Cloning and functional expression of the D-beta-hydroxybutyrate dehydrogenase gene of Rhodobacter sp. DSMZ 12077.
Nucleotide sequence and biochemical analysis of D-beta-hydroxybutyrate dehydrogenase (EC 1.1.1.30), isolated from Rhodobacter sp., indicate functional oligomers composed of subunits of 257 amino acids with a calculated M(r) of 26,800 and a pI of 5.90. Compared to mammalian short-chain alcohol dehydrogenases, the bacterial enzyme lacks a C-terminal lipid anchor domain and was found to be highly active upon expression in Escherichia coli even without lipid supplement. The recombinant enzyme could be highly enriched using a single chromatography step and was shown to be stable over a broad range of pH and temperature.
Krüger
K
K
Roche Diagnostics GmbH, R&D Rare Reagents, D-82377 Penzberg, Germany.
Lang
G
G
Weidner
T
T
Engel
A M
AM
D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism; D001483:Base Sequence; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005798:Genes, Bacterial; D006801:Humans; D006884:Hydroxybutyrate Dehydrogenase / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D051381:Rats; D016992:Rhodobacter / Q000201:enzymology / Q000235:genetics*; D016415:Sequence Alignment
D001426:Bacterial Proteins; D006884:Hydroxybutyrate Dehydrogenase