CHAINIA PENICILLIN-V ACYLASE - STRAIN CHARACTERISTICS, ENZYME IMMOBILIZATION, AND KINETIC-STUDIES

Aerobic cultures of an actinomycete were found to produce penicillin V acylase (PVA) (PA, EC-3.5.1.11) extracellularly. The presence of L-2- 3 diamino-propionic acid in cell wall and formation of sclerotia on cu lture media led to its identification as Chainia, a sclerotial Strepto myces. Partially purified acylase was adsorbed on kieselguhr and entra pped in polyacrylamide gel. The immobilized preparation proved effecti ve with respect to retention of enzyme and enzyme activity even after 15 successful cycles. The pH optimum for crude enzyme was in the range of pH 7.5-8.0, and for the (NH4)(2)SO4 fraction it was pH 8.5. The im mobilized enzyme showed maximal activity at pH 9.5. The optimum temper ature for acylase activity was at 55 degrees C. The crude enzyme, ammo nium sulfate fraction, and immobilized enzyme showed K-m value for pen icillin V of 6.13 mM, 14.3 mM, and 17.1 mM, respectively.