Functional analyses of cytochrome P450 genes responsible for the early steps of brassicicene C biosynthesis
Functional analyses of cytochrome P450 genes responsible for the early steps of brassicicene C biosynthesis
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DOI：
10.1016/j.bmcl.2009.08.026
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发表时间：
2009-10-01
影响因子：
2.7
通讯作者：
Dairi, Tohru
中科院分区：
文献类型：
--
作者：
Hashimoto, Makoto;Higuchi, Yusuke;Dairi, Tohru
我们在之前的研究中发现，Orf8和Orf6分别为fusicoccadiene （FD）合成酶和16- o -甲基转移酶，Orf8和Orf6分别在甘蓝交替菌ATCC96836的芸苔菌C生物合成基因簇中被鉴定出来。本文研究了FD形成后的早期生物合成步骤。构建了携带FD合酶基因、5个细胞色素P450基因（orf1、orf2、orf5、orf7、orf11）中的1个（或2个）基因和从ATCC96836菌株中克隆的细胞色素P450还原酶基因的质粒，并将其导入酵母中。根据转化体产生的化合物的结构，Orf1被认为是一种8 - β -羟基化酶，产生FD 8 - β -醇(4)，随后Orf7进行16-羟基化，产生FD 8 - β - 16-二醇(5)。(C) 2009 Elsevier Ltd.。版权所有。
We previously revealed that Orf8 and Orf6, which were identified in the brassicicene C biosynthetic gene cluster in Alternaria brassicicola strain ATCC96836, were fusicoccadiene (FD) synthase and 16-O-methyltransferase, respectively. In the present Letter, the early biosynthetic steps after the formation of FD were investigated. Plasmids carrying the FD synthase gene, one (or two) of five cytochrome P450 genes (orf1, orf2, orf5, orf7, and orf11) identified in the cluster and a cytochrome P450 reductase gene cloned from strain ATCC96836 were constructed and introduced into Saccharomyces cerevisiae. Based on the structures of the compounds produced by the transformants, Orf1 is suggested to be an 8 beta-hydroxylation enzyme that yields FD 8 beta-ol ( 4), followed by 16-hydroxylation by Orf7 to produce FD 8 beta 16-diol ( 5). (C) 2009 Elsevier Ltd. All rights reserved.