草生欧文氏菌中的crtE基因编码香叶基香叶基二磷酸合酶。
The crtE gene in Erwinia herbicola encodes geranylgeranyl diphosphate synthase.
作者信息
Math S K, Hearst J E, Poulter C D
机构信息
Department of Chemistry, University of Utah, Salt Lake City 84112.
出版信息
Proc Natl Acad Sci U S A. 1992 Aug 1;89(15):6761-4. doi: 10.1073/pnas.89.15.6761.
A cluster of genes essential for the biosynthesis of carotenoids in Erwinia herbicola has been isolated and characterized [Armstrong, G.A., Alberti, M. & Hearst, J. E. (1990) Proc. Natl. Acad. Sci. USA 87, 9975-9979]. Related gene clusters are found in other carotenoid-producing bacteria. Two of these genes, crtB and crtE, have been assigned to enzymes responsible for conversion of geranylgeranyl diphosphate (GGPP) to prephytoene diphosphate and prephytoene diphosphate to phytoene, respectively. We isolated crtE from the Er. herbicola cluster by PCR amplification and cloned the coding region into the Escherichia coli expression vector pARC306N. Es. coli JM101 was transformed with the expression plasmid, and transformants were assayed for GGPP synthase and phytoene synthase activity. Extracts from JM101/pSM145 accumulated [14C]GGPP when incubated with [14C]isopentenyl diphosphate and farnesyl diphosphate, whereas similar incubations with [3H]GGPP did not yield prephytoene diphosphate or phytoene. Thus, crtE encodes GGPP synthase.
已从草生欧文氏菌中分离并鉴定出一组对类胡萝卜素生物合成至关重要的基因[阿姆斯特朗，G.A.，阿尔贝蒂，M.和赫斯特，J.E.（1990年）《美国国家科学院院刊》87，9975 - 9979]。在其他产类胡萝卜素的细菌中也发现了相关的基因簇。其中两个基因，crtB和crtE，分别被指定为负责将牻牛儿基牻牛儿基二磷酸（GGPP）转化为前八氢番茄红素二磷酸以及将前八氢番茄红素二磷酸转化为八氢番茄红素的酶。我们通过聚合酶链反应（PCR）扩增从草生欧文氏菌基因簇中分离出crtE，并将编码区克隆到大肠杆菌表达载体pARC306N中。用该表达质粒转化大肠杆菌JM101，并对转化子进行GGPP合酶和八氢番茄红素合酶活性检测。当JM101/pSM145提取物与[14C]异戊烯基二磷酸和法呢基二磷酸一起孵育时，会积累[14C]GGPP，而与[3H]GGPP进行类似孵育时，并不会产生前八氢番茄红素二磷酸或八氢番茄红素。因此，crtE编码GGPP合酶。
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