Cis-GPRA synthesis by enzymatic removal of a p-methoxybenzyl protecting group from p-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 aqueous– organic solvents. No activity was detected for 12 commercial enzymes. However, the crude cell extract showed significant 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 μM h⁻¹ under the optimized reaction conditions.

Keywords::

• p-Methoxybenzyl protecting group
• cefprozil
• p-methoxybenzyl-7-phenylacetylamino-3-propenyl-3-cephem-4-carboxylate (GPRE)
• 7-phenylacetamido-3-propenyl-cephalosporanic acid (GPRA)
• esterase

Declaration of interest: The authors are grateful to the National Natural Science Foundation of China (grant no. 51078147), Guangdong Provincial Science and Technology Program (grant nos. 2010B031700022 and 2010A 010500005) and Guangzhou Science and Technology Program (grant no. 2008A1-E4041). The authors report no conflicts of interest. The authors alone are responsible for the content and writing of the paper.