BACILLUS-SPHAERICUS PENICILLIN-V ACYLASE - PURIFICATION, SUBSTRATE-SPECIFICITY, AND ACTIVE-SITE CHARACTERIZATION

Penicillin V acylase from Bacillus sphaericus was purified to homogene ity with an overall yield of 15%. The enzyme exhibited comparatively h igh specificity for penicillin V, penicillin G, and other related comp ounds being hydrolyzed at less than 10% of the rate of penicillin V. M oreover, the high rate of hydrolysis was observed when the side chain of the substrate molecule was unsubstituted. Lysine-modifying reagents inactivated the enzyme rapidly. Kinetics and titration studies indica ted the involvement of lysine in the catalytic activity of the enzyme.