Preparation and evaluation of a polymer–metal–enzyme hybrid nanowire for the immobilization of multiple oxidoreductases

Abstract

BACKGROUND: The immobilization of multiple oxidoreductases has attracted a lot of research interest for its industrial application in eco-friendly production. Here, a novel immobilization strategy was developed using manganese ion chelated linear polyethyleneimine as artificial hybrid scaffold that allowed self-assembly of bifunctional oxidoreductase (glycerol dehydrogenase-NADH oxidase) via multipoint coordination interaction.RESULT: The immobilization formed a highly ordered polymer-metal-enzyme complex structure shaped like a leaf vein with 200-nm-wide nanowires. Preliminary results showed that the immobilization could not only retain but substantially increased the biological activity of the immobilized enzyme by 8.2-fold. The equilibrium concentration of the reaction product DHA also was increased by 4.5-fold compared with that of the free enzyme, exhibiting higher elimination efficiency of product inhibition compared with other enzyme immobilization strategy using nanoparticles or activated agarose beads. At the same time, linear polyethyleneimine (LPEI)-manganese scaffolds could prevent enzyme subunit disassociation and stabilize the active multimeric form, which improved catalytic condition adaption and thermal stability of target enzyme. CONCLUSION: These results indicated a great potential of polymer-metal-enzyme hybrid in the enzyme immobilization and have provided a simple and efficient strategy for the immobilization of multiple oxidoreductases.

Fast Immobilization of Human Carbonic Anhydrase II on Ni-Based Metal-Organic Framework Nanorods with High Catalytic Performance

“…However, the poor stability and reusability of carbonic anhydrase limits its application in practice [6,8]. Enzyme immobilization on solid supports has been developed to overcome these shortcomings [9][10][11][12][13]. Previous immobilization studies mainly focused on commercial carbonic anhydrase on a wide range of solid supports such as: polyurethane foam [14], SBA-15 [15], mesoporous silica [16], MIL-160 [9] and ZIF-8 [17].…”

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

“…PEI‐CLEAs (cross‐linked enzyme aggregates) have been applied to hydrolyze the fish oil and enrich polyunsaturated fatty acids efficiently with organic solvent tolerance . PEI‐metal complexes acted as coating reagents to encapsulate oxidoreductases was also proved to be an efficient way .…”

Efficient synthesis of an antiviral drug intermediate using overexpressed short‐chain dehydrogenase and cross‐linked enzyme aggregates stabilization

“…However, immobilization of an enzyme on a carrier often leads to the loss of more than 50% native activity, especially at high enzyme loadings 9 . Based on previous reports, various support materials were utilized to prepare immobilized alcohol dehydrogenase due to the lack of an universal material suitable for all enzymes, such as titania nanoparticles, silica gel, magnetic Fe 3 O 4 @SiO 2 nanoparticles, porous glass beads, polyvinyl alcohol fiber, and derivatized resin 11–19 . However, only the magnetic Fe 3 O 4 @SiO 2 nanoparticles demonstrated a high activity recovery of 92%.…”

“…9 Based on previous reports, various support materials were utilized to prepare immobilized alcohol dehydrogenase due to the lack of an universal material suitable for all enzymes, such as titania nanoparticles, silica gel, magnetic Fe 3 O 4 @SiO 2 nanoparticles, porous glass beads, polyvinyl alcohol fiber, and derivatized resin. [11][12][13][14][15][16][17][18][19] However, only the magnetic Fe 3 O 4 @SiO 2 nanoparticles demonstrated a high activity recovery of 92%. Most reported carrierbound immobilization techniques only obtained an activity recovery between 40% and 60%.…”