MILLON氏反應によるProteaseの比色定量法

概要

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A few colorimetric methods are hitherto used for the determination of the proteolytic activity, but not simple and inexpensive.In this paper, a simple colorimetric method, using the color reaction between tyrosine and MILLON's reagent, is described.The procedure is as follows : To each volume of the digestion mixture add an equal volume of the solutions of trichloracetic acid and phosphotungstic acid, and filter after 30 min. To 2 cc of the filtrate add 1 cc of MILLON's reagent A (15% HgSO_4 in 7N H_2SO_4) and 0.15 cc of the same B (0.8% NaNO_2 solution). The color is read in photoelectric colorimeter (filter 530 mμ) after 30 min at 37℃. The concentration of tyrosine is calculated from the regression line or formula.Digestion curves obtained by plotting the tyrosine-value of the digestion mixtures, freed of unaltered protein, against the time of digestion by the enzymes extracted from wheat bran-koji may be classified into two types : the one is sigmoidtype and the other is a type bearing resemblance to monomolecular reaction curve. The difference in types may be due to the difference in the strains employed for the preparation of koji.
社団法人日本生物工学会の論文
1954-03-15