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荚膜红细菌光合作用基因簇中的开放阅读框176编码异戊烯基二磷酸异构酶基因idi。

Open reading frame 176 in the photosynthesis gene cluster of Rhodobacter capsulatus encodes idi, a gene for isopentenyl diphosphate isomerase.

作者信息

Hahn F M, Baker J A, Poulter C D

机构信息

Department of Chemistry, University of Utah, Salt Lake City 84112, USA.

出版信息

J Bacteriol. 1996 Feb;178(3):619-24. doi: 10.1128/jb.178.3.619-624.1996.

Abstract

Isopentenyl diphosphate (IPP) isomerase catalyzes an essential activation step in the isoprenoid biosynthetic pathway. A database search based on probes from the highly conserved regions in three eukaryotic IPP isomerases revealed substantial similarity with ORF176 in the photosynthesis gene cluster in Rhodobacter capsulatus. The open reading frame was cloned into an Escherichia coli expression vector. The encoded 20-kDa protein, which was purified in two steps by ion exchange and hydrophobic interaction chromatography, catalyzed the interconversion of IPP and dimethylallyl diphosphate. Thus, the photosynthesis gene cluster encodes all of the enzymes required to incorporate IPP into the ultimate carotenoid and bacteriochlorophyll metabolites in R. capsulatus. More recent searches uncovered additional putative open reading frames for IPP isomerase in seed-bearing plants (Oryza sativa, Arabidopsis thaliana, and Clarkia breweri), a worm (Caenorhabditis elegans), and another eubacterium (Escherichia coli). The R. capsulatus enzyme is the smallest of the IPP isomerases to be identified thus far and may consist mostly of a fundamental catalytic core for the enzyme.

摘要

异戊烯基二磷酸(IPP)异构酶催化类异戊二烯生物合成途径中的一个关键激活步骤。基于来自三种真核生物IPP异构酶高度保守区域的探针进行数据库搜索,发现其与荚膜红细菌光合作用基因簇中的ORF176有显著相似性。该开放阅读框被克隆到大肠杆菌表达载体中。编码的20 kDa蛋白通过离子交换和疏水相互作用色谱两步法纯化,催化IPP和二甲基烯丙基二磷酸的相互转化。因此,光合作用基因簇编码了将IPP整合到荚膜红细菌最终类胡萝卜素和细菌叶绿素代谢产物中所需的所有酶。最近的搜索在种子植物(水稻、拟南芥和布鲁氏克拉克花)、一种蠕虫(秀丽隐杆线虫)和另一种真细菌(大肠杆菌)中发现了更多假定的IPP异构酶开放阅读框。荚膜红细菌的这种酶是迄今为止鉴定出的最小的IPP异构酶,可能主要由该酶的基本催化核心组成

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