High-level extracellular production of penicillin acylase by genetic engineering of Escherichia coli

The extracellular production of penicillin acylase (PAC) in genetically eng ineered Escherichia coli by coexpression of the brp gene encoding bacterioc in release protein (BRP) and the pac gene was demonstrated. Cell physiology was affected while PAC was released into the medium, depending on the stra tegy for brp expression. The performance for the production and release of PAC was optimized by taking several culture parameters, including host, ind ucer (mitomycin C) concentration, and induction timing for brp expression, into consideration. The effect of PAC release on inclusion body formation w as also investigated. It was observed that the amount of inclusion bodies w as significantly affected by brp expression. A reason for the limitation of PAC production and a strategy for resolving this problem are proposed. (C) 2001 Society of Chemical Industry.