Purification and characterization of the protease enzymes of Streptomyces thermovulgaris grown in rapemeal-derived media

• Published in: Journal of applied microbiology Vol. 82; no. 2; pp. 149 - 156
• Main Authors: YEOMAN, K. H, EDWARDS, C
• Format: Journal Article
• Language: English
• Published: Oxford Blackwell Science 01.02.1997
• Subjects: Biological and medical sciences; Biology of microorganisms of confirmed or potential industrial interest; Biotechnology; Chromatography, Ion Exchange; Culture Media; Endopeptidases - biosynthesis; Endopeptidases - isolation & purification; Endopeptidases - metabolism; Enzyme Stability; Fundamental and applied biological sciences. Psychology; Isoelectric Point; Kinetics; Miscellaneous; Mission oriented research; Streptomyces - enzymology; Streptomyces - growth & development; Substrate Specificity; Culture medium; Streptomycetaceae; Purification; Enzyme; Thermal stability; Actinomycetales; Characterization; Peptidases; Enzymatic activity; Production; Microorganism culture; Bacteria; Hydrolases; Actinomycetes; Rapeseed meal; Physicochemical properties
• ISSN: 1364-5072; 1365-2672
• DOI: 10.1111/j.1365-2672.1997.tb02845.x

When grown in a particulate-free, protein-rich medium derived from rapemeal (termed medium B), Streptomyces thermovulgaris produced multiple protease enzymes. The main protease activity was attributed to two types of serine protease, denoted as SV1 and SV2. A metallo protease component (SV3) and an azocaseinase component (SV4) were also present. Protease SV1 had a molecular weight of 30 kDa and a pI of 5.8. Protease SV2 was characterized by a high thermostability in the presence of calcium ions and had a pI of 8.4. This enzyme had a molecular weight of 60 kDa, but we suggest that this is the dimeric form, with 30 kDa being the monomer unit. The method chosen for initial downstream processing influenced both the yield and type of protease purified. When cell-free supernatant fluid was concentrated using ultrafiltration, rather than acetone precipitation, a higher percentage and a greater range of proteases were recovered. The medium used for the growth of Strep. thermovulgaris also appeared to affect the type of protease produced. A more diverse range of proteases were produced on rapemeal-derived medium when compared to yeast extract medium.