Cloning L-lactate Dehydrogenase Gene from Pediococcus acidilactici and Overexpression of It in Recombination Strain E.coli JH12

Abstract

Abstract: Gene ldhL encoding L-lactate dehydrogenase was amplified by PCR technique using genome DNA of Escherichia coli SZ85 as temple. The PCR product was cloned into pUcm-T vector and double digested with restriction endonucleases, and then the DNA fragment of ldhL was inserted into pET-28a(+). The recombinants expression plasmid pET-ldhL was transformed into high purity L-lactate production of E. coli. JH12, which inserted ldhL of P. acidilactici into chromosome. The overexpression system of ldhL of Pediococcus acidilactici gene E. coli JH12(pET-28a-ldhL)could product L-lactic acid using 7% xylose. Overexpression of ldhL led to L-lactic acid yield of 64.86 g/L, which is 10 g/L higher than control without Overexpression of ldhL. Meanwhile, high sugar-acid ratio of 96% was achieved by overexpression of ldhL.

Key words: Recombination, E.coli, Overexpression, L-lactic acid

Luo Xuan, Xu Liyuan, Wang Yongze, Zhao Jinfang, Zhao xiao, Wang Jinhua,. Cloning L-lactate Dehydrogenase Gene from Pediococcus acidilactici and Overexpression of It in Recombination Strain E.coli JH12[J]. Biotechnology Bulletin, 2014, 0(4): 121-126.

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