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Characterization of thermostable Xyn10A enzyme from mesophilic Clostridium acetobutylicum ATCC 824

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Journal of Industrial Microbiology and Biotechnology

Abstract

A thermostable xylanase gene, xyn10A (CAP0053), was cloned from Clostridium acetobutylicum ATCC 824. The nucleotide sequence of the C. acetobutylicum xyn10A gene encoded a 318-amino-acid, single-domain, family 10 xylanase, Xyn10A, with a molecular mass of 34 kDa. Xyn10A exhibited extremely high (92%) amino acid sequence identity with Xyn10B (CAP0116) of this strain and had 42% and 32% identity with the catalytic domains of Rhodothermus marinus xylanase I and Thermoascus aurantiacus xylanase I, respectively. Xyn10A enzyme was purified from recombinant Escherichia coli and was highly active toward oat-spelt and Birchwood xylan and slightly active toward carboxymethyl cellulose, arabinogalactouronic acid, and various p-nitrophenyl monosaccharides. Xyn10A hydrolyzed xylan and xylooligosaccharides larger than xylobiose to produce xylose. This enzyme was optimally active at 60°C and had an optimum pH of 5.0. This is one of a number of related activities encoded on the large plasmid in this strain.

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Acknowledgements

We thank Prof. Kazuo Sakka, Mie University, Japan, for supplying pQE-30T. This research was supported by the United States Department of Agriculture, Grant 00-35500-926, and the Robert A. Welch Foundation, grants C-1268 and C-1372.

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  1. Department of Biochemistry and Cell Biology, Rice University, Houston, TX 77005, USA

    Mursheda K. Ali, Frederick B. Rudolph & George N. Bennett

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  1. Mursheda K. Ali
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  2. Frederick B. Rudolph
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  3. George N. Bennett
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Correspondence to George N. Bennett.

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Ali, M.K., Rudolph, F.B. & Bennett, G.N. Characterization of thermostable Xyn10A enzyme from mesophilic Clostridium acetobutylicum ATCC 824. J IND MICROBIOL BIOTECHNOL 32, 12–18 (2005). https://doi.org/10.1007/s10295-004-0192-z

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