The glucose oxidase and catalase immobilized in polyvinyl alcohol and sodium alginate matrix as biocatalyst for conversion of glucose to gluconic acid in bubbling reactor coupled with in situ product removal was investigated. The conversion was inhibited by glucose acid. Using the D335 resin, which has good adsorption capacity of gluconic acid, could relieve the inhibition. By the fed batch- in situ removal, gluconic acid conversion can reach 95.5%. The system was stable, it could successively convert for four cycles, and the average conversion was more than 85.7%.