Optimization of IspS_ib stability through directed evolution to improve isoprene production.

ABSTRACT: Enzyme stability is often a limiting factor in the microbial production of high-value-added chemicals and commercial enzymes. A previous study by our research group revealed that the unstable isoprene synthase from Ipomoea batatas (IspS_ib) critically limits isoprene production in engineered Escherichia coli. Directed evolution was, therefore, performed in the present study to improve the thermostability of IspS_ib. First, a tripartite protein folding system designated as lac'-IspS_ib-'lac, which could couple the stability of IspS_ib to antibiotic ampicillin resistance, was successfully constructed for the high-throughput screening of variants. Directed evolution of IspS_ib was then performed through two rounds of random mutat

SUBMITTER: Li M 

PROVIDER: S-EPMC10617563 | biostudies-literature | 2023 Oct

REPOSITORIES: biostudies-literature