Identification of thermostable glyoxalase I in the fission yeast Schizosaccharomyces pombe.
Glyoxalase I is a ubiquitous enzyme that detoxifies methylglyoxal, which is derived from glycolysis but inhibits the growth of cells from microorganisms to mammals. Here, the structural gene for glyoxalase I ( glo1(+)) from the fission yeast Schizosaccharomyces pombe was identified. Disruption of glo1(+) enhanced susceptibility to methylglyoxal, while expression of glo1(+) in a Delta glo1 mutant of Saccharomyces cerevisiae restored tolerance to this aldehyde. The glo1(+) gene product was purified. The glyoxalase I of S. pombe was a monomeric enzyme with a molecular weight of 34000 and the k(cat)/ K(m) value for methylglyoxal was 4.3 x 10(7) M(-1) x min(-1). Treatment of purified enzyme with EDTA in imidazole buffer completely abolished enzyme activity, whereas the EDTA-treated enzyme was reactivated by several divalent metal ions, such as Zn(2+), Co(2+), Ni(2+) and Mn(2+). The glyoxalase I of S. pombe exhibited fairly high thermal stability, and almost 100% activity was retained after incubating the enzyme at 60 degrees C for 4 h.
Takatsume
Yoshifumi
Y
Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Uji, Kyoto 611-0011, Japan.
Izawa
Shingo
S
Inoue
Yoshiharu
Y
D001483:Base Sequence; D020134:Catalytic Domain; D004271:DNA, Fungal / Q000235:genetics; D004795:Enzyme Stability; D005800:Genes, Fungal; D006358:Hot Temperature; D007700:Kinetics; D007791:Lactoylglutathione Lyase / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D008670:Metals / Q000494:pharmacology; D008970:Molecular Weight; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics; D012568:Schizosaccharomyces / Q000201:enzymology* / Q000235:genetics
D004271:DNA, Fungal; D008670:Metals; D011994:Recombinant Proteins; D007791:Lactoylglutathione Lyase