Changes in proteinase activities and subcellular distribution during inactivation of alcohol oxidase in Candida boidinii.
Adaptation of methanol-grown Candida boidinii to ethanol utilization was accompanied by an increase in proteolytic activities, which behaved like known vacuolar enzymes. Degradation of alcohol oxidase protein was partially prevented by the serine proteinase inhibitor phenylmethanesulphonyl fluoride, but not by the carboxyl proteinase inhibitor pepstatin. Fractionation of cell-free extracts, by high-speed zonal centrifugation, of methanol-grown C. boidinii showed non-sedimentable and sedimentable proteolytic activities. Naturally occurring inhibitors of vacuolar proteinases were non-sedimentable. Fractionation of extracts prepared from methanol-grown cells which had been adapted to ethanol utilization for 5 h revealed significant changes in the sedimentability and distribution of proteolytic and acid phosphatase activities. These results suggest the possible involvement of a vacuolar process during alcohol oxidase degradation.
D000429:Alcohol Oxidoreductases / Q000037:antagonists & inhibitors*; D002175:Candida / Q000187:drug effects / Q000201:enzymology*; D002499:Centrifugation, Density Gradient; D002738:Chloroquine / Q000494:pharmacology; D010450:Endopeptidases / Q000378:metabolism*; D000431:Ethanol / Q000378:metabolism; D000432:Methanol / Q000378:metabolism; D010436:Pepstatins / Q000494:pharmacology; D010664:Phenylmethylsulfonyl Fluoride / Q000494:pharmacology; D013347:Subcellular Fractions / Q000201:enzymology
D010436:Pepstatins; C014195:Streptomyces pepsin inhibitor; D000431:Ethanol; D010664:Phenylmethylsulfonyl Fluoride; D002738:Chloroquine; D000429:Alcohol Oxidoreductases; C019643:alcohol oxidase; D010450:Endopeptidases; C031375:pepstatin; D000432:Methanol