1887

Abstract

Alcohol oxidase, a major peroxisomal protein of methanol-utilizing yeasts, may possess two different forms of flavin adenine dinucleotide, classical FAD and so-called modified FAD (mFAD). Conversion of FAD into mFAD was observed both in purified preparations of the enzyme and in cells grown in batch and continuous culture. The relative amount of mFAD in the enzyme varied from 5 to 95 %, depending on the growth or storage conditions. The presence of mFAD led to a slight decrease in and a significant (about one order) decrease in the of alcohol oxidase with respect to methanol. The kinetics of modification measured in purified preparations of the enzyme obeyed first-order kinetics (=0·78 h). The modification process was strongly inhibited by methanol, formaldehyde or hydroxylamine. Modification observed in continuous culture under steady state conditions depended on the dilution rate and could also be described as a spontaneous first-order reaction ( = 0·27 h). FAD modification could only be detected in alcohol oxidase and not in other yeast peroxisomal flavoenzymes, such as -amino acid oxidase from .

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-137-10-2381
1991-10-01
2021-08-02
Loading full text...

Full text loading...

/deliver/fulltext/micro/137/10/mic-137-10-2381.html?itemId=/content/journal/micro/10.1099/00221287-137-10-2381&mimeType=html&fmt=ahah

References

  1. Brooke A. G., Dijkhuizen L., Harder W. 1986; Regulation of flavin biosynthesis in the methylotrophic yeast Hansenula polymorpha. Archives of Microbiology 145:62–70
    [Google Scholar]
  2. Bystrykh L. V., Romanov V. P., Steczko J., Trotsenko Y. A. 1989a; Catalytic variability of alcohol oxidase from the methylotrophic yeast Hansenula polymorpha. Biotechnology and Applied Biochemistry 11:184–192
    [Google Scholar]
  3. Bystrykh L. V., Dvorakova J., Volfova O. 1989b; Alcohol oxidase of methylotrophic thermo- and acidotolerant yeast Hansenula sp. Folio Microbiologica 34:233–237
    [Google Scholar]
  4. Bystrykh L. V., Kellogg R. M., Kruizinga W., Dijkhuizen L., Harder W., Vervoort J., van Berkel W. J. H. 1991; Structural and kinetic analysis of flavin adenine dinucleotide modification in alcohol oxidase from methylotrophic yeasts. In Flavins and Flavoproteins 1990Proceedings of the 10th International Symposium197–200 Curti B, Ronchi S., Zanetti G. Berlin: Walter de Gruyter;
    [Google Scholar]
  5. Egli Th., Lindley N., Quayle J. R. 1983; Regulation of enzyme synthesis and variation of residual concentration during carbon-limited growth of Kloeckera sp. 2201 on mixtures of methanol and glucose. Journal of General Microbiology 129:1269–1281
    [Google Scholar]
  6. Egli Th., Bosshard C., Hamer G. 1986; Simultaneous utilization of methanol-glucose mixtures by Hansenula polymorpha in a chemostat: influence of dilution rate and mixture composition on utilization pattern. Biotechnology and Bioengineering 28:1735–1741
    [Google Scholar]
  7. Geissler J., Hemmerich P. 1981; Yeast methanol oxidases: an unusual type of flavoprotein. FEBS Letters 126:152–156
    [Google Scholar]
  8. Geissler J., Ghisla S., Kroneck P. H. 1986; Flavin-dependent alcohol oxidase from yeast. Studies on the catalytic mechanism and inactivation during turnover. European Journal of Biochemistry 160:93–100
    [Google Scholar]
  9. Hopkins T., Muller F. 1987; Biochemistry of alcohol oxidase. In Proceedings of the 5th International Symposium on Microbial Growth on Cl compounds150–157 van Verseveld H. W., Duine J. A. Dordrecht: Martinus Nijhoff;
    [Google Scholar]
  10. van der Klei I. J., Bystrykh L. V., Harder W. 1990; Alcohol oxidase from Hansenula polymorpha CBS 4732. Methods in Enzymology 188:420–427
    [Google Scholar]
  11. Mincey T., Taylern G., Mildvan A. S., Abeles R. H. 1980; Presence of a flavin semiquinone in methanol oxidase. Biochemistry 77:7099–7101
    [Google Scholar]
  12. Roggenkamp R., Janowicz Z., Stanikowski B., Hollenberg C. 1984; Biosynthesis and regulation of the peroxisomal methanol oxidase from the methylotrophic yeast Hansenula polymorpha. Molecular and General Genetics 194:489–493
    [Google Scholar]
  13. Sherry B., Abeles R. H. 1985; Mechanism of action of methanol oxidase, reconstitution of methanol oxidase with 5-deazaflavin, and inactivation of methanol oxidase by cyclopropanol. Biochemistry 24:2594–2605
    [Google Scholar]
  14. Woodward J. R. 1990; Biochemistry and applications of alcohol oxidase from methylotrophic yeasts. In Advances in Autotrophic Microbiology and One-Carbon Metabolism 1205–238 Codd G. A., Dijkhuizen L., Tabita F. R. Dordrecht: Martinus Nijhoff;
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-137-10-2381
Loading
/content/journal/micro/10.1099/00221287-137-10-2381
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error