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【目的】利用比较代谢组学的分析方法,研究不同发酵培养基中阿维链霉菌的胞内代谢差异,揭示合成阿维菌素的关键代谢物和代谢途径,再通过理性优化添加主要关键代谢物,提高阿维菌素产量。【方法】对M1和M2培养基中生长的菌体进行基于GC-MS的胞内代谢组学分析,通过理性添加强化前体代谢物,确定阿维菌素高产培养基。【结果】GC-MS共检测到232种物质,能够精确匹配70种胞内代谢物,通过PCA和PLS分析,最终确定了21种已知的胞内代谢物与阿维菌素的生物合成密切相关。其中乳酸、丙酮酸、琥珀酸、苏氨酸、异亮氨酸、缬氨酸和油脂类物质对阿维菌素的产量影响较为显著。通过单独或组合优化添加这些前体,阿维菌素的产量从5.36 g/L提高到了5.92 g/L,增加了10.4%。【结论】基于比较代谢组学分析的理性优化培养基的方法可有效提高阿维菌素的产量,并为提高当下生物基产品的产量提供了新思路。
【Objective】 The aim of the present study was to compare the intracellular metabolic differences of S. avermitilis in different fermentation media by using comparative metabolomics methods and to reveal the key metabolites and metabolic pathways of avermectin synthesis and to add the key metabolites through rational optimization Materials to improve avermectin production. 【Method】 The intracellular metabolomic analysis based on GC-MS was carried out on the mycelia grown on the medium of M1 and M2, and the precursor metabolites of the precursor were determined by rational addition to determine the high-yield avermectin medium. 【Result】 A total of 232 substances were detected by GC-MS, which can exactly match 70 kinds of intracellular metabolites. Through the analysis of PCA and PLS, the biosynthesis of 21 known intracellular metabolites and abamectin was finally confirmed Related. Among them, lactic acid, pyruvic acid, succinic acid, threonine, isoleucine, valine and oil substances have a significant impact on the yield of abamectin. By adding these precursors individually or in combination, the yield of avermectin increased from 5.36 g / L to 5.92 g / L, an increase of 10.4%. 【Conclusion】 The method of rational optimization of media based on comparative metabolomics analysis can effectively increase the yield of abamectin and provide a new idea for improving the yield of current bio-based products.