雪白根霉脂肪酶基因组DNA的克隆及其在酿酒酵母中的表达。

Cloning of genomic DNA of Rhizopus niveus lipase and expression in the yeast Saccharomyces cerevisiae.

作者信息

Kohno M, Enatsu M, Kugimiya W

机构信息

Central Research Institute, Tsukuba R&D Center, Fuji Oil Co., Ltd., Ibaraki, Japan.

出版信息

Biosci Biotechnol Biochem. 1998 Dec;62(12):2425-7. doi: 10.1271/bbb.62.2425.

DOI:10.1271/bbb.62.2425

PMID:9972270

Abstract

Genomic DNA encoding Lipase I was cloned from Rhizopus niveus strain IFO4759. For expression of this gene in S. cerevisiae, DBY746 was transformed with YEp352PLipS, which had the cloned lipase gene under the control of a PGK promoter. This strain secreted the lipase at a high level (350 U/ml). The strain ND-12B was produced by a mating of DBY746, harboring YEp352PLipS, and NA74-3A, and dissection of asci. This new strain secreted the lipase up to 530 U/ml. Moreover, the lipase was produced most effectively in a medium containing Bacto-yeast extract, soy-peptide, and sucrose.

摘要

编码脂肪酶I的基因组DNA是从雪白根霉IFO4759菌株中克隆得到的。为了在酿酒酵母中表达该基因，用YEp352PLipS转化DBY746，该质粒中的克隆脂肪酶基因受PGK启动子控制。该菌株能高水平分泌脂肪酶（350 U/ml）。菌株ND - 12B是通过将携带YEp352PLipS的DBY746与NA74 - 3A进行交配并对孢子囊进行解剖而产生的。这个新菌株分泌的脂肪酶高达530 U/ml。此外，在含有细菌酵母提取物、大豆肽和蔗糖的培养基中，脂肪酶的产量最高。