Regulatory mechanism of C4-dicarboxylates in cyclo (Phe-Pro) production

文献类型： 外文期刊

第一作者： Xu, Xinyan

作者： Xu, Xinyan;Liu, Liu;Zhang, Yang;Hafeez, Rahila;Ijaz, Munazza;Ahmed, Temoor;Li, Bin;Xu, Lihui;Ali, Hayssam M.;Shahid, Muhammad Shafiq;Ahmed, Temoor;Ahmed, Temoor;Ondrasek, Gabrijel

作者机构：

关键词： Dct system; Burkholderia seminalis; Anti-fungus; C4-dicarboxylates; Cyclo (Phe-Pro)

期刊名称：MICROBIAL CELL FACTORIES （ 影响因子：4.9； 五年影响因子：6.1 ）

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年卷期： 2024 年 23 卷 1 期

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收录情况： SCI

摘要： Cyclo (Phe-Pro) (cFP), a cyclic dipeptide with notable antifungal, antibacterial, and antiviral properties, shows great promise for biological control of plant diseases. Produced as a byproduct by non-ribosomal peptide synthetases (NRPS), the regulatory mechanism of cFP biosynthesis remains unclear. In a screening test of 997 Tn5 mutants of Burkholderia seminalis strain R456, we identified eight mutants with enhanced antagonistic effects against Fusarium graminearum (Fg). Among these, mutant 88's culture filtrate contained cFP, confirmed through HPLC and LC-MS, which actively inhibited Fg. The gene disrupted in mutant 88 is part of the Dct transport system (Dct-A, -B, -D), responsible for C4-dicarboxylate transport. Knockout mutants of Dct genes exhibited higher cFP levels than the wild type, whereas complementary strains showed no significant difference. Additionally, the presence of exogenous C4-dicarboxylates reduced cFP production in wild type R456, indicating that these substrates negatively regulate cFP synthesis. Given that cFP synthesis is related to NRPS, we previously identified an NRPS cluster in R456, horizontally transferred from algae. Specifically, knocking out gene 2061 within this NRPS cluster significantly reduced cFP production. A Fur box binding site was predicted upstream of gene 2061, and yeast one-hybrid assays confirmed Fur protein binding, which increased with additional C4-dicarboxylates. Knockout of the Fur gene led to up-regulation of gene 2061 and increased cFP production, suggesting that C4-dicarboxylates suppress cFP synthesis by enhancing Fur-mediated repression of gene 2061.

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