A genetically amenable platensimycin- and platencin-overproducer as a platform for biosynthetic explorations: a showcase of PtmO4, a long-chain acyl-CoA dehydrogenase.
A genetically amenable platensimycin- and platencin-overproducer as a platform for biosynthetic explorations: a showcase of PtmO4, a long-chain acyl-CoA dehydrogenase.
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DOI：
10.1039/c5mb00303b
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发表时间：
2015-10
影响因子：
--
通讯作者：
Shen B
中科院分区：
文献类型：
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作者：
Rudolf JD;Dong LB;Huang T;Shen B
铂霉素（Platensimycin， PTM）和铂霉素（platencin， PTN）是一类新的靶向细菌和哺乳动物脂肪酸合酶的有前途的药物先导物。我们之前克隆并测序了PTM和PTN基因簇，发现了另外六个PTM-PTN双产菌株，并通过灭活四种不同菌株的通路特异性调节因子ptmR1或ptnR1，证明了PTM和PTN的显著过量生产。我们利用这些PTM-PTN双重高产菌株的能力受到其缺乏遗传适应性的限制。在这里，我们报道了platptomyces SB12029的构建，这是一种遗传可适应的，框架内ΔptmR1双PTM-PTN过量生产菌株。为了突出该菌株在未来PTM和PTN生物合成研究中的潜力，我们创建了ΔptmR1 ΔptmO4双突变体S. platensis SB12030。从SB12030中分离到14个PTM和PTN同源基因，其中10个为新基因，为PTM和PTN的生物合成提供了新的见解。PtmO4是一种长链酰基辅酶a脱氢酶，与催化二萜中间体β-氧化形成PTM和PTN支架密切相关。SB12029为PTM和PTN家族天然产物的未来生物合成和生物工程研究奠定了基础。
Platensimycin (PTM) and platencin (PTN) are members of a new class of promising drug leads that target bacterial and mammalian fatty acid synthases. We previously cloned and sequenced the PTM and PTN gene clusters, discovered six additional PTM-PTN dual producing strains, and demonstrated the dramatic overproduction of PTM and PTN by inactivating the pathway-specific regulators ptmR1 or ptnR1 in four different strains. Our ability to utilize these PTM-PTN dual overproducing strains was limited by their lack of genetic amenability. Here we report the construction of Streptomyces platensis SB12029, a genetically amenable, in-frame ΔptmR1 dual PTM-PTN overproducing strain. To highlight the potential of this strain for future PTM and PTN biosynthetic studies, we created the ΔptmR1 ΔptmO4 double mutant S. platensis SB12030. Fourteen PTM and PTN congeners, ten of which were new, were isolated from SB12030, shedding new insights into PTM and PTN biosynthesis. PtmO4, a long-chain acyl-CoA dehydrogenase, is strongly implicated to catalyze β-oxidation of the diterpenoid intermediates in to the PTM and PTN scaffolds. SB12029 sets the stage for future biosynthetic and bioengineering studies of the PTM and PTN family of natural products.