Extracellular acid proteases produced by Saccharomycopsis lipolytica.
Saccharomycopsis lipolytica CX161-1B produced at least three extracellular acid proteases during exponential growth in medium containing glycerol, Difco Proteose Peptone, and mineral salts at pH 3.4 (Difco Laboratories, Detroit, Mich.). Little extracellular acid protease activity was produced with glutamic acid as the sole nitrogen source, somewhat higher levels were obtained with peptone, and much higher levels were obtained with Difco Proteose Peptone. The relative amounts of the three proteases varied during growth on Difco Proteose Peptone, which suggested that the proteases were not coordinately regulated. The proteases were purified to near homogeneity (as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis) by use of ultrafiltration, gel filtration, and DEAE-Sephacel and hydroxylapatite chromatography. Protease I had a molecular weight near 28,000, an isoelectric point of pH 4.9, and a pH optimum of 3.5. Protease II had a molecular weight near 32,000 and a pH optimum of 4.2. Protease III had a molecular weight near 36,000, an isoelectric point of 3.8, and a pH optimum of 3.1. All three proteases were glycoproteins; proteases I, II, and III contained 25, 12, and 1.2% carbohydrate, respectively. The proteases were inhibited by pepstatin and 1,2-epoxy-3-(4-nitrophenoxy) propane and were largely insensitive to diazoacetyl-DL-norleucine methylester and to compounds which inhibit the serine, sulfhydryl, or metallo-proteases.
D001203:Ascomycota / Q000201:enzymology*; D016282:Aspartic Acid Endopeptidases; D002241:Carbohydrates / Q000032:analysis; D010450:Endopeptidases / Q000096:biosynthesis* / Q000302:isolation & purification; D005110:Extracellular Space / Q000201:enzymology*; D006863:Hydrogen-Ion Concentration; D007526:Isoelectric Point; D008970:Molecular Weight; D011480:Protease Inhibitors; D012442:Saccharomycopsis / Q000201:enzymology* / Q000254:growth & development
D002241:Carbohydrates; D011480:Protease Inhibitors; D010450:Endopeptidases; D016282:Aspartic Acid Endopeptidases