Analysis of the lysozyme-catalyzed hydrolysis and transglycosylation of N-acetyl-d-glucosamine oligomers by high-pressure liquid chromatography

Although the enzyme lysozyme is one of the most thoroughly studied enzymes, quantitative kinetic studies of the lysozyme-catalyzed hydrolysis can only be achieved in a limited set of circumstances. The use of chromophoric substrates is beset by a number of difficulties and no low molecular weight substrate has gained acceptance in a standard assay. This report describes an efficient and rapid technique for monitoring the lysozyme-catalyzed hydrolysis and transglycosylation of β(1 → 4)-linked N-acetyl-d-glucosamine oligomers that utilizes high-pressure liquid chromatography to separate the products and liquid scintillation counting to ascertain their concentrations.