Immobilized whole-cell fermentation has been proven to be an effective method to improve the performance and cost-effectiveness of Gluconobacter oxydans ATCC 621. In the bio-oxidation of xylose to xylonic acid, the oxygen supply through the immobilized beads is a well-known factor that limits the biocatalytic performance of Gluconobacter oxydans as obligate aerobic bacteria. The activity of immobilized cells could be efficiently improved by execution of pressurized pure oxygen supply strategy. Subsequently, in order to further enhance the production efficiency of xylonic acid and reduce end-product inhibition, online-electrodialysis was employed. Finally, a design of pressurized oxygen supply bioreactor combining with online-electrodialysis was put forward for implementing successive production of xylonic acid. The central features of this a highly integrated design are feasible and thus might enable cost-competitive bacterial xylonic acid production.