A Modular Enzyme Cascade for Coenzyme Regeneration: A Simple Approach to Master the Circumstance of Leaching

Originating from three enzymes immobilized onto the surface of mesoporous silicas a modular enzyme cascade was constructed. The major drawbacks concerning the utilization of immobilized enzymes are frequently associated with a pronounced leaching behavior of the enzyme, as well as its dependency on a distinct coenzyme. Accordingly, to obtain more efficient enzyme catalysis, a universal approach ensuring that the coenzyme is recycled is preferable. Herein, the three adsorptively immobilized enzymes were affected by extensive leaching. Thus, a principle was developed to re-immobilize the amounts of leached enzyme preventing the contamination of the substrate solution and a reliable coenzyme recycling step was considered. The entire enzyme cascade was continuously performed over several cycles and the stability, or rather the efficiency, of the enzyme cascade was investigated in detail, with a special focus on a sophisticated coenzyme recycling procedure.