枯草芽胞杆菌168不对称转化产生磷霉素的蛋白质组学分析

发布时间：2018-04-01 22:32

本文选题：枯草芽胞杆菌　切入点：顺丙烯磷酸　出处：《生物工程学报》2013年06期

【摘要】：旨在用蛋白质组学方法揭示枯草芽胞杆菌Bacillus subtilis 168将顺丙烯磷酸转化成磷霉素的机理。B.subtilis 168能够将顺丙烯磷酸不对称转化成磷霉素。气相色谱分析发现在转化培养基发酵液中的磷霉素的含量达816.6μg/mL,转化率为36.05%。将分别培养在含有底物和不含底物的培养基中的B.subtilis 168的胞质蛋白进行双向凝胶电泳。对两种条件下的电泳图谱进行比较,发现有98个差异表达蛋白。其中在有底物存在时,表达量下调的点有20个,表达量上调的点52个,底物特异性表达的点有26个。对差异表达蛋白进行质谱鉴定,共鉴定到80个蛋白点,其中下调的点17个,上调的点45个,底物特异性表达的点18个。这些蛋白分别参与胁迫反应、氧化还原反应、物质转运、核苷酸代谢、糖代谢、氨基酸和蛋白质代谢等。根据上述对B.subtilis 168蛋白质组学分析结果,推测菌株是通过两步将顺丙烯磷酸转化成磷霉素的。第一步是水化反应,第二步是脱氢反应。
[Abstract]:The aim of this study was to reveal the mechanism of conversion of cispropene phosphoric acid to fosfomycin by Bacillus subtilis Bacillus subtilis 168 by proteomics. B. subtilis 168 was able to convert propene phosphoric acid asymmetrically to phosphomycin. The content of fosfomycin in fermentation broth was 816.6 渭 g / mL and the conversion rate was 36.05.The cytoplasmic proteins of B.subtilis 168 cultured in medium containing substrate and without substrate were analyzed by two-dimensional gel electrophoresis. 98 differentially expressed proteins were found, of which 20 were down-regulated, 52 were up-regulated and 26 were substrate-specific. Differential expression proteins were identified by mass spectrometry. A total of 80 protein spots were identified, of which 17 were down-regulated, 45 up-regulated and 18 substrate-specific. These proteins were involved in stress response, redox reaction, substance transport, nucleotide metabolism and glucose metabolism, respectively. Amino acid and protein metabolism, etc. According to the results of proteomic analysis of B.subtilis 168, it is speculated that the strain transformed cispropene phosphoric acid into phosphomycin through two steps. The first step is hydration reaction, the second step is dehydrogenation reaction.
【作者单位】：中国科学院微生物研究所
【基金】：中国科学院知识创新工程重要方向项目(No.KSCX2-YW-G-051)资助~~
【分类号】：R37

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