Escherichia coli engineered to synthesize isopentenyl diphosphate and dimethylallyl diphosphate from mevalonate:: a novel system for the genetic analysis of the 2-C-methyl-D-erythritol 4-phosphate pathway for isoprenoid biosynthesis
Escherichia coli engineered to synthesize isopentenyl diphosphate and dimethylallyl diphosphate from mevalonate:: a novel system for the genetic analysis of the 2-C-methyl-D-erythritol 4-phosphate pathway for isoprenoid biosynthesis
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DOI：
10.1042/bj3530059
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发表时间：
2001-01-01
影响因子：
4.1
通讯作者：
Boronat, A
中科院分区：
文献类型：
--
作者：
Campos, N;Rodríguez-Concepción, M;Boronat, A
异戊烯基二磷酸 (IPP) 及其异构体二甲基烯丙基二磷酸 (DMAPP) 构成类异戊二烯的基本组成部分，类异戊二烯是结构和功能极其多样的化合物家族。 IPP和DMAPP可以通过两条独立的途径合成：甲羟戊酸途径和最近发现的2-C-甲基-D-赤藓糖醇4-磷酸(MEP)途径。尽管 MEP 途径在大多数真细菌、藻类和植物中至关重要，并且具有巨大的生物技术意义，但仅确定了其中的一些步骤。我们设计了一个适合大肠杆菌MEP途径遗传分析的系统，编码酵母5-二磷酸甲羟戊酸脱羧酶、人5-磷酸甲羟戊酸激酶、酵母甲羟戊酸激酶和大肠杆菌异戊烯基二磷酸异构酶的合成操纵子被整合到该细菌的染色体中。该操纵子的表达允许从外源添加的甲羟戊酸合成IPP和DMAPP并补充MEP途径的致死突变体。我们使用该系统证明了 ygbP、ychB 和 ygbB 基因在大肠杆菌中是必需的，并且这些基因产物催化的步骤属于 MEP 途径的主干线。
Isopentenyl diphosphate (IPP) and its isomer dimethylallyl diphosphate (DMAPP) constitute the basic building block of isoprenoids, a family of compounds that is extraordinarily diverse in structure and function. IPP and DMAPP can be synthesized by two independent pathways: the mevalonate pathway and the recently discovered 2-C-methyl-D-erythritol 4-phosphate (MEP) pathway. Although the MEP pathway is essential in most eubacteria, algae and plants and has enormous biotechnological interest, only some of its steps have been determined. We devised a system suitable for the genetic analysis of the MEP pathway in Escherichia coli, A synthetic operon coding for yeast 5-diphosphomevalonate decarboxylase, human 5-phosphomevalonate kinase, yeast mevalonate kinase and E. coli isopentenyl diphosphate isomerase was incorporated in the chromosome of this bacterium. The expression of this operon allowed the synthesis of IPP and DMAPP from mevalonate added exogenously and complementation of lethal mutants of the MEP pathway. We used this system to show that the ygbP, ychB and ygbB genes are essential in E. coli and that the steps catalysed by the products of these genes belong to the trunk line of the MEP pathway.