TY - JOUR
T1 - Purification and characterization of formaldehyde dehydrogenase from rat liver cytosol
AU - Tsuboi, Seiji
AU - Kawase, Michi
AU - Takada, Aya
AU - Hiramatsu, Mio
AU - Wada, Yumiko
AU - Kawakami, Yasuhiko
AU - Ikeda, Mikiko
AU - Ohmori, Shinji
PY - 1992/4
Y1 - 1992/4
N2 - Formaldehyde dehydrogenase was purified to electrophoretic and column chromatographic homogeneity from rat liver cytosolic fraction by a procedure which includes ammonium sulfate precipitation, DEAE-cellulose-, hydroxyapatite-, Mono Q-chromatography, and gel filtration. Its molecular mass was estimated to be 41 kDa by gel filtration and SDS-PAGE, suggesting that it is a monomer. It utilized neither methylglyoxal nor aldehydes except formaldehyde as a substrate. It has been reported that liver class in alcohol dehydrogenase and formaldehyde dehydrogenase are the same enzyme and oxidize formaldehyde and long chain primary alcohols. However, the enzyme examined here did not use n-octanoi as a substrate. The Km values for formaldehyde and NAD+ were 5.09 and 2.34 μM at 25°C, respectively. The amino acid sequences of 10 peptides obtained from the purified enzyme after digestion with either Vs protease or lysyl endopeptidase were determined. From these results, the enzyme was proved to be different from the previously described mammalian formaldehyde dehydrogenase and is the first true formaldehyde dehydrogenase to be isolated from a mammalian source.
AB - Formaldehyde dehydrogenase was purified to electrophoretic and column chromatographic homogeneity from rat liver cytosolic fraction by a procedure which includes ammonium sulfate precipitation, DEAE-cellulose-, hydroxyapatite-, Mono Q-chromatography, and gel filtration. Its molecular mass was estimated to be 41 kDa by gel filtration and SDS-PAGE, suggesting that it is a monomer. It utilized neither methylglyoxal nor aldehydes except formaldehyde as a substrate. It has been reported that liver class in alcohol dehydrogenase and formaldehyde dehydrogenase are the same enzyme and oxidize formaldehyde and long chain primary alcohols. However, the enzyme examined here did not use n-octanoi as a substrate. The Km values for formaldehyde and NAD+ were 5.09 and 2.34 μM at 25°C, respectively. The amino acid sequences of 10 peptides obtained from the purified enzyme after digestion with either Vs protease or lysyl endopeptidase were determined. From these results, the enzyme was proved to be different from the previously described mammalian formaldehyde dehydrogenase and is the first true formaldehyde dehydrogenase to be isolated from a mammalian source.
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U2 - 10.1093/oxfordjournals.jbchem.a123781
DO - 10.1093/oxfordjournals.jbchem.a123781
M3 - Article
C2 - 1618737
AN - SCOPUS:0026523978
SN - 0021-924X
VL - 111
SP - 465
EP - 471
JO - Journal of Biochemistry
JF - Journal of Biochemistry
IS - 4
ER -