Solanapyrone Synthase, a Possible Diels-Alderase and Iterative Type I Polyketide Synthase Encoded in a Biosynthetic Gene Cluster from Alternaria solani
Solanapyrone Synthase, a Possible Diels-Alderase and Iterative Type I Polyketide Synthase Encoded in a Biosynthetic Gene Cluster from Alternaria solani
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DOI：
10.1002/cbic.201000173
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发表时间：
2010-06-14
期刊：
影响因子：
3.2
通讯作者：
Fujii, Isao
中科院分区：
文献类型：
--
作者：
Kasahara, Ken;Miyamoto, Takanori;Fujii, Isao
克隆了番茄早疫病菌（Alternaria solani）的茄尼吡喃酮生物合成基因簇。它由六个基因sol 1 -6组成，分别编码聚酮合酶、O-甲基转移酶、脱氢酶、转录因子、黄素依赖性氧化酶和细胞色素p450。由sol 1编码的prosolanapyrone合酶（PSS）在Aspergillus sp.中表达，其产物被鉴定为去甲基prosolanapyronel（8）。虽然PSS与洛伐他汀和康伐他汀生物合成的PKSs/Diels-Alderases LovB和云母密切相关，但它不催化环加成。Sol 5编码一种黄素依赖性氧化酶（solanapyronesynthase，SPS），在毕赤酵母中表达并纯化。纯化的重组SPS显示出从非手性前茄吡喃酮II（2）形成（-）-茄吡喃酮A（1）的活性，这表明该单一酶催化氧化和随后的环加成反应，可能是Diels-Alder酶。
The solanapyrone biosynthetic gene cluster was cloned from Alternaria solani. It consists of six genes-sol1-6-coding for a polyketide synthase, an O-methyltransferase, a dehydrogenase, a transcription factor, a flavin dependent oxidase, and cytochrome p450. The prosolanapyrone synthase (PSS) encoded by sol1 was expressed in Aspergillus oryzae and its product was identified as desmethyprosolanapyronel (8). Although PSS is closely related to the PKSs/Diels-Alderases LovB and MIcA of lovastatin and compactin biosynthesis, it did not catalyze cycloaddition. Sol5 encoding a flavin-dependent oxidase (solanapyrone synthase, SPS), was expressed in Pichia pastoris and purified. The purified recombinant SPS showed activity for the formation of (-)-solanapyrone A (1) from achiral prosolanapyrone II (2) establishing that this single enzyme catalyzes both the oxidation and the subsequent cycloaddition reaction, possibly as a Diels-Alder enzyme.