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International Journal of Biological Macromolecules
Volume 167, 2021, Pages 1297-1307

One pot clarification and debittering of grapefruit juice using co-immobilized enzymes@chitosanMNPs

Mayur R.Ladolea, Pravin B.Pokaleb, Vrushali R.Varudea, Prasad G.Belokarc, Aniruddha B.Pandita

Department of Chemical Engineering, Institute of Chemical Technology, Mumbai, India.


In the present work, enzymes pectinase and naringinase were simultaneously co-immobilized on an eco-friendly chitosan coated magnetic nanoparticles (chitosanMNPs) by cross-linking using chitosan as a macro-molecular cross-linker. The maximum activity recovery of both enzymes in the co-immobilized form was obtained at chitosanMNPs to enzymes ratio of 1:3, 3% cross-linker concentration and 150 min cross-linking time. The synthesized MNPs before and after co-immobilization were characterized using different techniques. The prepared biocatalyst was found spherical with an average size below 200 nm and showed supermagnetic property with saturation magnetization of 38.28 emu/g. The optimum pH and temperature of both enzymes in co-immobilized form was found at 5.5 and 65°C. The prepared biocatalyst exhibited an improved thermal stability with 1.8-fold increase in the half-life. The secondary structural analysis revealed that, prepared co-immobilized biocatalyst undergone changes in the conformational and structural rigidity due to macro-molecular cross-linker. The co-immobilized biocatalysts were evaluated for one pot clarification and debittering of grapefruit juice and found ~52% reduction in turbidity and ~85% reduction in the naringin content. The co-immobilized enzymes were recycled up to 7th cycle and can be easily stored at room temperature for 30 days retaining up to 64% and 86% residual activities respectively.

Keywords: Naringinase, Co-immobilization, Debittering.

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