(406a) Ampicillin Synthesis Using a Two-Enzyme Cascade System with Both &alpha-Amino Ester Hydrolase and Penicillin G Acylase

Deaguero, A. L., Georgia Institute of Technology
Blum, J., Georgia Institute of Technology
Perez, C. V., Georgia Institute of Technology
Bommarius, A. S., Georgia Institute of Technology

The current enzymatic production of semi-synthetic penicillin antibiotics requires isolation and purification of the intermediate 6-aminopenicillanic acid (6-APA), which adds cost and complexity to the manufacturing process.  In this work we take advantage of the unique substrate specificity of α-amino ester hydrolases  (AEH, EC, which are specific for α-amino containing β-lactam antibiotics, to perform a purely aqueous one-pot production of ampicillin directly from penicillin G, catalyzed by both AEH and penicillin G acylase (PGA, EC  The synthesis was performed in both a one-pot, one-step synthesis with 39% maximum conversion to ampicillin, and a one-pot, two-step process resulting in a maximum of 47% conversion to ampicillin.  This two-enzyme cascade reaction to semi-synthetic antibiotics is a promising alternative to the current two-step, two-pot enzymatic process with intermittent isolation of 6-APA.