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Characterization of thermostable Xyn10A enzyme from mesophilic Clostridium acetobutylicum ATCC 824

  • Mursheda K. Ali
  • Frederick B. Rudolph
  • George N. BennettEmail author
Original Paper

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.

Keywords

Xylanase Clostridium acetobutylicum Biomass Thermostable Hemicellulose Xylose 

Notes

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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Copyright information

© Society for Industrial Microbiology 2005

Authors and Affiliations

  • Mursheda K. Ali
    • 1
  • Frederick B. Rudolph
    • 1
  • George N. Bennett
    • 1
    Email author
  1. 1.Department of Biochemistry and Cell BiologyRice UniversityHoustonUSA

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