title: Purification and properties of extracellular carboxyl proteinase secreted by Candida pulcherrima.
abstract: An extracellular proteinase secreted by Candida pulcherrima KSY 188-5 was purified about 60-fold to electrophoretical homogeneity from its culture supernatant, by ammonium sulfate fractionation, anion-exchange chromatography, and gel filtration. The proteinase had a molecular weight of approximately 36,500 and an isoelectric point of pH 4.7. The enzyme had an optimum pH of around 2.5-3.5 for activity and 3.0-5.0 for stability. The optimum temperature was around 45 degrees C at pH 3.0. The enzyme showed a broad substrate specificity for a variety of proteins to hydrolyze casein, BSA, hemoglobin keratin, and collagen. Among several proteinase inhibitors, pepstatin A completely abolished the enzyme activity; indicating that the extracellular proteinase from C. pulcherrima KSY 188-5 was classified in the group of carboxyl proteinases.
authors.affiliation: Department of Materials Engineering and Applied Chemistry, Mining College, Akita University, Japan.
authors.forename: T
authors.initials: T
authors.lastname: Gotoh
authors.forename: K
authors.initials: K
authors.lastname: Kikuchi
authors.forename: K
authors.initials: K
authors.lastname: Kodama
authors.forename: H
authors.initials: H
authors.lastname: Konno
authors.forename: T
authors.initials: T
authors.lastname: Kakuta
authors.forename: T
authors.initials: T
authors.lastname: Koizumi
authors.forename: K
authors.initials: K
authors.lastname: Nojiro
mesh_terms: D000596:Amino Acids / Q000032:analysis; D016282:Aspartic Acid Endopeptidases / Q000737:chemistry / Q000302:isolation & purification*; D002175:Candida / Q000201:enzymology*; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D010450:Endopeptidases / Q000737:chemistry / Q000302:isolation & purification*; D005285:Fermentation; D005656:Fungal Proteins / Q000032:analysis / Q000302:isolation & purification; D006863:Hydrogen-Ion Concentration; D007525:Isoelectric Focusing; D008970:Molecular Weight; D011480:Protease Inhibitors / Q000737:chemistry / Q000494:pharmacology; D013696:Temperature
chemical_list: D000596:Amino Acids; D005656:Fungal Proteins; D011480:Protease Inhibitors; D010450:Endopeptidases; D016282:Aspartic Acid Endopeptidases; C093457:extracellular carboxyl proteinase, Candida pulcherrima