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生防放线菌 Bikiniensis sp. HD-087 对稻瘟病菌的抑菌活性及其聚酮合酶基因 pksL 的生物信息学分析

Anti-Magnaporthe oryzae Activity of Streptomyces bikiniensis HD-087 In Vitro and Bioinformatics Analysis of Polyketide Synthase Gene pksL.

机构信息

Engineering Research Center of Agricultural Microbiology Technology, Ministry of Education & Heilongjiang Provincial Key Laboratory of Plant Genetic Engineering and Biological Fermentation Engineering for Cold Region & Heilongjiang Provincial Key Laboratory of Ecological Restoration and Resource Utilization for Cold Region & Key Laboratory of Microbiology, College of Heilongjiang Province & School of Life Sciences, Heilongjiang University, Harbin, 150080, China.

出版信息

Curr Microbiol. 2024 Sep 28;81(11):379. doi: 10.1007/s00284-024-03898-0.

Abstract

Streptomyces bikiniensis HD-087 is capable of synthesizing various antimicrobial substances to counter the detrimental effects of hazardous microorganisms. To elucidate whether it produces polyketide antibiotics and the synthesis mechanism of antibiotic substances, the metabolites and related genes of S. bikiniensis HD-087 were analyzed through LC-MS, anti-Magnaporthe oryzae activity detection, and bioinformatics approaches. The result indicated that the strain HD-087 could produce erythromycin, a polyketide antibiotic. The inhibitory zones of the fermentation supernatant of strain HD-087 and methanol solution of erythromycin extract against M. oryzae were 40.84 ± 0.68 mm and 33.18 ± 0.81 mm, respectively. The IC50 value of erythromycin extract for inhibiting spore germination of erythromycin extract was 220.43 μg/mL. There are two polyketide synthesis gene clusters in the genome of strain HD-087, namely t1pks-nrps and t3pks-lantipeptide-t1pks-nrps. The key gene pksL in the t3pks-lantipeptide-t1pks-nrps gene cluster was predicted. The results suggested that it encodes a stable, hydrophilic, and acidic protein, mainly composed of α-helix and random coil. The PksL protein contains dehydrogenase (DH), ketone reductase (KR), acyl carrier protein (ACP), and ketone synthase (KS) domains. Moreover, it can form interaction networks with 11 proteins containing domains, such as polyketide synthase and ACP synthase. The molecular docking between PksL and acetyl-CoA is stable and strong, suggesting that PksL protein could catalyze the synthesis of polyketides with CoA as a substrate. This study provides a theoretical basis for further exploring the polyketides synthesis mechanism and developing antifungal metabolites in S. bikiniensis HD-087.

摘要

生酮基古龙沙链霉菌 HD-087 能够合成各种抗菌物质,以抵御有害微生物的不利影响。为了阐明其是否产生聚酮类抗生素以及抗生素物质的合成机制,通过 LC-MS、抗稻瘟霉活性检测和生物信息学方法分析了 S. bikiniensis HD-087 的代谢物和相关基因。结果表明,该菌株能够产生红霉素,一种聚酮类抗生素。菌株 HD-087 发酵上清液和红霉素提取物甲醇溶液对稻瘟霉的抑菌圈分别为 40.84±0.68mm 和 33.18±0.81mm。红霉素提取物对孢子萌发的 IC50 值为 220.43μg/mL。在 HD-087 基因组中存在两个聚酮合成基因簇,即 t1pks-nrps 和 t3pks-lantipeptide-t1pks-nrps。预测了 t3pks-lantipeptide-t1pks-nrps 基因簇中的关键基因 pksL。结果表明,它编码一种稳定、亲水、酸性的蛋白质,主要由α-螺旋和无规卷曲组成。PksL 蛋白含有脱氢酶(DH)、酮还原酶(KR)、酰基载体蛋白(ACP)和酮合酶(KS)结构域。此外,它可以与包含结构域的 11 种蛋白质形成相互作用网络,如聚酮合酶和 ACP 合酶。PksL 和乙酰辅酶 A 之间的分子对接稳定且牢固,表明 PksL 蛋白可以催化以 CoA 为底物的聚酮类化合物的合成。本研究为进一步探索生酮古龙沙链霉菌 HD-087 中聚酮类化合物的合成机制和开发抗真菌代谢物提供了理论依据。

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