Biotechnology Bulletin ›› 2014, Vol. 0 ›› Issue (9): 157-163.

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Cloning,Expression and Characterization of Gluconate 5-dehydrogenase from Gluconobacter oxydans

Li Ling, Xu Lin,Wei Miao, Li Yan, Yan Ming   

  1. College of Biotechnology and Pharmaceutical Engineering,Nanjing Tech University,Nanjing 211816
  • Received:2014-02-08 Online:2014-09-15 Published:2014-09-07

Abstract: Guconate 5-dehydrogenase gene from Gluconobacter oxydans H24 was amplified with primes designed based on NCBI database published sequence information. Ga5DH was cloned and expressed in Escherichia coli Rosetta strain. Results showed that the molecular mass of the enzyme was 26.5 kD by SDS-PAGE gel analysis. After purification, the enzyme activity could reach 7.83 U/mg. It was suggested that the recombinant protein had a maximum activity at 40℃, pH 11. The characterization of purified enzyme indicated that Ga5DH was highly resistant to organic solvents.

Key words: Gluconate 5-dehydrogenase, Gluconobacter oxydans, Expression, Enzymatic properties