Isolation and characterization of Xylanase producing strain of Bacillus cereus from soil
Abstract
Background and Objectives: Research on Xylanase has markedly increased due to its potential applications in pulping and bleaching processes using cellulose free preparations, textile processes, the enzymatic saccharification of lignocellulosic materials and waste treatment. The present study was aimed at isolation and characterization of xylan degrading strain of Bacillus cereus from soil for production of xylanase.
Materials and Methods: Twelve isolates were obtained from soil samples of different areas in the Rajshahi University campus and studied for detection of xylanase activity. One of the strains was identified as Bacillus cereus on the basis of the nucleotide sequence of the 16S rRNA gene which produces xylanase extracellularly. We purified xylanase to homogeneity by a combination of ammonium-sulphate precipitation, DEAE-sepharose, Phenyl-5PW and Hydroxyapatite column chromatography using culture supernatant.
Results: The SDS-PAGE gave a single band at 32 kDa. The optimum temperature and pH of the purified enzyme was 40oC and 6.0, respectively. The xylanase hydrolyzed oat spelt xylan, birch wood xylan and beech wood xylan efficiently but showed no activity towards cellulose, CM-cellulose and Avicell pH 101.
Conclusion: Thus it was a true and neutral xylanase. The isolation of xylanase from Bacillus cereus is rare.
Whistler RL, Richard EL (1970). Hemicelluloses in the carbohydrates, 2nd ed. Academic Press, New York, pp.447-469.
Gilbert HJ, Hazlewood GP. Bacterial celluloses and xylanases. J Gen Microbiol 1993; 139: 187-194.
Sunna A, Antranikian G. Xylanolytic enzymes from fungi and bacteria. Crit Rev Biotechnol 1997; 17: 39-67.
Prade RA. Xylanases: from biology to biotechnology.Biotechnol Gene Eng Rev 1995; 13: 101-129.
Viikari V, Kantelinen J, Linko M. Xylanases in bleaching: from an idea to the industry FEMS. Microbiol Rev 1994; 13: 335-350.
Wong K, Saddler JN. Multiplicity of 1,4 xylanase in micro- organism: functions and application. Microbial Rev 1988; 52: 305-317.
Sneath PHA (1986). Endospore-forming gram-positive rods and cocci, Bergey’s manual of systematic bacteriology. The William and Wilkinss, Co., Baltimore, pp.1104-1139.
Lowry OH, Rosebrough NJ; Randal RJ. Protein measurement with folin-phenol reagent. J Biol Chem 1951; 193: 265-275.
Bailey MJ, Bielpy P, Poutanen K. Interlaboratory testing of methods for assay of xylanases activity. J Biotech 1992; 25: 257-270.
Miller G., Blum LR, Burton AI. Use of dinitosalisalic acid reagent for determination of reducing sugars. Anal Chem 1960; 31: 426-428.
Gallardo O, Diaz P, Pastor FI. Cloning and characterization of xylanase A from the strain Bacillus sp. BP-7: Comparison with Alkaline pI-low molecular weight xylanases of family 11. Current Micribiol 2003; 48: 276-779.
Duarte MCT, Pellegrino ACA, Portugal EP; Ponezi AN, Franco TT. Characterization of alkaline xylanaes from Bacillus pulmilus. Braz J Microbiol 2000; 31: 1-9.
Roy N, Okai N, Tomita T, Muramoto K, Kamio Y. Purification and some properties of high-molecular-weight sylanases, the xylanases 4 and 5 of Aeromonas caviae W-61. Biosci Biotechnol Biochem 2000; 64: 408-413.
Dung NV, Vetayasuporn S, Kamio Y, Abe N, Kaneko N; Izaki K. Purification and properties of β-1 ,4-xylanases 2 and 3 from A. caviae W-61. Biosci Biotech Biochem 1993;57: 1708-1712.
Dung NV, Kamio, Y, Abe N, Kaneko J; Izaki K. Purification and characterization of β-1, 4 xylanase from A. caciae W-61. Appl Environ Microbiol 1991; 57: 445-449.
Panbangred W, Shinmyo A, Kinoshita S, Okada H. Purification and properties of Endoxylanase produced by Bacillus pumilus. Agric Biol Chem 1983; 47: 957-963.
Bernier R, Desrochers M, Jurasek ML, Paice MG. Isolation and characterization of a xylanase from Bacillus subtilis. Appl Environ Microbiol 1983; 46:511-514.
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Issue | Vol 1 No 2 (2009) | |
Section | Articles | |
Keywords | ||
Xylan Xylanases Bacillus cereus |
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