Pei Guangsheng, Sun Tao, Chen Shuo, Chen Lei, Zhang Weiwen
Laboratory of Synthetic Microbiology, School of Chemical Engineering & Technology, Tianjin University, Tianjin, 300072 People's Republic of China.
Key Laboratory of Systems Bioengineering, Ministry of Education of China, Tianjin, 300072 People's Republic of China.
Biotechnol Biofuels. 2017 Mar 7;10:57. doi: 10.1186/s13068-017-0743-y. eCollection 2017.
The unicellular model cyanobacterium sp. PCC 6803 is considered a promising microbial chassis for biofuel production. However, its low tolerance to biofuel toxicity limits its potential application. Although recent studies showed that bacterial small RNAs (sRNAs) play important roles in regulating cellular processes in response to various stresses, the role of sRNAs in resisting exogenous biofuels is yet to be determined.
Based on genome-wide sRNA sequencing combined with systematic analysis of previous transcriptomic and proteomic data under the same biofuel or environmental perturbations, we report the identification of 133 -encoded sRNA transcripts with high-resolution mapping of sRNAs in , including 23 novel sRNAs identified for the first time. In addition, according to quantitative expression analysis and sRNA regulatory network prediction, sRNAs potentially involved in biofuel tolerance were identified and functionally confirmed by constructing sRNA overexpression or suppression strains of . Notably, overexpression of sRNA Nc117 revealed an improved tolerance to ethanol and butanol, while suppression of Nc117 led to increased sensitivity.
The study provided the first comprehensive responses to exogenous biofuels at the sRNA level in and opens an avenue to engineering sRNA regulatory elements for improved biofuel tolerance in the cyanobacterium s.
单细胞模式蓝细菌集胞藻PCC 6803被认为是用于生物燃料生产的一种有前景的微生物底盘。然而,其对生物燃料毒性的低耐受性限制了其潜在应用。尽管最近的研究表明细菌小RNA(sRNA)在响应各种胁迫时调节细胞过程中发挥重要作用,但sRNA在抵抗外源生物燃料中的作用尚未确定。
基于全基因组sRNA测序,并结合在相同生物燃料或环境扰动下对先前转录组和蛋白质组数据的系统分析,我们报告了在集胞藻PCC 6803中鉴定出133个编码的sRNA转录本,并对sRNA进行了高分辨率定位,其中包括首次鉴定出的23个新型sRNA。此外,根据定量表达分析和sRNA调控网络预测,鉴定出可能参与生物燃料耐受性的sRNA,并通过构建集胞藻PCC 6803的sRNA过表达或抑制菌株对其功能进行了确认。值得注意的是,sRNA Nc117的过表达显示出对乙醇和丁醇耐受性的提高,而Nc117的抑制导致敏感性增加。
该研究首次在集胞藻PCC 6803中提供了sRNA水平对外源生物燃料的全面响应,并为工程改造sRNA调控元件以提高蓝细菌对生物燃料的耐受性开辟了一条途径。