草菇重组β-葡萄糖苷酶的表达、纯化及特性分析
Expression, purification and characterization of a recombinant beta-glucosidase from Volvariella Volvacea.
作者信息
Li Xun, Pei Jianjun, Wu Guogan, Shao Weilan
机构信息
The Key Laboratory of Microbial Engineering, Nanjing Normal University, 210097, Nanjing, China.
出版信息
Biotechnol Lett. 2005 Sep;27(18):1369-73. doi: 10.1007/s10529-005-3683-8.
For the first time, a beta-glucosidase gene from the edible straw mushroom, Volvariella volvacea V1-1, has been over-expressed in E. coli. The gene product was purified by chromatography showing a single band on SDS-PAGE. The recombinant enzyme had a molecular mass of 380 kDa with subunits of 97 kDa. The maximum activity was at pH 6.4 and 50 degrees C over a 5 min assay. The purified enzyme was stable from pH 5.6-8.0, had a half life of 1 h at 45 degrees C. The beta-glucosidase had a K(m) of 0.2 mM: for p-nitrophenyl-beta-D: -glucopyranoside.
首次在大肠杆菌中过量表达了来自食用草菇草菇V1-1的β-葡萄糖苷酶基因。通过色谱法纯化基因产物，其在SDS-PAGE上显示为单一条带。重组酶的分子量为380 kDa，亚基为97 kDa。在5分钟的测定中，最大活性出现在pH 6.4和50℃。纯化后的酶在pH 5.6 - 8.0范围内稳定，在45℃下半衰期为1小时。该β-葡萄糖苷酶对对硝基苯基-β-D-吡喃葡萄糖苷的K(m)为0.2 mM。
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