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Identification of thermostable glyoxalase I in the fission yeast Schizosaccharomyces pombe

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Abstract

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 Δglo1 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 34,000 and the k cat/K m value for methylglyoxal was 4.3×107 M−1 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 Zn2+, Co2+, Ni2+ and Mn2+. The glyoxalase I of S. pombe exhibited fairly high thermal stability, and almost 100% activity was retained after incubating the enzyme at 60°C for 4 h.

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Acknowledgements

We thank Dr K. Shiozaki for S. pombe strains and Dr J. Kohli for plasmid pCG1. This study was partially supported by grants from BRAIN.

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Authors and Affiliations

  1. Laboratory of Molecular Microbiology, Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Uji, Kyoto, 611-0011, Japan

    Yoshifumi Takatsume, Shingo Izawa & Yoshiharu Inoue

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  1. Yoshifumi Takatsume
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  2. Shingo Izawa
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  3. Yoshiharu Inoue
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Correspondence to Yoshiharu Inoue.

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Takatsume, Y., Izawa, S. & Inoue, Y. Identification of thermostable glyoxalase I in the fission yeast Schizosaccharomyces pombe . Arch Microbiol 181, 371–377 (2004). https://doi.org/10.1007/s00203-004-0666-4

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  • DOI: https://doi.org/10.1007/s00203-004-0666-4

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