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嗜热嗜酸古菌酸栖热硫化叶菌S5硫氧化酶/还原酶基因在大肠杆菌中的克隆与异源表达

Cloning and heterologous expression of a sulfur oxygenase/reductase gene from the thermoacidophilic archaeon Acidianus sp. S5 in Escherichia coli.

作者信息

He Z, Li Y, Zhou P, Liu S

机构信息

State Key Laboratory of Institute of Microbiology, Chinese Academy of Sciences, 100080, Beijing, PR China.

出版信息

FEMS Microbiol Lett. 2000 Dec 15;193(2):217-21. doi: 10.1111/j.1574-6968.2000.tb09427.x.

Abstract

A thermoacidophilic, obligately chemolithotrophic, facultatively aerobic archaebacterium, Acidianus sp. S5, was isolated from acidothermal springs in southwest China. The sulfur oxygenase/reductase (SOR) gene of Acidianus sp. S5 was cloned and expressed in Escherichia coli. Several primers were designed and successfully applied for detection and cloning of the sor gene. A 3.7-kb EcoRI fragment containing the sor gene and three neighboring open reading frames was sequenced. Sequence analysis indicated that the sor gene of Acidianus sp. S5 showed 81% identity to the sor gene of Acidianus ambivalens. E. coli cells carrying the sor gene on pBV220SOR were able to overproduce SOR upon a temperature shift from 30 to 42 degrees C. SOR produced in E. coli catalyzes the oxidation of elemental sulfur and concomitant production of sulfite, thiosulfate and hydrogen sulfide. The recombinant enzyme exhibits the same catalytic properties as the one from Acidianus S5.

摘要

从中国西南部的酸性温泉中分离出了一种嗜热嗜酸、专性化能无机营养、兼性好氧的古细菌——嗜酸硫化叶菌(Acidianus sp. S5)。克隆了嗜酸硫化叶菌S5的硫氧化酶/还原酶(SOR)基因,并在大肠杆菌中进行表达。设计了几条引物并成功用于sor基因的检测和克隆。对包含sor基因和三个相邻开放阅读框的3.7 kb EcoRI片段进行了测序。序列分析表明,嗜酸硫化叶菌S5的sor基因与兼性嗜酸硫化叶菌的sor基因有81%的同一性。携带pBV220SOR上的sor基因的大肠杆菌细胞在温度从30℃转变为42℃时能够过量产生SOR。在大肠杆菌中产生的SOR催化元素硫的氧化并伴随产生亚硫酸盐、硫代硫酸盐和硫化氢。重组酶表现出与来自嗜酸硫化叶菌S5的酶相同的催化特性。

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