Fang Lei, Guell Marc, Church George M, Pfeifer Blaine A
Dept. of Chemical and Biological Engineering, University at Buffalo, The State University of New York, Buffalo, NY.
Dept. of Genetics and Biological and Biomedical Sciences Program, Harvard Medical School, Boston, Massachusetts and Wyss Institute for Biologically Inspired Engineering, Harvard University, Cambridge, MA.
Biotechnol Prog. 2018 Jan;34(1):271-276. doi: 10.1002/btpr.2567. Epub 2017 Oct 13.
The establishment of erythromycin production within the heterologous host E. coli marked an accomplishment in genetic transfer capacity. Namely, over 20 genes and 50 kb of DNA was introduced to E. coli for successful heterologous biosynthetic reconstitution. However, the prospect for production levels that approach those of the native host requires the application of engineering tools associated with E. coli. In this report, metabolic and genomic engineering were implemented to improve the E. coli cellular background and the plasmid platform supporting heterologous erythromycin formation. Results include improved plasmid stability and metabolic support for biosynthetic product formation. Specifically, the new plasmid design for erythromycin formation allowed for ≥89% stability relative to current standards (20% stability). In addition, the new strain (termed LF01) designed to improve carbon flow to the erythromycin biosynthetic pathway provided a 400% improvement in titer level. © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 34:271-276, 2018.
在异源宿主大肠杆菌中建立红霉素生产标志着基因转移能力方面的一项成就。也就是说，超过20个基因和50 kb的DNA被引入大肠杆菌以成功实现异源生物合成重构。然而，要达到天然宿主的生产水平，需要应用与大肠杆菌相关的工程工具。在本报告中，实施了代谢和基因组工程以改善大肠杆菌的细胞背景以及支持异源红霉素形成的质粒平台。结果包括提高了质粒稳定性和对生物合成产物形成的代谢支持。具体而言，用于红霉素形成的新质粒设计相对于当前标准（20%稳定性）实现了≥89%的稳定性。此外，为改善碳流向红霉素生物合成途径而设计的新菌株（称为LF01）使滴度水平提高了400%。© 2017美国化学工程师学会生物技术进展，34:271 - 276，2018。
