AbstractXylooligosaccharides (XO), derived from the alkaline (NaOH) extractant of Mikania micrantha, were produced using multiple staged membrane separation and enzymatic xylanolysis. Staged nanofiltration (NMX), ultrafiltration (EUMX), and centrifugation (EMX) processes for the ethanol precipitates were conducted. NMX recovered 97.26% of total xylose and removed 73.18% of sodium ions. Concentrations of total xylose were raised from 10.98 to 51.85 mg/mL by the NMX process. Recovered xylan-containing solids were hydrolyzed by the recombinant Paenibacillus xylanase. 68% XO conversions from total xylose of NMX was achieved in 24 hours. Xylopentaose (DP 5) was the major product from NMX and EMX hydrolysis. Xylohexaose (DP 6) was the major product from EUMX hydrolysis. Results of the present study suggest the applicability for XO production by nanofiltration, as NMX gave higher XO yields compared to those from a conventional ethanol-related lignocellulosic waste conversion process.