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Immobilization of Cellulase Enzyme on Zinc Ferrite Nanoparticles in Increasing Enzymatic Hydrolysis on Ultrasound-Assisted Alkaline Pretreated Crotalaria Juncea Biomass


  • Department of Biotechnology, National Institute of Technology Warangal, Warangal − 506004, Telangana, India


Objectives: To synthesize ferrite nano particles; to measure size of the particles by FTIR and XRD studies and enzyme saccharification on ultra sound assisted alkaline pretreated biomass using free and immobilized enzyme. Methods/ Statistical Analysis: In the present work, Zinc ferrite particles were synthesized using co-precipitation method. Cellulase enzymes were immobilized on covalently activated ferrite nanoparticles via glutaraldehyde as a crosslinker. Biochemical characterization of free and immobilized enzyme activity were performed on CMC as a substrate. The efficiency of immobilized enzyme was evaluated based on its binding efficiency on the nanoparticles, thermostability, and reusability. Enzymatic hydrolysis was performed on ultrasound-assisted alkaline pretreated sunn hemp biomass using free and immobilized enzymes. Findings: Around 74% of binding was achieved at 4mg/ml of ferrite loading to enzyme concentration of 20 units. Comparative study on effects of pH and temperature was done on both free and immobilized enzyme and it was observed that the immobilized enzyme has maximum activity at pH 5 and temperature 60˚C. Also, the immobilized enzyme was stable at 60˚C while retaining its activity up to 3 recycles. The immobilized enzyme showed 53% hydrolysis yield on pretreated sunn hemp biomass. Application/Improvements: The research on the interaction between zinc ferrite and cellulase in immobilization and also the recovery of enzymes can determine an efficient approach for bioethanol production in industrial scale. The lab scale can be scaled-up to use at pilot and industrial scales.


Crotalaria Juncea Biomass, Enzymatic Hydrolysis, Immobilization, Nanoparticles, Ultrasound

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