DERA in Flow: Synthesis of a Statin Side Chain Precursor in Continuous Flow Employing Deoxyribose-5-Phosphate Aldolase Immobilized in Alginate-Luffa Matrix

Abstract

Statins, cholesterol-lowering drugs used for the treatment of coronary artery disease (CAD), are among the top 10 prescribed drugs worldwide. However, the synthesis of their characteristic side chain containing two chiral hydroxyl groups can be challenging. The application of deoxyribose-5-phosphate aldolase (DERA) is currently one of the most promising routes for the synthesis of this side chain. Herein, we describe the development of a continuous flow process for the biosynthesis of a side chain precursor. Design of experiments (DoE) was used to optimize the reaction conditions (pH value and temperature) in batch. A pH of 7.5 and a temperature of 32.5 °C were identified to be the optimal process settings within the reaction space considered. Additionally, an immobilization method was developed using the alginate-luffa matrix (ALM), which is a fast, simple, and inexpensive method for enzyme immobilization. Furthermore, it is non-toxic, biodegradable, and from renewable resources. The final continuous process was operated stable for 4 h and can produce up to 4.5 g of product per day.

Biocatalysis making waves in organic chemistry

“…Immobilisation has also given promising results, including applications in flow. 94,95 DERA is employed industrially. [96][97][98] For several years already the synthesis of the statin side chain is based on an intriguing double aldol reaction.…”

Immobilisation and flow chemistry: tools for implementing biocatalysis

“…8). 72 Fig. 8 Continuous EcDERA-C47M catalysed aldol reaction for the synthesis of (3R,5S)-6-chloro-2,4,6-trideoxyhexapyranoside in aqueous medium.…”

Current state of and need for enzyme engineering of 2-deoxy-D-ribose 5-phosphate aldolases and its impact

“…A continuous-flow process was also developed by Grabner et al for the synthesis of statin side-chain precursor. In this process, freeze-dried whole E. coli cells expressing the C47M DERA mutant were immobilized using alginate-luffa matrix (Grabner et al 2020).…”