Engineering Research Center of Agricultural Microbiology Technology, Ministry of Education, Heilongjiang University, Harbin, 150500, Heilongjiang, China.
Key Laboratory of Microbiology, College of Heilongjiang Province, School of Life Sciences, Heilongjiang University, Harbin, 150080, Heilongjiang, China.
Microb Cell Fact. 2022 Sep 10;21(1):187. doi: 10.1186/s12934-022-01913-2.
The lipopeptides produced by Streptomyces bikiniensis have a significant inhibitory effect on Magnaporthe oryzae, but the low yield limits its application. In this study, the anti-M. oryzae activity of the broth of S. bikiniensis HD-087 co-cultured with M. oryzae Guy11 mycelium has risen by 41.22% compared with pure culture, and under induction conditions of adding Guy11-inducer (cell-free supernatant of M. oryzae Guy11), the activity of strain HD-087 improved 61.76%. The result proved that the enhancement effect of Guy11 on the antimicrobial activity of HD-087 was mainly related to metabolites but mycelium cells. Under optimum induction conditions, NRPS gene expression levels of HD-087 were significantly increased by induction with Guy11-inducer, the biomass of HD-087 had no significant change, but crude extract of lipopeptide (CEL) production was 107.4% higher than pure culture, and TLC result under acid hydrolysis showed that the induced culture has one component more than pure culture. To clarify the regulation mechanism of improving lipopeptide production of HD-087 with Guy11-inducer, transcriptomic analysis was performed using RNAseq to compare the induced culture and pure culture. In the induced culture, 943 genes were up-regulated, while 590 genes were down-regulated in DEGs (differentially expressed genes). KEGG results showed that the expression of genes related to amino acid synthesis, fatty acid metabolism, TCA cycle and pyruvate metabolism pathway were significantly increased. The increased expression of genes related to these metabolic pathways provided sufficient precursors for lipopeptide synthesis. Accordingly, key enzyme genes responsible for the synthesis of lipopeptides Srf and NRPS was significantly increased. Quorum sensing related genes OppA and MppA were significantly up-regulated, and then ComP was activated and promoted lipopeptide synthesis. These results provided a scientific basis for using M. oryzae to induce the increase of the production of Streptomyces lipopeptides, and also laid a foundation for further exploring the co-culture mechanisms among different genera.
生酮糖脂肽是由泳泳短稳杆菌产生的,对稻瘟病菌具有显著的抑制作用,但由于产量低,限制了其应用。本研究中,泳泳短稳杆菌 HD-087 与稻瘟病菌 Guy11 菌丝共培养的发酵液对稻瘟病菌的抑菌活性比纯培养提高了 41.22%,在添加 Guy11 诱导剂(稻瘟病菌 Guy11 的无细胞上清液)的诱导条件下,菌株 HD-087 的活性提高了 61.76%。结果表明,Guy11 对 HD-087 抗菌活性的增强作用主要与代谢产物有关,而与菌丝细胞无关。在最佳诱导条件下,HD-087 的 NRPS 基因表达水平在 Guy11 诱导剂的诱导下显著增加,HD-087 的生物量没有明显变化,但脂肽(CEL)粗提物的产量比纯培养提高了 107.4%,酸水解 TLC 结果表明,诱导培养比纯培养多一个成分。为了阐明 Guy11 诱导剂提高 HD-087 脂肽产量的调控机制,采用 RNAseq 进行转录组分析,比较诱导培养和纯培养。在诱导培养中,有 943 个基因上调,590 个基因下调(差异表达基因)。KEGG 结果表明,与氨基酸合成、脂肪酸代谢、TCA 循环和丙酮酸代谢途径相关的基因表达显著增加。这些代谢途径相关基因表达的增加为脂肽合成提供了足够的前体。因此,负责合成脂肽 Srf 和 NRPS 的关键酶基因显著增加。群体感应相关基因 OppA 和 MppA 显著上调,然后 ComP 被激活并促进脂肽合成。这些结果为利用稻瘟病菌诱导泳泳短稳杆菌脂肽产量增加提供了科学依据,也为进一步探索不同属间共培养机制奠定了基础。