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红球菌属菌株YYL在四氢呋喃降解过程中的动态代谢和转录谱分析

Dynamic metabolic and transcriptional profiling of Rhodococcus sp. strain YYL during the degradation of tetrahydrofuran.

作者信息

He Zhixing, Yao Yanlai, Lu Zhenmei, Ye Yangfang

机构信息

College of Life Sciences, Zhejiang University, Hangzhou, China.

出版信息

Appl Environ Microbiol. 2014 May;80(9):2656-64. doi: 10.1128/AEM.04131-13. Epub 2014 Feb 14.

Abstract

Although tetrahydrofuran-degrading Rhodococcus sp. strain YYL possesses tetrahydrofuran (THF) degradation genes similar to those of other tetrahydrofuran-degrading bacteria, a much higher degradation efficiency has been observed in strain YYL. In this study, nuclear magnetic resonance (NMR)-based metabolomics analyses were performed to explore the metabolic profiling response of strain YYL to exposure to THF. Exposure to THF slightly influenced the metabolome of strain YYL when yeast extract was present in the medium. The metabolic profile of strain YYL over time was also investigated using THF as the sole carbon source to identify the metabolites associated with high-efficiency THF degradation. Lactate, alanine, glutarate, glutamate, glutamine, succinate, lysine, trehalose, trimethylamine-N-oxide (TMAO), NAD(+), and CTP were significantly altered over time in strain YYL grown in 20 mM THF. Real-time quantitative PCR (RT-qPCR) revealed changes in the transcriptional expression levels of 15 genes involved in THF degradation, suggesting that strain YYL could accumulate several disturbances in osmoregulation (trehalose, glutamate, glutamine, etc.), with reduced glycolysis levels, an accelerated tricarboxylic acid cycle, and enhanced protein synthesis. The findings obtained through (1)H NMR metabolomics analyses and the transcriptional expression of the corresponding genes are complementary for exploring the dynamic metabolic profile in organisms.

摘要

尽管降解四氢呋喃的红球菌属菌株YYL拥有与其他降解四氢呋喃细菌相似的四氢呋喃(THF)降解基因,但在菌株YYL中观察到了更高的降解效率。在本研究中,进行了基于核磁共振(NMR)的代谢组学分析,以探索菌株YYL暴露于THF后的代谢谱响应。当培养基中存在酵母提取物时,暴露于THF对菌株YYL的代谢组有轻微影响。还以THF作为唯一碳源研究了菌株YYL随时间的代谢谱,以鉴定与高效THF降解相关的代谢物。在以20 mM THF培养的菌株YYL中,乳酸、丙氨酸、戊二酸、谷氨酸、谷氨酰胺、琥珀酸、赖氨酸、海藻糖、氧化三甲胺(TMAO)、NAD(+)和CTP随时间显著变化。实时定量PCR(RT-qPCR)揭示了15个参与THF降解的基因转录表达水平的变化,表明菌株YYL可能在渗透调节(海藻糖、谷氨酸、谷氨酰胺等)方面积累了一些干扰,糖酵解水平降低,三羧酸循环加速,蛋白质合成增强。通过¹H NMR代谢组学分析和相应基因的转录表达获得的结果在探索生物体动态代谢谱方面具有互补性。

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