iTRAQ-BASED Proteomic Analysis of the Mechanism of Fructose on Improving Fengycin Biosynthesis in Bacillus Amyloliquefaciens.
iTRAQ-BASED Proteomic Analysis of the Mechanism of Fructose on Improving Fengycin Biosynthesis in Bacillus Amyloliquefaciens.
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基于 iTRAQ 的蛋白质组学分析果糖改善解淀粉芽孢杆菌风霉素生物合成的机制
DOI：
10.3390/molecules26206309
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发表时间：
2021-10-19
期刊：
影响因子：
--
通讯作者：
Zhao L
中科院分区：
文献类型：
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作者：
Lu H;Li R;Yang P;Luo W;Chen S;Bilal M;Xu H;Gu C;Liu S;Zhao Y;Geng C;Zhao L
丰霉素是枯草芽孢杆菌产生的一种脂肽，对黄曲霉、软腐菌等丝状真菌具有较强的抗氧化活性，在食品、农业、医药等领域具有广泛的应用前景。为了更好地阐明果糖对丰霉素生物合成的调控机制，我们利用itraq蛋白组学方法研究了解淀粉酵母菌fmb-60在ML（不含果糖）和MLF（含果糖）培养基中的差异表达蛋白。结果表明，共检测到811个蛋白，其中差异表达蛋白248个，其中上调162个（fold > 2），下调86个（fold < 0.5），大部分蛋白与细胞代谢、生物合成和生物调控过程有关。利用实时荧光定量PCR对目的基因的相对表达进行验证，验证蛋白质组学分析结果。根据蛋白质组学分析结果，果糖促进解淀粉芽孢杆菌fmb-60凤霉素生物合成的途径可归纳为改善代谢过程，包括细胞氨基酸和酰胺、脂肪酸生物合成、多肽和蛋白质、核苷酸和含核碱基化合物、药物/毒素、辅因子和维生素；肽/蛋白翻译、修饰、生物过程和对刺激的反应的强化。总之，本研究对果糖促进解淀粉芽孢杆菌合成凤霉素的机制进行了全面、系统的研究，为凤霉素或其同源物在丝状真菌防御中的潜在应用提供了思路。
Fengycin, as a lipopeptide produced by Bacillus subtilis, displays potent activity against filamentous fungi, including Aspergillus flavus and Soft-rot fungus, which exhibits a wide range of potential applications in food industries, agriculture, and medicine. To better clarify the regulatory mechanism of fructose on fengycin biosynthesis, the iTRAQ-based proteomic analysis was utilized to investigate the differentially expressed proteins of B. amyloliquefaciens fmb-60 cultivated in ML (without fructose) and MLF (with fructose) medium. The results indicated that a total of 811 proteins, including 248 proteins with differential expression levels (162 which were upregulated (fold > 2) and 86, which were downregulated (fold < 0.5) were detected, and most of the proteins are associated with cellular metabolism, biosynthesis, and biological regulation process. Moreover, the target genes’ relative expression was conducted using quantitative real-time PCR to validate the proteomic analysis results. Based on the results of proteome analysis, the supposed pathways of fructose enhancing fengycin biosynthesis in B. amyloliquefaciens fmb-60 can be summarized as improvement of the metabolic process, including cellular amino acid and amide, fatty acid biosynthesis, peptide and protein, nucleotide and nucleobase-containing compound, drug/toxin, cofactor, and vitamin; reinforcement of peptide/protein translation, modification, biological process, and response to a stimulus. In conclusion, this study represents a comprehensive and systematic investigation of the fructose mechanism on improving fengycin biosynthesis in B. amyloliquefaciens, which will provide a road map to facilitate the potential application of fengycin or its homolog in defending against filamentous fungi.