CHARACTERIZATION AND CATALYTIC PROPERTY OF XYLAN DEGRADING ENZYME FROM MICROBIAL SOURCE
Abstract
Objective: To study the catalytic property and characterization of xylan degrading enzyme from microbial source. Xylan degrading enzyme xylanase are extracted, purified and characterized from Bacillus megaterium SV1. Methods: The enzyme was purified to homogeneity by ammonium sulphate precipitation, gel permeation and ion exchange chromatographic techniques. Â Results: During the series of steps, 28.5 fold purification was obtained with 72.7Uper mg specific activity of xylanse and the corresponding molecular weight of the enzyme was identified as 24kDa. The optimum conditions for maximal enzyme activity were identified at pH8.0, temperature 40degreeC and with 5 of sodium chloride. It is observed that the reactants like calcium chloride, Dithiothreitol, beetamercaptoethanol wasfound to enhance the activity of enzyme and Mercuric chloride strongly inhibits the enzyme activity. Conclusion: Degradation of Brich wood Xylan was investigated to determine the kinetic parameters Km and Vmax values and was found to be 6.1mg per ml and 280micromol per minper mg respectively. It is also verified that the enzyme Xylanase extracted from Bacillus SV1 was identified as alkalophilic and halotolerant.
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Keywords: Purification, Characterization, xylanase, Bacillus megaterium SV1.
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Perez J, MunozDorado J, DeLaRubia T and Martinez J: Biodegradation and biological treatments of cellulose, hemicellulose and lignin: an overview. Int Microbiol 2002; 5: 5363.
Saha BC: Production, purification and properties of xylanase from a newly isolated Fusarium proliferatum. Bioresour Technol 2002; 37: 12791284.
Tuomela M, Vikman M, Hatakka A and Itavaara M: Biodegradation of lignin in a compost environment: a review. Bioresour Technol 2000; 72: 169183.
Prade RA: Xylanases: from biology to biotechnology. Biotechnol Genet Eng Rev 1995; 13: 101131.
Chivero ET, Mutukumira AN and Zvauya R: Partial purification and characterization of a xylanase enzyme produced by a microorganism isolated from selected indigenous fruits of Zimbabwe. Food Chem 2001; 72: 179183.
Beg QK, Kapoor M, Mahajan L and Hoondal GS: Microbial xylanases and their industrial applications: a review. Appl Microbiol Biotechnol 2001; 56: 326328.
Collins T, Gerday C and Feller G: Xylanases, xylanase families and extremophilic xylanases. FEMS Microbiol Rev 2005; 29: 323.
Subramaniyan S and Prema M: Cellulase free xylanases from Bacillus and other microorganisms. Microbiol Lett 2000; 183: 17.
Srinivasan MC and Rele MV: Microbial xylanases for pulp and paper industry. Curr Sci 1999; 77: 137142.
Biely P, Markovic O and Mislovikova D: Sensitive detection of endo 1,4_glucanases and endo1,4xylanases in gels. Anal Biochem 1985; 144: 147151.
Carmona EC, Fialho MB, Buchgnani EB, Coelho GD, BrochetoBraga MR and Jorge JA: Production, purification and characterization of a minor form of xylanase from Aspergillus versicolor. Proces Biochem 2005; 40: 359364.
Techapun C, Poosaran N, Watanabe M and Sasaki K: Thermostable and alkalinetolerant microbial cellulase free xylanases produced from agricultural wastes and the properties required for use in pulp bleaching bioprocess: a review. Proces Biochem 2003; 38: 13271240.
Badhan AK, Chadha BS, Jatinder K, Saini HS and Bhat MK: Production of multiple xylanolytic and cellulolytic enzymes by thermophilic fungus myceliophthora sp. IMI 387099. Bioresour Technol 2007; 98: 504510.
Kulkarni N, Shendye A and Rao M: Molecular and biotechnological aspects of xylanases. FEMS Microbiol Rev 1999; 23: 411456.
Bajpai P: Microbial xylanolytic enzyme system: properties and applications. Adv Appl Microbiol 1998; 43: 141149.
Morales P, Madarro A, Flors A, Sendra JM and Perez–Gonzalez JA: Purification and characterization of a xylanase and an arabinofuranosidase from Bacillus polymyxa. Enzym Microb Technol 1995; 17: 424429.
Breccia JD, Sineriz F, Baigori DM, Castro GR and Hatti–Kaul R: Purification and characterization of a thermostable xylanase from Bacillus amyloliquefaciens. Enzym Microb Technol 1998; 22: 4249.
Mamo G, HattiKaul R and Mattiasson B: A thermostable alkaline active endo_14xylanase from Bacillus halodurans S7: Purification and characterization. Enzym Microb Technol 2006; 39: 14921498.
Kortekaas S, Wijngaarde RE, Klomp JW, Lettinga G and Field JA: Anaerobic treatment of hemp thermomechanical pulping wastewater. Wat Res 1998; 32: 33623370.
Dhillon A, Gupta JK and Khanna S: Enhanced production, purification and characterization of a novel cellulasepoor, thermostable alkalitolerant xylanase from Bacillus circulans AB16. Proces Biochem 2000; 35: 210215.
Takahashi H, Nakai R and Nakamura S. Puriï¬cation and partial characterization of a basic xylanase produced by thermoalkalophilic Bacillus sp. strain TAR1. Biosci Biotechnol Biochem 2000; 64: 887890.
Archana B and Satyanarayana T: Puriï¬cation and characterization of a cellulasefree xylanase of a moderate thermophile Bacillus licheniformis A99. World J Microbiol Biotechnol 2003; 19: 5357.
Khandeparkar R and Bhosle NB: Application of thermoalkalophilic xylanase from Arthrobacter sp. MTCC 5214 in biobleaching of kraft pulp. Bioresour Technol 2007; 98: 897903.
Li L, Tian H, Cheng Y, Jiang Z and Yang S: Purification and characterization of a thermostable cellulasefree xylanase from the newly isolated Paecilomycesthermophila. Enzym Microb Technol 2006; 38: 780787.
Bastawade KB: Xylan structure, microbial xylanase, and their mode of action. World J Microbiol Biotechnol 1992; 8: 353368.
Kamble RD and Jadhav AR: Isolation, Purification, and Characterization of Xylanase Produced by a New Species of Bacillus in Solid State Fermentation. Int J Microbiol 2012; 1: 18.
Bajaj BK and Singh NP: Production of xylanase from an alkalitolerant Streptomyces sp. under solidstate fermentation, itspurification, and characterization. Appl Biochem Biotechnol 2010; 162: 18041818.
Sanghi A, Garg N, Gupta VK, Mittal A and Kuhad RC. One step purification and characterization of a cellulose free xylanase produced by alkalophilic Bacillus subtilis ASH. Braz J Microbiol 2010; 41: 467476.
Lee JW, Gwak KS, Kim SI, Kim M, Choi DH and Choi IG. Characterization of xylanase from Lentinus edodes M290 cultured on waste mushroom logs. J Microbiol Biotechnol 2007; 17: 18111817.
Franco PF, Ferreira HM and Filho EX: Production and characterization of hemicellulase activities from Trichoderma harzianum strain T4. Biotechnol Appl Biochem 2004; 40: 255259.
Lee JW, Park JY, Kwon M and Choi IG. Purification and characterization of a thermostable xylanase from the brownrot fungus Laetiporus sulphureus. J Biosci Bioeng 2009; 107: 3337.
Khandeparker R, Preeti Verma and Deepti Deobagkar: A novel halotolerant xylanase from marine isolate Bacillus subtilis cho40: gene cloning and sequencing. New Biotechnol 2011; 28: 814821.
Saha BC: Production, purification and properties of xylanase from a newly isolated Fusarium proliferatum. Proces Biochem 2002; 37: 12791284.
Heck JX, Flores SH, Hertz PF and Ayub MAZ: Optimization os cellulasefree xylanase activity produced by Bacillus coagulans BL69 in solidstate cultivation. Proces Biochem 2005; 40: 107112.
Bataillon M, Nunes Cardinali AP, Castillon N and Duchiron F: Purification and characterization of a moderately thermostable xylanase from Bacillus sp. strain SPS0. Enzym Microb Technol 2000; 26: 187192.
Wang CY, Chan H, Lin HT and Shyu YT: Production, purification and characterisation of a novel halostable xylanase from Bacillus sp. NTU06. Annal Appl Biol 2010; 156: 187197.
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