Enzymatic behavior of laccase following interaction with γ-CD and immobilization into PCL nanofibers

Abstract

This study examines the effects of CD use on enzymatic activity, following enzyme immobilization into nanofibers. There is almost no research available on the change in enzyme activity following interaction with cyclodextrin and electrospun nanofiber mats together. Laccase enzyme was immobilized into nanofibrous structures by various techniques, with and without γ-CD addition, and the enzymatic activity of the laccase was analyzed. SEM, XRD, and FTIR analyses were used for the characterization of the resulting structures. Our results showed that cyclodextrin use has a positive effect on the enzyme's activity, and increases its stability. The enzymes treated by cyclodextrin showed activation after complex formation trials, and no activation loss or enzyme denaturation was detected. Our conclusions were supported by the enzyme activity test results, which also showed that immobilization by encapsulation methods gave better activity results than layering methods. Another important finding concerned the laccase's stable characteristics that helped to maintain its enzyme activation after the freeze drying process. Among all test groups, the best activity result was recorded by laccase-γ-CD complex encapsulated PCL nanofibers with 96.48 U/mg.

Biocompatible Polymers Combined with Cyclodextrins: Fascinating Materials for Drug Delivery Applications

“…The positive influence of CDs on enzyme activity and the stability of the catalase was showed. Similar to the catalase enzyme, laccase was immobilized on γ-CD/PCL nanofibers and showed higher catalytic activity (96.48 U/mg), compared to enzymes immobilized on PCL nanofibers without CDs (23.2 U/mg) or γ-CD/laccase physical mixtures in PCL nanofibers (71.6 U/mg) [75]. During the formation of the CD-enzyme complex, the enzyme used did not lose activity and no denaturation was observed.…”

“…In addition, the CD enriched PCL fibers were used for encapsulation of small molecules like α-tocopherol [71], naproxen [72], sulfisoxazole [68], ciprofloxacin [73], as well as even large molecule (e.g., polymer or enzyme) [74,75,98,101,102]. Those studies were initiated to enhance the stability of active compounds against various environmental factors during delivery by entrapment them in PCL nanofibers.…”

“…The enzymes are a type of large biomacromolecules incorporated into CD enriched PCL fibers. The introduction of CD-ICs into electrospun nanofibers having high surface area and highly porous nanostructure make them suitable substrate for biocatalyst immobilization [74,75]. For instance, catalase, an anti-free radical enzyme, was successfully immobilized onto poly(ethylene oxide) nanofibers containing γ-CDs, sandwiched between PCL nanofibers [74].…”

Interfacial Biocatalytic Performance of Nanofiber-Supported β-Galactosidase for Production of Galacto-Oligosaccharides

“…Recent studies reported that enzymes could be immobilized on various nanofibers made from cellulose polymers [15], polystyrene [7], poly(ε-caprolactone) [16], polyaniline [17], chitosan [18], polyethylene oxide [19], polylactic acid [20], and polyacrylonitrile [21]. Nanofiber is fabricated using an electrospinning device, and the size of fibrous mats ranges from nanometers to a few micrometres.…”

Immobilization of laccase onto modified PU/RC nanofiber via atom transfer radical polymerization method and application in removal of bisphenol A

“…A critical challenge for enzyme immobilization is to develop high‐performance carriers . The characteristics of an ideal carrier for enzyme immobilization include inexpensive nature , being able to load a significant amount of enzyme per unit weight , low hydrophobicity , inertness after immobilization , microbial resistance , pH, thermal, and mechanical resistance . Conventionally, nanoparticles and microspheres have been used as carriers for immobilized enzymes because of their large specific surface area and porosity, resulting in higher enzyme loading of the immobilization process .…”