Assembly of graphene oxide‐formate dehydrogenase composites by nickel‐coordination with enhanced stability and reusability

Abstract

Featuring unique planar structure, large surface area and biocompatibility, graphene oxide (GO) has been widely taken as an ideal scaffold for the immobilization of various enzymes. In this regard, nickel‐coordinated graphene oxide composites (GO‐Ni) were prepared as novel supporters for the immobilization of formate dehydrogenase. The catalytic activity, stability and morphology were studied. Compared with GO, the enzyme loading capacity of GO‐Ni was enhanced by 5.2‐fold, besides the immobilized enzyme GO‐Ni‐FDH exhibited better thermostability, storage stability and reuse stability than GO‐FDH. GO‐Ni‐FDH retained 40.9% of its initial activity after 3 h at 60°C, and retained 31.4% of its initial relative activity after 20 days’ storage at 4°C. After eight times usages, GO‐Ni‐FDH maintained 63.8% of its initial activity. Mechanism insights of the multiple interactions of enzyme with the GO‐Ni were studied, considering coordination bonds, hydrogen bonds, electrostatic forces, coordination bonds, and etc. A practical and simple immobilization strategy by metal ions coordination for multimeric dehydrogenase was developed.

Immobilization of formate dehydrogenase on polyethylenimine‐grafted graphene oxide with kinetics and stability study

“…FDH activity was measured according to the methods reported . Concentration of NADH was detected at 340 nm using an Infinite 200 PRO microplate reader (TECAN).…”

“…FDH was immobilized on the GO and GO‐PEI, respectively to investigate the activities of the enzymes before and after immobilization . FDH solution was dispersed in 1 mg/mL GO‐PEI solution mentioned above.…”

“…The improved trypsin activity and stability indicated the PEGylated GO‐induced acceleration is substrate‐dependent . In our previous study, formate dehydrogenase (FDH) was assembled with graphene oxide‐nickel composites (GO‐Ni) with enhanced stability, the multiple interactions such as electrostatic forces and coordination bond revealed the immobilization mechanism .…”

Mechanistic Aspects for the Modulation of Enzyme Reactions on the DNA Scaffold

“…The reactions of individual or multienzymes have been conducted on a wide range of carriers, such as metal–organic frameworks (MOFs) [ 40 ], hydrogel [ 41 ], graphene oxide [ 42 ], liposome [ 43 ], polymersome [ 44 ], proteins [ 45 ] and DNA nanostructures [ 46 ]. These artificial scaffolds are suggested to enhance the enzyme stability, reusability or catalytic ability, expanding the applications of enzyme in different fields such as the biosynthesis of value-added chemicals.…”

Metal ions coordinated immobilization of phenylalanine dehydrogenase by GO‐PEI with high activity recovery and enhanced stability

“…Lin et al 9 studied the nickel-coordinated graphene oxide composites (GO-Ni) as a novel supporter for the immobilization of formate dehydrogenase with enhanced thermal stability. Liu et al 10 demonstrated the combination of amino Fe 3 O 4 nanoparticles with GO as magnetic GO (MGO) nanocomposites for the immobilization of alcohol dehydrogenase, showing an enhanced broader range of pH stability and storage thermostability.…”