Molecular Cloning and Gene Expression of Sinorhizobium meliloti Mannitol Dehydrogenase in Escherichia coli, and Its Enzymatic Characterization

Sinorhizobium meliloti 유래 Mannitol Dehydrogenase 유전자의 클로닝 및 대장균 내 발현과 효소특성 규명

  • Jang, Myoung-Uoon (Department of Food Science and Technology, Chungbuk National University) ;
  • Park, Jung-Mi (Department of Food Science and Technology, Chungbuk National University) ;
  • Kim, Min-Jeong (Department of Food Science and Technology, Chungbuk National University) ;
  • Lee, So-Won (Department of Food Science and Technology, Chungbuk National University) ;
  • Kang, Jung-Hyun (Department of Food Science and Technology, Chungbuk National University) ;
  • Kim, Tae-Jip (Department of Food Science and Technology, Chungbuk National University)
  • Received : 2013.02.06
  • Accepted : 2013.03.27
  • Published : 2013.06.28


A mannitol dehydrogenase (MDH; EC gene was cloned from the Sinorhizobium meliloti 1021 (KCTC 2353) genome and expressed in Escherichia coli. It was seen to have an open reading frame consisting of 1,485 bp encoding 494 amino acids (about 54 kDa), which shares approximately 35-55% of amino acid sequence identity with some known long-chain dehydrogenase/ reductase family enzymes. The recombinant S. meliloti MDH (SmMDH) showed the highest activity at $40^{\circ}C$, and pH 7.0 (D-fructose reduction) and pH 9.0 (D-mannitol oxidation), respectively. SmMDH could catalyze the oxidative/reductive reactions between D-mannitol and D-fructose in the presence of $NAD^+/NADH$ as a coenzyme, but not with NADP+/NADPH. These results indicate that SmMDH is a typical $NAD^+/NADH$-dependent mannitol dehydrogenase.


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