产环糊精葡萄糖苷转移酶的基因工程菌及其应用
申请公布号:CN105647847A
申请号:CN201610151808.9
申请日期:2016.03.17
申请公布日期:2016.06.08
发明人:马江锋;蒋羽佳;董维亮;吴若凡;陈娟;高有军;吴昊;姜岷
分类号:C12N1/21(2006.01)I;C12N15/70(2006.01)I;C12N9/10(2006.01)I;C12P19/60(2006.01)I;C12R1/19(2006.01)N
主分类号:C12N1/21(2006.01)I
代理人:王月霞
地址:210009 江苏省南京市浦口区浦珠南路30号
摘要:本发明公开了一种产环糊精葡萄糖苷转移酶的基因工程菌及其应用,所述基因工程菌是将环糊精葡萄糖苷转移酶基因通过载体导入到宿主细胞中得到的,其可将产生的环糊精葡萄糖苷转移酶留在细胞内;其中所述的载体为质粒载体pET28a。所述宿主细胞中导入有含分子伴侣Dnak-DnaJ-GrpE-GroES-GroEL的质粒pGKJE8。所述菌株将含有分子伴侣Dnak-DnaJ-GrpE-GroES-GroEL的质粒pGKJE8和pET28a-cgt一并导入宿主细胞中,实现共表达,可以有效提高蛋白的正确折叠率,从而减少包涵体的形成,提高酶的正确表达率,提高酶活。本发明构建的菌株酶活可提高到7.82U/mg DCW,通过酶活的提高可以有效提高2-O-α-D-吡喃葡萄糖基抗坏血酸产量。
主权项:一种产环糊精葡萄糖苷转移酶的基因工程菌,其特征在于,该基因工程菌是将环糊精葡萄糖苷转移酶基因通过载体导入到宿主细胞中得到的,其可将产生的环糊精葡萄糖苷转移酶留在细胞内;其中所述的载体为质粒载体pET28a。
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