Purification and characterization of pyridoxal 4-dehydrogenase from Aureobacterium luteolum.
A pyridoxal dehydrogenase was purified to homogeneity from Aureobacterium luteolum, which can use pyridoxine as a carbon and nitrogen source, and characterized. The enzyme was a dimeric protein with a subunit molecular weight of 38,000. It had several properties distinct from those of the partially purified enzyme from Pseudomonas MA-1. The optimum pH (8.0-8.5) was 0.8-1.3 lower than that of the Pseudomonas enzyme. The Aureobacterium enzyme showed much higher and lower affinities for NAD+ (Km, 0.140 +/- 0.008 mM) and pyridoxal (0.473 +/- 0.109 mM), respectively, than those of the Pseudomonas enzyme. The Aureobacterium enzyme could use NADP+ as a substrate: the reactivity was 6.5% of NAD+. The enzyme was much more tolerant to metal-chelating agents. Irreversibility of the enzymatic reaction was shared by the two enzymes. No aldehyde dehydrogenase showed similarity to the amino-terminal amino acid sequence of the enzyme.
Trongpanich
Yanee
Y
Department of Bioresources Science, Faculty of Agriculture, Kochi University, Japan.
Abe
Kinuyo
K
Kaneda
Yasuo
Y
Morita
Tomotake
T
Yagi
Toshiharu
T
D000429:Alcohol Oxidoreductases / Q000096:biosynthesis / Q000737:chemistry* / Q000302:isolation & purification; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D003067:Coenzymes / Q000378:metabolism; D003470:Culture Media; D004790:Enzyme Induction / Q000187:drug effects; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D007202:Indicators and Reagents; D007527:Isoenzymes / Q000737:chemistry / Q000302:isolation & purification; D008970:Molecular Weight; D009161:Mycobacterium / Q000201:enzymology* / Q000254:growth & development / Q000302:isolation & purification; D009243:NAD / Q000378:metabolism; D012988:Soil Microbiology; D013050:Spectrometry, Fluorescence; D013696:Temperature
D003067:Coenzymes; D003470:Culture Media; D004791:Enzyme Inhibitors; D007202:Indicators and Reagents; D007527:Isoenzymes; D009243:NAD; D000429:Alcohol Oxidoreductases; C042642:pyridoxal dehydrogenase