The Influence of Knockout of menA Gene in Escherichia coli on the Accumulation of CoQ

doi:10.13560/j.cnki.biotech.bull.1985.2015.12.030

Abstract

Abstract:

Knocking out menA gene of Escherichia coli by homologous recombination increases the synthetized amount of CoQ, which is used for constructing the strains of high-yield CoQ. Using pKD4 plasmid as template, the kan^r fragments were amplified by PCR. In the presence of auxiliary plasmid pKD46, the kan^r fragments were transformed into Escherichia coli, and the recombinant one was confirmed by antibiotic screening and verification of PCR；By contrast with uv-induced mutation, the strains with menA gene knocked out were fermented, and the types of CoQ and the changes of CoQ yield were analyzed. The results showed that the strains with menA gene knockout were successfully obtained, while the types of CoQ unchanged and the yield increased about 38%. In conclusion, this is the first time of knocking out menA gene, CoQ production of mutant strains is improved, and the desired goal is achieved, which lays a foundation for the further construction of high-yield CoQ strains.

Key words: Escherichia coli, gene knockout, menA genes, CoQ, homologous recombination

Liu Yongqing, Ren Lin, Zhang Zifeng. The Influence of Knockout of menA Gene in Escherichia coli on the Accumulation of CoQ[J]. Biotechnology Bulletin, 2015, 31(12): 207-213.

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