Facile immobilization of Trametes versicolor laccase on highly monodisperse superparamagnetic iron oxide...

The development of enzymatic conjugates with industrial applications require approaches with good scalability and batch-to-batch reproducibility. Hereof, nearly monodisperse iron oxide nanoparticles can be synthesized by thermal decomposition with high yields. A mixture of gallic and polyacrylic acid is used for the direct water transfer and later immobilization of laccase (Trametes versicolor).

Nanoparticles were synthesized by thermal decomposition (13.1 nm by TEM, 50 nm by DLS) and later transferred to water by a ligand exchange method with polyacrylic acid and a polyacrylic acid/gallic acid mixture. Laccase was immobilized on water dispersions of both nanoparticles via a carbodiimide coupling.

The nanoparticles exhibited superparamagnetic behavior with insignificant values of iH_c. The presence of gallic acid hindered the formation of multiple polyacrylic acid layers, therefore improving the colloidal stability of the nanoparticles (100 nm by DLS) after weeks of storage. Nanoparticles containing only polyacrylic acid showed poor activity (60% loading, 4.5% activity), while nanoparticles with both polyacrylic and gallic acids showed enzymatic activity values 4.4 times higher than the free enzyme (13% loading, 57% activity). The nanoparticles improved the storage stability (8 times) of the enzyme, its thermoresistance (4 times), and its reactivity against azo dyes Camalgite and Congo Red (21 and 27% increase, respectively). In addition to some improved catalytic properties in comparison to similar works, this is the first report of the use of gallic acid for both the direct transfer to water and enzyme immobilization on highly monodisperse, batch-to-batch reproducible superparamagnetic nanoparticles.