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SEQUENCE AND REGULATION OF VITAMIN K BIOSYNTHESIS GENES

维生素 K 生物合成基因的序列和调控

基本信息

批准号：
2444839
负责人：
RANGASWAMY MEGANATHAN
金额：
$ 14.35万
依托单位：
NORTHERN ILLINOIS UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
1994
资助国家：
美国
起止时间：
1994-07-01 至 1999-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/2444839
关键词：
Escherichia coli alpha ketoglutarate chimeric proteins enzyme activity enzyme biosynthesis genetic regulation high performance liquid chromatography menadione molecular cloning nucleic acid sequence operon protein purification vitamin K vitamin biosynthesis

项目摘要

The principal aim of this investigation is to understand the regulation of expression of the genes and enzymes involved in the vitamin K biosynthetic pathway. Vitamin K is an important fat soluble vitamin that is required for the well being of humans and animals. Vitamin K responsive hypoprothrombia is an important clinical problem under a number of conditions. The bacterial flora of the human intestine contribute from 50-100% of the human vitamin K requirement. Vitamin K is derived from the "shikimate pathway" and it is formed from isochorismate and 2-ketoglutarate. The intermediates that have been identified are 2-succinyl-6-hydroxy-2,4-cyclohexadiene-1-carboxylate (SHCHC), o-succinylbenzoate (OSB), a CoA ester of OSB (OSB-CoA), 1,4- dihydroxy-2-naphthoate (DHNA), and desmethylmenaquinone (DMK). The menD, C, E, and B genes located at 48.5 min of the chromosome have been cloned and sequenced. The sequencing of the menA gene at 89 min is in progress and we will identify and clone the gene for the conversion of DMK---> MK. To overcome the difficulty of working with small quantities of enzymes and intermediates, we will use operon and protein fusions to study regulation and we will introduce the genes into overexpression vectors to overproduce, purify and study the enzymes and intermediates of the pathway.

本次调查的主要目的是了解法规维生素 K 相关基因和酶的表达生物合成途径。维生素K是一种重要的脂溶性维生素是人类和动物福祉所必需的。维生素K 反应性低血栓症是一个重要的临床问题条件数。人体肠道的细菌菌群满足人体维生素 K 需求的 50-100%。维生素 K 源自“莽草酸途径”，由异分支酸和2-酮戊二酸。已经得到的中间体确定为 2-琥珀酰-6-羟基-2,4-环己二烯-1-羧酸酯 (SCHHC), 邻琥珀酰苯甲酸酯 (OSB), OSB 的 CoA 酯 (OSB-CoA), 1,4- 二羟基-2-萘甲酸酯（DHNA）和去甲基甲萘醌（DMK）。位于染色体 48.5 分钟的 menD、C、E 和 B 基因具有已被克隆并测序。 89 分钟时 menA 基因的测序结果为正在进行中，我们将识别并克隆用于转换的基因 DMK---> MK。克服与小企业合作的困难大量的酶和中间体，我们将使用操纵子和蛋白质融合来研究调控，我们将把基因引入过度表达载体来过度生产、纯化和研究酶和途径的中间体。