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Characterization of a Glucose-, Xylose-, Sucrose-, and d-Galactose-Stimulated beta-Glucosidase from the Alkalophilic Bacterium Bacillus halodurans C-125

文献类型： 外文期刊

作者： Xu, Hu 1 ; Xiong, Ai-Sheng 2 ; Zhao, Wei 2 ; Tian, Yong-Sheng 2 ; Peng, Ri-He 2 ; Chen, Jian-Min 1 ; Yao, Quan-Hong 2 ;

作者机构： 1.Yangzhou Univ, Coll Biosci & Biotechnol, Nanjing, Jiangsu, Peoples R China

2.Shanghai Acad Agr Sci, Shanghai Key Lab Agr Genet & Breeding, Biotechnol Res Inst, Shanghai, Peoples R China

关键词： magnesium ion: 22537-22-0;open reading frame: ORF;cadmium ion: 22537-48-0;zinc ion: 23713-49-7;ferric ion: 20074-52-6;o-nitrophenyl-beta-D-glucopyranoside;glucose: 58367-01-4;sugar;sucrose: 57-50-1;sugar;xylose: 25990-60-7;sugar;453-amino acid protein: glycoside hydrolase family 1;D-galactose: 59-23-4;sugar;enzyme activity;alkaline condition

期刊名称：CURRENT MICROBIOLOGY （ 影响因子：2.188； 五年影响因子：2.197 ）

ISSN：

年卷期：

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收录情况： SCI

摘要： The gene (Bhbgl) encoding a beta-glucosidase from the alkalophilic bacterium Bacillus halodurans C-125 was synthesized chemically via the PCR-based two-step DNA synthesis (PTDS) method and expressed in Escherichia coli. Bhbgl contained an open reading frame (ORF) of 1359 bp encoding a 453-amino acid protein belonging to glycoside hydrolase family 1 (GHF1), and the deduced molecular mass of recombinant Bhbgl (52,488 Da) was confirmed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The enzyme exhibited a high specific activity with o-nitrophenyl-beta-d-glucopyranoside (oNPGlu) and an apparent K (m) value of 0.32 mM. With oNPGlu as the substrate, Bhbgl displayed pH and temperature optima of similar to 7.0 and 50A degrees C, respectively. The enzyme was relatively stable under alkaline conditions and > 50% activity was retained after incubation at pH 9.5 for 24 h at 4A degrees C. Recombinant Bhbgl activity was inhibited by 5 mM Zn2+, Fe3+, or Cd2+, but was enhanced by 1 mM Mg2+ and other metal ions. Enzyme activity was also stimulated by at least four sugars (sucrose, d-galactose, xylose, glucose) at concentrations ranging from 50 to 800 mM.

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