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

BACKGROUND: The statin side-chain synthesis by the sequential aldol condensation catalyzed by DERA (EC 4.1.2.4) is the most known and promising 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. This article is protected by copyright. All rights reserved. This article has been accepted for publication and undergone full peer review but has not been through the copyediting, typesetting, pagination and proofreading process, which may lead to differences between this version and the Version of Record. Please cite this article as doi: 10.1002/jctb.5956 A cc ep te d A rti cl e RESULTS: DERA enzyme from Thermotoga maritima (DERA) was extensively investigated in this work. The influence of aldehydes on DERA stability was studied in detail. Based on experimentally determined kinetic parameters of all reactions included in the synthesis of statin side chain, mathematical models in different reactor configurations were developed. Models also include enzyme inactivation by all compounds that have negative impact on its stability. Mathematical model-based optimization enabled finding the optimal process conditions and choosing the best reactor configuration. The fed-batch reactor proved to be the most suitable choice. Under optimal conditions product concentration of 78 g/L, productivity of 56 g/(L day) and yield of 95% was achieved. CONCLUSION: The validated mathematical model and the used methodology can be exploited for further process improvement and for process design of similar systems.