The roles of genes associated with regulation, transportation, and macrocyclization in desotamide biosynthesis in Streptomyces scopuliridis SCSIO ZJ46
The roles of genes associated with regulation, transportation, and macrocyclization in desotamide biosynthesis in Streptomyces scopuliridis SCSIO ZJ46
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与调节、运输和大环化相关的基因在链霉菌 SCSIO ZJ46 去索酰胺生物合成中的作用
DOI：
10.1007/s00253-020-10414-4
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发表时间：
2020-01-31
影响因子：
5
通讯作者：
Li, Qinglian
中科院分区：
文献类型：
--
作者：
Ding, Wenjuan;Dong, Yuliang;Li, Qinglian
源自深海的微生物 Streptomyces scopuliridis SCSIO ZJ46 产生去索酰胺 A-D。值得注意的是，去索酰胺 A 和 B 对致病性革兰氏阳性肺炎链球菌 NCTC 7466、金黄色葡萄球菌 ATCC 29213 和耐甲氧西林临床分离株表皮葡萄球菌 (MRSE) shhs-E1 具有抗菌活性。 39-kb 去索酰胺生物合成基因簇 (dsa) 先前已在 S. coelicolor M1152 中被鉴定并异源表达，用于指定 dsa 基因功能。在这项工作中，我们鉴定了 dsa 簇中的 7 个基因，包括 3 个调节基因（dsaA、dsaM 和 dsaN）、2 个转运蛋白基因（dsaK 和 dsaL）以及另外两个基因 dsaB（注释为磷酸合酶）和 dsaJ（PBP 型硫酯酶）。 DsaA 和 DsaN 被明确证明是去索酰胺生物合成的正调节因子，并且与这些作用一致，任一基因的失活都完全消除了去索酰胺的产生。此外，dsaA 或 dsaN（彼此独立）的过度表达可提高去索酰胺滴度。 M1152/07-6H::dsaA 菌株中去索酰胺的产量比异源 dsa 表达菌株 M1152/07-6H 中的去索酰胺产量高 2.4 倍，而来自 M1152/07-6H::dsaN 菌株的去索酰胺效价约为 M1152/07-6H 的两倍。此外，dsaB 和 dsaJ 的失活（彼此独立）完全消除了去索酰胺的产生，表明它们对于去索酰胺的组装是必不可少的。这些研究为dsa生物合成基因簇内七个关键基因的功能和组合生物合成潜力提供了新的见解。这里报告的研究结果可能会促进旨在评估和开发去索酰胺及相关类似物以供未来应用的进一步努力。
The deep-sea-derived microbe Streptomyces scopuliridis SCSIO ZJ46 produces desotamides A-D. Notably, desotamides A and B display antibacterial activities against pathogenic Gram-positive Streptococcus pneumoniae NCTC 7466, Staphylococcus aureus ATCC 29213, and the methicillin-resistant clinical isolate Staphylococcus epidermidis (MRSE) shhs-E1. The 39-kb desotamide biosynthetic gene cluster (dsa) has previously been identified and heterologously expressed in S. coelicolor M1152 for the purposes of assigning dsa gene functions. In this work, we identified seven genes in the dsa cluster including three regulatory genes (dsaA, dsaM, and dsaN), two transporter genes (dsaK and dsaL), and two other genes, dsaB (annotated as a phosphate synthase) and dsaJ (a PBP-type thioesterase). The DsaA and DsaN were unambiguously shown to be positive regulators of desotamide biosynthesis, and consistent with these roles, inactivation of either gene completely abolished desotamide production. Moreover, overexpression of dsaA or dsaN (independent of each other) was shown to improve desotamide titers. Production of desotamides in M1152/07-6H::dsaA strain was 2.4-fold greater than that in the heterologous dsa expression strain M1152/07-6H whereas desotamide titers from the M1152/07-6H::dsaN strain were about twice that of M1152/07-6H. In addition, inactivation of dsaB and dsaJ (independent of each other) completely abolished desotamide production, indicating their indispensability for desotamide assembly. These studies provide new insights into the functions and combinatorial biosynthetic potentials of seven key genes within the dsa biosynthetic gene cluster. Findings reported here are likely to facilitate further efforts aimed at assessing and developing the desotamides and related analogs for future applications.