Carbapenems, a family of β-lactam antibiotics, are among the most powerful bactericidal compounds in clinical use. However, as rational engineering of native carbapenem-producing microbes is not currently possible, the present carbapenem supply relies upon total chemical synthesis of artificial carbapenem derivatives. To enable access to the full diversity of natural carbapenems, we have engineered production of a simple carbapenem antibiotic within Escherichia coli. By increasing concentrations of precursor metabolites and identifying a reducing cofactor of a bottleneck enzyme, we improved productivity by 60-fold over the minimal pathway and surpassed reported titers obtained from carbapenem-producing Streptomyces species. We stabilized E. coli metabolism against antibacterial effects of the carbapenem product by artificially inhibiting membrane synthesis, which further increased antibiotic productivity. As all known naturally occurring carbapenems are derived from a common intermediate, our engineered strain provides a platform for biosynthesis of tailored carbapenem derivatives in a genetically tractable and fast-growing species.

碳青霉烯是β-内酰胺类抗生素，在临床上是最有效的杀菌化合物之一。但是，由于目前尚无法对天然产碳青霉烯的微生物进行合理的工程设计，因此目前的碳青霉烯供应依赖于人工合成碳青霉烯衍生物的全部化学合成。为了能够获得天然碳青霉烯的全部多样性，我们在大肠杆菌中设计了一种简单的碳青霉烯类抗生素的生产方法。通过增加前体代谢物的浓度并确定瓶颈酶的还原辅助因子，我们将生产率提高了超过最小途径的60倍，并超过了从生产碳青霉烯的链霉菌种获得的滴度。我们稳定了大肠杆菌通过人为抑制膜合成来抑制碳青霉烯产物的抗微生物代谢作用，从而进一步提高了抗生素的生产率。由于所有已知的天然碳青霉烯类化合物均来自一个常见的中间体，因此我们的工程菌株为在遗传上易于控制且快速生长的物种中生物合成量身定制的碳青霉烯类化合物提供了平台。