1887

Abstract

The methylotrophic yeast exhibits -formylglutathione hydrolase activity (FGH, EC 3.1.2.12), which is involved in the glutathione-dependent formaldehyde oxidation pathway during growth on methanol as the sole carbon source. The structural gene, , was cloned from , and its predicted amino acid sequence showed more than 60 % similarity to those of FGHs from and , and human esterase D. FGH from contained a C-terminal tripeptide, SKL, which is a type I peroxisome-targeting signal, and a bimodal distribution of FGH between peroxisomes and the cytosol was demonstrated. The gene was disrupted in the genome by one-step gene disruption. The Δ strain was still able to grow on methanol as a carbon source under methanol-limited chemostat conditions with low dilution rates (<0·05 h), conditions under which a strain with disruption of the gene for formaldehyde dehydrogenase (another enzyme involved in the formaldehyde oxidation pathway) could not survive. These results suggested that FGH is not essential but necessary for optimal growth on methanol. This is believed to be the first report of detailed analyses of the gene in a methylotrophic yeast strain.

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2003-08-01
2020-07-07
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References

  1. Cryer D. R., Eccleshal R., Murmur J.. 1975; Isolation of yeast DNA. Methods Cell Biol12:39–44
    [Google Scholar]
  2. Cygler M., Schrag J. D., Sussman J. L., Harel M., Silman I., Gentry M. K.. 1993; Relationship between sequence conservation and three-dimensional structure in a large family of esterase, lipase, and related proteins. Protein Sci2:366–382
    [Google Scholar]
  3. Davis R. W., Thomas M., Cameron J., John T. P. S., Scherer S., Padgett R. A.. 1980; Rapid DNA isolation for enzymatic and hybridization analysis. Methods Enzymol65:404–411
    [Google Scholar]
  4. Degrassi G., Uotila L., Klima R., Venturi V.. 1999; Purification and properties of an esterase from the yeast Saccharomyces cerevisiae and identification of the encoding gene. Appl Environ Microbiol65:3470–3472
    [Google Scholar]
  5. Derewenda Z. S.. 1994; Structure and function of lipase. Adv Protein Chem45:1–52
    [Google Scholar]
  6. Eiberg H., Mohr J.. 1986; Identity of the polymorphisms for esterase D and S -formylglutathione hydrolase in red blood cells. Hum Genet139:913–920
    [Google Scholar]
  7. Elgersma Y., Vos A., van den Berg M., van Roermund C. W. T., van der Sluijs P., Distel B., Tabak H. F.. 1996; Analysis of the carboxy-terminal peroxisomal targeting signal 1 in a homologous context in Saccharomyces cerevisiae . J Biol Chem271:26375–26382
    [Google Scholar]
  8. Fernandez L., Beerthuyzen M. M., Brown J., Siezen R. J., Coolbear T., Holland R., Kuipers O. P.. 2000; Cloning, characterization, controlled overexpression, and inactivation of the major tributyrin esterase gene of Lactococcus lactis . Appl Environ Microbiol66:1360–1368
    [Google Scholar]
  9. Gellissen G.. 2000; Heterologous protein production in methylotrophic yeasts. Appl Microbiol Biotechnol54:741–750
    [Google Scholar]
  10. Harms N., Ras J., Reijnders W. N., van Spanning R. J. M., Stouthamer A. H.. 1996; S -Formylglutathione hydrolase of Paracoccus denitrificans is homologous to human esterase D: a universal pathway for formaldehyde detoxification?. J Bacteriol178:6296–6299
    [Google Scholar]
  11. Haslam R., Rust S., Pallett K., Cole D., Coleman J.. 2002; Cloning and characterisation of S -formylglutathione hydrolase from Arabidopsis thaliana : a pathway for formaldehyde detoxification. Plant Physiol Biochem40:281–288
    [Google Scholar]
  12. Horiguchi H., Yurimoto H., Kato N., Sakai Y.. 2001a; Antioxidant system within yeast peroxisome: biochemical and physiological characterization of CbPmp20 in the methylotrophic yeast Candida boidinii . J Biol Chem276:14279–14288
    [Google Scholar]
  13. Horiguchi H., Yurimoto H., Goh T.-K., Nakagawa T., Kato N., Sakai Y.. 2001b; Peroxisomal catalase in the methylotrophic yeast Candida boidinii : transport efficiency and metabolic significance. J Bacteriol183:6372–6383
    [Google Scholar]
  14. Kato N., Sakazawa C., Nishizawa T., Tani Y., Yamada H.. 1980; Purification and characterization of S -formylglutathione hydrolase from a methylotrophic yeast, Kloeckera sp. No. 2201. Biochim Biophys Acta611:323–332
    [Google Scholar]
  15. Laemmli U. K.. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature227:680–685
    [Google Scholar]
  16. Lee B., Yurimoto H., Sakai Y., Kato N.. 2002; Physiological role of the glutathione-dependent formaldehyde dehydrogenase in the methylotrophic yeast Candida boidinii . Microbiology148:2697–2704
    [Google Scholar]
  17. Neben I., Sahm H., Kula M.-R.. 1980; Studies on an enzyme, S -formylglutathione hydrolase, of the dissimilatory pathway of methanol in Candida boidinii . Biochim Biophys Acta614:81–91
    [Google Scholar]
  18. Nishikawa M., Hagishita T., Yurimoto H., Kato N., Sakai Y., Hatanaka T.. 2000; Primary structure and expression of peroxisomal acetylspermidine oxidase in the methylotrophic yeast Candida boidinii . FEBS Lett476:150–154
    [Google Scholar]
  19. Sakai Y., Tani Y.. 1992; Directed mutagenesis in an asporogenous methylotrophic yeast: cloning, sequencing, and one-step gene disruption of the 3-isopropylmalate dehydrogenase gene ( LEU2 ) of Candida boidinii to derive doubly auxotrophic marker strains. J Bacteriol174:5988–5993
    [Google Scholar]
  20. Sakai Y., Kazarimoto T., Tani Y.. 1991; Transformation system for an asporogenous methylotrophic yeast, Candida boidinii : cloning of the orotidine-5′-phosphate decarboxylase gene ( URA3 ), isolation of uracil auxotrophic mutants, and use of the mutants for integrative transformation. J Bacteriol173:7458–7463
    [Google Scholar]
  21. Sakai Y., Goh T. K., Tani Y.. 1993; High-frequency transformation of a methylotropic yeast, Candida boidinii , with autonomously replicating plasmids which are also functional in Saccharomyces cerevisiae . J Bacteriol175:3556–3562
    [Google Scholar]
  22. Sakai Y., Saigannji A., Yurimoto H., Takabe K., Saiki H., Kato N.. 1996; The absence of Pmp47, a putative yeast peroxisomal transporter, causes a defect in transport and folding of a specific matrix enzyme. J Cell Biol134:37–51
    [Google Scholar]
  23. Sakai Y., Murdanoto A. P., Konishi T., Iwamatsu A., Kato N.. 1997; Regulation of the formate dehydrogenase gene, FDH1 , in the methylotrophic yeast Candida boidinii and growth characteristics of an FDH1 -disrupted strain on methanol, methylamine, and choline. J Bacteriol179:4480–4485
    [Google Scholar]
  24. Sakai Y., Yurimoto H., Matsuo H., Kato N.. 1998; Regulation of peroxisomal proteins and organelle proliferation by multiple carbon sources in the methylotrophic yeast Candida boidinii . Yeast14:1175–1187
    [Google Scholar]
  25. Subramani S.. 1998; Components involved in peroxisome import, biogenesis, proliferation, turnover, and movement. Physiol Rev78:171–188
    [Google Scholar]
  26. Tani Y., Sakai Y., Yamada H.. 1985; Production of formaldehyde by a mutant of methanol yeast, Candida boidinii S2. J Ferment Technol63:443–449
    [Google Scholar]
  27. Towbin H., Staehelin T., Gordon J.. 1979; Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A76:4350–4354
    [Google Scholar]
  28. Ubiyvovk V. M., Nazarko T. Y., Stasyk O. G., Sohn M. J., Kang H. A., Sibirny A. A.. 2002; GSH2 , a gene encoding γ-glutamylcysteine synthetase in the methylotrophic yeast Hansenula polymorpha . FEMS Yeast Res2:327–332
    [Google Scholar]
  29. Uotila L., Koivusalo M.. 1974; Purification and properties of S -formylglutathione hydrolase from human liver. J Biol Chem279:7664–7672
    [Google Scholar]
  30. Yurimoto H., Hasegawa T., Sakai Y., Kato N.. 2000a; Physiological role of the d-amino acid oxidase gene, DAO1 , in carbon and nitrogen metabolism in the methylotrophic yeast Candida boidinii . Yeast16:1217–1227
    [Google Scholar]
  31. Yurimoto H., Komeda T., Lim C. R., Nakagawa T., Kondo K., Kato N., Sakai Y.. 2000b; Regulation and evaluation of five methanol-inducible promoters in the methylotrophic yeast Candida boidinii . Biochim Biophys Acta 1493;56–63
    [Google Scholar]
  32. Yurimoto H., Sakai Y., Kato N.. 2002; Methanol metabolism. In Hansenula polymorpha Biology and Applications pp61–75 Edited by Gellissen G. Weinheim: Wiley-VCH;
    [Google Scholar]
  33. Zwart K., Veenhuis M., van Dijken J. P., Harder W.. 1980; Development of amine oxidase containing peroxisomes in yeasts during growth of glucose in the presence of methylamine as the nitrogen source. Arch Microbiol126:117–126
    [Google Scholar]
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