Insoluble dye-protein complexes offer many advantages when used as substrates for proteolytie enzymes. The proteolytic split products can be separated from the starting substrat.e conveniently by filtration, and the concentration of the soluble colored products of hydrolysis can easily be measured a
Assay of proteolytic enzyme with a leucine-selective electrode
β Scribed by Paul T. Chien; Leslie W. Michael
- Publisher
- Elsevier Science
- Year
- 1975
- Tongue
- English
- Weight
- 293 KB
- Volume
- 68
- Category
- Article
- ISSN
- 0003-2697
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β¦ Synopsis
A potentiometric method utilizing a leucine-selective membrane electrode has been developed for the assay of proteolytic enzyme. Subtilisin Carlsberg was used for the enzymatic digestion of casein in solution. Responses on the reaction media were linear on a semilog plot for active enzyme concentrations ranging from 0.6-60 fig/ml. The system uses commercially available components as well as simplified methods of solution handling without precipitation or filtration steps. This method offers significant advantages in terms of simplicity, speed, and cost over the traditional techniques.
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Chymotrypsin, covalently immobilized to the surface of an IrO,-coated titanium electrode, responds potentiometrically to various substrates. A flow-injection system is described for assay of the activity of the immobilized enzyme with N-benzoyl-L-tyrosine ethyl ester as substrate and an ultraviolet