Abstract
A metagenomic library containing ca. 3.06 × 108 bp insert DNA was constructed from a rice straw degrading enrichment culture. A xylanase gene, umxyn10A, was cloned by screening the library for xylanase activity. The encoded enzyme Umxyn10A showed 58% identity and 73% similarity with a xylanase from Thermobifida fusca YX. Sequence analyses showed that Umxyn10A contained a glycosyl hydrolase family 10 catalytic domain. The gene was expressed in Escherichia coli, and the recombinant enzyme was purified and characterized biochemically. Recombinant Umxyn10A was highly active toward xylan. However, the purified enzyme could slightly hydrolyze β-1,3/4-glucan and β-1,3/6-glucan. Umxyn10A displayed maximal activity toward oat spelt xylan at a high temperature (75°C) and weak acidity (pH 6.5). The K m and V max of Umxyn10A toward oat spelt xylan were 3.2 mg ml−1 and 0.22 mmol min−1 mg−1 and were 2.7 mg ml−1 and 1.0 mmol min−1 mg−1 against birchwood xylan, respectively. Metal ions did not appear to be required for the catalytic activity of this enzyme. The enzyme Umxyn10A could efficiently hydrolyze birchwood xylan to release xylobiose as the major product and a negligible amount of xylose. The xylanase identified in this work may have potential application in producing xylobiose from xylan.
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This work was supported by grants from the Ministry of Science and Technology of China (2009DFA30700) and the National Hi-tech Research and Development Program of China (863 Program 2007AA021307).
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Mo, Xc., Chen, Cl., Pang, H. et al. Identification and characterization of a novel xylanase derived from a rice straw degrading enrichment culture. Appl Microbiol Biotechnol 87, 2137–2146 (2010). https://doi.org/10.1007/s00253-010-2712-2
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DOI: https://doi.org/10.1007/s00253-010-2712-2