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Immobilization of glycerol dehydrogenase and NADH oxidase for enzymatic synthesis of 1,3‐dihydroxyacetone with in situ cofactor regeneration
Author(s) -
Zhuang MengYao,
Jiang XiaoPing,
Ling XiaoMin,
Xu MengQiu,
Zhu YiHao,
Zhang YeWang
Publication year - 2018
Publication title -
journal of chemical technology and biotechnology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.64
H-Index - 117
eISSN - 1097-4660
pISSN - 0268-2575
DOI - 10.1002/jctb.5579
Subject(s) - cofactor , chemistry , immobilized enzyme , enzyme , dihydroxyacetone , formate dehydrogenase , alcohol dehydrogenase , dehydrogenase , nad+ kinase , biochemistry , oxidoreductase , glycerol , combinatorial chemistry
Abstract BACKGROUND Nicotinamide 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. RESULTS Glycerol 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. CONCLUSION The 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