title: Purification and characterization of intracellular proteinase from Lactobacillus casei ssp. casei LLG.
abstract: The intracellular proteinase of Lactobacillus casei ssp. casei LLG was isolated in the cytoplasmic fraction with 0.05 M Tris-HCl buffer (pH 7.5). The enzyme was purified by the fast protein liquid chromatography system equipped with ion-exchange and gel filtration chromatographies. This proteinase comprised a single monomeric form and had a molecular weight of about 55 kDa and an isoelectric point near pH 4.9. The optimum pH and temperature for the enzyme activity were determined to be pH 6.5 and 37 degrees C, respectively. The enzyme was inactivated by metal-chelating compounds (EDTA, 1,10-phenanthroline) and less affected by serine proteinase inhibitors (diisopropylfluorophosphate, phenylmethylsulfonyl fluoride). Proteinase activity was increased by Ca++, Mn++, and Co++, and inhibited by Cu++, Mg++, and Zn++. The activity of this enzyme to hydrolyze casein appeared to be more active on beta-casein than alphas1-casein and kappa-casein as monitored by polyacrylamide gel electrophoresis.
authors.affiliation: Department of Food Science and Agricultural Chemistry, McGill University, 21,111 Lakeshore Road, Ste-Anne-de-Bellevue, Quebec Canada H9X 3V9.
authors.forename: J Y
authors.initials: JY
authors.lastname: Shin
authors.forename: W M
authors.initials: WM
authors.lastname: Jeon
authors.forename: G-B
authors.initials: GB
authors.lastname: Kim
authors.forename: B H
authors.initials: BH
authors.lastname: Lee
mesh_terms: D002413:Cations, Divalent / Q000494:pharmacology*; D002611:Cheese / Q000382:microbiology; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D002853:Chromatography, Liquid; D004587:Electrophoresis, Agar Gel; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D007780:Lacticaseibacillus casei / Q000201:enzymology*; D008970:Molecular Weight; D010447:Peptide Hydrolases / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D011480:Protease Inhibitors / Q000494:pharmacology*; D013696:Temperature
chemical_list: D002413:Cations, Divalent; D011480:Protease Inhibitors; D010447:Peptide Hydrolases