Thioesterase domain swapping of a linear polyketide tautomycetin with a macrocyclic polyketide pikromycin in Streptomyces sp. CK4412.
Thioesterase domain swapping of a linear polyketide tautomycetin with a macrocyclic polyketide pikromycin in Streptomyces sp. CK4412.
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DOI：
10.1007/s10295-016-1790-2
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发表时间：
2016-08
影响因子：
3.4
通讯作者：
Kim ES
中科院分区：
文献类型：
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作者：
Tripathi A;Choi SS;Sherman DH;Kim ES
变构霉素（TMC）是由链霉菌CK4412产生的线性聚酮代谢物，据报道具有多种生物学功能，包括T细胞特异性免疫抑制和抗癌活性，这些活性通过差异抑制蛋白磷酸酶（如PP1、PP2A和SHP 2）的机制发生。我们以前报道的特征的多功能I型聚酮合酶（PKS）组件构成的整个TMC生物合成基因簇，并建议，线性形式的TMC可以通过一个狭窄的TMC硫酯酶（TE）口袋通过游离酸链终止产生。组装的模块化性质提供了改变或交换天然生物合成结构域以产生TMC的靶向变体的独特机会。在此，我们报告了TMC TE结构域序列与大环聚酮吡克罗霉素（PIK）TE的确切对应物的交换。PIK TE交换的链霉菌CK4412突变体不仅产生TMC，而且还产生TMC的环化形式，这意味着基于生物工程的体内定制构建体可以用于产生具有新结构功能的工程化大环内酯。
Tautomycetin (TMC) is a linear polyketide metabolite produced by Streptomyces sp. CK4412 that has been reported to possess multiple biological functions including T cell-specific immunosuppressive and anticancer activities that occur through a mechanism of differential inhibition of protein phosphatases such as PP1, PP2A, and SHP2. We previously reported the characterization of the entire TMC biosynthetic gene cluster constituted by multifunctional type I polyketide synthase (PKS) assembly and suggested that the linear form of TMC could be generated via free acid chain termination by a narrow TMC thioesterase (TE) pocket. The modular nature of the assembly presents a unique opportunity to alter or interchange the native biosynthetic domains to produce targeted variants of TMC. Herein, we report swapping of the TMC TE domain sequence with the exact counterpart of the macrocyclic polyketide pikromycin (PIK) TE. PIK TE-swapped Streptomyces sp. CK4412 mutant produced not only TMC, but also a cyclized form of TMC, implying that the bioengineering based in vivo custom construct can be exploited to produce engineered macrolactones with new structural functionality.