A novel method for preparation of MNP@CS-tethered coenzyme for coupled oxidoreductase system.
Journal of biotechnology
confidence
Key findings
MNP@CS platform immobilized NADH and NAD+; 150 µmol NADH and 50 µmol NAD+ attached to 1g MNP@CS in 120 min; good activity in alcohol dehydrogenase-catalyzed reaction.
View source on PubMed (PMID 25617681) ↗
- Population
- In vitro biocatalytic system
- Dosing
- 150 µmol NADH and 50 µmol NAD+ attached to 1g MNP@CS
- Duration
- 120 min
- Route
- Immobilization on magnetic nanoparticle platform
- Blinding
- not_reported
- Controls
- not_reported
- Drug class
- coenzyme
Full 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.