2012 ; 22(10):
|Author||Bin Liu, Ningning Zhang, Chao Zhao, Baixue Lin, Lianhui Xie, Yifan Huang|
|Affiliation||National Engineering Research Center of Juncao, Fuzhou, Fujian 350002, China,Institute of Plant Virology, Fujian Agriculture and Forestry University, Fuzhou, Fujian 350002, China|
|Title||Characterization of a Recombinant Thermostable Xylanase from Hot Spring Thermophilic Geobacillus sp. TC-W7|
J. Microbiol. Biotechnol.2012 ; 22(10):
|Abstract||A xylanase-producing thermophilic strain, Geobacillus sp.
TC-W7, was isolated from a hot spring in Yongtai
(Fuzhou, China). Subsequently, the xylanase gene that
encoded 407 amino acids was cloned and expressed. The
recombinant xylanase was purified by GST affinity
chromatography and exhibited maximum activity at 75oC
and a pH of 8.2. The enzyme was active up to 95oC and
showed activity over a wide pH range of 5.2 to 10.2.
Additionally, the recombinant xylanase showed high
thermostability and pH stability. More than 85% of the
enzyme’s activity was retained after incubation at 70oC
for 90 min at a pH of 8.2. The activity of the recombinant
xylanase was enhanced by treatment with 10 mM enzyme
inhibitors (DDT, Tween-20, 2-Me, or TritonX-100) and
was inhibited by EDTA or PMSF. Its functionality was
stable in the presence of Li+, Na+, and K+, but inhibited by
Hg2+, Ni2+, Co2+, Cu2+, Zn2+, Pb2+, Fe3+, and Al3+. The
functionality of the crude xylanase had similar properties
to the recombinant xylanase except for when it was
treated with Al2+ or Fe2+. The enzyme might be a promising
candidate for various industrial applications such as the
biofuel, food, and paper and pulp industries.|
|Keywords||Thermostable xylanase, Recombinant expression, Characterization, Stable pH, Geobacillus sp. TC-W7|
Ayyachamy, M. and T. M. Vatsala. 2007. Production and partial characterization of cellulase free xylanase by Bacillus subtilis C01 using agriresidues and its application in biobleaching of nonwoody plant pulps. Lett. Appl. Microbiol. 45: 467-472.
Bailey, M. J., P. Biely, and K. Poutanen. 1992. Interlaboratory testing of methods for assay of xylanase activity. J. Biotechnol. 23: 257-270.
Bajpai, P. 1999. Application of enzymes in the pulp and paper industry. Biotechnol. Prog. 15: 147-157.
Beg, Q. K., M. Kapoor, L. Mahajan, and G. S. Hoondal. 2001. Microbial xylanases and their industrial applications: A review. Appl. Microbiol. Biotechnol. 56: 326-338.
Cazemier, A. E., J. C. Verdoes, A. J. van Ooyen, and H. J. Camp. 1999. Molecular and biochemical characterization of two xylanase-encoding genes from Cellulomonas pachnodae. Appl. Environ. Microbiol. 65: 4099-4107.
Collins, T., C. Gerday, and G. Feller. 2005. Xylanases, xylanase families and extremophilic xylanases. FEMS Microbiol. Rev. 29:3-23.
Dutta, T., R. Sengupta, R. Sahoo, S. R. Sinha, A. Bhattacharjee, and S. Ghosh. 2007. A novel cellulase free alkaliphilic xylanase from alkali tolerant Penicillium citrinum: Production, purification and characterization. Lett. Appl. Microbiol. 44: 206-211.
Huang, J., G. Wang, and L. Xiao. 2006. Cloning, sequencing and expression of the xylanase gene from a Bacillus subtilis strain B10 in Escherichia coli. Bioresour. Technol. 97: 802808.
Laemmli, U. K. 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680685.
Linko, M., K. Poutanen, and L. Viikari. 1989. New developments in the application of enzymes for biomass processing, pp. 331-
In M. P. Coughlan (ed.). Enzyme Systems for Lignocellulose Degradation. Elsevier Applied Science, London.
Liu, B., Y. Wang, and X. Zhang. 2006. Characterization of a recombinant maltogenic amylase from deep sea thermophilic Bacillus sp. WPD616. Enzyme Microb. Technol. 39: 805-810.
Miller, G. L. 1959. Use of dinitrosalicylic acid reagent for determination of reducing sugar. Anal. Chem. 31: 426-428.
Savitha, S., S. Sadhasivam, and K. Swaminathan. 2007. Application of Aspergillus fumigatus xylanase for quality improvement of waste paper pulp. Bull. Environ. Contam. Toxicol. 78: 217-221.
Shah, A. R. and D. Madamwar. 2005. Xylanase production by a newly isolated Aspergillus foetidus strain and its characterization. Process Biochem. 40: 1763-1771.
Srivastava, P. and K. J. Mukherjee. 2001. Cloning, characterization, and expression of xylanase gene from Bacillus lyticus in Escherichia coli and Bacillus subtilis. Prep. Biochem. Biotechnol. 31: 389-400.
Subramaniyan, S. and P. Prema. 2002. Biotechnology of microbial xylanases: Enzymology, molecular biology, and application. Crit. Rev. Biotechnol. 22: 33-46.
Sunna, A. and G. Antranikian. 1996. Growth and production of xylanolytic enzymes by the extreme thermophilic anaerobic bacterium Thermotoga thermarum. Appl. Microbiol. Biotechnol. 45: 671-676.
Whistler, R. L. and E. L. Richards. 1970. Hemicelluloses, pp. 447-469. In W. Pigman and D. Horton (eds.). The Carbohydrates. Academic Press, New York.
Wood, P., J. D. Erfle, and R. M. Teather. 1989. Use of complex formation between Congo red and polysaccharide in detection and assay of polysaccharide hydrolases. Methods Enzymol. 160:59-74.
Wu, S., B. Liu, and X. Zhang. 2006. Characterization of a recombinant thermostable xylanase from deep-sea thermophilic Geobacillus sp. MT-1 in East Pacific. Appl. Microbiol. Biotechnol. 72: 1210-1216.
Xue, Y., A. Wu, H. Zeng, and W. Shao. 2006. High-level expression of an alpha-L-arabinofuranosidase from Thermotoga maritima in Escherichia coli for the production of xylobiose from xylan. Biotechnol. Lett. 28: 351-356.