来自嗜热栖热菌的耐热甘油激酶：该酶基因的克隆、测序及表达

Thermostable glycerol kinase from Thermus flavus: cloning, sequencing, and expression of the enzyme gene.

作者信息

Huang H S, Kabashima T, Ito K, Yin C H, Nishiya Y, Kawamura Y, Yoshimoto T

机构信息

School of Pharmaceutical Sciences, Nagasaki University, Japan.

出版信息

Biochim Biophys Acta. 1998 Feb 17;1382(2):186-90. doi: 10.1016/s0167-4838(97)00206-9.

DOI:10.1016/s0167-4838(97)00206-9

PMID:9540790

Abstract

The thermostable glycerol kinase (EC 2.7.1.30) gene from Thermus flavus was cloned and expressed in Escherichia coli DH5 alpha. An open reading frame of 1488 bp for the glycerol kinase gene (glpK) starting with an ATG methionine codon was found, which encodes a protein of 496 amino acid residues whose calculated molecular weight is 54,835. The amino acid sequence of T. flavus glycerol kinase is 80.6% and 64.1% identical with those of Bacillus subtilis and E. coli. Transformants of E. coli DH5 alpha harboring plasmid pGYK12 with a 1505 bp chromosomal DNA fragment containing the T. flavus glycerol kinase gene showed about 23.8-fold higher glycerol kinase activity than T. flavus.

摘要

从嗜热栖热菌（Thermus flavus）中克隆出热稳定甘油激酶（EC 2.7.1.30）基因，并在大肠杆菌DH5α中进行表达。发现了一个1488 bp的甘油激酶基因（glpK）开放阅读框，其起始密码子为ATG甲硫氨酸密码子，该基因编码一个由496个氨基酸残基组成的蛋白质，其计算分子量为54,835。嗜热栖热菌甘油激酶的氨基酸序列与枯草芽孢杆菌（Bacillus subtilis）和大肠杆菌（E. coli）的氨基酸序列分别具有80.6%和64.1%的同一性。携带含有嗜热栖热菌甘油激酶基因的1505 bp染色体DNA片段的质粒pGYK12的大肠杆菌DH5α转化子显示出比嗜热栖热菌高约23.8倍的甘油激酶活性。