Actinoallolides are anti-trypanosomal macrolides isolated from the secondary metabolites of two endophytic actinomycete strains, Actinoallomurus fulvus MK10-036 and K09-0307. A putative actinoallolide biosynthetic gene cluster was predicted from the genome sequence of the strain K09-0307. The gene cluster spans a contiguous 53 kb DNA region that comprises seven genes encoding three PKSs (aalA1, aalA2, and aalA3), cytochrome P450 (aalB), acyl-CoA dehydrogenase (aalC), crotonyl-CoA reductase (aalD), and TetR family regulator (aalR). The entire gene cluster was cloned into a plasmid pYIK1 by assembling DNA fragments, which were obtained from two cosmids containing left and right parts of the gene cluster. Following the introduction of an ermE* promoter at 100bp upstream from the start codon of aalA1, the gene cluster was introduced into Streptomyces coelicolor M1152. Subsequent LC-MS analysis revealed production of actinoallolide A in the culture broth. Thus, the actinoallolide biosynthetic gene cluster was identified by heterologous expression in Streptomyces.

中文翻译：

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放线肌醇生物合成基因簇的鉴定和异源表达。

放线菌素是从两个内生放线菌菌株Actinoallomurus fulvus MK10-036和K09-0307的次生代谢产物中分离的抗锥虫大环内酯类药物。从菌株K09-0307的基因组序列预测了推定的放线菌素生物合成基因簇。基因簇跨越一个连续的53 kb DNA区域，该区域包含七个编码三个PKS（aalA1，aalA2和aalA3），细胞色素P450（aalB），酰基辅酶A脱氢酶（aalC），巴豆酰基辅酶A还原酶（aalD）和TetR的基因。家庭管理者（aalR）。通过组装DNA片段，将整个基因簇克隆到质粒pYIK1中，这些片段是从两个含有基因簇左右部分的粘粒中获得的。在aalA1起始密码子上游100 bp处引入ermE *启动子后，将基因簇导入链霉菌M1152。随后的LC-MS分析表明在培养液中产生放线型A的生成。因此，通过链霉菌中的异源表达鉴定了放线菌素内酯生物合成基因簇。