A Multi-Enzyme Cascade for Efficient Production of Pyrrolidone from l-Glutamate.

ABSTRACT: Pyrrolidone is a high value-added monomer and an important active drug intermediate. However, the efficient enzymatic synthesis of pyrrolidone remains a challenge. Here, we developed and reconstructed a three-enzyme cascade pathway using Escherichia coli BL21(DE3) for the production of pyrrolidone from l-glutamate (l-Glu). The carnitine-CoA ligase from Escherichia coli (EcCaiC) at a low expression level and with a low activity is regarded as the rate-limiting enzyme. Here, we obtained the best EcCaiC^F380M/N430D double mutant with a k_cat/K_m value 1.5 times higher than that of the wild type via mechanism-based protein engineering. For this, we (i) eliminated the steric hindrance of the loop ring to improve the precatalytic conforma

SUBMITTER: Jiang X 

PROVIDER: S-EPMC10132116 | biostudies-literature | 2023 Apr

REPOSITORIES: biostudies-literature