Department of Chemical and Biochemical Engineering, College of Chemistry and Chemical Engineering, Xiamen University, Xiamen 361005, China.
Department of Chemical and Biochemical Engineering, College of Chemistry and Chemical Engineering, Xiamen University, Xiamen 361005, China; The Key Laboratory for Synthetic Biotechnology of Xiamen City, Xiamen University, Xiamen 361005, China.
J Biosci Bioeng. 2014 May;117(5):591-7. doi: 10.1016/j.jbiosc.2013.10.021. Epub 2013 Dec 4.
This first-attempt study provided liquid chromatography tandem mass (LC-MS/MS) proteomics approach to explore precursor effects on daptomycin synthesis from Streptomyces roseosporus NRRL 11379. Among all, 357 and 691 differential proteins from 601 proteins in precursor group (144 h+) and 935 proteins in non-precursor group (144 h-) were identified, respectively. Through the simulation of the 2D-protein mapping, most proteins were found in isoelectric points ranged of 4.5-10.0 as well as Mws ranged 10-100 kDa. As a result, LC-MS/MS analysis was consistence with the analytical results of two-dimensional electrophoresis (2DE) but provided much intact profiles of proteins by precursor effect on S. roseosporus. To have more insight exploration, differential proteins associated to Streptomyces spp. were defined into 14 groups of their functional classification. The major differential proteins were in transport/membrane functional group with an occupation of 12.4% for 144 h+ and 5.2% for 144 h-, respectively. LC-MS/MS results as a direct proteomic mapping approach reveal more daptomycin synthetic and regulation-related proteins from precursor group in terms of methyltransferase, ATP-binding cassette (ABC) transporters, resistance proteins and regulators.
本首次尝试性研究提供了液质联用（LC-MS/MS）蛋白质组学方法，以探讨前体对玫瑰孢链霉菌 NRRL 11379 合成达托霉素的影响。在所有前体组（144 h+）的 601 种蛋白和非前体组（144 h-）的 935 种蛋白中，分别鉴定出 357 种和 691 种差异蛋白。通过二维蛋白图谱模拟，大多数蛋白的等电点在 4.5-10.0 之间，分子量（Mw）在 10-100 kDa 之间。因此，LC-MS/MS 分析与二维电泳（2DE）的分析结果一致，但通过前体对玫瑰孢链霉菌的影响，提供了更完整的蛋白图谱。为了进行更深入的探索，将与链霉菌属相关的差异蛋白定义为 14 个功能分类组。主要差异蛋白存在于运输/膜功能组，144 h+的占有率为 12.4%，144 h-的占有率为 5.2%。LC-MS/MS 结果作为一种直接的蛋白质组学图谱方法，在前体组中揭示了更多与达托霉素合成和调节相关的蛋白，包括甲基转移酶、ATP 结合盒（ABC）转运蛋白、抗性蛋白和调节蛋白。
