K. Naskar, J.N. Saddler,
T.J. Tschaplinski, G.A. Tuskan, C.E. Wyman, Lignin valorization: improving lignin
processing in the biorefinery, Science 344 (2014) 1246843, https://doi.org/10.
1126/science.1246843.
[34] Y. Matsuhita, Conversion of technical lignins to functional materials with retained
polymeric properties, J. Wood Sci. 61 (2015) 230–250.
[35] P. Smith, M. Chen, S. Cline, Biorefinery value chain outputs, Final report, Northwest
advanced renewable Alliance (2016) (August 03, 2017) Available from: https://
nararenewables.org/documents/2017/02/155205-nara-biorefinery-value-chain-
outputs-vcea-b-p2.pdf.
[36] S.I. Mussatto, I.C. Roberto, Alternatives for detoxification of diluted-acid lig-
nocellulosic hydrolyzates for use in fermentative processes - a review, Bioresour.
Technol. 93 (2004) 1–10.
[37] M.J. Taherzadeh, L. Gustafsson, C. Niklasson, G. Lidén, Conversion of furfural in
aerobic and anaerobic batch fermentation, J. Biosci. Bioeng. 87 (1999) 169–174.
[38] M.G.A. Felipe, D.C. Vieira, M. Vitolo, S.S. Silva, I.C. Roberto, I.M. Manchilha, Effect
of acetic acid on xylose fermentation to xylitol by Candida guilliermondii, J. Basic
Microbiol. 35 (1995) 171–177.
[39] N. Murali, K. Srinivas, B.K. Ahring, Biochemical production and separation of
carboxylic acids for biorefinery applications, Fermentation 3 (2017) 22 https://doi.
org/10.3390/fermentation3020022.
[40] J.A. Kocal, Uop Llc. Acetic acid production from biomass pyrolysis. US Patent n.
US8716522B2, 06 may 2014, https://patents.google.com/patent/US20120172622.
[41] S.R. Ceccato-Antonini, C.B. Codato, C. Martini, R.G. Bastos, S.M. Tauk-Tornisielo,
Yeast for pentose fermentation: isolation, screening, performance, manipulation,
and prospects, in: M.S. Buckeridge, A.P. de Souza (Eds.), Advances of Basic Science
for Second Generation Bioethanol from Sugarcane, Springer International
Publishing, 2017, pp. 133–157.
[42] A.A. Stuchi, Second Generation Ethanol. Ethanol Summit, 26-27/jun/2017, Sao
Paulo Brazil (August 03, 2017) Available from: http://ethanolsummit.com.br/wp-
content/uploads/2017/07/27.06-palco4-10h45-Antonio-Alberto-Stuchi.pdf.
[43] R. Bossle, Exclusivo: Custo de produção estimado do etanol celulósico nas 6 maiores
usinas do mundo, Novacana, 2016 (August 03, 2017) Available from: http://www.
novacana.com/n/etanol/2-geracao-celulose/custo-producao-etanol-celulosico-
usinas-mundo-150316/.
[44] U. Sauer, Evolutionary engineering of industrially important microbial phenotypes,
Adv. Biochem. Eng. Biotechnol. 73 (2001) 129–169.
[45] V. Narayanan, V.S. Nogué, E.W.J. van Niel, M.F. Gorwa-Grauslund, Adaptation to
C.K. Yamakawa et al.
low pH and lignocellulosic inhibitors resulting in ethanolic fermentation and
growth of Saccharomyces cerevisiae, Amb. Express 6 (2016) 59 https://doi.org/10.
1186/s13568-016-0234-8.
[46] P. Morales, J.C. Gentina, G. Aroca, S.I. Mussatto, Development of an acetic acid
tolerant Spathaspora passalidarum strain through evolutionary engineering with
resistance to inhibitors compounds of autohydrolysate of Eucalyptus globulus, Ind.
Crop. Prod. 106 (2017) 5–11.
[47] R. Koppram, E. Albers, L. Olsson, Evolutionary engineering strategies to enhance
tolerance of xylose utilizing recombinant yeast to inhibitors derived from spruce
biomass, Biotechnol. Biofuels 5 (2012) 32 https://doi.org/10.1186/1754-6834-
5-32.
[48] N.P. Nghiem, S. Kleff, S. Schwegmann, Succinic acid: technology development and
commercialization, Fermentation 3 (2017) 26 https://doi.org/10.3390/
60
Biomass and Bioenergy 119 (2018) 54–60
fermentation3020026.
[49] N.J. Gallage, B.L. Møller, Vanillin – bioconversion and bioengineering of the most
popular plant flavor and its de novo biosynthesis in the vanilla orchid, Mol. Plant 8
(2015) 40–57.
[50] A.L. Meadows, K.M. Hawkins, Y. Tsegaye, E. Antipov, Y. Kim, L. Raetz, R.H. Dahl,
A. Tai, T. Mahatdejkul-Meadows, L. Xu, L. Zhao, M.S. Dasika, A. Murarka,
J. Lenihan, D. Eng, J.S. Leng, C.-L. Liu, J.W. Wenger, H. Jiang, L. Chao, P. Westfall,
J. Lai, S. Ganesan, P. Jackson, R. Mans, D. Platt, C.D. Reeves, P.R. Saija,
G. Wichmann, V.F. Holmes, K. Benjamin, P.W. Hill, T.S. Gardner, A.E. Tsong,
Rewriting yeast central carbon metabolism for industrial isoprenoid production,
Nature 537 (2016) 694–697.
[51] H. Kawaguchi, C. Ogino, A. Kondo, Microbial conversion of biomass into bio-based
polymers, Bioresour. Technol. 245 (2017) 1664–1673.