52] Production of 6-aminopenicillanic acid with immobilized Escherichia coli acylase

Abstract

This chapter presents the production of 6-aminopenicillanic acid with immobilized Escherichia coli acylase. A new procedure for purification of the enzyme, adapted for work on a production scale, has been developed recently. This purification method gives higher yields, as well as a method for producing the enzyme covalently bound (CVB) to Sephadex G-200 and a basic production method for 6-APA using this Sephadex G-200 CVB-E. coli acylase.. Assay methods have been developed based on either of these products. Photometric methods using hydroxylamine s or p-dimethylaminobenzaldehyde as reagents for the determination of the amount of 6-APA formed appear to be best suited for analysis of large series of samples—for example, automatic equipment. The method preferred to determine the phenylacetic acid by titration at constant pH with a pH star. It is well suited for single analyses, is rapid and easy to carry out, and gives high accuracy as well as reproducibility.