Optimized Production of Cellulase using Different Agrowaste Biomass Substrates

Abeer Zahra; Misbah Zafar; Anam Mubarak; Zahid Anwar; Taimoor Hassan; Khizar Hayat Bhatti; Noshia Arshad; Muhammad Zeeshan Baqir; Muahmmad Ishtiaq; Tanveer Hussain

Authors

Abeer Zahra Department of Chemistry, University of Gujrat, Hafiz Hayat Campus, Gujrat, Pakistan
Misbah Zafar Department of Plant Sciences, Quaid-i-Azam University Islamabad Pakistan
Anam Mubarak Department of Botany, University of Gujrat, Hafiz Hayat Campus, Gujrat-Pakistan
Zahid Anwar Department of Biochemistry and Biotechnology, University of Gujrat, H. H. Campus, Gujrat, Pakistan
Taimoor Hassan Department of Botany, University of Gujrat, Hafiz Hayat Campus, Gujrat-Pakistan
Khizar Hayat Bhatti Department of Botany, University of Gujrat, Hafiz Hayat Campus, Gujrat, Pakistan
Noshia Arshad Department of Botany, University of Gujrat, Hafiz Hayat Campus, Gujrat, Pakistan
Muhammad Zeeshan Baqir Department of Botany, University of Gujrat, Hafiz Hayat Campus, Gujrat, Pakistan
Muahmmad Ishtiaq Department of Botany, University of Bhimber, AJK, Pakistan.
Tanveer Hussain Department of Botany, University of Bhimber, AJK, Pakistan.

Keywords:

Cellulase, High Production of Cellulase, Mixture of Substrates, Optimization, RSM, Solid State Fermentation, Spectrophotometry

Abstract

Cellulase is a crucial industrial enzyme, with developing countries expending significant resources on its import for various industrial and scientific applications. A major challenge in cellulase production is the lack of affordable technology and suitable substrates for cultivating enzyme-producing microbes. This study optimized a substrate mixture to enhance cellulase production using solid-state fermentation with the Aspergillus Niger strain. Five agro-industrial substrates—sugarcane bagasse, corn cobs, rice straw, orange peel, and wheat straw—were individually inoculated with A. Niger spore suspension, and their cellulase activity was compared to that of a substrate mixture. The enzyme activity from individual substrates was notably lower compared to the mixture. Response Surface Methodology (RSM) was employed to identify the optimal substrate combination, which consisted of equal amounts of sugarcane bagasse, corn cobs, orange peel, and wheat straw, with rice straw in double the amount of the other substrates. The study also optimized fermentation parameters, including temperature, pH, incubation time, substrate concentration, moisture content, urea, MgSO4, and inoculum size of A. Niger. Maximum cellulase activity was achieved at 50°C, 80% moisture content, pH 4.0, 120 hours incubation, with 6.5 g of the substrate mixture, 2% w/w urea, 0.2% w/w MgSO4, and 4 ml of A. Niger spore suspension. Optimization resulted in cellulase activity of 0.205 IU/ml, significantly higher than the 0.025 IU/ml from individual substrates. Given its key role in industries such as pulp and paper, textiles, food and beverages, detergents, and agriculture, the demand for cellulase is expected to surge, particularly with the rise in biofuel production.

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