Miniaturizing Biocatalysis: Enzyme‐Catalyzed Reactions in an Aqueous/Organic Segmented Flow Capillary Microreactor

Abstract

A segmented flow capillary microreactor was used to perform the enzyme-catalyzed conversion of 1-heptaldehyde to 1-heptanol in a two liquidliquid phase system. These reactor formats are established for chemical reactions but so far data describing the relevant system parameters for enzymatic catalysis are lacking. This work now addresses the impact of important parameters such as capillary diameter, flow velocity, phase ratio, and enzyme as well as substrate concentration on the performance of the enzymatic reaction under segmented flow conditions. All key parameters governing reaction performance have been correlated in a novel operational window for an easy assessment of the various system constraints.

A peculiar segmented flow microfluidics for isoquercitrin biosynthesis based on coupling of reaction and separation

“…Dore developed more efficient mixing patterns using water plugs in water/ionic liquid segmented flow in micro-channels (Dore et al, 2012). A segmented flow capillary microreactor was used to perform the enzyme-catalyzed conversion of 1-heptaldehyde to 1-heptanol in a two-phase liquid-liquid system, indicating it as a promising tool to perform multiphasic enzymatic catalysis (Karande et al, 2011). In this system, the problem of product degradation could not have been solved by a gas/liquid or water/ionic liquid system.…”

Biotechnical Micro-Flow Processing at the EDGE – Lessons to be learnt for a Young Discipline

“…By far the most contributions so far were given from researchers experienced in biocatalysis and thus chemistry. The following diagram shows operational windows determined for enzymatic segmented flow as described in 10 (and in this paper below in more detail) (Fig. 2).…”

“…10 To fill that gap a generic study was done for the enzyme-catalyzed conversion of 1-heptaldehyde to 1-heptanol in a liquid-liquid phase system (Scheme 2). Relevant parameters for segmented flow such as capillary diameter, flow velocity, phase ratio, and enzyme as well as substrate concentration were varied to show their impact on the enzymatic reaction.…”

Integrated lipase-catalyzed isoamyl acetate synthesis in a miniaturized system with enzyme and ionic liquid recycle

“…This influenced in situ product extraction to the n-heptane phase and, thereby, reaction rate. A recent study of the influence of fluid flow ratio of an aqueous/ organic two-phase system on enzyme-catalyzed conversion of 1-heptaldehyde to 1-heptanol using segmented flow microreactor also revealed a huge impact of this parameter on volumetric productivity [30]. A detailed investigation of the effect of [bmpyr][dca] and n-heptane flow rates and their ratio on fluid flow pattern, as well as substrate and enzyme concentration on product synthesis within microfluidic system, is foreseen in order to optimize the integrated biotransformation process.…”

“…Enzymatic microreactors could operate as homogeneous systems with multiphase parallel [27][28][29] or segmented [30][31][32] flow and as a heterogeneous system with immobilized enzymes [17,33,34]. The first report on an enzymatic reaction in an ionic-liquid (IL)/organic solvent two-phase system within a microfluidic device was published by Pohar et al [31], where a continuously operated Ψ-shaped microreactor was used for lipase-catalyzed synthesis of isoamyl acetate in the 1-butyl-3-methylpyridinium dicyanamide ([bmpyr][dca])/n-heptane two-phase system.…”