Department of Bioengineering, Rice University, Houston, TX, 77005, USA.
Technology Holding LLC, West Valley City, UT, 84119, USA.
J Ind Microbiol Biotechnol. 2020 Aug;47(8):609-620. doi: 10.1007/s10295-020-02303-6. Epub 2020 Aug 26.
This study addressed the functionality of genetic circuits carrying natural regulatory elements of Clostridium acetobutylicum ATCC 824 in the presence of the respective inducer molecules. Specifically, promoters and their regulators involved in diverse carbon source utilization were characterized using mCherryOpt or beta-galactosidase as a reporter. Consequently, most of the genetic circuits tested in this study were functional in Clostridium acetobutylicum ATCC 824 in the presence of an inducer, leading to the expression of reporter proteins. These genetic sensor-regulators were found to be transferable to another Clostridium species, such as Clostridium beijerinckii NCIMB 8052. The gradual expression of reporter protein was observed as a function of the carbohydrates of interest. A xylose-inducible promoter allows a titratable and robust expression of a reporter protein with stringency and efficacy. This xylose-inducible circuit was seen to enable induction of the expression of reporter proteins in the presence of actual sugar mixtures incorporated in woody hydrolysate wherein glucose and xylose are present as predominant carbon sources.
本研究探讨了在存在相应诱导分子的情况下，携带天然调控元件的 Clostridium acetobutylicum ATCC 824 基因回路的功能。具体来说，使用 mCherryOpt 或β-半乳糖苷酶作为报告基因，对涉及多种碳源利用的启动子及其调控因子进行了表征。因此，本研究中测试的大多数基因回路在存在诱导物的情况下在 Clostridium acetobutylicum ATCC 824 中均具有功能，导致报告蛋白的表达。这些遗传传感器-调节剂被发现可转移到另一种梭菌属物种，如 Clostridium beijerinckii NCIMB 8052。作为感兴趣的碳水化合物的函数，观察到报告蛋白的逐渐表达。木糖诱导型启动子允许具有严格性和功效的报告蛋白的可滴定和稳健表达。该木糖诱导型回路可在含有实际糖混合物的木质水解物中诱导报告蛋白的表达，其中葡萄糖和木糖是主要的碳源。
