A Simple Strategy for Co‐Immobilization of NADH‐Dependent Enzymes and NADH for Efficient Biocatalysis in Continuous Flow Reactors
Year: 2025
Type: article
Abstract: The application of nicotinamide adenine dinucleotide (NAD)(H)‐dependent enzymes in continuous flow reactors poses significant challenges due to the high cost of NAD(H) and its inevitable loss during reactor operation. To overcome these limitations, a novel platform comprising Cry3Aa‐enzyme fusion particles modified with polyethylenimine (PEI) is developed and its application for two catalytic processes is demonstrated. In the first system, two NAD(H)‐dependent enzymes, formate dehydrogenase (FDH), and leucine dehydrogenase (LDH) are genetically co‐immobilized within Cry3Aa particles and then modified with PEI to facilitate NADH co‐immobilization. To adapt the system for continuous flow reactors, PEI‐modified particles are entrapped within agarose beads, loaded with NADH, and then used to catalyze L‐tert leucine production under continuous flow conditions for 30 days. This configuration achieved LDH and NADH turnover numbers of 22,196 and 7,202, respectively, and a space‐time yield (STY) of 0.0262 g L−1 h−1. This platform is then validated for another NAD(H)‐dependent multienzyme system comprised of FDH and alcohol dehydrogenase (ADH) and used for the continuous conversion of ethyl acetoacetate to ethyl‐(R)‐3‐hydroxybutyrate. ADH and NADH turnover numbers of 15,074 and 3,256 are obtained after 30 days with a STY of 0.02 g L−1 h−1. These examples illustrate significant potential of this technology for facilitating NADH‐dependent enzyme‐mediated biocatalysis in continuous flow reactors.
Source: ChemSusChem
Institution: Chinese University of Hong Kong
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: hybrid
Estimated APC paid: $4,550
OpenAlex ID: W4410145639