Emerging Trends in Glycoside Hydrolases-Based Innovations for Sustainable Stubble Transformation into Biofuel

The worldwide demand for renewable energy alternatives with agricultural stubble has driven the advancement of second-generation (2G) biofuels. Despite the abundance of stubble, it remains underexploited due to the complexity of plant cell walls, posing a noteworthy challenge to efficient biotransformation. The review explores recent advancements in extremophilic glycoside hydrolases (GHs) as promising biocatalysts to overcome biomass complexity. The study analyses recent innovations revealing how these robust enzymes, integrated with computational and biotechnology tools, unlock new ways to convert agricultural waste into bioenergy. The novel interdisciplinary convergence of microbiology, protein engineering, and artificial intelligence is highlighted for emerging innovations in enzyme design and formulation. Furthermore, recently discovered lytic polysaccharide monooxygenases (LPMOs) with GHs in enzyme cocktails synergistically enhance saccharification, facilitating the breakdown of complex polysaccharides. This synergy, combined with innovations in advanced recombinant DNA technology, synthetic biology, machine learning, and artificial intelligence, offers novel strategies for designing robust enzyme cocktails. Computational analysis of extremophilic endoglucanases identified conserved glycine, proline, and tryptophan residues as key contributors to structural integrity and catalysis in extreme environments. The review identifies emerging business opportunities in enzyme-based biomass valorization and outlines prospective directions for future research, including challenges in enzyme stability, process scalability, and integration into biorefineries.