Bacillus siamensis和Bacillus subtilis中2个β-D-葡萄糖苷酶编码基因克隆、表达及酶学研究

Cloning,Expression and Enzymology of Two Genes Encoding β-D-glucosidases from Bacillus siamensis XY18 and Bacillus subtilis XY20

采用RACE技术扩增2株Bacillus属菌株XY18、XY20的β-D-葡萄糖苷酶编码基因bgl全长,构建重组载体pET28a(+)/bgl、pET28b(+)/bgl,导入E.coli BL21诱导表达,采用Ni亲和层析纯化蛋白。经酶学性质测定发现:2个蛋白均为弱酸性蛋白,温度为40℃时酶活力达到最大,具有一定耐热性。以上结果表明,为优化2株Bacillus属菌株参与香草兰发酵工艺,可提供适当弱酸性条件、适宜温度。

The full length of bgl of Bacillus sp.XY18 and Bacillus sp.XY20 was amplified by RACE.The recombinant vectors pET28a(+)/bgl and pET28b(+)/bgl were constructed,then transferred into E.coli BL21 for induction expression,and the protein was purified by Ni affinity chromatography.The enzymatic properties of the two proteins were determined.The two proteins were weak acidic proteins,and the enzyme activity reached the maximum at 40℃ with certain heat resistance,suggesting that appropriate weak acid condition and temperature could be provided to optimize the vanilla curing process.