title: A novel method for preparation of MNP@CS-tethered coenzyme for coupled oxidoreductase system.
abstract: The immobilized cofactor NAD(H) is easily recovered from the reaction bulk, which is essential for repeated use of NAD(H) in the bioprocess catalyzed by NAD(H)-dependent oxidoreductase. Here, a magnetic nanoparticle platform was designed to immobilize both of the NADH and the NAD(+). The design was based on chitosan-coated magnetic nanoparticles (MNP@CS) which was activated by the EDC/NHS with the aid of azelaic acid as spacer. Interestingly, the succinimide group at the end of spacer arm catalyzed direct coupling of a carboxyl-terminal to the 6-amino group of the adenine residue of NAD(H). Our results indicated that 150 μmol NADH and 50 μmol NAD(+) was effectively attached to 1g MNP@CS at 25°C in 120 min and the prepared MNP@CS-NAD(H) showed good activity according to the coupling reaction of benzyl alcohol and acetaldehyde catalyzed by alcohol dehydrogenase.
authors.affiliation: Department of Biotechnology and Bioengineering, Huaqiao University, 668 Jimei Road, Xiamen 361021, Fujian, PR China. Electronic address: chenguo@hqu.edu.cn.
authors.forename: Guo
authors.initials: G
authors.lastname: Chen
authors.affiliation: Department of Biotechnology and Bioengineering, Huaqiao University, 668 Jimei Road, Xiamen 361021, Fujian, PR China.
authors.forename: Zhichao
authors.initials: Z
authors.lastname: Wu
authors.affiliation: Department of Biotechnology and Bioengineering, Huaqiao University, 668 Jimei Road, Xiamen 361021, Fujian, PR China.
authors.forename: Yunhui
authors.initials: Y
authors.lastname: Ma
mesh_terms: D000426:Alcohol Dehydrogenase / Q000378:metabolism; D048271:Chitosan / Q000737:chemistry / Q000378:metabolism; D004800:Enzymes, Immobilized / Q000737:chemistry / Q000378:metabolism; D058185:Magnetite Nanoparticles / Q000737:chemistry*; D009243:NAD / Q000378:metabolism*
chemical_list: D004800:Enzymes, Immobilized; D058185:Magnetite Nanoparticles; D009243:NAD; D048271:Chitosan; D000426:Alcohol Dehydrogenase
keywords: Coenzyme immobilization; Cofactor regeneration; Magnetic nanoparticles; Oxidoreductase