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Current Microbiology

, Volume 61, Issue 1, pp 7–12 | Cite as

Structural and Catalytic Properties of the D-3-Hydroxybutyrate Dehydrogenase from Pseudomonas aeruginosa

  • Driss Mountassif
  • Pierre Andreoletti
  • Mustapha Cherkaoui-Malki
  • Norbert LatruffeEmail author
  • M’hammed Saïd El Kebbaj
Article

Abstract

To put forward BDH from Pseudomonas aeruginosa’s enzymatic properties, we report a two-step purification of BDH and its gene sequencing allowing the investigation of its structural properties. Purification of BDH was achieved, using ammonium sulfate fractionation and Blue Sepharose CL-6B affinity chromatography. SDS–PAGE analysis reveals a MM of 29 kDa, whereas the native enzyme showed a MM of 120 kDa suggesting a homotetrameric structure. BDH encoding gene sequence shows a nucleotide open reading frame sequence of 771 bp encoding a 265 amino acid residues polypeptide chain. The modeling analysis of the three dimensional structure fits with the importance of amino acids in the catalysis reaction especially a strictly conserved tetrad. Amino-acid residues in interaction with the coenzyme NAD+ were also identified.

Keywords

Ammonium Sulfate Fractionation Rhodospirillum Rubrum Acidovorax Paracoccus Denitrificans Native Molecular Weight 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Notes

Acknowledgments

Supported by the Regional Council of Burgundy and IFR no. 100, the Programme Thématique d’Appui à la Recherche Scientifique-Morocco, Biologie No.134, and the AI franco-marocaine MA/05/134.

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Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  • Driss Mountassif
    • 1
    • 2
  • Pierre Andreoletti
    • 1
  • Mustapha Cherkaoui-Malki
    • 1
  • Norbert Latruffe
    • 1
    Email author
  • M’hammed Saïd El Kebbaj
    • 2
  1. 1.INSERM U866; Université de Bourgogne; LBMC (Biochimie Métabolique et Nutritionnelle)Faculté des SciencesDijon cedexFrance
  2. 2.Laboratoire de Biochimie et Biologie Moléculaire, Université Hassan II - Aïn ChockFaculté des Sciences Aïn ChockMaarif, CasablancaMorocco

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