title: Extracellular proteinase from Enterococcus faecalis subsp. liquefaciens. II. Partial purification and some technological important properties.
abstract: An extracellular proteinase from Enterococcus faecalis subsp. liquefaciens has been purified 780-fold by a method including gel filtration on Sephadex G-50 and affinity chromatography with gramicidin J as ligand. Approximately 15% of the original enzyme activity was recovered. A purification of 14,800-fold, with 11.4% yield, may be reached using chromatofocusing as final step in the purification procedure. The molar mass of the enzyme has been estimated to be approximately 30 kDa by Sephadex gel filtration and approximately 26 kDa by SDS-PAGE. The isoelectric point has been found to be 4.6. Maximum enzyme activity of the proteinase has been observed at pH 7.5 and 45 degrees C. The enzyme hydrolyzed bovine serum albumin, alpha-lactoalbumin, beta-lactoglobulin, casein and pork myofibrillar and sarcoplasmic proteins. The extracellular proteinase was very stable; the enzyme maintained its activity in cell-free extracts over a very wide range of temperatures (-25 to 37 degrees C) for at least 2 months. At 12 degrees C, it was stable in the pH range of 5.5 to 8.0.
authors.affiliation: Departamento de Nutrición y Bromatología III (Higiene y Tecnología de los Alimentos), Facultad de Veterinaria, Universidad Complutense, Madrid, Spain.
authors.forename: G D
authors.initials: GD
authors.lastname: García de Fernando
authors.forename: P E
authors.initials: PE
authors.lastname: Hernández
authors.forename: J
authors.initials: J
authors.lastname: Burgos
authors.forename: B
authors.initials: B
authors.lastname: Sanz
authors.forename: J A
authors.initials: JA
authors.lastname: Ordóñez
mesh_terms: D002611:Cheese / Q000382:microbiology; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D010450:Endopeptidases / Q000737:chemistry / Q000302:isolation & purification*; D013293:Enterococcus faecalis / Q000201:enzymology* / Q000302:isolation & purification; D004795:Enzyme Stability; D005110:Extracellular Space / Q000201:enzymology; D005516:Food Microbiology; D005524:Food Technology; D006863:Hydrogen-Ion Concentration; D007526:Isoelectric Point; D008970:Molecular Weight
chemical_list: D010450:Endopeptidases