西索米星是一种氨基糖苷类广谱抗生素,具有广阔的应用前景。目前,因缺少理性指导,提高西索米星的微生物发酵水平仍面临困难。该研究以课题组前期选育的西索米星高产菌株和出发菌株为研究对象,基于其发酵特性差异,利用非靶向代谢组学技术对其代谢差异进行分析。KEGG功能通路分析发现,绝大部分代谢物属于代谢类,最多的是氨基酸代谢,其次是脂质代谢和次级代谢物的生物合成以及辅因子类的代谢。进一步分析发现,2株菌在不同发酵时期的代谢物差异显著,中心代谢途径中的主要差异代谢物为黄素单核苷酸;氨基酸代谢途径中的主要差异代谢物为赖氨酸和组氨酸;与西索米星合成途径相关的主要差异代谢物N-乙酰氨基葡萄糖的丰度显著下调。基于上述分析结果,发现赖氨酸的添加强化了西索米星的生物合成,较不添加时提高了25.8%。该研究为基于代谢调控的西索米星生产菌株选育及发酵优化提供了指导和参考。
Sisomicin is a kind of aminoglycoside broad-spectrum antibiotic, which has broad application prospect.At present, it is still difficult to improve the microbial fermentation level of sisomicin due to the lack of rational guidance.In this study, the sisomicin high-yielding strain and the original strain selected by the research group were selected as the research object.Based on the differences in their fermentation characteristics, the untargeted metabolomics technology was used to analyze the metabolic differences.The KEGG functional pathway analysis found that most metabolites belonged to metabolism, and the most were amino acid metabolism, which was followed by lipid metabolism and biosynthesis of secondary metabolites and metabolism of cofactors.Further analysis showed that the metabolites of the two strains were significantly different in different stages of fermentation, and the main metabolite in the central metabolic pathway was flavin mononucleotide;The main metabolites in amino acid metabolism pathway were lysine and histidine;The abundance of N-acetylglucosamine, a major differential metabolite associated with the sisomicin synthesis pathway, was significantly down-regulated.Based on the above analysis results, it was found that the addition of lysine enhanced the biosynthesis of sisomicin, which increased by 25.8% compared with no addition.This study provided guidance and references for the breeding and fermentation optimization of sisomicin production strains based on metabolic regulation.
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