Rice husk (RH) is the major agricultural waste obtained during rice hulling process, which can be a sustainable source of xylooligosaccharide (XOS). The current study deals with the production of XOS from Thai rice husk using alkaline pretreatment and enzyme hydrolysis method. The response surface methodology consisted of central composite design and Box-Behnken design was employed to achieve the maximum response in alkaline pretreatment and XOS production, respectively. The optimum conditions for alkaline pretreatment to recover maximum xylan yield were 12-18% of alkaline concentration, the temperature at 110-120 °C, and steaming time for 37.5-40 min. The FTIR results suggested that the extracted sample was the xylan fraction. The maximum XOS production of 17.35 ± 0.31 mg XOS per mL xylan was observed in the run conditions of 6.25 mg enzyme per g xylan, 9 h of incubation time, and 5% of xylan. The results revealed that the xylan extracted from RH by using an effective base couple with the steam application and the enzymatic hydrolysis help to maximize the yield of XOS, which can be further used in functional foods and dietary supplements.