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首页> 外文期刊>Journal of molecular catalysis, B. Enzymatic >Overexpression and characterization of a glycoside hydrolase family 1 enzyme from Cellulosimicrobium cellulans sp 21 and its application for minor ginsenosides production
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Overexpression and characterization of a glycoside hydrolase family 1 enzyme from Cellulosimicrobium cellulans sp 21 and its application for minor ginsenosides production

机译：Cellulosimicrobium cellulans sp 21的糖苷水解酶家族1酶的过表达和表征及其在少量人参皂苷生产中的应用

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摘要

A novel beta-glucosidase gene (ccbgl1a) was cloned from the ginsenosides-transforming strain Cellulosimicrobium cellulans sp. 21. This enzyme was overexpressed in Escherichia coli, the recombinant beta-glucosidase (CcBgl1A) containing N-terminal His-tag was sufficiently purified by nickel metal affinity chromatography with purification factor of 1.9-fold and specific activity of 31.5 U/mg. The molecular mass of recombinant CcBgl1A was estimated to be approximately 46 kDa. CcBgl1A exhibited optimal activity at 35 degrees C and pH 5.5. However, above 40 degrees C, the enzyme stability significantly decreased. The enzyme showed high bioconversion ability on protopanaxadiol-type ginsenosides mixture (PPDGM), which could hydrolyze the outer C-3 glucose moieties of ginsenosides Rb1, Rb2, Rc and Rd into the rare ginsenosides Gypenoside XVII (Gyp XVII), compound 0, ginsenoside Mb and ginsenoside F2. Scaled-up production using 1 g of the PPDGM resulted in 292 mg Gyp XVII, 134 mg CO, 184 mg Mb, and 62 mg F2, with chromatographic purities. These results suggest that CcBgl1A would be potentially useful in the preparation of pharmacologically active minor ginsenosides Gyp XVII, CO, Mb and F2. (C) 2015 Elsevier B.V. All rights reserved.
机译：一个新的β-葡萄糖苷酶基因（ccbgl1a）从人参皂苷转化菌株纤维素微纤维素纤维素sp。克隆。 21.该酶在大肠杆菌中过表达，用镍金属亲和色谱法以1.9倍的纯化因子和31.5U / mg的比活度充分纯化了含有N-末端His-标签的重组β-葡糖苷酶（CcBgl1A）。估计重组CcBgl1A的分子量约为46 kDa。 CcBgl1A在35摄氏度和pH 5.5下表现出最佳活性。然而，在40℃以上，酶稳定性显着降低。该酶对原人参二醇型人参皂苷混合物（PPDGM）具有很高的生物转化能力，可将人参皂苷Rb1，Rb2，Rc和Rd的外部C-3葡萄糖部分水解为稀少的人参皂苷人参皂苷XVII（Gyp XVII），化合物0，人参皂苷Mb和人参皂甙F2。使用1 g PPDGM进行放大生产，得到292 mg Gyp XVII，134 mg CO，184 mg Mb和62 mg F2，且具有色谱纯度。这些结果表明，CcBgl1A在制备具有药理活性的次要人参皂苷Gyp XVII，CO，Mb和F2方面可能会很有用。 （C）2015 Elsevier B.V.保留所有权利。

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