Optimization, Isolation and Characterization of Cellulase–Free Thermostable Xylanase from Paenibacillus sp.
American Journal of Life Sciences
Volume 4, Issue 4, August 2016, Pages: 93-98
Received: Aug. 2, 2016; Accepted: Aug. 13, 2016; Published: Aug. 29, 2016
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Authors
Md. Abdul High Siddiqui, Department of Biochemistry and Molecular Biology, University of Rajshahi, Rajshahi, Bangladesh
Mrityunjoy Biswas, Department of Biochemistry and Molecular Biology, University of Rajshahi, Rajshahi, Bangladesh
Md. Omar Faruk, Department of Biochemistry and Molecular Biology, University of Rajshahi, Rajshahi, Bangladesh
Manoranjan Roy, Department of Biochemistry and Molecular Biology, University of Rajshahi, Rajshahi, Bangladesh
A. K. M. Asaduzzaman, Department of Biochemistry and Molecular Biology, University of Rajshahi, Rajshahi, Bangladesh
Subed Chandra Dev Sharma, Department of Biochemistry and Molecular Biology, University of Rajshahi, Rajshahi, Bangladesh
Topodeb Biswas, Department of Botany, Jahangirnagar University, Savar, Dhaka, Bangladesh
Narayan Roy, Department of Biochemistry and Molecular Biology, University of Rajshahi, Rajshahi, Bangladesh
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Abstract
Xylanases are hydrolytic enzymes that cleave the β-1,4-linkage ofwheat bran xylan. For screening of xylanase producing bacteria soil samples were diluted by serial dilution and cultured on selective wheat bran xylan agar media. Two bacterial strains showing clear transparent zone around the colony on xylan agar plate were selected as xylanase producing bacteria. The strain Paenibacillus sp. showed highest xylanolytic activity. The strain was thermophile and produced highly active cellulase free xylanase. The enzyme secretion was enhanced when the medium was supplemented with 0.5% wheat bran xylan, peptone and Ca2+ salt. The peak in xylanase production was achieved within 48-60 hours at temperature 50°-55°C and at pH 7.0.The cellulase free xylanase was partially purified by ammonium sulfate fractionation and heat treatment at 50°C. The xylanase was optimally active at pH 7.0 and 55°C; and showed high substrate activity to wheat bran xylan but no activity towards carboxymethylcellulose, cellulose and starch.In future we want to know the structure function relationship of the purified enzyme and also want to known the molecular biological study using highly purified xylanase. For this purpose we have to determine the N-terminal & C-terminal amino acid sequence.
Keywords
Xylanases, Paenibacillus sp., Cellulose, Xylan, Thermophile, N-terminal & C-terminal Amino Acid Sequence
To cite this article
Md. Abdul High Siddiqui, Mrityunjoy Biswas, Md. Omar Faruk, Manoranjan Roy, A. K. M. Asaduzzaman, Subed Chandra Dev Sharma, Topodeb Biswas, Narayan Roy, Optimization, Isolation and Characterization of Cellulase–Free Thermostable Xylanase from Paenibacillus sp., American Journal of Life Sciences. Vol. 4, No. 4, 2016, pp. 93-98. doi: 10.11648/j.ajls.20160404.11
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Copyright © 2016 Authors retain the copyright of this article.
This article is an open access article distributed under the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
References
[1]
Chavez, R., Schaether, K., Navarro, C., Alenssandra, Peirrano, Bull, P., and Eyzaguirre, J. (2004). The acetyl xylan esterase II gene from Penicillium porpurogenum is differentially expressed in several carbon sources, and tightly regulated by pH.
[2]
Blanco, A., Diaz, P. Zueco, J. Parascandola, P. and Pastor, F. I. J. A. (1999). A multi domain xylanase from a Bacillus sp. with a region homologous to thermo-stabilizing domains of thermophilic enzymes. Microbiol. 145:2163- 2170:
[3]
Reilly, P. J. (1981). Xylanases: Structure and function. Basic life Sci.18: 11-129.
[4]
Iwamoto, T. T., Sasaki and Inaoka, M. (1973). Purification and some properties of xylanase from Aspergillus niger. Mem Coll. Agric. Ehime. Univ. 17: 185-197.
[5]
Takenishi, S. and Tsujisaka, Y. (1975). On the mode of action of three xylanases produced by a strain of Aspergillus niger. Van. Tieghem. Agric. Biol. Chem. 39: 2315-2323.
[6]
Dekker, R. F. H. (1985). Biodegradation of the hemicellulases. 505-533. In T. Higuchi (ed) Biosynthesis and Biodegradation of wood components. Academic Press Inc. Orlando, Fla.
[7]
Fontes, C. M., Gilbert, H. J., Hazlewood, G. P., Clarke, J. H., Prates, J. A., McKie, V. A., Nagy, T., Fernandes, T. H., Ferreira, L. M. (2000) A novel Cellvibrio mixtus family 10 xylanase that is both intracellular and expressed under non-inducing conditions. Microbiology 146 (Pt 8), 1959–1967.
[8]
Shulami, S., Gat, O., Sonenshein, A. L., Shoham, Y. (1999) The glucuronic acid utilization gene cluster from Bacillus stearothermophilus T-6. J. Bacteriol. 181, 3695–3704.
[9]
Teplitsky, A., Shulami, S., Moryles, S., Shoham, Y., Shoham, G. (2000) Crystallization and preliminary X-ray analysis of an intracellular xylanase from Bacillus stearothermophilus T-6. Acta Crystallogr. D: Biol. Crystallogr. 56 (Pt 2), 181–184. Links.
[10]
Biely, P., Mackenzic, C. R., Puls, J. and Schneider, H. (1986). Cooperatively of esterases and xylanases in the enzymatic degradation of acetylxylan. Biotechnology 4: 731-733.
[11]
Lee, S. F. and Forsberg, C. W. (1987). Purification and characterization of an -2-arabinofuranosidase from Clostridium acetobutylicum ATCC 824. Can. J. Microbiol. 33: 644-650.
[12]
Puls, J., Schmidt, O. and Granzow, C. (1987). Glucuronidase in microbial xylanolytic syatems. Enzyme Microbial Technology. 9: 83-88.
[13]
Greve, L. C., Labavitch, J. M. and Hunggate, R. E. (1984). -L-Arabinofuranosidase from Ruminococcus albus. Purification and possible role in hydrolysis of alfalfa cell wall. Al. Environ. Microbiol.47: 1135-1140.
[14]
Lee, H., To, R. J. B., Latta, R. K. Biely, P. and Schneider H. (1987). Some properties of extra-cellular acetyl xylan esterase produced by the yeast Rhodotorula mucilaginosa. Al. Environ. Microbiol. 53: 283-2834.
[15]
Singh, S., Madlala, A. M., Prior, B. A. (2003). Thermomyces lanuginosus: properties of strains and their hemicellulases. FEMS Microbiol. Rev. 27, 3–16.
[16]
Biely, P. (1985) Microbial xylanolytic systems. Trends Biotechnol. 3, 286–290.
[17]
Defaye, J., Guillot, J. M., Biely, P., Vrsanska, M. (1992) Positional isomers of thioxylobiose, their synthesis and inducing ability for d-xylan-degrading enzymes in the yeast Cryptococcus albidus. Carbohydr. Res. 228, 47–64.
[18]
Michael, J., Pelezar, Jr and Chan, E. C. S. and Noel R., Krieg, (1993). Microbiology, concepts and applications, Biotechnology: The industrial application of microbiology, International Edition; 878.
[19]
Rolf, A. and Prade, (1995). Xylanases: from biology to biotechnology. Biotechnology and Genetic Engineering Reviews-Vol 13: 101-129.
[20]
Nguyen, U. D., Kamio, Y., Abe, N., Kaneko, Y. and Izaki, K. (1993). Purification and properties of - 1,4-xylanases 2 & 3 from Aesomonas caviae W-61. Biosci. Biotechnol. Biochem., 57 P 1708-1712.
[21]
Sabini, E., Wilson, K. S., Danielsen, S., Schulein, M., Davies, G. J. (2001) Oligosaccharide binding to family 11 xylanases: both covalent intermediate and mutant product complexes display (2, 5) B conformations at the active. centre. Acta Crystallogr. D: Biol. Crystallogr. 57, 1344–1347.
[22]
Wakarchuk, W. W., Campbell, R. L., Sung, W. L., Davoodi, J., Yaguchi, M. (1994) Mutational and crystallographic analyses of the active site residues of the Bacillus circulans xylanase. Protein Sci. 3, 467–475.
[23]
Oakley, A. J., Heinrich, T., Thompson, C. A., Wilce, M. C. (2003) Characterization of a family 11 xylanase from Bacillus subtillis B230 used for paper bleaching. Acta Crystallogr. D: Biol. Crystallogr. 59, 627–636.
[24]
Esteban, R., Chordi, A. and Villa, T. G. (1983). Some aspects of a 1,4 β-D- xylosidase selected by Bacillus coagulans. Microbiol. Lett. 17: 163-166.
[25]
Teplitsky, A., Shulami, S., Moryles, S., Shoham, Y., Shoham, G. (2000) Crystallization and preliminary X-ray analysis of an intracellular xylanase from Bacillus stearothermophilus T-6. Acta Crystallogr. D: Biol. Crystallogr. 56 (Pt 2), 181–184. Links.
[26]
Canals, A., Vega, M. C., Gomis-Ruth, F. X., Diaz, M., Santamaria, R. R., Coll, M. (2003) Structure of xylanase Xys1delta from Streptomyceshalstedii. Acta Crystallogr. D: Biol. Crystallogr. 59, 1447–1453.
[27]
Derewenda, U., Swenson, L., Green, R., Wei, Y., Morosoli, R., Shareck, F., Kluepfel, D., Derewenda, Z. S. (1994) Crystal structure, at 2.6-Å resolution, of the Streptomyces lividans xylanase A, a member of the F family of beta-1,4-d-glycanases. J. Biol. Chem. 269, 20811–20814.
[28]
Pell, G., Szabo, L., Charnock, S. J., Xie, H., Gloster, T. M., Davies, G. J., Gilbert, H. J. (2004) Structural and biochemical analysis of Cellvibrio japonicus xylanase 10C: how variation in substrate-binding cleft influences the catalytic profile of family GH-10 xylanases. J. Biol. Chem. 279, 11777–11788.
[29]
Fushinobu, S., Ito, K., Konno, M., Wakagi, T., Matsuzawa, H. (1998) Crystallographic and mutational analyses of an extremely acidophilic and acid-stable xylanase: biased distribution of acidic residues and importance of Asp37 for catalysis at low pH. Protein Eng. 11, 1121–1128.
[30]
Krengel, U., Dijkstra, B. W. (1996) Three-dimensional structure of Endo-1,4-beta-xylanase I from Aspergillus niger, molecular basis for its low pH optimum. J. Mol. Biol. 263, 70–78.
[31]
Campbell, R. L., Rose, D. R., Wakarchuk, W. W., To, R. J., Sung, Z., Yagushi, M. (1993) High resolution structures of xylanases from Bacillus circulans and Trichoderma harzianum identify a new folding pattern and implications for the atomic basis of the catalysis. Foundation for biotechnical and industrial fermentation research. In: Trichoderma reesei Cellulases and Other Hydrolases (Souminen, P., Reikainen, T., Eds.), pp.63–72 Espoo, Finland.
[32]
Torronen, A., Rouvinen, J. (1995) Structural comparison of two major endo-1,4-xylanases from Trichoderma reesei. Biochemistry 34, 847–856.
[33]
Hashimoto, S., Muramatsu, and Funatsu, M. (1971). Studies of xylanase from trialu derma-viride part, isolation & some properties of crystalline xylanase. Biol. Chem. 35: 501-508
[34]
Schmidt, A., Schlacher, A., Steiner, W., Schwab, H., Kratky, C. (1998) Structure of the xylanase from Penicillium simplicissimum. Protein Sci. 7, 2081–2088.
[35]
Takenishi, S. and Tsuijisaka, Y. (1973). Purification and some properties of three xylanase from Pernicillium janthinellum Bioarge. J. Ferment. Technol. 51: 458-463.
[36]
Sreenath, H. K. and Joseph, R. (1982). Purification and properties of extracellular xylan hydrolases of Streptomyces exfoliatus. Folia Microbio. 27: 107-155.
[37]
Izuka, H. and Kawaminami, T. (1965). Studies on the xylanase from Streptomyces sp., Part-I purification and some properties of xylanase from Streptomyces xylophagus. Agric Biol. Chem. 29: 520-524.
[38]
Bernier, R. Jr., Desrochers, M., Jurasek, L. and Paice, M. G. (1983). Isolation and characterization of a xylanase from Bacillus subtilis. Al. Environ. Microbiol. 46: 511-514.
[39]
Puls, J., Schmidt, O. and Granzow, C. (1987). Glucuronidase in microbial xylanolytic syatems. Enzyme Microbial Technology. 9: 83-88.
[40]
Roy, N., Rana, M. M. and Uddin, A. T. M. S. (2003). Isolation and some properties of new xylanase from intestine of a herbivorous insect (Samia cynthia pryeri) Jounal of biological Sciences 4 (1): 27-33.
[41]
Roy, N. 2004. Characterization and identification of xylanase producting bacterial strains isolated from soil and water PJBS 7 (5): 711-716.
[42]
Lowry, O. H., Rose brough, N. J., Farr, A. L. and Randall, R. J. (1951). Protein measurement with the Folin ciocalteu’s reagent. J. Biol. Chem. 193: 265-275.
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