Cellular content of the Krebs cycle keto acids in yeasts grown on different nitrogen sources, including hydroxylamine.

The cellular pool of Krebs cycle keto acids was followed as a function of growth in three yeasts. The keto acids were analyzed as silylated methoximes by quantitative gas chromatography with capillary glass columns. The 2-oxoglutaric acid content was strikingly high in the hydroxylamine (HA)-tolerant, HA-utilizing Endomycopsis lipolytica when compared to that in the nitrate-utilizing yeast Cryptococcus albidus and Saccharomyces cerevisiae, requiring fully reduced nitrogen for growth. The content of E. lipolytica increased throughout the log phase to maxima of about 200-250 microgram per g dry weight in HA and ammonia media. These amounts are 20-25 times greater than those attained in the two other yeasts. The cellular content of pyruvic acid was at a maximum early in the log phase, amounting to 50-70 microgram per g dry weight for all yeasts. The oxalacetic acid content never exceeded 9 microgram per g dry weight in any of the yeasts. Oximeformation, for which keto acid production is a prerequisite, is discussed as part of the HA-tolerance.