A hemicellulose-first strategy of acetic acid pretreatment was applied for the protein-rich paper mulberry biowaste (PMB) to produce amino acid-rich xylo-oligosaccharide (XOS). In addition, the pretreated PMB was further used to produce fermentable monosaccharides by enzymatic hydrolysis. Results showed a yield of 18.2% (based on xylan in raw PMB) was achieved for XOS in hydrolyzate under the optimal pretreatment conditions (168 °C, 68 min, 1% acetic acid). Besides, it’s found that the obtained hydrolyzate contains 14.3% (based on amino acids in raw PMB) amino acids degraded from the protein in PMB. Furthermore, an additional enzymatic hydrolysis process with endo-xylanase was used to upgrade the XOS with DP of 2–6 in the hydrolysate, which could increase XOS yield to 29.8%. Amino acid contents were unchanged after post-enzymatic hydrolysis. Besides, a glucose yield of 51.7% (11.5 g/L glucose) could be achieved for the pretreated PMB after enzymatic hydrolysis. This work serves as proof of concept for creating higher-value XOS from biorefinery processes considering protein-enriched low-value biomass feedstocks.
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