Browse > Article
http://dx.doi.org/10.5352/JLS.2005.15.5.734

Molecular Cloning and Nucleotide Sequence of Xylanase gene (xynT) from Bacillus alcalophilus AX2000.  

Park Young-Seo (Division of Biotechnology, Kyungwon University)
Publication Information
Journal of Life Science / v.15, no.5, 2005 , pp. 734-738 More about this Journal
Abstract
A gene coding for xylanase from alkali-tolerant Bacillus alcalophilus AX2000 was cloned into Escherichia coli $DH5\alpha$ using pUC19. Among 2,000 transformants, one transformant showed clear zone on the detection agar plate containing oat-spells xylan. Its recombinant plasmid, named pXTY99, was found to carry 7.0 kb insert DNA fragment. When the nucleotide sequence of the cloned xylanase gene (xynT) was determined, xynT gene was found to consist of 1,020 base-pair open reading frame coding for a poly-peptide of 340 amino acids with a deduced molecular weight of 40 kDa. The coding sequence was preceded by a putative ribosome binding site, and the transcription initiation signals. The deduced amino acid sequence of xylanase is similar to those of the xylanases from Bacillus sp. Nl37 and B. stearothermophilus 21 with $61\%$ and $59\%$ identical residues, respectively.
Keywords
xylanase gene; cloning; nucleotide sequence; Bacillus alcalophilus;
Citations & Related Records
연도 인용수 순위
  • Reference
1 Velikodvorskaya, T. V., I. Y. Volkov, V. T. Vasilevko, V. V. Zverlov and E. S. Piruzian. 1997. Purification and some properties of Thermotoga neapolitana thermostable xylanase B expressed in E. coli cells. Biochemistry 62, 66-70
2 Wakarchuk, W. W., R. J. Campbell, W. J. Sung, J. Davoodi, M. Taguchi M. 1994. Mutational and crystallographic analyses of the active site residues of the Bacillus circulans xylanase. Protein Sci. 3, 467-475   DOI   ScienceOn
3 Warren, R. A. J. 1996. Microbial hydrolysis of polysaccharides. Annu. Rev. Microbial. 50, 183-212   DOI   ScienceOn
4 Wong, K. K. Y. and J. N. Saddler. 1992. Applications of hemicellulases in the food, feed, and pulp and paper industries, In: Hemicellulose and hemicellulases (Coughlen, P. P. and G. P. Hazlewood, eds), pp. 127-143, Portland Press, London
5 Wong, K. K. Y., J. U. J. Tan and J. N. Saddler. 1988. Multiplicity of ${\beta}$-1,4-xylanase in microorganism: Functions and applications. Microbial. Rev. 52, 305-317
6 Xie, H., H. J. Gilbert, S. J. Charnock, G. J. Davies, M. P. Williamson, P. J. Simpson, S. Raghothama, C. M. Fontes, F. M. Dias, J. M. Ferreira and D. N. Bolam. 2001. Clostridium thermocellum Xyn10B carbohydrate-binding module 22-2: the role of conserved amino acids in ligand binding. Biochemistry 40, 9167-9176   DOI   ScienceOn
7 McCarthy, A. A., D. D. Morris, P. J. Bergquist and E. N. Baker. 2000. Structure of XynB, a highly thermostable beta-1,4-xylanase from Dictyogiomus thermophilum Rt46B.1, at 1.8 A resolution. Acta Crystallogr. D. BioI. Crystallogr. 11, 1367-1375   DOI
8 Moran C. P. Jr, N. Lang, S. F. LeGrice, G. Lee, M. Stephens, A. J. Sonenshein, J. Pero and R. Losick. 1982. Nucleotide sequences that signal the initiation of transcription and translation in Bacillus subtilis. Mol. Gen. Genet. 186, 339-346   DOI   ScienceOn
9 Park, Y. S. and T. Y. Kim. 2003. Isolation of Bacillus alcalaphilus AX2000 producing alkaline xylanase and its enzyme production, Korean J. Microbol. Biotechnol. 31, 157-164
10 Sambrook, J. and D. Russell. 2001. Molecular cloning - A laboratory manual. 3rd eds., CSHL Press. Cold Spring Harbor, New York
11 Shorenstein, R. G., R. Losick. 1973. Comparative size and properties of the sigma subunits of ribonucleic acid polymerase from Bacillus subtilis and Escherichia coli. J. Biol. Chem. 248, 6170-6173
12 Kulkarni, N., A. Shendye, M. Rao. 1999. Molecular and biotechological aspects of xylanases. FEMS Microbiol. Rev. 23, 411-456   DOI   ScienceOn
13 Somogyi, M. 1952. Notes on sugar determinations. J. Biol. Chem. 195, 19-23
14 Timell, T. E. 1965. Wood hemicelluloses: Part II. Carbohydr. Chem. 20, 409-483
15 Timell. T. E. 1967. Recent progress in the chemistry of wood hemicelluloses. Wood Sci. Technol. 1, 45-70   DOI