The present application belongs to the field of
enzyme activity determination method, aiming at the problems of existing
pectinase activity determination technology, such as complicated operation, low sensitivity, and the problems of existing MBTH method for determining
pectinase activity, such as a large amount of precipitate in the
chromogenic product solution and the high background value of the
chromogenic product, the present application provides a kind of determination method of
pectinase activity: a series of
concentration gradient galacturonic acid standard solution is prepared by using substrate solution, the precipitate is provided by using alkaline
reagent to provide
high concentration alkaline environment, and the
precipitation of precipitated material is accelerated by using low temperature condition, then the supernatant is obtained by
centrifugation, MBTH reaction is carried out, the
absorbance value is determined, and the standard corresponding relationship curve is drawn;
Pectinase enzymolysis reaction; preparation of enzymolysis blank; the initial speed of the concentration of reducing
end group produced by the enzymolysis of the substrate per unit time is determined to calculate the
enzyme activity of pectinase. The method is accurate, reliable, good in
repeatability, high in sensitivity and stable in
chromogenic product.