[Purification and properties of the riboflavin kinase of the yeast Pichia guilliermondii].
Riboflavin kinase (E.C.2.7.1.26) was isolated from the cells of the yeast Pichia guilliermondii. The enzyme was 680-fold purified uzing ammonium sulphate fractionation, chromatography on DEAE-Sephadex A-50 and CM-Sephadex C-50 and gel-filtration through Sephadex G-75. Purified enzyme preparation was free from phosphatases and FAD-synthetase. The pH optimum was 8,7, the temperature optimum-45 degrees C. The enzyme was activated by Zn2+, Mg2+ and Co2+ ions. Km for riboflavin was 1,0x10(-5) M, for ATP -- 6,7X10(-6) M. Riboflavin kinase catalyzed the phosphorylation of riboflavin analogues with the substitution of methyl groups at positions 7 and 8. UTP, GTP, ADP and CTP, besides ATP, were phosphate donors. AMP inhibited the enzyme activity. Molecular weight of the enzyme was 28000, as estimated by gel-filtration through Sephadex G-150. Purified riboflavin kinase was stable under storage.
D001203:Ascomycota / Q000201:enzymology*; D002384:Catalysis; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D010770:Phosphotransferases / Q000302:isolation & purification; D010843:Pichia / Q000201:enzymology*; D013696:Temperature
D010770:Phosphotransferases
Ochistka i svoĭstva riboflavinkinazy drozhzheĭ Pichia guilliermondii