Ren Kang, Zhao Yiqing, Chen Guo-Qiang, Ao Xiang, Wu Qiong
MOE Key Laboratory of Bioinformatics, Center for Synthetic and Systems Biology, School of Life Sciences, Tsinghua University, Beijing 100084, China.
Beijing No.12 High School, Beijing 100071, China.
ACS Synth Biol. 2024 Jan 19;13(1):61-67. doi: 10.1021/acssynbio.3c00622. Epub 2023 Dec 15.
is a halophilic bacterium capable of efficiently producing polyhydroxyalkanoates and other valuable chemicals through high salinity open fermentation, offering an appealing platform for next-generation industrial biotechnology. Various techniques have been developed to engineer , each with its inherent shortcomings. Genome editing methods often entail complex and time-consuming processes, while flexible expression systems relying on plasmids necessitate the use of antibiotics. In this study, we developed a stable recombinant plasmid vector, pHbPBC, based on a novel / toxin-antitoxin system found within the endogenous plasmid of . Remarkably, pHbPBC exhibited exceptional stability during 7 days of continuous subculture, eliminating the need for antibiotics or other selection pressures. This stability even rivaled genomic integration, all while achieving higher levels of heterologous expression. Our research introduces a novel approach for genetically modifying and harnessing nonmodel halophilic bacteria, contributing to the advancement of next-generation industrial biotechnology.
是一种嗜盐细菌，能够通过高盐度开放式发酵高效生产聚羟基脂肪酸酯和其他有价值的化学品，为下一代工业生物技术提供了一个有吸引力的平台。已经开发了各种技术来改造，每种技术都有其固有的缺点。基因组编辑方法通常需要复杂且耗时的过程，而依赖质粒的灵活表达系统需要使用抗生素。在本研究中，我们基于在的内源质粒中发现的一种新型/毒素-抗毒素系统，开发了一种稳定的重组质粒载体pHbPBC。值得注意的是，pHbPBC在连续传代培养7天期间表现出非凡的稳定性，无需抗生素或其他选择压力。这种稳定性甚至可与基因组整合相媲美，同时实现了更高水平的异源表达。我们的研究引入了一种对非模式嗜盐细菌进行基因改造和利用的新方法，为下一代工业生物技术的发展做出了贡献。
