Structural and Catalytic Properties of the D-3-Hydroxybutyrate Dehydrogenase from Pseudomonas aeruginosa
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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.
KeywordsAmmonium Sulfate Fractionation Rhodospirillum Rubrum Acidovorax Paracoccus Denitrificans Native Molecular Weight
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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