产朊假丝酵母α-淀粉酶基因在酿酒酵母中的克隆与表达。

Cloning and Expression of a Schwanniomyces occidentalis alpha-Amylase Gene in Saccharomyces cerevisiae.

机构信息

Culture Collection and Research Center, Food Industry Research and Development Institute, P.O. Box 246, Hsinchu, Taiwan, Republic of China.

出版信息

Appl Environ Microbiol. 1989 Dec;55(12):3167-72. doi: 10.1128/aem.55.12.3167-3172.1989.

DOI:10.1128/aem.55.12.3167-3172.1989

PMID:16348077

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC203241/

Abstract

An alpha-amylase gene (AMY) was cloned from Schwanniomyces occidentalis CCRC 21164 into Saccharomyces cerevisiae AH22 by inserting Sau3AI-generated DNA fragments into the BamHI site of YEp16. The 5-kilobase insert was shown to direct the synthesis of alpha-amylase. After subclones containing various lengths of restricted fragments were screened, a 3.4-kilobase fragment of the donor strain DNA was found to be sufficient for alpha-amylase synthesis. The concentration of alpha-amylase in culture broth produced by the S. cerevisiae transformants was about 1.5 times higher than that of the gene donor strain. The secreted alpha-amylase was shown to be indistinguishable from that of Schwanniomyces occidentalis on the basis of molecular weight and enzyme properties.

摘要

从施氏假丝酵母 CCRC 21164 中克隆了α-淀粉酶基因（AMY），并通过将 Sau3AI 生成的 DNA 片段插入 YEp16 的 BamHI 位点，将其导入酿酒酵母 AH22。该 5kb 插入片段可指导α-淀粉酶的合成。在筛选出含有各种长度受限片段的亚克隆后，发现供体菌株 DNA 的 3.4kb 片段足以进行α-淀粉酶的合成。与基因供体菌株相比，酿酒酵母转化体产生的发酵液中α-淀粉酶的浓度约高 1.5 倍。根据分子量和酶特性，分泌的α-淀粉酶与施氏假丝酵母的α-淀粉酶无法区分。