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Xylanases and their industrial applications: A review

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Biochem. Cell. Arch. Vol. 17, No. 1, pp. 353-360, 2017 www.connectjournals.com/bca ISSN 0972-5075

XYLANASES AND THEIR INDUSTRIAL APPLICATIONS : A REVIEW


Dushyant Kumar1, Smita S. Kumar2, Jagdeesh Kumar3, Om Kumar4, Shashi Vind Mishra4,
Rahul Kumar5 and Sandeep K. Malyan4*
1
Environmental Research Laboratory, Department of Paper Technology, IIT,Roorkee,Saharanpur Campus, Saharanpur - 247 001,
India.
2
Department of Environ.Science and Engineer., Guru Jambheshwar University of Science and Technology,Hisar-125 001, India
3
Department of Hydrology, Indian Institute of Technology, Roorkee - 247 667, India
4
Centre for Environment Science and Climate Resilient Agriculture, ICAR-Indian Agricultural Research Institute,
New Delhi - 110 012, India
5
Division of Vegetable Science, ICAR-Indian Agricultural Research Institute, New Delhi - 110 012, India
*e-mail : sandeepkmalyan@gmail.com
(Accepted 3 April 2017)

ABSTRACT : Global population is increasing at rapid rate which result in high demand of life supporting product. For the
production of daily required product such as food, cloths, paper and pharmaceuticals larger amounts of raw material is
consumed. During the production huge amount of chemical and other waste is generated which result in environmental
problem. Therefore, Industries around the world are thus looking for eco-friendly alternatives technologies for fulfilling the
product demand without polluting environment. The use of enzymatic processes have been implanted in broad range of industries
in recent decades because they are specific, fast in action and often save raw materials, energy and chemical compared to
conventional processes. Only in past two decades, however, have microbial enzymes been used commercially in many industries
such as pulp and paper, food industry, textile industry and feed industry. The aim of this study was to explore the application of
xylanases enzyme at different industrial level. Xylanases was produced from different microorganisms (fungi and bacteria) and
it is widely used in pulp and paper industries. Apart from pulp and paper industry xylanase have potential for application in fruit
juice processing and bakery processes. Xylanase enzyme was found to be effective in getting enhanced sugar extraction from
fruit juices, clarification of fruit juices, and substantial dough-raising in bakery. This review will focus on some properties of
xylan, as well as regulation and production and their potential industrial applications.
Key words : Enzyme, xylanase, pulp and paper industry, bioleaching.

INTRODUCTION commercial Xylanases has been growing since the


Xylanase is the class of enzymes produced by beginning of the 1990s. Several Xylanases with different
microorganisms which break down hemicellulose. trade name and properties are now commercially
Hemicellulose (major part of plant cell wall) consists of available. With the research and development of industrial
polymer glucose known as xylan and after cellulose xylan biotechnology, protein engineering, structural biology and
was second most abundant renewable biomass present molecular biology significant progress has been made in
on earth after. Xylanases (3.2.1.8) catalyze the the area on structures and functions of xylanases (Bajpai
breakdown of xylan into xylooligosaccharides and D- 1999). An increasing concerned on environmental
xylose. Xylan is a heterogeneous polysaccharide and pollution and sustainable development has enforce the
consists of a linear backbone of b-1, 4-D-xylopyranoside application of xylanase in different industries such as
residues and short side-chain branches. Xylan has pulp and paper, food and feed etc, which reduce the use
complex structure so it required several enzymes for its of hazards chemicals in their processes ( Thomas et al,
complete hydrolysis, but endo b-1, 4-xylanase play 2015). Wastewater generated from different sources
important role in depolymerization of xylan (Thomas et such as industry effluent (Sharma and Dhaka 2008;
al, 2015; Bajpai 2009). Xylanases application in different Sharma et al, 2012; Kumar et al, 2016) need proper
sectors has increased markedly in the past decade. Due management. It has application in agricultural silage and
to its hydrolysis and other properties Xylanases is widely grain feed and in combination with cellulose and pectinase,
used in many industrial such as –pulp and paper, food, xylanases was used for clarification of fruit juices.
textiles, bio-fuel, animal feed and drinks. The market for Recently Awalgonkar et al. (2015) reported that xylanase
also aid quality of papads, an Indian traditional food based
354 Dushyant Kumar et al
on black gram. Xylanases for industrial use are produced fungi are most widely used for production of this enzyme.
mainly from filamentous fungi (Sunna and Antranikian Several types of Xylanase is obtained from fungi including
1997) and bacteria (Bajaj and Singh 2010) in solid state Aspergillus, Disporotrichum, Penicillium,
fermentation (SSF) or submerged fermentation (SmF). Trichoderma, Neurospora, Neocallimastix,
Fungi known to produce Xylanase include Aspergillus, Coniothyrium, T. lanuginosus, Fusarium, and so on.
Neurospora, Penicillium, Neocallimastix, Apart from xylanase fungi produce different xylanolytic
Disporotrichum, Trichoderma, Fusarium, enzymes which help in de-branching of xylan present in
Coniothyrium, T. lanuginosus, and so on. In this study cell wall. Xylanases may be produced from some extreme
we synthesized the information on xylanases enzyme environments bacteria species, which may be more
production, and it application in different industries. suitable in industrial applications in future. Characteristics
Xylanases structures of some xylanases produced by different microorganisms
were summarized in table 1.
Both â-xylosidases and Endo-â-1,4-xylanases belong
to enzyme class of glycoside hydrolyses (GH). GHs are In economics point of view the cost of bio-catalytic
classified into families and families are further grouped production of xylanases enzymes can be reduced by
in clans. â-Xylosidases are grouped into five families, 3, finding optimum conditions for their production by the
39, 43, 52 and 54, while Endo-xylanases are found in construction feasible producers through genetic
families 5, 7, 8, 10, 11, 26 and 43 of GH (Juturu and Wu engineering and by the of isolation of hyperproducing
2012). Xylanase from the family GH10 or family G have mutants. A rational approach to these targets requires
isoelectric point (pI) between 8 to 9.5 with low molecular knowledge of the regulatory mechanisms governing
mass, while those from family F or GH11 have a high enzyme production. Studies of the regulation of xylanolytic
molecular mass and lower pI values (Buchert et al, 1995; enzymes have largely focused on the induction of enzyme
Ahmed et al, 2009). The structure of family 10 endo- activities under various conditions rather than on gene
xylanases has been explained taking the crystal structure regulation (Bajpai 2009). Xylanases is produced in both
of thermostable xylanase from Thermoascus submerged liquid and solid substrate, but out of total
aurantiacus as an example (Fig. 1). production about 90 % in produced in liquid submerged
culture. Table 2 summarizes data on different xylanases
It is a 32.5 kDa polypeptide chain and assumes (á/
produced worldwide.
â)8 TIM-barrel fold, which consists of eight major parallel
â-strands arranged side-by-side forming a cylinder in the Application of xylanases
center followed by eight major á helices (Juturu and Wu In recent published studies, the biotechnological use
2012). The structure of family 11 xylanases has explained of xylonlytic enzymes has grown increased for the last
taking Bacillus subtilis (generally known as the XynA few decades. The important applications of xylanases
or xylanase A) as an example (Fig. 2) determined by x- are in the paper industries, feed industries, food industries,
ray crystallography. It contains two twisted b-sheets pharmaceutical industries, and bio-fuel ( Bajpai 1999;
forming a so called “jellyroll” fold which looks remarkably Ahlawat et al, 2007, Pedersen et al, 2015). The
like a right hand, in which the individual b-strands thread application in major industries was described below:
back and forth to form the “finger” and “palm” domains Pulp and Paper industry
(shown as the regions F and P in the diagram on the
Process of lignin removal from pulps to produce more
right. GH11 xylanases members display several
brightness in paper or to get almost white finished paper
interesting properties, such as high catalytic efficiency
from pulp is called ‘bleaching’. Bleaching of pulp can be
and high substrate selectivity, a variety of optimum
achieved by both chemical and biological process.
temperature values and pH values, makes them suitable
Process of lignin removal from pulp by the application of
for various applications. GH11 family composed only of
alkaline chemical at the stage of cooking of pulp at 170
Xylanases (EC3.2.1.8), leading to their consideration as
ÚC was kwon as chemical leaching. Chemical bleaching
“true xylanases” as they are exclusively active on D-
process requires chlorine based agents such as ClO2, Cl2,
xylose-containing substrates (Motta et al, 2013).
and hypochlorite, which results in the environmental
Production of xylanases pollution (Bajpai 2012). In order to reduce environmental
Xylanases can be produced different living organisms pollution load role of bio-leaching came into light. The
such as fungi, yeast, bacteria, marine algae, snails, application of microorganism or their derivatives
crustaceans, insect, seeds, and even from protozoans (enzymes) in the process of bleaching of pulp in the paper
also. In economical production point of view, filamentous industry is known as biobleaching (Fig. 3). During the
Xylanases and their industrial applications 355

Fig. 1 : Structural views of endo-xylanase from T. aurantiacus (A): Fig. 2 : Topology diagram of typical GH11 family xylaneses (Source:
A front view showing the (á/â) 8 TIM-barrel fold; (B): A Paes et al, 2012).
salad bowl view exposing the substrate binding cleft (Source:
Juturu and Wu, 2012).

Fig. 3 : General flow-diagram of biobleaching process (Kumar et al, 2016).

past few years, the use of xylanases enzymes in paper Xylanases enzymes have been used in pulp bleaching
and pulp industries has caught the attention of researchers primarily to reduce lignin and increase the brightness of
and industries all over the world. A number of scientific the pulp (Sunna and Antranikian 1997). It reduces the
research on xylanases and their application in paper pulp usage of chemical chlorine as the bleaching agent as
bio-bleaching support the feasibility of their industrial Xylanases in pulp and paper industries is related to the
application (Sharma et al, 2015; Bajpai 2004). Enzymes hydrolysis of xylan, which enhances the process of lignin
xylanases application improve pulp fibrillation and water removal from paper pulp.
retention time, reduction of beating duration, restoration Xylanases are commercially used in bleaching of kraft
of bonding and increases freeness in recycled fibers etc. pulp and xylanases are gaining global importance as eco-
356 Dushyant Kumar et al
Table 1 : Characteristics of some Xylanases produced by different microorganisms.
Microorganisms Optimal pH Optimal temperature (°C) Reference
Cryptococcus albidus 5 25 Morosoli et al (1986)
Aureobasidium pullulansY-2311-1 4.4 54 Li et al (1993)
Aspergillus sydowii 4 50 Ghosh & Nanda (1994)
Aspergillus oryzae 5 60 Fernández-Espinar et al (1994)
Aspergillus terreus 4.5 45 Kimura et al (1995)
Thermotoga maritime MSB8 6.5 55 Winterhalter & Liebel (1995)
Aspergillus fischeri 6 60 Raj & Chandra (1996)
Paecilomyces varioti 5 60 Cesar & Mrsa (1996)
Acidobacterium capsulatum 5 65 Inagaki et al, 1998
Aspergillus versicolor 6 55 Carmona et al (1998)
Trichosporon cutaneumSL 409 6.5 50 Liu et al (1998)
Acrophialophora nainiana 7 55 Martinez-Anaya & Jimenez (1998)
Aspergillus oryzae 6 50 Kitamoto et al (1999)
Chaetomium cellulolyticum 6.5 50 Baraznenok et al (1999)
Acrophialophora nainiana 5 50 Ximenes et al (1999)
Aspergillus terreus 7 50 Ghanen et al (2000)
Aspergillus nidulans 6 56 Salles et al (2000)
Bacillus circulans 6-7 80 Dhillon et al (2000)
Aspergillus nidulans 8 55 Taneja et al (2002)
Penicillium capsulatum 3.8 48 Ryan et al (2003)
Myceliophthora sp. 6 75 Chadha et al (2004)
Schizophyllum commune 5.5 50 Kolenova et al (2005)
Aspergillus oryzae 4-6 50 Szendefy et al (2006)
Bacillus pumilus 8.0 37 Battan et al (2007)
Thermomyces lanuginosus 6.5 65 Ziaie-Shirkolaee et al (2008)
Penicillium oxalicum 9 55 Dwivedi et al (2009)
Aspergillus sydowii 2-12 30 Nair et al (2010)
Promicromonospora sp MARS 8 65 Kumar et al (2011)
Bacillus lichenformis 7.5 50 Liu et al (2012)
Aspergillus niger 7.5 60 Ahmad et al (2013)
Paenibacillus terrae HPL-003 4-11 55 Song et al (2013)
Aspergillus terreus 6 50 Moreira et al (2013)
Aspergillus lentulus 5.3 50 Kaushik et al (2014)
Geobacillus stearothermophilus KIBGE IB29 6 60 Bibi et al (2015)

friendly alternatives to chlorine-containing chemicals Xylanase applications in paper industry are often
which were toxic in environment. Viikari (1986) was the termed pre-bleaching as it boost bleaching by enhancing
first to report that application of xylanase in pulp and the chemical bleaching process rather than to remove
paper enhance the process of bleaching. The Finnish lignin directly. Application of xylanases does not attack
forest companies were the first in the global level to start lignin-based chromophores directly (Bajpai 2009; 2004).
mill-scale experimentation in 1988 (Bajpai 2009). In A limited break down of the xylan network is often enough
laboratory studies results shown about 30-40 % and 10- to facilitate the subsequent chemical attack on lignin with
25% reduction in active chlorine application at chlorination various bleaching chemicals, without effecting yield.
stage for hardwoods and softwoods respectively, whereas Therefore, a xylanases application for pre-bleaching of
in mill trials total active chlorine was found to be 20-25% pulp appears to be the most suitable step to facilitate
and 10-15% for hardwoods and softwoods respectively bleaching boosting of pulp. Xylanases catalyze the
(Bajpai 1999; Garg et al, 1998). Endo-xylanase was the hydrolysis of xylan complex chain and solubilize only some
key enzyme required to enhance the delignification of fraction of total available xylan. The main enzyme needed
kraft pulp but additions hemicellulolytic enzymes alone to enhance the process of bioleaching of kraft pulp is
with xylanase improved to be more in bioleaching process. endo-beta- xylanase. The actual mechanism in bleaching
Xylanases and their industrial applications 357
Table 2 : Xylanases and their industrial application.
Trade name Supplier Application
Bleachzyme F Bicon, India Paper industry
Irgazyme 40 Nalco-Genencor, Ciba, -Geigy Paper industry
Pulpzyme Novozymes, Denmark Paper industry
Ecopulp Rohn Enzyme 0Y, Primalco, Finland Paper industry
Ecozyme Thomas Swan, UK Paper industry
VAI Xylanase Voest Alpine, Austria Paper industry
Ecopulp AlkoRajamaki, Finland Paper industry
Cartazyme Sandoz, Charlotte, N.C. and Basle, Switzerland Paper industry
Optipulp L-8000 Solvay Interox, USA Paper industry
GS-35, HS70 Iogen, Canada Paper industry
Rholase 7118 Rohm, Germany Food industry
Multifect XL Genencor, Netherlands Food industry
Sternzym HC 46 Stern-Enzym, Mexico Food industry
Sanzyme X Sankyo, Japan Food industry
Ceremix Novo Nordisk, Denmark Food industry
Solvay pentonase Solvay Enzymes, Canada Food industry
Sanzyme PX, Alpelase F Sankyo, Japan Feed industry
Bio-Feed Plus Novo Nordisk, Denmark Feed industry
Enzekoxylanase Enzyme Development, USA Feed industry
Ecosane Biotec, Thailand Feed industry
Grindazym GP e GV Danisco Ingredients, India Feed industry
Biofeed Novo Nordisk, Denmark Feed industry
Allzym PT Alltech, America Feed industry
Xylanase Meito Sankyo, Nogaya Japan Research
Amano 90 Amano, Japan Pharmaceutical industry
Xylanase250 HankyoBioindustryCo.Ltd, Japan Baking industry
Grindazym GP 5000 Danisco Ingredients, Denmark Feed industry
Gammazym X4000L Gamma Chemie GmbH, Germany Brewing industry

process is not fully understood. But the application of dough to grow. Butt et al (2008) reported that the
xylanases reduces chemical load into the environment application of xylanases increased the volume of bread
and it is economical feasible process in pulp and paper and also improved the resistance potential of bread against
industry. fermentation. Xylanases was widely used in juice and
Xylanases in food industry wine industries also. The production of vegetable and
fruit juices requires three main methods namely:
From some recent published study it was clear that
extraction, clearing and stabilization of juices. In the early
the application of xylanases in food industries such as in
nineteen’s when the manufacture of juices (citrus fruit)
bread, juice, papad industries have important role (Polizeli
began; there was a low yield due to filtration and turbidly
et al, 2005; Butt et al. 2008; Harris and Ramalingam,
problem. The application of microbe based enzyme helped
2010; Goswani and Rawat, 2015). Xylanses along with
to solve these problems. Recently, xylanases, in
malting amylase, á-amylase, proteases, and glucose
conjunction with amylases, pectinases and cellulases lead
oxidase were use in food industries. The study reports
to an improved quality and quantity of juice by means of
that the application xylanases improve bread quality by
liquefaction of vegetables and fruit; stabilization of the
increases the bread volume and it also reported that
fruit pulp; increased recovery of aromas, essential
amylase and xylanases gave better result in bread
vitamins, oils, mineral salts, pigments, edible dyes, etc.,
production. Application of xylanases breaks down the
reduction of viscosity, hydrolysis of substances that hinder
wheat-flour hemicelluloses content which result in
the physical or chemical clearing of the juice, or that may
redistribution of water and enhances the softness of
cause cloudiness in the concentrate. Xylanases, along
dough (Harris and Ramalingam, 2010). During the bread-
with endoglucanase, takes part in the breakdown of
baking process, they delay crumb formation, allowing the
arabinoxylan and starch, separating and isolating the
358 Dushyant Kumar et al
gluten from the starch in the wheat flour. Due to Production of thermostable pectinase and xylanase for their
maximum activity at acidic pH and due to its high stability potential application in bleaching of kraft pulp. J. Industrial
Microbiol. Biotechnol. 34, 763–770.
enzyme xylanases have achieved wide application in food
Ahmad Z, Butt M S and Riaz M (2013) Paritial purification and
industries.
characterization of Xylanase produced from Aspergillus niger
Xylanases in bio-fuel production using wheat bran. Pakistan J. Agricul. Sci. 50, 433-437.
Xylanases with other hydrolytic enzymes may be used Ahmed S, Riaz S and Jamil A (2009) Molecular cloning of fungal
xylanases: an overview. App. Microbiol. Biotechnol. 84, 19-35.
for bio-fuels production, from lignocellulosic biomass
(Dood and Cann, 2009; Motta et al, 2013; Garg 2015). Awalgaonkar G, Sarkar S, Bankar S and Singhal R S (2016) Xylanase
as a processing aid for papads, an Indian traditional food based
Lignocellulosic biomass ere primary used in the production on black gram. Food Sci. Technol. 62, 1148-1153.
of second generation bio-fuels (Parsad et al, 2012).
Bajaj B K and Singh N P (2010) Production of xylanase from analkali
Biomass is de-lignified followed by de-polymerization and tolerant Streptomyces sp.7bunder solid-state fermentation, its
fermentation to generate biofuel such as ethanol and purification, and characterization. App. Biochem. Biotechnol. 162,
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Bajpai P (2012) Environmentally benign approaches for pulp bleaching.
CONCLUSION 2nd edn. Elsevier, New York
Xylanases have wide range of application across Baraznenok V A, Becker E G, Ankudimova N and Okunev N N (1999)
several industries such as pulp and paper, food, animal Characterization of neutral xylanases from Chaetomium
feed, and pharmaceutical. Due to its biodegradable cellulolyticum and their biobleaching effect on eucalyptus pulp.
Enzyme Microb. Technol. 25, 651–659.
property xylanases can be used different industries
including food without any toxicity. Xylanases use in food Battan B, Sharma J, Dhiman S S and Kuhad R C (2007) Enhanced
production of cellulase-free thermostable xylanase by Bacillus
industries should be promoted to reduce load of chemical pumilus ASH and its potential application in paper industry.
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moving toward safer industrial production. Lager scale Production of xylan degrading endo-1, 4-b-xylanase from
production of xylanases to reduce their cost is still big thermophilic Geobacillus stearothermophilus KIBGE-IB29. J.
challenging task. The knowledge gathered about the Rad. Res. App. Sci. 7, 478-485.
powerful xylanases enzymes to meet the industrial Buchert J, Tenkanen M, Kantelinen A and Viikari L (1995) Application
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50, 65-72.
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Butt M S, Tahir-Nadeem M and Ahmad Z (2008). Xylanases and
chemicals etc, has greatly promoted the rapid application their applications in baking industry. Food Technol. Biotechnol
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ACKNOWLEDGMENTS
Cesar T and Mrsa V (1996) Purification and properties of xylanase
First and last author were thankful for University produced by Thermomyceslanuginosus. Enzyme Microb.
Grants Commission, New Delhi, India for providing Technol. 19, 289-296.
fellowship. We were thankful to P.G School IARI, New Chadha B S, Ajay B K, Mellon F and Bhat M K (2004) Two
Delhi-110012 for providing all necessary facility provided. endoxylanases active and stable at alkaline pH from the newly
isolated thermophilic fungus, Myceliophthora sp. IMI 387099.
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