State Key Laboratory of Microbial Metabolism, School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai 200240, China; Department of Chemical and Biological Engineering, The State University of New York at Buffalo, Buffalo, NY 14260, USA.
Metab Eng. 2013 Sep;19:42-9. doi: 10.1016/j.ymben.2013.05.005. Epub 2013 Jun 5.
The heterologous production of the complex antibiotic erythromycin through Escherichia coli provides a unique challenge in metabolic engineering. In addition to introducing the 19 foreign genes needed for heterologous biosynthesis, E. coli metabolism must be engineered to provide the propionyl-CoA and (2S)-methylmalonyl-CoA substrates required to allow erythromycin formation. In this work, three different pathways to propionyl-CoA were compared in the context of supporting E. coli erythromycin biosynthesis. The comparison revealed that alternative citramalate and threonine metabolic pathways (both starting from exogenous glycerol) were capable of supporting final compound formation equal to a proven pathway reliant upon exogenous propionate. Furthermore, two pathways to (2S)-methylmalonyl-CoA were compared in the production of a novel benzyl-erythromycin analog. A pathway dependent upon exogenous methylmalonate improved selectivity and facilitated antibiotic assessment of this new analog.
通过大肠杆菌异源生产复杂抗生素红霉素在代谢工程方面提出了一个独特的挑战。除了引入 19 个用于异源生物合成的外源基因外，还必须对大肠杆菌的代谢进行工程改造，以提供用于红霉素形成的丙酰辅酶 A 和（2S）-甲基丙二酰辅酶 A 底物。在这项工作中，在支持大肠杆菌红霉素生物合成的背景下比较了三种不同的丙酰辅酶 A 途径。比较结果表明，替代的柠檬酸和苏氨酸代谢途径（均从外源甘油开始）能够支持最终化合物的形成，与依赖外源丙酸盐的已有途径相当。此外，在新型苄基-红霉素类似物的生产中比较了两种（2S）-甲基丙二酰辅酶 A 途径。依赖于外源甲基丙二酸的途径提高了选择性，并促进了对这种新类似物的抗生素评估。
