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

作　　者：陈永敢 谷风林[2,3,4] 蔡莹莹[2] 徐飞 朱科学 CHEN Yonggan;GU Fenglin;CAI Yingying;XU Fei;ZHU Kexue(Hainan Tropical Ocean University,Sanya,Hainan 572022,China;Spice and Beverage Research Institute,Chinese Academy of Tropical Agricultural,Wanning,Hainan 571533,China;National Center of Important Tropical Crops Engineering and Technology Research,Wanning,Hainan 571533,China;Hainan Provincial Engineering Research Center of Tropical Spice and Beverage Crops,Wanning,Hainan 571533,China)

机构地区：[1]海南热带海洋学院,海南三亚572022 [2]中国热带农业科学院香料饮料研究所,海南万宁571533 [3]国家重要热带作物工程技术研究中心,海南万宁571533 [4]海南省热带香料饮料作物工程技术研究中心,海南万宁571533

出　　处：《热带作物学报》2020年第2期300-304,共5页Chinese Journal of Tropical Crops

基　　金：国家自然科学基金项目(No.31671848);中国热带农业科学院基本科研业务费专项资金(No.1630142017019)。

摘　　要：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.

关 键 词：BACILLUS β-D-葡萄糖苷酶 基因克隆 

分 类 号：Q939.124[生物学—微生物学]