Biosynthesis of terpenes:: Studies on 1-hydroxy-2-methyl-2-(E)-butenyl 4-diphosphate reductase
Biosynthesis of terpenes:: Studies on 1-hydroxy-2-methyl-2-(E)-butenyl 4-diphosphate reductase
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DOI：
10.1073/pnas.182412599
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发表时间：
2002-09-17
影响因子：
11.1
通讯作者：
Rohdich, F
中科院分区：
文献类型：
--
作者：
Adam, P;Hecht, S;Rohdich, F
早期的体内研究表明，IspH蛋白参与1-羟基-2-甲基-2-（E）-丁烯基4-二磷酸转化为异戊烯基二磷酸（IPP）和二甲基烯丙基二磷酸（DMAPP）。我们现在已经证明，为ispH（lytB）基因的过表达而工程化的大肠杆菌菌株的细胞提取物催化1-羟基-2-甲基-2-（E）-丁烯基4-二磷酸体外转化为IPP和DMAPP。该反应需要NADH、FAD、二价阳离子（优选Co2+），以及可能的一种或多种尚未鉴定的蛋白质。野生型E.通过添加纯化的重组IspH蛋白增强了红辣椒（Capsicum annuum）的大肠杆菌细胞提取物和分离的色质体。
Earlier in vivo studies showed the involvement of IspH protein in the conversion of 1-hydroxy-2-methyl-2-(E)-butenyl 4-diphosphate into isopentenyl diphosphate (IPP) and dimethylallyl diphosphate (DMAPP). We have demonstrated now that cell extract of an Escherichia coli strain engineered for hyperexpression of the ispH (lytB) gene catalyzes the in vitro conversion of 1-hydroxy-2-methyl-2-(E)-butenyl 4-diphosphate into IPP and DMAPP. The reaction requires NADH, FAD, divalent cations (preferably Co2+), and probably one or more as-yet-unidentified proteins. The low intrinsic catalytic activities of wild-type E. coli cell extract and isolated chromoplasts of red pepper (Capsicum annuum) are enhanced by the addition of purified recombinant IspH protein.