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  • Title: Biosynthesis of poly-γ-glutamic acid in Escherichia coli by heterologous expression of pgsBCAE operon from Bacillus.
    Author: Liu CL, Dong HG, Xue K, Yang W, Liu P, Cai D, Liu X, Yang Y, Bai Z.
    Journal: J Appl Microbiol; 2020 May; 128(5):1390-1399. PubMed ID: 31837088.
    Abstract:
    AIMS: Poly-γ-glutamic acid (γ-PGA) is an excellent water-soluble biosynthesis material. To confirm the rate-limiting steps of γ-PGA biosynthesis pathway, we introduced a heterologous Bacillus strain pathway and employed an enzyme-modulated dismemberment strategy in Escherichia coli. METHODS AND RESULTS: In this study, we heterologously introduced the γ-PGA biosynthesis pathway of two laboratory-preserved strains-Bacillus amyloliquefaciens FZB42 and Bacillus subtilis 168 into E. coli, and compared their γ-PGA production levels. Next, by changing the plasmid copy numbers and supplying sodium glutamate, we explored the effects of gene expression levels and concentrations of the substrate l-glutamic acid on γ-PGA production. We finally employed a two-plasmid induction system using an enzyme-modulated dismemberment of pgsBCAE operon to confirm the rate-limiting genes of the γ-PGA biosynthesis pathway. CONCLUSION: Through heterologously over-expressing the genes of the γ-PGA biosynthesis pathway and exploring gene expression levels, we produced 0·77 g l-1 γ-PGA in strain RSF-EBCAE(BS). We also confirmed that the rate-limiting genes of the γ-PGA biosynthesis pathway were pgsB and pgsC. SIGNIFICANCE AND IMPACT OF THE STUDY: This work is beneficial to increase γ-PGA production and study the mechanism of γ-PGA biosynthesis enzymes.
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