西方许旺酵母α-淀粉酶基因分析及其基因产物在不同酵母属转化子中的分泌

Analysis of the alpha-amylase gene of Schwanniomyces occidentalis and the secretion of its gene product in transformants of different yeast genera.

作者信息

Strasser A W, Selk R, Dohmen R J, Niermann T, Bielefeld M, Seeboth P, Tu G H, Hollenberg C P

机构信息

Rhein Biotech GmbH, Düsseldorf, Federal Republic of Germany.

出版信息

Eur J Biochem. 1989 Oct 1;184(3):699-706. doi: 10.1111/j.1432-1033.1989.tb15069.x.

DOI:10.1111/j.1432-1033.1989.tb15069.x

PMID:2806251

Abstract

We have cloned and characterized the alpha-amylase gene (AMY1) of the yeast Schwanniomyces occidentalis. A cosmid gene library of S. occidentalis DNA was screened in Saccharomyces cerevisiae for alpha-amylase secretion. The positive clone contained a DNA fragment harbouring an open reading frame of 1536 nucleotides coding for a 512-amino-acid polypeptide with a calculated Mr of 56,500. The deduced amino acid sequence reveals significant similarity to the sequence of the Saccharomycopsis fibuligera and Aspergillus oryzae alpha-amylases. The AMY l gene was found to be expressed from its original promoter in S. cerevisiae, Kluyveromyces lactis and Schizo-saccharomyces pombe leading to an active secreted gene product and thus enabling the different yeast transformants to grow on starch as a sole carbon source.

摘要

我们已经克隆并鉴定了西方许旺酵母（Schwanniomyces occidentalis）的α-淀粉酶基因（AMY1）。在酿酒酵母（Saccharomyces cerevisiae）中筛选西方许旺酵母DNA的黏粒基因文库以寻找α-淀粉酶分泌情况。阳性克隆包含一个DNA片段，该片段带有一个1536个核苷酸的开放阅读框，编码一个512个氨基酸的多肽，计算所得的分子量为56,500。推导的氨基酸序列显示与扣囊复膜孢酵母（Saccharomycopsis fibuligera）和米曲霉（Aspergillus oryzae）的α-淀粉酶序列有显著相似性。发现AMY1基因在酿酒酵母、乳酸克鲁维酵母（Kluyveromyces lactis）和粟酒裂殖酵母（Schizosaccharomyces pombe）中从其原始启动子开始表达，产生一种有活性的分泌型基因产物，从而使不同的酵母转化体能以淀粉作为唯一碳源生长。