Immobilization of glycerol dehydrogenase and NADH oxidase for enzymatic synthesis of 1,3‐dihydroxyacetone with in situ cofactor regeneration

Abstract

BACKGROUNDNicotinamide cofactor‐dependent oxidoreductases have been widely applied to the bioproduction of varieties of useful compounds. Efficient cofactor regeneration is often required for these oxidoreductase‐catalyzed reactions. Herein, enzymatic production of 1, 3‐dihydroxyacetone (DHA) via immobilized enzymes with in situ cofactor regeneration was reported.RESULTSGlycerol dehydrogenase (GDH) and NADH oxidase (Nox) were immobilized on two carriers: macroporous resin and epoxy functionalized magnetic nanoparticles (EFMN). Higher enzyme loading capacity and activity recovery were achieved on EFMN. A high immobilization yield of 90% and >70% recovery activity were obtained for both enzymes using EFMN as carriers. Besides, both immobilized enzymes showed enhanced pH stability and temperature tolerance. Co‐immobilized and mixed immobilized enzymes were evaluated for synthesis of DHA, and mixed immobilized enzymes showed higher catalytic rate. The mixed reaction system also was investigated in view of various operational factors. Under the optimal conditions, a high DHA concentration with mixed immobilized enzymes system can reach up to 3.5 mmol L–1, which was about four times higher than that without cofactor regeneration system.CONCLUSIONThe immobilized GDH and Nox on magnetic nanoparticles showed enhanced stabilities. A cofactor regeneration system with immobilized enzymes could be utilized for efficient production of DHA from inexpensive substrates. © 2018 Society of Chemical Industry

Rapid Immobilization of Cellulase onto Graphene Oxide with a Hydrophobic Spacer

“…Immobilization of an enzyme involves the interactions between the enzyme and the carrier [8][9][10][11][12]. The properties of the carrier play an important role and normally the carrier should have large surface area [13][14][15].…”

Bioglycerol (C3) upgrading to 2,3‐butanediol (C4) by cell‐free extracts of Enterobacter aerogenes NCIM 2695

“…The intermediate (DHA) is also an important chemical in the food and pharmaceutical sectors. DHA is then converted to pyruvate, which results in 2,3‐BDO, acetoin, etc., as the metabolic end products 16,51 . The formation of 2,3‐BDO suggests the presence of glycerol dehydrogenase enzyme in the cell‐free based system, and this was confirmed by studying glycerol dehydrogenase assays, as discussed below.…”

Recent Developments in Carriers and Non-Aqueous Solvents for Enzyme Immobilization

“…Numerous covalent bond-based methods have been directly used in nanomaterials recently [44,45]. These nanomaterials can be tailored to the target enzyme, and their large surface area and selectivity of porosity provide great advantages in industrial and pharmaceutical products [6,[46][47][48]. For example, immobilization of urease on epichlorohydrin cross-linked carboxymethyl cellulose beads with polyacrylamide resulted in both the optimum pH and temperature shifting higher to 8 and 45 • C, respectively, and enabled 88% of the enzyme activity to be maintained after 10 cycles, and the modified support helped improve the stability of the urease structure [49].…”