Yang Gaohua, Jia Dechen, Jin Lin, Jiang Yuqian, Wang Yong, Jiang Weihong, Gu Yang
Key Laboratory of Synthetic Biology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences , Shanghai 200032, China.
Department of Biochemistry and Molecular Medicine, University of California at Davis , Sacramento, California 95817, United States.
ACS Synth Biol. 2017 Sep 15;6(9):1672-1678. doi: 10.1021/acssynbio.7b00155. Epub 2017 Jun 16.
Engineering solventogenic clostridia, a group of important industrial microorganisms, to realize their full potential in biorefinery application is still hindered by the absence of plentiful biological parts. Here, we developed an effective approach for rapid generation of a synthetic promoter library in solventogenic clostridia based on a dual-reporter system (catP-lacZ) and a widely used strong thl promoter. The yielded artificial promoters, spanning 2 orders of magnitude, comprised two modular components (the core promoter region and the spacer between RBS and the translation-initiating code), and the strongest promoter had an over 10-fold-higher activity than the original expression part P. The test of these synthetic promoters in controlled expression of sadh and danK in saccharolytic C. acetobutylicum and gas-fermenting C. ljungdahlii, respectively, gave the expected phenotypes, and moreover, showed good correlation between promoter activities and phenotypic changes. The presented wide-strength-range promoters here will be useful for synthetic biology application in solventogenic clostridia.
工程改造产溶剂梭菌（一类重要的工业微生物），以充分发挥它们在生物炼制应用中的潜力，仍然受到缺乏丰富生物元件的阻碍。在此，我们基于双报告系统（catP-lacZ）和广泛使用的强thl启动子，开发了一种在产溶剂梭菌中快速生成合成启动子文库的有效方法。产生的人工启动子跨度为2个数量级，由两个模块组件组成（核心启动子区域以及核糖体结合位点与翻译起始密码之间的间隔区），并且最强的启动子比原始表达元件P的活性高10倍以上。这些合成启动子分别在解糖丁酸梭菌中对sadh和在嗜糖嗜热栖热放线菌中对danK的可控表达测试中，产生了预期的表型，此外，还显示出启动子活性与表型变化之间具有良好的相关性。这里展示的具有广泛强度范围的启动子将有助于产溶剂梭菌在合成生物学中的应用。
