来自淡紫链霉菌的α-淀粉酶基因的克隆与特性分析

Cloning and characterization of an alpha-amylase gene from Streptomyces lividans.

作者信息

Tsao L S, Lin L L, Chen J C, Chen J H, Hsu W H

机构信息

Culture Collection and Research Center, Food Industry Research and Development Institute, Hsinchu, Taiwan, R.O.C.

出版信息

Biochim Biophys Acta. 1993 Jan 23;1171(3):255-62. doi: 10.1016/0167-4781(93)90063-j.

DOI:10.1016/0167-4781(93)90063-j

PMID:8424949

Abstract

The alpha-amylase gene (amy) of Streptomyces lividans TK24 was cloned in an amylase deficient mutant strain S. lividans M2. The cloned gene contained an open reading frame (ORF) of 2757 nucleotides (919 amino acids) coding for a protein of 100 kDa. Sequencing of the amino terminus of the extracellular alpha-amylase protein revealed the presence of a signal peptide of 33 amino acid residues. The transcriptional initiation site was mapped by the primer extension method with T4 DNA polymerase and was found to be transcribed from an unique promoter. The alpha-amylase protein produced by S. lividans was larger than those derived from other origins. It also contained the four common conserved regions characteristic of other alpha-amylase proteins.

摘要

将天蓝色链霉菌TK24的α-淀粉酶基因（amy）克隆到淀粉酶缺陷型突变株天蓝色链霉菌M2中。克隆的基因包含一个2757个核苷酸（919个氨基酸）的开放阅读框（ORF），编码一种100 kDa的蛋白质。对细胞外α-淀粉酶蛋白的氨基末端进行测序，发现存在一个33个氨基酸残基的信号肽。通过用T4 DNA聚合酶的引物延伸法确定转录起始位点，发现它是从一个独特的启动子转录而来的。天蓝色链霉菌产生的α-淀粉酶蛋白比其他来源的α-淀粉酶蛋白更大。它还包含其他α-淀粉酶蛋白特有的四个常见保守区域。