engleski

2-Deoxyribose-5-phosphate aldolase from Thermotoga maritima in the synthesis of a statin side-chain precursor: characterization, modeling and optimization

BACKGROUND: Producing statin side-chains by sequential aldol condensation catalyzed by 2- deoxyribose-5-phosphate aldolase (DERA) (EC 4.1.2.4) is the best-known and most promising synthetic route. The advantage of this process is the formation of two stereocenters in a single step starting from inexpensive achiral components, acetaldehyde and chloroacetaldehyde. The limitation for the industrial-scale application is enzyme inactivation caused by substrates as well by the intermediate produced by single aldol addition. RESULTS: The DERA enzyme from Thermotoga maritima (DERATm) was investigated extensively in this work. The influence of aldehydes on DERATm stability was studied in detail. Mathematical models in different reactor configurations were developed based on experimentally determined kinetic parameters of all reactions included in the synthesis of statin side-chain. Models also included enzyme inactivation by all compounds with negative impact on its stability. Mathematical model-based optimization enabled optimization of process conditions and reactor configuration. The fed-batch reactor proved to be the most suitable choice achieving product concentration of 78 g L−1, productivity of 56 g L day−1 and yield of 95% under an optimal condition. CONCLUSION: The validated mathematical model and applied methodology can be exploited for further process improvement and for process design of similar systems.

biocatalysis ; kinetics ; mathematical modeling ; process intensification ; enzymes ; optimization

This work has received funding from the EU Horizon 2020 research and innovation programme under grant agreement 635595 "CarbaZymes".