Molecular cloning and characterization of the novel acidic xylanase XYLD from Bispora sp MEY-1 that is homologous to family 30 glycosyl hydrolases

文献类型： 外文期刊

第一作者： Luo, Huiying

作者： Luo, Huiying;Yang, Jun;Shi, Pengjun;Huang, Huoqing;Bai, Yingguo;Yao, Bin;Li, Jiang;Fan, Yunliu

作者机构：

关键词： Acidic xylanase;Glycosyl hydrolase family 30;Bispora sp.;MEY-1;Pichia pastoris

期刊名称：APPLIED MICROBIOLOGY AND BIOTECHNOLOGY （ 影响因子：4.813； 五年影响因子：4.697 ）

ISSN： 0175-7598

年卷期： 2010 年 86 卷 6 期

页码：

收录情况： SCI

摘要： We cloned and sequenced a xylanase gene named xylD from the acidophilic fungus Bispora sp. MEY-1 and expressed the gene in Pichia pastoris. The 1,422-bp full-length complementary DNA fragment encoded a 457-amino acid xylanase with a calculated molecular mass of 49.8 kDa. The mature protein of XYLD showed high sequence similarity to both glycosyl hydrolase (GH) families 5 and 30 but was more homologous to members of GH 30 based on phylogenetic analysis. XYLD shared the highest identity (49.9%) with a putative endo-1,6-beta-d-glucanase from Talaromyces stipitatus and exhibited 21.1% identity and 34.3% similarity to the well-characterized GH family 5 xylanase from Erwinia chrysanthemi. Purified recombinant XYLD showed maximal activity at pH 3.0 and 60 A degrees C, maintained more than 60% of maximal activity when assayed at pH 1.5-4.0, and had good thermal stability at 60 A degrees C and remained stable at pH 1.0-6.0. The enzyme activity was enhanced in the presence of Ni2+ and beta-mercaptoethanol and inhibited by some metal irons (Hg2+, Cu2+, Pb2+, Mn2+, Li+, and Fe3+) and sodium dodecyl sulfate. The specific activity of XYLD for beechwood xylan, birchwood xylan, 4-O-methyl-d-glucuronoxylan, and oat spelt xylan was 2,463, 2,144, 2,020, and 1,429 U mg(-1), respectively. The apparent K (m) and V (max) values for beechwood xylan were 5.6 mg ml(-1) and 3,622 mu mol min(-1) mg(-1), respectively. The hydrolysis products of different xylans were mainly xylose and xylobiose.

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