Soluble, dye-labelled substrates for a micro-plate assay of proteinase activity

• Published in: Journal of microbiological methods Vol. 25; no. 3; pp. 337 - 342
• Main Authors: Wolf, Gerhard A., Wirth, Stephan J.
• Format: Journal Article
• Language: English
• Published: Shannon Elsevier B.V 1996; Elsevier Science
• Subjects: Bacteriological methods and techniques used in bacteriology; Bacteriology; Biological and medical sciences; Casein; Collagen; Fundamental and applied biological sciences. Psychology; Gelatin; Micro-plate assay; Microbiology; Plate-clearing assay; Proteinase; Remazol Brilliant Blue R; Casein; Gelatin; Remazol Brilliant Blue R; Collagen; Proteinase; Micro-plate assay; Plate-clearing assay; Enzyme; Glycoproteins; Colorimetry; Method; Fungi; Enzymatic activity; Proteolysis; Hydrolases; Bacteria; Detection; Proteinases; Thallophyta
• ISSN: 0167-7012; 1872-8359
• DOI: 10.1016/0167-7012(96)00005-X

Gelatin, collagen and casein derivatives were covalently linked with Remazol Brilliant Blue R (RBB). New dye-labelled, soluble substrates were obtained, suitable for the detection and assay of proteinase activity. A reliable colorimetric micro-plate assay was designed, based on the precipitability of gelatin-RBB, collagen-RBB and casein-RBB from buffered solutions with hydrochloric acid. Additionally, a sensitive plate-clearing assay was developed for the detection and isolation of proteolytic microorganisms from mixed populations, or for the screening of pure cultures.