Simultaneous assay of dihydroxyacetone synthase and transketolase in a methylotrophic yeast grown in continuous culture. A cautionary note.
The methylotrophic yeast Candida boidinii CBS 5777 was grown in continuous culture under carbon limitation on glucose, glucose plus methanol, and methanol as carbon and energy sources. During adaptation from glucose to methanol there was a rapid rise in the specific activities of triokinase, fructose-1,6-bisphosphatase and dihydroxyacetone synthase, which are key enzymes of the xylulose phosphate cycle of formaldehyde fixation. The specific activity of classical transketolase fell during this adaptation. Extracts from carbon-limited C. boidinii contained an enzyme which catalysed oxidation of NADH when some preparations or ribose 5-phosphate were added, which was not a transketolase. This enzyme activity was dependent on an impurity in such ribose 5-phosphate preparations and can be confused with transketolase activity.
D019880:Aldehyde-Ketone Transferases; D002175:Candida / Q000201:enzymology*; D006597:Fructose-Bisphosphatase / Q000378:metabolism; D005947:Glucose / Q000378:metabolism; D006358:Hot Temperature; D000432:Methanol / Q000378:metabolism; D008828:Microbiological Techniques; D010428:Pentosephosphates / Q000378:metabolism; D012267:Ribosemonophosphates / Q000378:metabolism; D013997:Time Factors; D014166:Transferases / Q000032:analysis* / Q000378:metabolism; D014174:Transketolase / Q000032:analysis* / Q000378:metabolism
D010428:Pentosephosphates; D012267:Ribosemonophosphates; C031625:xylulose-5-phosphate; D014166:Transferases; D019880:Aldehyde-Ketone Transferases; D014174:Transketolase; C031222:formaldehyde transketolase; D006597:Fructose-Bisphosphatase; D005947:Glucose; D000432:Methanol