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Applied Microbiology and Biotechnology

, Volume 80, Issue 4, pp 597–604 | Cite as

Cloning and expression of the l-1-amino-2-propanol dehydrogenase gene from Rhodococcus erythropolis, and its application to double chiral compound production

  • M. KataokaEmail author
  • T. Ishige
  • N. Urano
  • Y. Nakamura
  • E. Sakuradani
  • S. Fukui
  • S. Kita
  • K. Sakamoto
  • S. Shimizu
Biotechnologically Relevant Enzymes and Proteins

Abstract

The gene encoding NADP+-dependent l-1-amino-2-propanol dehydrogenase (AADH) of Rhodococcus erythropolis MAK154 was cloned and sequenced. A 780-bp nucleotide fragment was confirmed to be the gene encoding AADH by agreement of the N-terminal and internal amino acid sequences of the purified AADH. The gene (aadh) codes a total of 259 amino acid residues, and the deduced amino acid sequence shows similarity to several short-chain dehydrogenase/reductase family proteins. An expression vector, pKKAADH, which contains the full length aadh was constructed. Escherichia coli cells possessing pKKAADH exhibited a 10.4-fold increase in specific activity as to catalysis of the reduction of (S)-1-phenyl-2-methylaminopropan-1-one (MAK), as compared with that of R. erythropolis MAK154 induced by 1-amino-2-propanol (1 mg/ml). Coexpression of aadh with a cofactor regeneration enzyme (glucose dehydrogenase) gene was also performed, and a system for sufficient production of d-pseudoephedrine from racemic MAK was constructed.

Keywords

Rhodococcus erythropolis Asymmetric reduction l-1-amino-2-propanol dehydrogenase Ephedrine 

Notes

Acknowledgements

This work was supported in part by a Grant-in-Aid for Scientific Research from the Japan Society for the Promotion of Science. This work was carried out as a part of the Project for the “Development of a Technological Infrastructure for Industrial Bioprocesses on R&D of New Industrial Science and Technology Frontiers” of the Ministry of Economy, Trade & Industry (METI), and of the New Energy and Industrial Technology Development Organization (NEDO).

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Copyright information

© Springer-Verlag 2008

Authors and Affiliations

  • M. Kataoka
    • 1
    Email author
  • T. Ishige
    • 1
  • N. Urano
    • 1
  • Y. Nakamura
    • 1
  • E. Sakuradani
    • 1
  • S. Fukui
    • 1
  • S. Kita
    • 2
  • K. Sakamoto
    • 2
  • S. Shimizu
    • 1
  1. 1.Division of Applied Life Sciences, Graduate School of AgricultureKyoto UniversityKyotoJapan
  2. 2.Technical DepartmentDaiichi Fine Chemical Co.ToyamaJapan

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