Measurement of casein digestion by a fluorometric method.
A qualitative and quantitative method to assay proteolytic degradation of casein with a spectrofluorometer was developed. Proteolysis produced by different pure or mixed proteinases in a pH range 2 to 7.4 quenches the fluorescence emitted at a wavelength of 350 nm by casein excited at 300 nm in less than 5 min. This method is very sensitive, fast, and requires minimal sample preparation. Proteinases that do not generate peptides appropriate for fluorescence quenching cannot be detected with this assay and proteinases with intrinsic fluorescence may require special adjustments of the spectrofluorometer. This method monitors the disappearance of intact substrate proteins continuously, omitting the separation step necessary in other methods to measure product peptides.
Ostoa-Saloma
P
P
Departamento de Microbiología, Universidad Nacional Autónoma de México, Mexico, D.F.
Ramírez
J
J
Perez-Montfort
R
R
D000818:Animals; D002364:Caseins / Q000378:metabolism*; D002417:Cattle; D003013:Clostridium / Q000201:enzymology; D010450:Endopeptidases / Q000032:analysis; D005470:Fluorometry / Q000379:methods; D008892:Milk / Q000378:metabolism; D013025:Glycine max / Q000201:enzymology; D013305:Streptomyces griseus / Q000201:enzymology
D002364:Caseins; D010450:Endopeptidases