Nanobiocatalysis is a new frontier of emerging nanosized material support in enzyme immobilization application. This paper is about a comprehensive review on cellulose nanofibers (CNF), including their structure, surface modification, chemical coupling for enzyme immobilization, and potential applications. The CNF surface consists of mainly -OH functional group that can be directly interacted weakly with enzyme, and its binding can be improved by surface modification and interaction of chemical coupling that forms a strong and stable covalent immobilization of enzyme. The knowledge of covalent interaction for enzyme immobilization is important to provide more efficient interaction between CNF support and enzyme molecule. Enzyme immobilization onto CNF is having potential for improving enzymatic performance and production yield, as well as contributing toward green technology and sustainable sources.
Immobilisation of cyclodextrin glucanotransferase (CGTase) on nanofibres was demonstrated. CGTase solution (1% v/v) and PVA (8 wt%) solution were mixed followed by electrospinning (-9 kV, 3 h). CGTase/PVA nanofibres with an average diameter of 176 ± 46 nm were successfully produced. The nanofibres that consist of immobilised CGTase were crosslinked with glutaraldehyde vapour. A CGTase/PVA film made up from the same mixture and treated the same way was used as a control experiment. The immobilised CGTase on nanofibres showed superior performance with nearly a 2.5 fold higher enzyme loading and 31% higher enzyme activity in comparison with the film.
In this study, the potential of electrohydrodynamic atomization or electrospraying to produce nanometer-order CGTase particles from aqueous suspension was demonstrated. CGTase enzyme was prepared in acetate buffer solution (1% v/v), followed by electrospraying in stable Taylor cone-jet mode. The deposits were collected on aluminium foil (collector) at variable distances from the tip of spraying needle, ranging from 10 to 25 cm. The Coulomb fission that occurs during electrospraying process successfully transformed the enzyme to the solid state without any functional group deterioration. The functional group verification was conducted by FTIR analysis. Comparison between the deposit and the as-received enzyme in dry state indicates almost identical spectra. By increasing the distance of the collector from the needle tip, the average particle size of the solidified enzyme was reduced from 200±117 nm to 75±34 nm. The average particle sizes produced from the droplet fission were in agreement with the scaling law models. Enzyme activity analysis showed that the enzyme retained its initial activity after the electrospraying process. The enzyme particles collected at the longest distance (25 cm) demonstrated the highest enzyme activity, which indicates that the activity was controlled by the enzyme particle size.
The present study developed the formulation of active bionanocomposites films endowed with the abilities of high biodegradability and antimicrobials for active packaging applications. The aim of this work was to prepare poly (lactic acid)/poly (butylene succinate) (PLA/PBS) blended films reinforced with different concentrations of nanofibrillated cellulose (NFC) and 9 % of thymol essential oil (EO) using the casting method. The active films were further evaluated through Fourier transform infrared spectroscopy (FTIR); as well as mechanical, physical, water vapour permeability (WVP), thermal analysis (TGA), biodegradation, morphological, and antimicrobial (% reduction of bacteria) testing. The tensile strength (TS) of PLA/PBS blend films increased by 12 % with the incorporation of 2 wt% of NFC. The PLA/PBS/NFC with 9 % thymol EO has a good water barrier performance with its tensile strength, elongation at break, and tensile modulus was 13.2 MPa, 13.1 %, and 513 MPa respectively. The presence of NFC promoted the disintegration of PLA/PBS films by 70.5 %. These films promoted the antibacterial activity against S. aureus and E. coli. The study demonstrates that the developed films improved the qualities of chicken fillets and have great potential to be used as active bionanocomposites in food packaging applications.