米曲霉固态发酵（SSF）中表达的glaB启动子的改进。

Improvement of the glaB promoter expressed in solid-state fermentation (SSF) of Aspergillus oryzae.

作者信息

Ishida Hiroki, Hata Yoji, Kawato Akitsugu, Abe Yasuhisa

机构信息

Research Institute, Gekkeikan Sake Co., Ltd, Kyoto, Japan.

出版信息

Biosci Biotechnol Biochem. 2006 May;70(5):1181-7. doi: 10.1271/bbb.70.1181.

DOI:10.1271/bbb.70.1181

PMID:16717420

Abstract

The glucoamylase-encoding gene (glaB) promoter should be very useful for recombinant protein production in solid-state fermentation (SSF) of Aspergillus oryzae. A 97-bp fragment containing the cis-element of the glaB promoter was inserted into the glaA promoter, which was little expressed in SSF. The chimeric promoter showed about a 24-fold increase in promoter activity in SSF. Eight copies of the 97-bp fragment were tandemly fused with the glaB promoter. The improved promoter showed about a 4.6-fold increase in promoter activity in SSF. The glaB gene was overexpressed under control of the improved glaB promoter in SSF. Recombinant glucoamylase production reached about 1524 mg/kg-broth for 2 d. The improved glaB promoter should be very useful for overproduction of a recombinant protein in SSF of A. oryzae.

摘要

葡糖淀粉酶编码基因（glaB）启动子对于米曲霉固态发酵（SSF）生产重组蛋白应该非常有用。将包含glaB启动子顺式元件的97 bp片段插入到在SSF中表达很少的glaA启动子中。嵌合启动子在SSF中的启动子活性显示出约24倍的增加。97 bp片段的八个拷贝与glaB启动子串联融合。改进后的启动子在SSF中的启动子活性显示出约4.6倍的增加。在SSF中，glaB基因在改进后的glaB启动子控制下过表达。重组葡糖淀粉酶产量在2天内达到约1524 mg/kg-发酵液。改进后的glaB启动子对于米曲霉SSF中重组蛋白的过量生产应该非常有用。