Utilization potential of low-cost media: Recombinant exoglucanase production in Pichia pastoris

The accumulation of cellulosic waste is a global concern, necessitating the development of environmentally friendly methods for waste degradation. Despite its high cost, exoglucanase emerges as a pivotal enzyme, recognized for its role in hydrolyzing cellulose. The research aims to reduce exoglucanase production costs without compromising activity and productivity. The initial research involved the production of exoglucanase in high-cost and low-cost medium on the Erlenmeyer scale. Following the scale-up production of the enzyme using 5 L of low-cost medium in a bioreactor, the production increased by up to five times compared to the Erlenmeyer scale. The exoglucanase activity reached 54,29 U/mL in 24 h incubation. Significance analysis was performed using one-way ANOVA in GraphPad Prism 10.4.1. The micrograph from SEM analysis demonstrated the effectiveness of enzyme-degraded cellulose. As the incubation time increased, the crystalline structure of the avicel became disrupted. Furthermore, the interaction between the enzyme and the avicel substrate was also analyzed using docking analysis. The interaction between exoglucanase and microcrystalline cellulose and cellulose substrates was found to have binding energy values of −7.797 kcal/mol and −7.962 kcal/mol, respectively. Collectively, these findings show the success of the biocatalyst production process utilizing low-cost medium.