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Applied Biochemistry and Biotechnology

, Volume 158, Issue 2, pp 343–351 | Cite as

Cloning, Protein Sequence Clarification, and Substrate Specificity of a Leucine Dehydrogenase from Bacillus sphaericus ATCC4525

  • Hongmei Li
  • Dunming ZhuEmail author
  • Brooke A. Hyatt
  • Fahad M. Malik
  • Edward R. Biehl
  • Ling Hua
Article

Abstract

Although an X-ray model sequence of a leucine dehydrogenase from Bacillus sphaericus ATCC4525 was reported, the amino acid sequence of this enzyme has not been confirmed. In the current study, this leucine dehydrogenase gene was cloned, sequenced, and over-expressed in Escherichia coli, and the protein sequence has been clarified. This leucine dehydrogenase is not identical with that of B. sphaericus IFO3525 because there are 16 different amino acid residues between these two proteins. Since the information on the catalytic properties of leucine dehydrogenase from B. sphaericus ATCC4525 has been limited, the recombinant enzyme was purified as His-tagged protein and further studied. This enzyme showed activity toward aliphatic substrates for both oxidative deamination and reductive amination and is an effective catalyst for the asymmetric synthesis of α-amino acids from the corresponding α-ketoacids.

Keywords

Leucine dehydrogenase Gene cloning Bacillus sphaericus Reductive amination Synthesis of α-amino acids 

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Copyright information

© Humana Press 2008

Authors and Affiliations

  • Hongmei Li
    • 1
  • Dunming Zhu
    • 1
    • 2
    Email author
  • Brooke A. Hyatt
    • 1
  • Fahad M. Malik
    • 1
  • Edward R. Biehl
    • 1
  • Ling Hua
    • 1
    • 3
  1. 1.Department of ChemistrySouthern Methodist UniversityDallasUSA
  2. 2.Tianjin Industrial Biotechnology Research and Development CenterChinese Academy of SciencesTianjinChina
  3. 3.China Research CenterGenencor InternationalShanghaiChina

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