We demonstrated the nano-immobilization of fungal enzymes through their encapsulation in cellulose nanoparticles (CNPs). An extracellular enzyme cocktail (a mixture of amylase, protease, lipase, and cellulose) was produced from Aspergillus niger and Phanerochaete chrysosporium through submerged fermentation. The process of encapsulation was carried out through a microemulsion nanoprecipitation method in the presence of a lipid, a surfactant, and a co-surfactant. The morphology of CNPs was determined by field-emission scanning electron microscopy and transmission electron microscopy; CNPs were less than 100 nm in diameter. Fourier transform infrared spectroscopy (FTIR) and energy dispersive spectroscopy demonstrated the successful encapsulation of the fungal enzyme cocktail and revealed C and O as its major components. FTIR peaks of CNPs with encapsulated enzymes occurred at 3421.80, 2828.91, 1649.29, 1450.24, and 1061.61 cm−1 as well as in the range of 1050–1150 cm−1. Encapsulated enzymes showed excellent stability with a peak at −70.91 mV in zeta potential studies. Thermogravimetric analysis proved that the CNP-encapsulated enzymes had an initial weight loss at 250C. The encapsulated fungal enzyme cocktail exhibited higher catalytic performance and stability than the free enzymes. The encapsulated fungal enzyme cocktail derived from A. niger at the concentration of 100 µg/mL, showed the highest amylase activity with a clear zone of 2.5 cm. Overall, the results of this research reveal the enhancement in the activity of fungal extracellular enzyme cocktail through nanoencapsulation.
