Cephalosporin C acylase from Pseudomonas species: Production and enhancement of its activity by optimization of process parameters

The bacterium, Pseudomonas species was isolated from rhizosphere soil and screened out for its ability to produce cephalosporin C acylase (CCA). Cephalosporin C acylase has the potential to transform cephalosporin C to 7- aminocephalosporanic acid which is an important intermediate for production of broad-spectrum semi-synthetic cephalosporins. This article describes the selection of medium components and parameters like temperature, pH, carbon sources, nitrogen sources and inoculum level for the optimal production of cephalosporin C acylase by Pseudomonas species. The biotransformation was conducted utilizing all optimized parameters for the production of cephalosporin C acylase at the end of study. The amount of product 7- aminocephalosporanic acid formed was measured by glutaryl p-nitroanilide (GpNA), p-Dimethyl aminobenzadehyde (p-DAB) and HPLC assays. The CCA activity in Pseudomonas species was measured 33.1 and 35.5 U/mL using substrate Cephalosporin C (CPC) and glutaryl p-nitroanilide (GpNA) respectively at optimized parameters. The maximum CCA acylase activity was detected at 72 h of incubation. The highest CCA activity for Pseudomonas species 42.9 U/mL was detected at 72 h of incubation by HPLC.