1887

Abstract

A method has been developed for the continuous spectrophotometric assay of dihydroxy-acetone synthase present in methanol-grown yeasts. This enzyme catalyses the condensation of formaldehyde with a pentose phosphate (most probably xylulose 5-phosphate) to give dihydroxyacetone and glyceraldehyde 3-phosphate. The assay is based on the NADH-and ATP-linked formation of -glycerol 3-phosphate from dihydroxyacetone via the added coupling enzymes glycerol kinase and glycerol-3-phosphate dehydrogenase, and the reaction was followed by the decrease in absorbance at 340 nm. Using this assay, the pH optimum of the dihydroxyacetone synthase was shown to be 7.4 to 7.6. The enzyme, which catalyses a transketolase-like reaction, is a separate enzyme from the classical transketolase because their specific activities varied independently in and grown on different substrates and the two enzyme activities could be separated by ion-exchange chromatography.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-120-1-219
1980-09-01
2021-07-27
Loading full text...

Full text loading...

/deliver/fulltext/micro/120/1/mic-120-1-219.html?itemId=/content/journal/micro/10.1099/00221287-120-1-219&mimeType=html&fmt=ahah

References

  1. Babel W., Loffhagen N. 1979; Assimilation of methanol by yeasts, a new approach. Zeitschrift für allgemeine Mikrobiologie 19:299–302
    [Google Scholar]
  2. Dawson R. M. C., Elliott D. C., Elliott W. H., Jones K. M. 1969 Data for Biochemical Research, 2nd edn.. Oxford: Clarendon Press;
    [Google Scholar]
  3. Dickens F., Williamson D. H. 1958; Formaldehyde as an acceptor aldehyde for transketolase, and the biosynthesis of triose. Nature, London 161:1790
    [Google Scholar]
  4. Gancedo C., Gancedo J. M., Sols A. 1968; Glycerol metabolism in yeasts.Pathways of utilization and production. European Journal of Biochemistry 5:165–172
    [Google Scholar]
  5. De La Haba G., Leder I. G., Racker E. 1955; Crystalline transketolase from bakers’ yeast: isolation and properties. Journal of Biological Chemistry 214:409–426
    [Google Scholar]
  6. Hers H. G. 1962; Triokinase. Methods in Enzymology 5:362
    [Google Scholar]
  7. Kato N., Nishizawa T., Sakazawa C., Tani Y., Yamada H. 1979; Xylulose 5-phosphate dependent fixation of formaldehyde in a methanolutilizing yeast Kloeckera sp. no. 2201. Agricultural and Biological Chemistry 43:2013–2015
    [Google Scholar]
  8. Lindley N. D., Waites M. J., Quayle J. R. 1980; A modified pulse labelling technique for the detection of early intermediates in microbial metabolism: detection of [14C]dihydroxyacetone during assimilation of [uC]methanol by Hansenulapolymorpha . FEMS Microbiology Letters 8:13–16
    [Google Scholar]
  9. O’Connor M., Quayle J. R. 1979; Mutants of Hansenulapolymorpha and Candida boidinii impaired in their ability to grow on methanol. Journal of General Microbiology 113:203–208
    [Google Scholar]
  10. Van Dijken J. P., Otto R., Harder W. 1976; Growth of Hansenulapolymorpha in a methanol- limited chemostat.Physiological responses due to the involvement of methanol oxidase as a key enzyme in methanol metabolism. Archives of Microbiology 111:137–144
    [Google Scholar]
  11. Van Dijken J. P., Harder W., Beardsmore A. J., Quayle J. R. 1978; Dihydroxyacetone: an intermediate in the assimilation of methanol by yeasts?. FEMS Microbiology Lettrs 4:97–102
    [Google Scholar]
  12. Waites M. J., Quayle J. R. 1980; Dihydroxyacetone: a product of xylulose 5-phosphate- dependent fixation of formaldehyde by methanol-grownCandida boidinii . Journal of General Microbiology 118:321–327
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-120-1-219
Loading
/content/journal/micro/10.1099/00221287-120-1-219
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