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Cell surface display of xylanase on Escherichia coliwas used for the hydrolysis of hemicellulose from pineapple pomace. The feasibility of bioconversion of lignocellulosic biomass into xylooligosaccharides (XOS) was investigated. In this study, pineapple pomace was pretreated, and the hemicellulose fraction was obtained for reaction with the whole-cell biocatalyst. FESEM and FTIR analyses were used to observe morphological and compositional changes of pineapple pomace respectively after pretreatment. Factors affecting hydrolysis reaction were investigated and optimized using the Box-Behnken Design. The highest amount of reducing sugar was produced at pH 7.5, cell loading of 100 g/L wet cell weight, and temperature of 30 °C. The amount of reducing sugar produced was 2.129 mg/ml. HPLC analysis indicated that the XOS produced were xylobiose and xylotriose with a total yield of 5.4 mg/g of pineapple hemicellulose. FESEM analysis on the surface structure of pineapple pomace after the hydrolysis reaction showed clear signs of degradation by xylanase. Based on the results presented, it can be deduced that the application of cell surface display on E. colifor degradation of lignocellulosic biomass is possible and should be explored as it offers great potential for the production of XOS in industry.