# Statical optimization of cellulase enzymes production by Trichoderma harzianum PP400831 using response surface methodology and their application in production of 2G bioethanol

## TL;DR

This study improves the production of cellulase enzymes by optimizing conditions for a Trichoderma harzianum strain, enhancing 2G bioethanol production from sugarcane bagasse.

## Contribution

A novel strain of Trichoderma harzianum is optimized using BBD-RSM to produce high cellulase activity for cost-effective 2G bioethanol.

## Key findings

- Maximal endoglucanase and exoglucanase activities of 4.01 IU/mL and 2.64 IU/mL were achieved after 9 days.
- Crude enzymes produced 25.63 g/L ethanol from pretreated sugarcane bagasse using S. cerevisiae MN901244.
- Optimized conditions included 2% cellulose and 0.15% tween 80 for enzyme production.

## Abstract

The development of second-generation (2G) bioethanol from lignocellulosic sources, such as sugarcane bagasse, is very important as a viable alternative to conventional fossil fuels. However, the high cost associated with enzymatic hydrolysis, which breaks down cellulose into fermentable sugars, poses a key challenge. This study focused on enhancing cellulase enzyme production by a novel, locally isolated strain, Trichoderma harzianum PP400831, using statistical optimization BBD-RSM to improve enzyme activity. Optimization efforts resulted in maximal endoglucanase and exoglucanase activities of 4.01 IU/mL and 2.64 IU/mL, respectively after 9 days at 2% cellulose mixture concentration and 0.15% tween 80. After saccharification of pretreated (SCB) by the crude enzymes and fermentation of produced reduced sugar by S. cerevisiae MN901244 yielded an ethanol concentration of 25.63 g/L. This work represents a significant step toward developing a cost-effective, sustainable, and high-performing cellulase production process for second-generation bioethanol.