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As predominant components of hemicelluloses in grasses, methylglucuroarabinoxylans (MeGAXn) are sources for the production of acidic xylooligosaccharides (U-XOS). Bacillus subtilis MR44, an engineered biocatalyst to secrete only the XynC xylanase and Axh43 arabinoxylan hydrolase is capable of processing MeGAXn to exclusively U-XOS. The present studies are directed at the explosion on direct alkaline extraction serving for production of U-XOS. Response Surface Methodology was used to optimize xylan extraction conditions on the sweet sorghum bagasse to achieve maximum hemicelluloses yield. The optimized condition was as follows: extraction time of 3.91 h, extraction temperature of 86.1°C, and NaOH concentration (w/w) of 12.33%. Crude xylan extracted with NaOH revealed a compositional analysis of xylose (79.0%), arabinose (5.3%), glucose (1.7%), lignin and ash (5.6%). After neutralization this xylan preparation supported growth of MR44, processing MeGAXn from sweet sorghum and accumulating U-XOS. The quality of U-XOS produced by MR44 using alkaline-treated sweet sorghum bagasse was comparable to that obtained from purified MeGAXn. Overall, the present study demonstrates that direct alkaline treatment of sweet sorghum bagasse is useful to improve the bioavailability of MeGAXn for MR44-mediated conversion to U-XOS with average degrees of polymerization of 11–12, providing alternative resources with applications in nutrition and human and veterinary medicine. [ABSTRACT FROM AUTHOR]