Efficiency of a Bienzyme Sequential Reaction System Immobilized on Polyelectrolyte Multilayer-Coated Colloids

Abstract

We assembled multilayer films of glucose oxidase (GOx) and horseradish peroxidase (HRP) coimmobilized together with polyelectrolyte layers on the surface of silica microparticles. The influence of different polyelectrolyte combinations on the immobilization and functionality of the enzymes was examined for several multilayer configurations. Precomplexation of the enzymes with a polyvinylpyridine-based polyamine allowed the stable adsorption of enzyme layers without affecting their catalytic activity. The efficiency of the sequential reaction between GOx and HRP on the surface of the colloids was quantitatively analyzed and rationalized in terms of the kinetic parameters of both enzymes and the reaction-diffusion kinetics of the system. In the optimized configuration, with GOx and HRP coimmobilized in the same layer, the overall rate of hydrogen peroxide conversion was around 2.5 times higher than for GOx and HRP in separate layers or for equivalent amounts of both enzymes free in solution.

Quantitative Collection and Enzymatic Activity of Glucose Oxidase Nanotubes Fabricated by Templated Layer-by-Layer Assembly

“…1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 Owing to the complexity of protein structure and the patchy nature of their distribution of surface charge, the assembly of proteins with oppositely charged polyelectrolytes is not always possible. 13,34,35 For instance, it was reported that only branched polyethyleneimine (bPEI) or poly-L-arginine (PLA) could be combined with HSA to produce stable nanotubes by LbL deposition in PC templates. 26 Likewise, we failed to build stable GOX nanotubes when paired with chitosan because the addition of chitosan desorbed heavily the previously-adsorbed GOX as was checked by ellipsometry on flat multilayers.…”

Capsosomes with Multilayered Subcompartments: Assembly and Loading with Hydrophobic Cargo

“…[7,[13][14][15][16][17][18] To establish a multifunctional advanced therapeutic platform, we recently introduced capsosomes, polymer carrier capsules containing liposomes. [19,20] This combination is a promising new approach which exploits the benefits of both systems while improving the flexibility in their design and application.…”

Enzyme:Nanoparticle Bioconjugates with Two Sequential Enzymes: Stoichiometry and Activity of Malate Dehydrogenase and Citrate Synthase on Au Nanoparticles

“…7−9 For example, the sequential enzymes horseradish peroxidase (HRP) and glucose oxidase (GOx) have been coimmobilized in polyelectrolyte multilayers on silica microparticles. (8) A set of three sequential enzymes was immobilized on phospholipid polymer particles to facilitate the overall reaction. (9) Multiactivity bioconjugates such as these are attractive as model systems for biological enzyme assemblies, in bioanalytical sensors, and for industrial processes.…”