Abstract
A multi-enzyme route for the production of an atorvastatin side-chain was proposed. The approach includes the consecutive double aldol addition of acetaldehyde to the phenylacetamide amino-protected propanal by 2-deoxyribose-5-phosphate aldolase (DERA), the oxidation by ketoreductase, the lipase-catalysed acylation and amino group deprotection by penicillin G-acylase. To explore the feasibility of the process, the DERA reaction was studied into detail. Based on the kinetic and stability studies, a mathematical model was developed and validated in a batch and fed-batch reactor. The highest productivity was obtained in repetitive batch reactor (229.1 g/(L day)). The highest final product concentration of 124 g/L was obtained in fed-batch reactor. The mathematical model-based optimisation provided insight into the possibilities for process metrics improvement. Further, the second step was explored. The results showed that the DERA reaction and the oxidation reaction can be carried out as the multi-enzyme one pot process in the sequential manner.
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