The aim of industrial enzymic amoxycillin production: Characterization of a novel carbamoylase enzyme in the form of a crude, cell-free extract

Amoxycillin production involves the generation of a racemic mixture of hydroxyphenylhydantoin by means of the Bucherer synthesis, The hydantoin is enzymically cleaved by a D-(-)-specific hydantoinase to form D-(-)-N-carbamoylhydroxyphenylglycine. This is subsequently hydrolysed by a carbamoylase enzyme that generates the amino acid derivative, D-(-)-hydroxyphenylglycine. The remaining L-(+)hydroxyphenylhydantoin spontaneously racemizes, allowing continual cleavage to continue (by the hydantoinase action) and giving a potential 100% yield of end product from the two-enzyme-catalysed process, rather than 50%, A novel carbamoylase from an Agrobacterium species has been studied in a crude (cell-free extract) form, The temperature stability was shown to be remarkable, with no loss of activity detected after 4 h at 50 degrees C. Kinetic values were calculated for the enzyme, with a variety of implications for a future industrial process, The substrate specificity, isoelectric point, pH and temperature activity profiles were elucidated, with the aim of generating a commercially beneficial method of purifying the enzyme, In addition, the concentration-dependent toxicities of several metal ions were studied; all bivalent ions studied were found to have some detrimental effect on activity.