title: Enzymatic cleavage of the C-glucosidic bond of puerarin by three proteins, Mn(2+), and oxidized form of nicotinamide adenine dinucleotide.
abstract: We previously isolated the human intestinal bacterium, strain PUE, which can cleave the C-glucosidic bond of puerarin to yield its aglycone daidzein and glucose. In this study, we partially purified puerarin C-glucosidic bond cleaving enzyme from the cell-free extract of strain PUE and demonstrated that the reaction was catalyzed by at least three proteins, Mn(2+), and oxidized form of nicotinamide adenine dinucleotide (NAD(+)). We completely purified one of the proteins, called protein C, by chromatographic separation in three steps. The molecular mass of protein C was approximately 40 kDa and the amino acid sequence of its N-terminal region shows high homology to those of two putative proteins which belong to Gfo/Idh/MocA family oxidoreductase. Protein C catalyzed hydrogen-deuterium exchange reaction of puerarin to 2"-deuterated puerarin in D(2)O condition, which closely resembles those of glycoside hydrolase family 4 and 109.
authors: Faculty of Pharmaceutical Sciences, Suzuka University of Medical Science, 3500-3 Minamitamagaki, Suzuka, Mie 513-8670, Japan.
authors: Kenichi
authors: K
authors: Nakamura
authors: Katsuko
authors: K
authors: Komatsu
authors: Masao
authors: M
authors: Hattori
authors: Makoto
authors: M
authors: Iwashima
mesh_terms: D001419:Bacteria / Q000302:isolation & purification / Q000378:metabolism; D005243:Feces / Q000382:microbiology; D005960:Glucosides / Q000378:metabolism*; D006801:Humans; D007529:Isoflavones / Q000378:metabolism*; D008345:Manganese / Q000378:metabolism*; D009243:NAD / Q000378:metabolism*; D010084:Oxidation-Reduction; D011506:Proteins / Q000378:metabolism
chemical_list: D005960:Glucosides; D007529:Isoflavones; D011506:Proteins; D009243:NAD; D008345:Manganese; C033607:puerarin