Journal of Environmental Biology

A study on critical associations of media components on enhanced cellulase production from wild Trichoderma viride and cellulase immobilization on iron-oxide magnetic nanoparticles

D. Revathi and S. Ramalingam*   

Centre for Biotechnology, Anna University, Chennai-600 025, India

*Corresponding Author Email : ramabioprocess@annauniv.edu                  *ORCiD: https://orcid.org/0000-0002-7784-1196

Received: 03.05.2022                                                                                               Revised: 01.07.2022                                                                                            Accepted: 23.08.2022

Abstract

Aim: The current study is a preliminary step towards enhancing the cellulase productivity in wild Trichoderma viride which will enable robust valorization of non-edible lignocellulosic biomass through co-generative enzymatic saccharification, specifically concentrating on influence of individual media components on biomass growth and cellulase productivity. Further, cellulase immobilization on iron-oxide magnetic nanoparticles was also achieved that can increase the shelf life of the enzyme.

Methodology: The cellulase production in the wild Trichoderma viride was enhanced using media design and formulation. EDC {1-Ethyl-3-(3-dimethylaminopropyl) carbodiimide} functionalized iron-oxide nanoparticles were chosen to act as carriers for cellulase immobilization. The binding efficiency and relative activity were measured in addition to optimal pH and temperature for cellulase bound iron-oxide nanoparticles. Further, the hydrolysis efficiency of immobilized cellulases was also measured after which it was subjected to consecutive hydrolytic cycles to calculate the recycle rate.

Results: A maximum growth rate of 60 PCV (Packed cell volume) and total cellulase activity of 7.4 U ml^-1 was obtained on media design and formulation. 82.5% binding efficiency was achieved on EDC functionalized iron-oxide magnetic nanoparticles which showed good stability at 5pH and 50⁰C. There was 44.4% activity loss after 5 consecutive hydrolytic cycles which showed steady decline with increased cycle number and finally at the end of the 10^th hydrolytic cycle, 22.2% of total relative activity was retained.

Interpretation: Unprecedented total cellulase activity from a wild strain was obtained through media design. The stability of cellulases was further enhanced using iron-oxide magnetic nanoparticle immobilization.

Key words: Cellulases, Immobilization, Iron-oxide magnetic nanoparticles, Submerged fermentation, Trichoderma viride

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