title: Tauropine dehydrogenase from the sandworm Arabella iricolor (Polychaeta: Errantia): purification and characterization.
abstract: This is the first report of the purification of tauropine dehydrogenase (NAD: tauropine oxidoreductase) from a polychaete worm. In the sandworm Arabella iricolor Montagu (Polychaeta: Errantia), two forms of TaDH were detected: major component (pl = 7.5) and minor one (pl = 6.4). The major TaDH component was purified to homogeneity by means of (NH4)2SO4 precipitation, anion-exchange, affinity, chromatofocusing and hydrophobic chromatography, and characterized. From the molecular mass of 43.7 kDa obtained by rapid gel-filtration and that of 43.5 kDa by SDS-PAGE, the sandworm enzyme appeared to be a monomeric protein. Maximum rates of reduction of pyruvate and oxidation of tauropine were observed at pH 7.0 and 8.5, respectively. Pyruvate and taurine were preferred substrate for the enzyme. Apparent K(m) values determined using constant co-substrate concentrations were: 35.7 mM, 0.34 mM, and 0.036 mM for taurine, pyruvate and NADH, respectively, in the tauropine synthesizing reaction; and 4.8 mM and 0.051 mM for tauropine and NAD+, respectively, in the tauropine oxidizing reaction. The tauropine synthesizing reaction was subject to substrate inhibition by pyruvate: maximum rate was observed at 2.5-3.0 mM (inhibitory range of pyruvate concentration producing half-maximal rate was 26.8 mM).
authors.affiliation: School of Fisheries Sciences, Kitasato University, Iwate, Japan.
authors.forename: N
authors.initials: N
authors.lastname: Kanno
authors.forename: M
authors.initials: M
authors.lastname: Sato
authors.forename: E
authors.initials: E
authors.lastname: Nagahisa
authors.forename: Y
authors.initials: Y
authors.lastname: Sato
mesh_terms: D000594:Amino Acid Oxidoreductases / Q000737:chemistry / Q000302:isolation & purification*; D000595:Amino Acid Sequence; D000818:Animals; D004591:Electrophoresis, Polyacrylamide Gel; D006863:Hydrogen-Ion Concentration; D007525:Isoelectric Focusing; D007700:Kinetics; D008969:Molecular Sequence Data; D008970:Molecular Weight; D011077:Polychaeta / Q000201:enzymology*; D013379:Substrate Specificity
chemical_list: D000594:Amino Acid Oxidoreductases; C050564:tauropine dehydrogenase