CAPILLARY ELECTROPHORESIS IN THE ASSAY OF THE HYDROLYSIS OF GLYCINE-CONTAINING PEPTIDES BY A PROTEASE FROM PSEUDOMONAS-AERUGINOSA

A rapid and simple capillary zone electrophoresis (CZE) method for mea suring the activity of a minor protease from Pseudomonas aeruginosa is described. When glycine-containing oligopeptides were used as substra tes, it was possible to separate and quantify substrate and products. Moreover oligopeptide hydrolysates were analysed by fast atom bombardm ent mass spectrometry providing the sites of splitting of the substrat es. By comparison with CZE calibration curves constructed with referen ce peptides, the initial rates of hydrolysis were calculated. The meth od, validated for pentaglycine hydrolysis, was also used for the analy sis of reaction mixtures and for monitoring the enzymic hydrolysis of various peptides in order to investigate enzyme specificity. (C) 1997 Elsevier Science B.V.