Characterization of Cellulolytic and Xylanolytic Enzymes of Bacillus licheniformis JK7 Isolated from the Rumen of a Native Korean Goat
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A facultative bacterium producing cellulolytic and hemicellulolytic enzymes was isolated from the rumen of a native Korean goat. The bacterium was identified as a Bacillus licheniformis on the basis of biochemical and morphological characteristics and 16S rDNA sequences, and has been designated Bacillus licheniformis JK7. Endoglucanase activities were higher than those of ß-glucosidase and xylanase at all temperatures. Xylanase had the lowest activity among the three enzymes examined. The optimum temperature for the enzymes of Bacillus licheniformis JK7 was 70C for endoglucanase (0.75 U/ml) and 50C for ß-glucosidase and xylanase (0.63 U/ml, 0.44 U/ml, respectively). All three enzymes were stable at a temperature range of 20 to 50C. At 50C, endoglucanse, ß-glucosidase, and xylanase had 90.29, 94.80, and 88.69% residual activity, respectively. The optimal pH for the three enzymes was 5.0, at which their activity was 1.46, 1.10, and 1.08 U/ml, respectively. The activity of all three enzymes was stable in the pH range of 3.0 to 6.0. Endoglucanase activity was increased 113% by K+, while K+, Zn+, and tween 20 enhanced ß-glucosidase activity. Xylanase showed considerable activity even in presence of selected chemical additives, with the exception of Mn2+ and Cu2+. The broad range of optimum temperatures (20 to 40C) and the stability under acidic pH (4 to 6) suggest that the cellulolytic enzymes of Bacillus licheniformis JK7 may be good candidates for use in the biofuel industry.
Abstract
A facultative bacterium producing cellulolytic and hemicellulolytic enzymes was isolated from the rumen of a native Korean goat. The bacterium was identified as a Bacillus licheniformis on the basis of biochemical and morphological characteristics and 16S rDNA sequences, and has been designated Bacillus licheniformis JK7. Endoglucanase activities were higher than those of ß-glucosidase and xylanase at all temperatures. Xylanase had the lowest activity among the three enzymes examined. The optimum temperature for the enzymes of Bacillus licheniformis JK7 was 70C for endoglucanase (0.75 U/ml) and 50C for ß-glucosidase and xylanase (0.63 U/ml, 0.44 U/ml, respectively). All three enzymes were stable at a temperature range of 20 to 50C. At 50C, endoglucanse, ß-glucosidase, and xylanase had 90.29, 94.80, and 88.69% residual activity, respectively. The optimal pH for the three enzymes was 5.0, at which their activity was 1.46, 1.10, and 1.08 U/ml, respectively. The activity of all three enzymes was stable in the pH range of 3.0 to 6.0. Endoglucanase activity was increased 113% by K+, while K+, Zn+, and tween 20 enhanced ß-glucosidase activity. Xylanase showed considerable activity even in presence of selected chemical additives, with the exception of Mn2+ and Cu2+. The broad range of optimum temperatures (20 to 40C) and the stability under acidic pH (4 to 6) suggest that the cellulolytic enzymes of Bacillus licheniformis JK7 may be good candidates for use in the biofuel industry.
참고문헌 (45)
- Archana, A., "Xylanase production by thermophilic Bacillus licheniformis A99 in solid-state fermentation", Enzyme and Microb. Technol, 21, pp.12~17 (1997)
- Ariffin, H., "Production of bacterial endoglucanase from pretreated oil palm empty fruit bunch by bacillus pumilus EB3", J. Biosci. Bioeng, 106, pp.231~236 (2008)
- Bajaj, B. K., "Partial purification and characterization of a highly thermostable and pH stable endoglucanase from a newly isolated Bacillus strain M-9", Indian J. Chem. Technol, 16, pp.382~387 (2009)
- Bischoff, K. M., "Cloning and characterization of a recombinant family 5 endoglucanase from Bacillus licheniformis strain B-41361", Process Biochem, 42, pp.1150~1154 (2007)
- Choi, I., "Characterization and application of recombinant -glucosidase (BglH) from Bacillus licheniformis KCTC 1918", J. Wood Sci, 55, pp.329~334 (1918)
- Christakopoulos, P., "Purification and mode of action of an alkali-resistant endo-1,4--glucanase from Bacillus pumilus", Arch. Biochem. Biophys, 364, pp.61~66 (1999)
- Damiano, V. B., "Application of crude xylanase from Bacillus licheniformis 77-2 to the bleaching of eucalyptus Kraft pulp", World J. Microbiol. Biotechnol, 19, pp.139~144 (2003)
- Junli Dong, "Molecular Cloning, Purification, and Characterization of a Novel, Acidic, pH-Stable Endoglucanase from Martelella mediterranea", The Journal of Microbiology, 48(3), pp.393~398 (2010), 한국미생물학회
- Gao, J., "Production and characterization of cellulolytic enzymes from the thermoacidophilic fungal Aspergillus terreus M11 under solid-state cultivation of corn stover", Bioresour. Technol, 99, pp.7623~7629 (2008)
- K. Geetha, "Optimization of Nutrient Medium Containing Agricultural Waste for Xylanase Production by Bacillus pumilus B20", Biotechnology and Bioprocess Engineering, 15(5), pp.882~889 (2010), 한국생물공학회
- Ghose, T. K., "Measurement of cellulase activities", Pure Appl. Chem, 59, pp.257~268 (1987)
- Hong, J., "Cloning of a gene encoding a highly stable endo-b-1,4-glucanase from Aspergillus niger and its expression in yeast", J. Biosci. Bioeng, 92, pp.434~441 (2001)
- Karnchanatat, A., "Purification and biochemical characterization of an extracellular β-glucosidase from the wood-decaying fungus Daldinia eschscholzii (Ehrenb.:Fr.) Rehm", FEMS Microbiol. Lett, 270, pp.162~170 (2007)
- Ko, C. -H., "Xylanase production by Paenibacillus campinasensis BL11 and its pretreatment of hardwood kraft pulp bleaching", Int. Biodeterior. Biodegradation, 64, pp.13~19 (2010)
- Ko, C. -H., "Identification of Paenibacillus sp. 2S-6 and application of its xylanase on biobleaching", Int. Biodeterior. Biodegradation, 65, pp.334~339 (2011)
- Kui, H., "Gene cloning, expression, and characterization of a thermostable xylanase from Nesterenkonia xinjiangensis CCTCC AA001025", Appl. Biochem. Biotechnol, 162, pp.953~965
- Kumar, R., "Bioconversion of lignocellulosic biomass: biochemical and molecular perspectives", J. Ind. Microbiol. Biotechnol., 35, pp.377~391 (2008)
- Lee, Y. -J., "Purification and characterization of cellulase produced by Bacillus amyoliquefaciens DL-3 utilizing rice hull", Bioresour. Technol, 99, pp.378~386 (2008)
- Li, W., "Simultaneous cloning and expression of two cellulase genes from Bacillus subtilis newly isolated from Golden Takin (Budorcas taxicolor Bedfordi).", Biochem. Biophys. Res. Commun, 383, pp.397~400 (2009)
- Liang, Y., "Study of cellulases from a newly isolated thermophilic and cellulolytic Brevibacillus sp. strain JXL", J. Ind. Microbiol. Biotechnol, 36, pp.961~970 (2009)
- Maki, M., "The prospects of cellulase-producing bacteria for the bioconversion of lignocellulosic biomass", Int. J. Biol. Sci, 5, pp.500~516 (2009)
- Mamo, G., "Athermostable alkaline active endo--1,4-xylanase from Bacillus halodurans S7: purification and characterization", Enzyme Microb. Biotechnol, 39, pp.1492~1498 (2006)
- Mawadza, C., "Purification and characterization of cellulases produced by two Bacillus strains", J. Biotechnol, 83, pp.177~187 (2000)
- Moaledh, K, "Comparison of Gram-stainig and alternate methods, KOH test and aminopeptidase activity in aquatic bacteria: their application to numerical taxonomy", J. Microbiol. Methods, 5, pp.303~310 (1986)
- Peixoto, S. B., "Cellulase-producing Bacillus strains isolated from the intestine of Amazon basin fish", Aquac. Res, 42, pp.887~891 (2011)
- Percival Zhang, Y. H., "Outlook for cellulase improvement: Screening and selection strategies", Biotechnol. Adv, 24, pp.452~481 (2006)
- Rastogi, G., "Characterization of thermostable cellulases produced by Bacillus and Geobacillus strains", Bioresour. Technol, 101, pp.8798~8806 (2010)
- Rastogi, G., "Isolation and characterization of cellulose-degrading bacteria from the deep subsurface of the Homestake gold mine, Lead, South Dakota, USA", J. Ind. Microbiol. Biotechnol, 36, pp.585~598 (2009)
- Rizzatti, A. C., "Purification and properties of a thermostable extracellular beta-D-xylosidase produced by a thermotolerant Aspergillus phoenicis", J. Ind. Microbiol. Biotechnol, 26, pp.156~160 (2001)
- Roboson, L. M., "Celluases of bacterial origin", Enzyme Microb. Technol, 11, pp.626~644 (1989)
- Samiullah, T. R., "Isolation, Purification and characterization of extracellular -glucosidase from Bacillus sp", Adv. Environ. Biol, 3, pp.269~277 (2009)
- Saratale, G. D., "Production of thermotolerant and alkalotolerant cellulolytic enzymes by isolated Nocardiopsis sp. KNU. Biodegradation" (2011), Biodegradation
- SAS., "SAS user’s guide: Statistics, Version 6.12th edition" (1996), SAS Institute Inc
- Scott, H. W., "Vitamin requirements of several cellulolytic rumen bacteria", J. Bacteriol, 89, pp.1169~1175 (1965)
- Shallmey, M., "Developments in the use of Bacillus species for industrial production", Can. J. Microbiol, 50, pp.1~17 (2004)
- Sharma, A., "Alkali-thermostable and cellulase free xylanase production by an extreme thermophile Geobacillus thermoleovorans", World J. Microbiol. Biotechnol, 23, pp.483~490 (2007)
- Sneath, P. H. A., "Bergy’s Manual of Systematic Bacteriology, vol 2" (1986), Williams & Wilkins
- Son, Y. S., "Production and uses of Korean Native Black Goat", Small Rumin. Res, 34, pp.303~308 (1999)
- Trivedi, N., "An alkali-halotolerant cellulase from Bacillus flexus isolated from green seaweed Ulva lactuca", Carbohydr. Polym, 83, pp.891~897 (2011)
- van Dyk, J. S., "The cellulolytic and hemi-cellulolytic system of Bacillus licheniformis SVD1 and the evidence for production of a large multi-enzyme complex", Enzyme Microb. Technol, 45, pp.372~378 (2009)
- van Dyk, J. S., "Identification of endoglucanases, xylanases, pectinases and mannanases in the multi-enzyme complex of Bacillus licheniformis SVD1", Enzyme Microb. Technol. In Press, Corrected Proof, 47, pp.112~118 (2010)
- Wang, C. M., "Characterization of a novel thermophilic, cellulose-degrading bacterium Paenibacillus sp. strain B39", Lett. Appl. Microbiol, 47, pp.46~53 (2008)
- Williams, A. G., "Bacillus spp. in the rumen ecosystem. Hemicellulose depolymerases and glycoside hydrolases of Bacillus spp. and rumen isolates grown under anaerobic conditions", J. Appl. Bacteriol, 55, pp.283~292 (1983)
- Yang, V. W., "Alkaline-active xylanase produced by an alkaliphilic Bacillus sp. isolated from kraft pulp", J. Ind. Microbiol, 15, pp.434~441 (1995)
- Yin, L. -J., "Bioproperties and Purification of Xylanase from Bacillussp. YJ6", J. Agric. Food Chem, 58, pp.557~562 (2010)