Jayaraman Premkumar, Holowko Maciej B, Yeoh Jing Wui, Lim Sierin, Poh Chueh Loo
Department of Biomedical Engineering, Faculty of Engineering, National University of Singapore , Singapore.
NUS Synthetic Biology for Clinical and Technological Innovation (SynCTI), Life Sciences Institute, National University of Singapore , Singapore.
ACS Synth Biol. 2017 Jul 21;6(7):1403-1415. doi: 10.1021/acssynbio.7b00058. Epub 2017 May 5.
New strategies to control cholera are urgently needed. This study develops an in vitro proof-of-concept sense-and-kill system in a wild-type Escherichia coli strain to target the causative pathogen Vibrio cholerae using a synthetic biology approach. Our engineered E. coli specifically detects V. cholerae via its quorum-sensing molecule CAI-1 and responds by expressing the lysis protein YebF-Art-085, thereby self-lysing to release the killing protein Art-085 to kill V. cholerae. For this report, we individually characterized YebF-Art-085 and Art-085 expression and their activities when coupled to our previously developed V. cholerae biosensing circuit. We show that, in the presence of V. cholerae supernatant, the final integrated sense-and-kill system in our engineered E. coli can effectively inhibit the growth of V. cholerae cells. This work represents the first step toward a novel probiotic treatment modality that could potentially prevent and treat cholera in the future.
迫切需要控制霍乱的新策略。本研究利用合成生物学方法，在野生型大肠杆菌菌株中开发了一种体外概念验证的感知并杀灭系统，以靶向致病性病原菌霍乱弧菌。我们构建的大肠杆菌通过群体感应分子CAI-1特异性检测霍乱弧菌，并通过表达裂解蛋白YebF-Art-085做出反应，从而自我裂解以释放杀伤蛋白Art-085来杀死霍乱弧菌。在本报告中，我们分别对YebF-Art-085和Art-085的表达及其与我们之前开发的霍乱弧菌生物传感电路偶联时的活性进行了表征。我们表明，在霍乱弧菌上清液存在的情况下，我们构建的大肠杆菌中的最终整合感知并杀灭系统能够有效抑制霍乱弧菌细胞的生长。这项工作代表了朝着一种新型益生菌治疗方式迈出的第一步，这种方式未来有可能预防和治疗霍乱。
