Cellulose Carriers from Spent Coffee Grounds for Lipase Immobilization and Evaluation of Biocatalyst Performance

In line with the circular economy approach and the pursuit of sustainable solutions for spent coffee grounds, this study investigates the valorization of spent coffee grounds as a source of cellulose-based enzyme immobilization carriers. Considering that global coffee consumption generates approximately 6.9 million tonnes of spent coffee grounds annually, their disposal represents both an environmental challenge and an opportunity for value-added applications. A multistep extraction process, including Soxhlet extraction followed by sequential subcritical extraction with ethanol and water, and alkaline treatment, led to the production of cellulose-enriched carriers. The carriers obtained were characterized by their morphology, porosity and surface properties and subsequently used for the two lipases immobilization, Burkholderia cepacia (BCL) and Pseudomonas fluorescens (PFL), using three techniques: adsorption and covalent binding via direct and indirect methods. The immobilized lipases were analyzed for key biochemical and operational properties and compared with each other and with their free enzymes. Based on their stability, catalytic activity, and reusability, the lipases immobilized by adsorption were identified as the most efficient biocatalysts. These immobilized enzymes were then used in two selected reactions to demonstrate their practical utility: cocoa butter substitute synthesis using PFL and the enzymatic pretreatment of wastewater from the oil processing industry using BCL. Both immobilized lipases showed excellent catalytic performance and maintained their high activity over four consecutive reuse cycles.