Refactoring the Concise Biosynthetic Pathway of Cyanogramide Unveils Spirooxindole Formation Catalyzed by a P450 Enzyme
Refactoring the Concise Biosynthetic Pathway of Cyanogramide Unveils Spirooxindole Formation Catalyzed by a P450 Enzyme
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重构氰基酰胺的简洁生物合成途径揭示了 P450 酶催化的螺吲哚形成
DOI：
10.1002/anie.202004978
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发表时间：
2020
期刊：
影响因子：
--
通讯作者：
Zhang Changsheng
中科院分区：
文献类型：
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作者：
Zhu Yiguang;Zhang Qingbo;Fang Chunyan;Zhang Yingli;Ma Liang;Liu Zhiwen;Zhai Shilan;Peng Jing;Zhang Liping;Zhu Weiming;Zhang Changsheng
来自海洋放线菌Actinoalloteichus cyanogriseus WH 1 - 2216 - 6的氰酰胺（1）具有独特的螺吲哚骨架，并表现出显著的生物活性，可有效逆转肿瘤细胞的耐药性。1的生物合成基因簇。通过启动子工程改造蓝灰链霉菌WH 1 - 2216 - 6，使其在天蓝色链霉菌YF 11中异源表达，从而使1和5个新衍生物的生产成为可能。有趣的是，其中四个，包括1，被确定为不同比例的对映体混合物。通过基因失活和饲养实验证实了剪裁酶（包括两种甲基转移酶（CyaEF）和三种细胞色素P450单加氧酶（CyaGHI））的功能，从而阐明了1的简洁生物合成途径。值得注意的是，CyaH被生物化学证实通过不寻常的碳阳离子介导的半频哪醇型重排反应催化1中螺羟吲哚骨架的形成。
Cyanogramide (1) from the marine actinomycete Actinoalloteichus cyanogriseus WH1‐2216‐6 features a unique spirooxindole skeleton and exhibits significant bioactivity to efficiently reverse drug resistance in tumor cells. The biosynthetic gene cluster of1in A. cyanogriseus WH1‐2216‐6 was identified and refactored by promoter engineering for heterologous expression in Streptomyces coelicolor YF11, thereby enabling the production of1and five new derivatives. Interesting, four of them, including1, were identified as enantiomeric mixtures in different ratios. The functions of tailoring enzymes, including two methyltransferases (CyaEF), and three cytochrome P450 monooxygenases (CyaGHI) were confirmed by gene inactivation and feeding experiments, leading to the elucidation of a concise biosynthetic pathway for1. Notably, CyaH was biochemically verified to catalyze the formation of the spirooxindole skeleton in1through an unusual carbocation‐mediated semipinacol‐type rearrangement reaction.