Cis-GPRA synthesis by enzymatic removal of ap-methoxybenzyl protecting group fromp-methoxybenzyl-7-phenylacetylamino-3-propenyl-3-cephem-4-carboxylate

Abstract

Twelve typical commercial hydrolases and crude cell extract of Bacillus subtilis ATCC6633 were tested for their activity to synthesize cis -7-phenylacetamido-3-propenyl-cephalosporanic acid ( cis -GPRA) by enzymatic removal of a p -methoxybenzyl protecting group from p -methoxybenzyl-7-phenylacetylamino-3-propenyl-3-cephem-4-carboxylate (GPRE) in aqueousorganic solvents. No activity was detected for 12 commercial enzymes. However, the crude cell extract showed signifi cant activity for removing the p -methoxybenzyl protecting group from GPRE to produce cis -GPRA. The yield of cis -GPRA reached 73% with the initial reaction rate of 10.2 μ M h -1 under the optimized reaction conditions.