由糖化酵母葡糖淀粉酶I信号序列指导的酵母中芽孢杆菌α-淀粉酶的分泌。

Secretion of Bacillus alpha-amylase from yeast directed by glucoamylase I signal sequence of Saccharomyces diastaticus.

作者信息

Kang D O, Hwang I K, Kim B Y, Ahn S C, Mheen T I, Ahn J S, Byun S M

机构信息

Microbial Chemistry Research Group, Korea Reseach Institute of Bioscience and Biotechnology, Taejon, Korea.

出版信息

Biochem Mol Biol Int. 1996 May;39(1):181-90. doi: 10.1080/15216549600201181.

DOI:10.1080/15216549600201181

PMID:8799340

Abstract

For the secretion of Bacillus stearothermophilus alpha-amylase from yeast, a recombinant plasmid pGAT17 was constructed by fusing B. stearothermophilus alpha-amylase structural gene in frame to the promoter and signal sequence of Saccharomyces diastaticus glucoamylase I gene (STA1). The secretion of the heterologous alpha-amylase from S. diastaticus transformed with pGAT17 was confirmed by the halo formation around colonies on selective starch agar medium. About 80% of the total alpha-amylase activity was detected in the extracellular culture medium. The secreted alpha-amylase was glycosylated and its molecular weight increased from 61 kDa to 75 kDa. The thermostability of the the glycosylated alpha-amylase was markedly enhanced, compared with that of the non-glycosylated enzyme from E. coli.

摘要

为了实现嗜热脂肪芽孢杆菌α-淀粉酶在酵母中的分泌，构建了重组质粒pGAT17，方法是将嗜热脂肪芽孢杆菌α-淀粉酶结构基因与糖化酵母葡糖淀粉酶I基因（STA1）的启动子和信号序列框内融合。用pGAT17转化的糖化酵母分泌异源α-淀粉酶，这通过在选择性淀粉琼脂培养基上菌落周围形成晕圈得以证实。约80%的总α-淀粉酶活性在细胞外培养基中检测到。分泌的α-淀粉酶被糖基化，其分子量从61 kDa增加到75 kDa。与来自大肠杆菌的非糖基化酶相比，糖基化α-淀粉酶的热稳定性显著增强。