Measuring Enzyme Kinetics of Glycoside Hydrolases Using the 3, 5-Dinitrosalicylic Acid Assay

• Use of the 3,5-dinitrosalicylic acid reagent allows the simple, rapid quantification of reducing sugars. The method can be used for analysis of biological samples or in characterization of enzyme reactions, as new reducing ends are generated when a polysaccharide substrate undergoes hydrolytic cleavage. Presented here is an application of the method in measuring the kinetics of a glycoside hydrolase reaction, including the optimization of the DNSA reagent, and the production of a standard curve of absorbance versus sugar concentration.

Ämnesord

NATURVETENSKAP  -- Biologi -- Biokemi och molekylärbiologi (hsv//swe)

NATURAL SCIENCES  -- Biological Sciences -- Biochemistry and Molecular Biology (hsv//eng)

Nyckelord

Reducing sugars

Enzyme kinetics

Glycoside hydrolase

UV/visible spectrophotometry

3

5-dinitrosalicylic acid

Publikations- och innehållstyp

ref (ämneskategori)

kap (ämneskategori)