• Microbiology and Biotechnology Letters
• /
• v.15 no.6
• /
• pp.420-424
• /
• 1987

Isocitrate lyase from crude extract of Saccharomycopsis lipolytica ATCC44601 and MX9-11RX8 temperature-sensitive mutant was purified about 54 times and 87 times, respectively by ammonium sulfate fractionation, Toyo peal HW-55F gel filtration and DEAE-Cellulose ion exchange chromatography, The molecular weight of the purified isocitrate lyase from this yeast was estimated to be 230, 000 by gel filtration on Sephadex G-200, and SDS-polyacrylamide Eel electrophoresis showed that the enzyme consisted of four identical or similar subunits with a molecular weight of 59, 000 and the enzyme showed optimum activity at pH 6.9.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.17 no.3
• /
• pp.277-281
• /
• 1988

Effect of itaconate upon citrate and isocitrate production of Saccharomycopsis lipolytica were investigated. The percent inhibition of isocitrate lyase activity was about 80% at 0.8 mM itaconate concentration, with Ki value of 0.17 mM in the cleavage reaction. Inhibitory effect of itaconate at 20 mM on S. lipolytica growth was significant on n-hexadecane medium, whereas almost no inhibitory effect on glucose medium. Increasing itaconate concentration in n-hexadecane medium improved isocitrate production up to 80% but no difference was found in glucose medium.

• Journal of Microbiology and Biotechnology
• /
• v.1 no.2
• /
• pp.130-135
• /
• 1991

The effects of media composition on citric acid fermentation using surface immobilized Saccharomycopsis lipolytica were studied. The use of the standard medium for these organisms resulted in rapid decrease of citric acid production and a transformation of immobilized cell morphologies from a yeast-type to a mycelium-type. When the standard medium was enriched with vitamins, trace minerals, a growth factor and ammonium to form a Vigorous Stationary Phase (VSP) fermentation type medium, relatively stable citric acid production (10 mg/lㆍh) was obtained. Using the VSP type medium, the surface immobilized cells also retained their yeast-type form.

• The Korean Journal of Food And Nutrition
• /
• v.1 no.1
• /
• pp.25-32
• /
• 1988

Properties of purified isocitrate lyase from Saccharomycopsis lipolytica ATCC 44601 and MX9-11RX8 temperature-sensitive mutant were investigated. Purified isocitrate lyase from temperature-sensitive mutant was indistinguishable from the wild type enzyme with respect to the isoelectric pH(5.3), the thermostability and Km value for threo-Ds-Isocitrate(about 0.2 mM). When isocitrate lyase induced by acetate minimal medium at 33$^{\circ}C$, MX9-11RX8 mutant did not express enzyme activity but did synthesize polypeptide chain whose electrophoretic mobilities were equal to those of the purified enzymes.

• Microbiology and Biotechnology Letters
• /
• v.13 no.3
• /
• pp.179-184
• /
• 1985

Sexually compatible heterothallic haploids and diploids therefrom of Saccharomycopsis lipolytica were compared with respect to production. Diploids constructed through mating were confirmed by random spore analysis, whereas those constructed through protoplast fusion were confirmed by haploidization. ATCC 44601 and IFO 1209 produced larger amount of citrate and isocitrate than IFO 1550 and IFO 1551. A mode of citrate production by diploids was intermediate of the parental haploid strains. The specific activities of citrate synthase and isocitrate lyase in IFO 1550 and IFO 1551 were higher than those in ATCC 44601 and IFO 1209, indicating little correlation between citrate production and specific activities of these enzymes.

• Applied Biological Chemistry
• /
• v.29 no.1
• /
• pp.3-9
• /
• 1986

A yeast strains, CJ 2, CJ 7 and CJ 8, isolated from soil and contaminated choose, mated with authentic strains of Saccharomycopsis lipolytica and were identified as Saccharomycopsis lipolytica with mating A, B and B, respectively. The strain CJ 7 produced large amount of isocitric acid in glucose and n-hexadecane medium as compared with another strains. All strains produced larger amount of citric acid in n-hexadecane medium as compared with glucose medium, and citric acid production of diploids was greater than that of the parental haploid strains. The specific activity of isocitrate lyase in n-hexadecane grown cells was $15{\sim}20$ times greater than that in glucose-grown cells, but the specific activity of citrate synthetase was not so influenced by carbon source. Little correlation between citric acid production and the specific acitivity of these enzymes was noticed irrespective of strains and ploidy.

• Microbiology and Biotechnology Letters
• /
• v.15 no.6
• /
• pp.414-419
• /
• 1987

Temperature-sensitive revertant could grow on acetic acid at 23$^{\circ}C$ but not at 33$^{\circ}C$, MX9-11RX8, isolated from mutant deficient in the activity of isocitrate lyase and its properties were investigated. The activity of isocitrate lyase and specific rate of isocitrate lyase synthesis decreased according to in-crease culture temperature from 23 to 33 $^{\circ}C$ in acetic acid as carbon source. A rapid cessation of in-crease enzyme activity observed when the temperature was shift up from 23 to 33$^{\circ}C$ but cell growth was continued. On the other hand, the revertant also exhibited temperature-sensitive in n-hexade-cane medium as carbon source, and the amount of isocitric acid was nearly equal produced to that at 23 $^{\circ}C$ when the temperature shift up from 23 to 33 $^{\circ}C$.

• Applied Biological Chemistry
• /
• v.31 no.2
• /
• pp.137-142
• /
• 1988

The analysis of condensation and cleavage reaction was carried out at $30^{\circ}C$ and pH 7.0 with purified isocitrate lyase from Saccharomycopsis lipolytica ATCC 44601. The Km values for condensation reaction of glyoxylate and succinate were 0.06 and 0.21 mM, respectively. In the cleavage reaction, glyoxylate was a linear competitive inhibitor with a Ki of 0.22 mM and succinate was a linear noncompetitive inhibitor with a Ki of 0.82 mM. Therefore, these kinetic analyses showed that the enzyme functioned in a ordered reaction with glyoxylate binding before succinate in the condensation reaction. 3-Bromopyruvate(BrP) was found to be irreversibly inactivation showing saturation kinetics, the inactivation half-time was 0.15 min and $K_{BrP}$ was 0.032 mM, and substrate or reactant protected against the inactivation.

• Microbiology and Biotechnology Letters
• /
• v.17 no.6
• /
• pp.624-628
• /
• 1989

A study on the citric acid production using Saccharomycopsis lipolytica (NRRL Y7576) was carried out in shake-flasks, air-lift and membrane recycle bioreactors. The cells entrapped in Ca-alginate beads were used in shake-flasks and air-lift reactor. Repeated batch fermentation in shake-flasks was successfully performed for 34 days and resulted in a yield of 54%. Increased yield (63%) was obtained in the air-lift reactor operation using nitrogen deficient medium (NDM). In the membrane recycle bioreactor operation, the maximal dry cell mass concentration was 39 g/1 at a dilution rate of 0.02 h$^{-1}$ and the yield with NDM was higher than that with growth medium. In addition, the yield and volumetric productivity with pure oxygen supply were greatly improved compared with those with air supply.

• Korean Journal of Microbiology
• /
• v.34 no.3
• /
• pp.75-81
• /
• 1998

The yeast strain 504D, isolated from salted shrimp soup, showed the best proteolytic activity under alkaline condition. The yeast formed vegetative cells in almost optimal media for yeasts, but formed only pseudohyphae in the MM medium containing citric acid and true hyphae in the MM medium containing N-acetylglucosamin and ${\beta}$-D-glucose. The yeast was classified as hemiascomycetes to form ascospores by sexual reproduction, and formed blastospores and athrospores by asexual reproduction. The yeast strain did not assimilate almost of the carbon sources, nitrate and nitrite, but some organic acids and alcohols. The fatty acids of whole cells were composed of 53.67% unsaturated fatty acids and 14.58% saturated, and, especially, C17:1 was observed in this strain but not in two control yeasts. However, almost of all results were very similar to the morphological and physiological characteristics of Yarrowia lipolytica KCCM 12495 and KCCM 35426, except for a little differences which are the composition of fatty acids and the manner of mycellium formation. Therefore, the isolated yeast strain 504D is identified as a Yarrowia lipolytica.