Comparative Enzymatic Production of Xylooligosaccharides from Wheat, Rice, Barley, and Oat Straw Using Xylanase from Bacillus sonorensis

The growing amounts of agricultural residues require sustainable solutions for their use. Here, wheat, rice, barley, and oat straw were evaluated as renewable feedstocks for the enzymatic production of xylooligosaccharides (XOS). Hydrolysis used recombinant xylanase from Bacillus sonorensis T6 under optimized conditions (40 °C, pH 7.0), with stepwise enzyme addition. Subsequently, hydrolysis efficiency was found to vary by substrate, with wheat straw producing the highest reducing sugar yield (up to 40.1 g kg−1), followed by barley, oat, and rice straw. As hydrolysis progressed, the influence of enzyme concentration became less pronounced, suggesting that substrate accessibility and the accumulation of hydrolysis products may increasingly affect the overall hydrolysis efficiency. FTIR, NMR, SEM, and TLC analyses confirmed substantial structural changes in the biomass and the formation of carbohydrate-rich hydrolysis products. TLC analysis indicated the presence of low-degree polymerization oligosaccharides with migration behavior similar to X2 and X3 standards, while FTIR and NMR spectra were consistent with β-(1→4)-linked carbohydrate structures. The xylanase from Bacillus sonorensis T6 hydrolyzed all substrates, revealing broad specificity and suitability for diverse lignocellulosic feedstocks, despite differences in biomass structure. Overall, the results highlight the importance of substrate-dependent factors in enzymatic hydrolysis and demonstrate that xylanase from Bacillus sonorensis T6 converts cereal straw into value-added oligosaccharide-rich products, thereby supporting the development of cost-effective, region-specific biorefinery strategies.