Purification and characterization of carboxymethyl cellulase from Bacillus sp. isolated from a paddy field

• Autorzy: Vijayaraghavan P. , Vincent S. G. P.
• Języki publikacji: EN

Abstrakty

EN

A microorganism hydrolyzing carboxymethyl cellulose was isolated from a paddy field and identified as Bacillus sp. Production of cellulase by this bacterium was found to be optimal at pH 6.5, 37°C and 150 rpm of shaking. This cellulase was purified to homogeneity by the combination of ammonium sulphate precipitation, DEAE cellulose, and sephadex G-75 gel filtration chromatography. The cellulase was purified up to 14.5 fold and had a specific activity of 246 U/mg protein. The enzyme was a monomeric cellulase with a relative molecular mass of 58 kDa, as determined by SDS-PAGE. The enzyme exhibited its optimal activity at 50°C and pH 6.0. The enzyme was stable in the pH range of 5.0 to 7.0 and its stability was maintained for 30 min at 50°C and its activity got inhibited by Hg2+, Cu2+, Zn2+, Mg2+, Na2+, and Ca2+.

• Wydawca: -
• Czasopismo: Polish Journal of Microbiology
• Rocznik: 2012
• Tom: 61
• Numer: 1
• Opis fizyczny: p.51-55,fig.,ref.

Twórcy

autor

Vijayaraghavan P.

• Centre for Marine Science and Technology, Manonmaniam Sundaranar University, Rajakkamangalam-629 502, Kanyakumari District, Tamilnadu, India

autor

Vincent S. G. P.

Bibliografia

• Ando S., H. Ishida, Y. Kosugi and K. Ishikawa. 2002. Hyperthermostable endoglucanase from Pyrococcus horikoshii. Appl. Environ. Microbiol. 68: 430–433.
• Camassola M. and A.J.P. Dillon. 2007. Production of cellulases and hemicellulases by Penicillium echinulatum grown on pretreated sugarcane bagasse and wheat bran in solid-state fermentation. J. Appl. Microbiol. 103: 2196–2204.
• Dhillon N., S. Chhibber, M. Sexena, S. Pajni and D.V. Vadehra. 1985. A constitutive endoglucanase (CMCase) from Bacillus licheniformis-1. Biotechnol. Lett. 7: 695–697.
• Henriette C., S. Zinebi, M.F. Aumaitre and E. Petitdemange. 1993. Protease and lipase production by strain of Serratia marcescens. J. Industrial. Microbiol. 12: 129–135.
• Jones D. and M.D. Collins. 1984. Irregular, nonsporeforming Grampositive rods. pp. 1261–1434. In: Bergey’s Manual of Systematic Bacteriology. (eds). P.H.A. Sneath, Baltimore, MD: Williams and Wilkins. Kasana R.C., R. Salwan, H. Dhar, S. Dutt, A. Gulati. 2008. A Rapid and Easy Method for the Detection of Microbial Cellulases on Agar Plates Using Gram’s Iodine. Curr. Microbiol. 57: 503–507.
• Kotchoni S.O., E.W. Gachomo, B.O. Omafuvbe and O.O. Shonukan. 2006. Purification and Biochemical Characterization of Carboxymethyl Cellulase (CMCase) from a Catabolite Repression Insensitive Mutant of Bacillus pumilus. Int. J. Agri. Biol. 8: 286–292.
• Laemmli U.K. 1970. “Cleavage of structural proteins during the assembly of the head of bacteriophage T4”. Nature 227: 680–685.
• Lee K.D, J. Kim and H. Kim. 1996. Isolation and Characterization of Bacillus sp. KD1014 Producing Carboxymethyl-Cellulase. J. Microbiol. 34: 305–310.
• Lee Y.J., B.K. Kim, B.H. Lee, K.I. Jo, N.K. Lee, C.H. Chung, Y.C. Lee and J.W. Lee. 2008. Purification and characterization of cellulase produced by Bacillus amyoliquefaciens DL-3 utilizing rice hull. Bioresour. Technol. 99: 378–386.
• Lowry O.H., N.J. Rosenbrough A.L. Fare and R.J. Randal. 1951. Protein measurement with the Folin phenol reagent. J. Biol. Chem. 194: 265–275.
• Lynd L.R., P.J. Weimer, W.H. van Zyl and I.S. Pretorius. 2002. Microbial cellulose utilization: fundamentals and biotechnology. Microbiol. Mol. Biol. Rev. 66: 506–577.
• Mawadza C., R. Hatti-Kaul, R. Zvauya and B. Mattiasson. 2000. Purification and characterization of cellulases produced by two Bacillus strains. J. Biotechnol. 83: 177–187.
• Nelson. 1944. A photometric adaption of the somogyi method for the determination of glucose. J. Biol. Chem. 153: 375–380.
• Nishida Y., K.I. Suzuki, Y. Kumagai, H. Tanaka, A. Inoue and T. Ojima. 2007. Isolation and primary structure of a cellulase from the Japanese sea urchin Strongylocentrotus nudus. Biochimie 89: 1002–1011.
• Odeniyi1, O.A., A.A. Onilude and M.A. Ayodele. 2009. Production characteristics and properties of cellulase/polygalacturonase by a Bacillus coagulans strain from a fermenting palm-fruit industrial residue. Afr. J. Microbiol. Res. 3: 407–417.
• Perez J., J. Munoz-Dorado, T. Rubia and J. Martinez. 2002. “Biodegradation and biological treatments of cellulose, hemicellulose and lignin:an overview.” Int. Microbiol. 5: 53–63.
• Singh V.K. and A. Kumar. 1998. Production and purification of an extracellular cellulase from Bacillus brevis VS-1. Biochem. Mol. Biol. Int. 45: 443–452.
• Yin L.J., H.H. Lin and Z.R. Xiao. 2010 Purification and Characterization of a cellulase from Bacillus subtilis YJ1. J. Marine Sci. Tech. 18: 466–471.

Uwagi

PL

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