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Volume 151, Issue 6

Pectin utilization by the methylotrophic yeast Free

Abstract

The methylotrophic yeast was able to grow on pectic compounds, pectin and polygalacturonate, as sole carbon sources. Under the growth conditions used, exhibited increased levels of pectin methylesterase, and pectin-depolymerizing and methanol-metabolizing enzyme activities. On the other hand, has two alcohol oxidase (AOD) genes, and . On growth on pectin, the Δ and ΔΔ strains showed a severe defect in the growth yield, although the Δ strain could grow on polygalacturonate to the same extent as the wild-type strain. The expression of was detected in pectin-grown cells, but the -gene expression detected by pectin was much lower than that of . Moreover, pectin could induce peroxisome proliferation in , like methanol and oleic acid. These findings showed that was able to utilize the methylester moiety of pectin by means of methanol-metabolic enzymes in peroxisomes, and that the functional AOD subunit for pectin utilization was Mod1p in .

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Keyword(s): AOD, alcohol oxidase , DE, degree of esterification , DHAS, dihydroxyacetone synthase , FDH, formate dehydrogenase , FLD, formaldehyde dehydrogenase , GFP, green fluorescent protein , PAL, pectate lyase , PG, polygalacturonase , PME, pectin methylesterase and PNL, pectin lyase
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2005-06-01
2024-10-05
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References

  1. Ashin V. V., Trotsenko Y. A. 1998; Formation and distribution of modified FAD between isozymes of alcohol oxidase in the methylotrophic yeast Pichia methanolica . Biochemistry 63:1407–1413
     [Google Scholar]
  2. Bradford M. M. 1976; A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein–dye binding. Anal Biochem 72:248–254 [CrossRef]
     [Google Scholar]
  3. Cregg J. M. 1993; Recent advances in the expression of foreign genes in Pichia pastoris. Bio/Technology 11:905–910 [CrossRef]
     [Google Scholar]
  4. Gellissen G. 2000; Heterologous protein production in methylotrophic yeasts. Appl Microbiol Biotechnol 54:741–750 [CrossRef]
     [Google Scholar]
  5. Goodman J. M., Scott C. W., Donahue P. N., Atherton J. P. 1984; Alcohol oxidase assembles post-translationally into the peroxisome of Candida boidinii . J Biol Chem 259:8485–8493
     [Google Scholar]
  6. Gruzman M. B., Titorenko V. I., Ashin V. V., Lusta K. A., Trotsenko Y. A. 1996; Multiple molecular forms of alcohol oxidase from the methylotrophic yeast Pichia methanolica . Biochemistry 61:1537–1544
     [Google Scholar]
  7. Hollenberg C. P., Gellissen G. 1997; Production of recombinant proteins by methylotrophic yeasts. Curr Opin Biotechnol 8:554–560 [CrossRef]
     [Google Scholar]
  8. Ishii S., Yokotsuka T. 1972; Purification and properties of pectin trans-eliminase from Aspergillus sojae. Agric Biol Chem 36:146–153 [CrossRef]
     [Google Scholar]
  9. Laemmli U. K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685 [CrossRef]
     [Google Scholar]
  10. Lee J. D., Komagata K. 1980; Taxonomic study of methanol-assimilating yeasts. J Gen Appl Microbiol 26:133–158 [CrossRef]
     [Google Scholar]
  11. MacDonald R. C., Fall F. 1993; Detection of substrational emission of methanol from plants to the atmosphere. Atmos Environ 27A:1709–1713
     [Google Scholar]
  12. Nakagawa T., Uchimura T., Komagata K. 1996; Isozymes of methanol oxidase in a methanol-utilizing yeast, Pichia methanolica IAM 12901. J Ferment Bioeng 81:498–503 [CrossRef]
     [Google Scholar]
  13. Nakagawa T., Mukaiyama H., Yurimoto H., Sakai Y., Kato N. 1999; Alcohol oxidase hybrid oligomers formed in vivo and in vitro . Yeast 15:1223–1230 [CrossRef]
     [Google Scholar]
  14. Nakagawa T., Miyaji T., Yurimoto H., Sakai Y., Kato N., Tomizuka N. 2000; Methylotrophic pathway participates in pectin utilization by Candida boidinii . Appl Environ Microbiol 66:4253–4257 [CrossRef]
     [Google Scholar]
  15. Nakagawa T., Sakai Y., Mukaiyama H., Mizumura T., Miyaji T., Yurimoto H., Kato N., Tomizuka N. 2001; Analysis of alcohol-oxidase isozymes in gene-disrupted strains of the methylotrophic yeast Pichia methanolica . J Biosci Bioeng 91:225–227 [CrossRef]
     [Google Scholar]
  16. Nakagawa T., Mizumura T., Mukaiyama H., Miyaji T., Yurimoto H., Kato N., Sakai Y., Tomizuka N. 2002; Physiological role of the second alcohol oxidase gene MOD2 in the methylotrophic growth ofPichia methanolica . Yeast 19:1067–1073 [CrossRef]
     [Google Scholar]
  17. Nakagawa T., Ito T., Fujimura S. 7 other authors 2004; Molecular characterization of the glutathione-dependent formaldehyde dehydrogenase gene FLD1 from the methylotrophic yeastPichia methanolica . Yeast 21:445–453 [CrossRef]
     [Google Scholar]
  18. Nakagawa T., Nagaoka T., Miyaji T., Tomizuka N. 2005; Cold-active polygalacturonase from psychrophilic-basidiomycetous yeast Cystofilobasidium capitatum strain PPY-1. Biosci Biotechnol Biochem 69:419–421 [CrossRef]
     [Google Scholar]
  19. Nash T. 1953; The colorimetric estimation of formaldehyde by means of the Hantzsch reaction. Biochem J 55:416–421
     [Google Scholar]
  20. Nelson N. 1944; A photometric adaptation of the Somogyi method for the determination of glucose. J Biol Chem 153:375
     [Google Scholar]
  21. Nemecek-Marshall M., MacDonald R. C., Franzen J. J., Wojciechowski C. L., Fall R. 1995; Methanol emission from leaves, enzymatic detection of gas-phase methanol and regulation of methanol fluxes to stomatal conductance and leaf development. Plant Physiol 108:1359–1368
     [Google Scholar]
  22. Raymond C. K. 1999; Recombinant protein expression in Pichia methanolica. In Gene Expression Systems, Using Nature of the Art of Expression pp 193–209 Edited by Fernandez J. M., Hoeffler J. P. New York: Academic Press;
     [Google Scholar]
  23. Raymond C. K., Bukowski T., Holderman S. D., Ching A. F. T., Vanaja E., Stamm R. 1998; Development of the methylotrophic yeast Pichia methanolica for the expression of the 65 kilodalton isoform of human glutamate decarboxylase. Yeast 14:11–23 [CrossRef]
     [Google Scholar]
  24. Sakai T., Sakamoto T., Hallaert J., Vandamme E. J. 1993; Pectin, pectinase and protopectinase, production, properties, and applications. Adv Appl Microbiol 39:213–294
     [Google Scholar]
  25. Sakai Y., Rogi T., Takeuchi R., Kato N., Tani Y. 1995; Expression of Saccharomyces adenylate kinase gene in Candida boidinii under the regulation of its alcohol oxidase promoter. Appl Microbiol Biotechnol 42:860–864 [CrossRef]
     [Google Scholar]
  26. Sakai Y., Nakagawa T., Shimase M., Kato N. 1998; Regulation and physiological role of the DAS1 gene encoding dihydroxyacetone synthase in the methylotrophic yeastCandida boidinii . J Bacteriol 180:5885–5890
     [Google Scholar]
  27. Sakai Y., Tani Y., Kato N. 1999; Biotechnological application of cellular functions of the methylotrophic yeast. J Mol Catal B Enzym 6:161–174 [CrossRef]
     [Google Scholar]
  28. Schütte H., Flossdorf J., Sahm H., Kula M.-R. 1976; Purification and properties of formaldehyde dehydrogenase and formate dehydrogenase from Candida boidinii. Eur J Biochem 62:151–160 [CrossRef]
     [Google Scholar]
  29. Somogyi M. 1952; Notes on sugar determination. J Biol Chem 159:19–23
     [Google Scholar]
  30. Stratilova E., Breierova E., Vadkertiova R., Machova E., Malovikova A., Slavikova E. 1998; The adaptability of the methylotrophic yeast Candida boidinii on media containing pectic substances. Can J Microbiol 44:116–120 [CrossRef]
     [Google Scholar]
  31. Tani Y., Kato N., Yamada H. 1978; Utilization of methanol by yeasts. Adv Appl Microbiol 24:165–186
     [Google Scholar]
  32. Tani Y., Sakai Y., Yamada H. 1985; Isolation and characterization of a mutant of a methanol yeast, Candida boidinii S2, with higher formaldehyde productivity. Agric Biol Chem 49:2699–2706 [CrossRef]
     [Google Scholar]
  33. Yanase H., Okuda M., Kita K., Sato Y., Shibata K., Sakai Y., Kato N. 1995; Enzymatic preparation of [1,3-13C] dihydroxyacetone phosphate from [13C]methanol and hydroxypyruvate using the methanol-assimilating system of methylotrophic yeasts. Appl Microbiol Biotechnol 43:228–234 [CrossRef]
     [Google Scholar]
/content/journal/micro/10.1099/mic.0.27895-0
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Pectin utilization by the methylotrophic yeast Pichia methanolica
Microbiology 151, 2047 (2005); https://doi.org/10.1099/mic.0.27895-0
/content/journal/micro/10.1099/mic.0.27895-0
/content/journal/micro/10.1099/mic.0.27895-0
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