The kinetic mechanism of penicillin V acylase from Streptomyces lavendulae

Penicillin V acylase isolated from Streptomyces lavendulae is an extracellu lar enzyme with interesting properties for the industrial production of 6-A PA. The enzyme hydrolysed penicillin V and synthetic analogues such as 2-ni tro-5-(phenoxyacetamido)-benzoic acid (NIPOAB). The corresponding kinetic p arameters were K-M = 3 mM and V-max = 92 mu mol min(-1) mg(-1) for penicill in V K-M = 15.3 mM and V-max = 14.5 mu mol.min(-1) mg(-1) for NIPOAB. Studi es of inhibition by products, alternative products and dead-end inhibition supported an ordered uni bi mechanism that could involve an acyl-enzyme int ermediate as has been described for penicillin G acylases and other amidase s.