Disruption of the plr1+ gene encoding pyridoxal reductase of Schizosaccharomyces pombe.
Pyridoxal (PL) reductase encoded by the plr1(+) gene practically catalyzes the irreversible reduction of PL by NADPH to form pyridoxine (PN). The enzyme has been suggested to be involved in the salvage synthesis of pyridoxal 5'-phosphate (PLP), a coenzyme form of vitamin B(6), or the excretion of PL as PN from yeast cells. In this study, a PL reductase-disrupted (plr1 Delta) strain was constructed and its phenotype was examined. The plr1 Delta cells showed almost the same growth curve as that of wild-type cells in YNB and EMM media. In EMM, the plr1 Delta strain became flocculent at the late stationary phase for an unknown reason. The plr1 Delta cells showed low but measurable PL reductase activity catalyzed by some other protein(s) than the enzyme encoded by the plr1(+) gene, which maintained the flow of "PL --> PN --> PNP --> PLP" in the salvage synthesis of PLP. The total vitamin B(6) and pyridoxamine 5'-phosphate contents in the plr1 Delta cells were significantly lower than those in the wild-type ones. The percentages of the PLP amount as to the other vitamin B(6) compounds were similar in the two cell types. The amount of PL in the culture medium of the disruptant was significantly higher than that in the wild-type. In contrast, PN was much higher in the latter than the former. The plr1 Delta cells accumulated a 6.1-fold higher amount of PL than the wild-type ones when they were incubated with PL. The results showed that PL reductase encoded by the plr1(+ )gene is involved in the excretion of PL after reducing it to PN, and may not participate in the salvage pathway for PLP synthesis.
Morita
Tomotake
T
Department of Bioresources Science, Faculty of Agriculture, Kochi University, Nankoku, Kochi 783-8502.
Takegawa
Kaoru
K
Yagi
Toshiharu
T
D000429:Alcohol Oxidoreductases / Q000235:genetics* / Q000378:metabolism; D001483:Base Sequence; D002384:Catalysis; D002478:Cells, Cultured; D003001:Cloning, Molecular; D015151:Immunoblotting; D009154:Mutation; D009249:NADP / Q000378:metabolism; D010641:Phenotype; D011730:Pyridoxal / Q000378:metabolism*; D011731:Pyridoxal Kinase / Q000235:genetics / Q000378:metabolism; D011732:Pyridoxal Phosphate / Q000096:biosynthesis / Q000494:pharmacology; D011733:Pyridoxamine / Q000378:metabolism; D011736:Pyridoxine / Q000096:biosynthesis / Q000494:pharmacology; D012568:Schizosaccharomyces / Q000201:enzymology* / Q000235:genetics; D025101:Vitamin B 6 / Q000494:pharmacology
D011730:Pyridoxal; D009249:NADP; D011732:Pyridoxal Phosphate; D011733:Pyridoxamine; D025101:Vitamin B 6; D000429:Alcohol Oxidoreductases; C022495:pyridoxine 4-dehydrogenase; D011731:Pyridoxal Kinase; D011736:Pyridoxine