Bacillus altitudinis SYBC hb4碱性β-葡萄糖苷酶基因的克隆表达及酶学性质的研究
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Cloning,Experssion and Characterization of Alkali-Stable β-Glucosidase from Bacillus altitudinis SYBC hb4
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    摘要:

    为获得耐碱性β-葡萄糖苷酶,进一步研究碱性β-葡萄糖苷酶的酶学性质。从实验室已有蜂蜜中筛选出一株耐碱性的高地芽孢杆菌Bacillus altitudinis SYBC hb4,根据Bacillus altitudinis SYBC hb4中β-葡萄糖苷酶(bglA)基因序列设计一对引物,通过PCR扩增技术,获得β-葡萄糖苷酶基因(bglA),将扩增后的bglA与质粒pColdII构建重组表达载体pColdII-bglA,并转化至大肠杆菌E. coli BL21(DE3)中表达。重组表达的β-葡萄糖苷酶酶活可达12.40 U/mL;该重组β-葡萄糖苷酶最适反应温度为60 ℃;最适反应pH值为pH 8.0;5 mmol/L的Mg2+可使酶活提高50%左右。本实验所获得的碱性β-葡萄糖苷酶比已报道的重组酶在碱性条件下稳定性更好。

    Abstract:

    In order to obtain alkali-stable β-glucosidase and study its enzyme characterization,an alkali-stable strain Bacillus altitudinis SYBC hb4 was screened and isolated from native honey. Based on the β-glucosidasecoding sequences from Bacillus altitudinis SYBC hb4,design up and downstream oligonucleotide primers were designed,and the target gene(bglA) was amplified by using PCR.The expression vector pColdII-bglA was constructed by subcloning the target gene into plasmid pColdII,and then transformed into E. coli BL21(DE3)for heterogeneous expression.The expression β-glucosidase acticity reached to 12.40 U/mL. The optimum temperature and pH of the recombinant β-glucosidase were 60 ℃ and 8.0,respectively. And 5mmol/L Mg2+ could increased 50% β-glucosidase activity.

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刘群,管政兵,蔡宇杰,廖祥儒. Bacillus altitudinis SYBC hb4碱性β-葡萄糖苷酶基因的克隆表达及酶学性质的研究[J].食品与生物技术学报,2017,36(11):1203-1209.

LIU Qun, GUAN Zhengbing, CAI Yujie, LIAO Xiangru. Cloning,Experssion and Characterization of Alkali-Stable β-Glucosidase from Bacillus altitudinis SYBC hb4[J]. Journal of Food Science and Biotechnology,2017,36(11):1203-1209.

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  • 在线发布日期: 2018-01-08
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