The DNSA reducing sugar assay: a how-to guide for glycoside hydrolases.

What is it about?

This chapter introduces the concepts behind the 3,5-dinitrosalicylic acid assays for reducing sugar quantification, and describes how it can be used to study the kinetics of glycoside hydrolase reactions. It provides step by step instructions, tips on how to optimise the method, and health and safety considerations.

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Why is it important?

The DNSA method was first described by GL Miller in 1959, in an excellent and thorough assessment of the technique and its limitations. The method is very widely applied in the characterisation of carbohydrate active enzymes, with a lot of variation in how reagents and samples are prepared. This chapter makes recommendations on how to optimise the various reagent components, and specifically describes the use of the method as applied to glycoside hydrolases.