Biotechnology Bulletin ›› 2014, Vol. 0 ›› Issue (4): 121-126.

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

Luo Xuan1, Xu Liyuan2, Wang Yongze2, Zhao Jinfang2, Zhao xiao2, Wang Jinhua2,   

  1. (1. College of Food Science and Biotechnology,Huazhong Agricultural University Chutian College,Wuhan 430205; 2. Key Laboratory of Fermentation Engineering(Ministry of Education),Hubei Provincial Cooperative Innovation Center of Industrial Fermentation,College of Bioengineering,Hubei University of Ttechnology,Wuhan 430068)
  • Received:2013-11-12 Online:2014-04-29 Published:2014-04-29

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