Cyclodextrin can be produced from the degradation of starch using the enzyme cyclodextrin glucanotransferase (CGTase). Its favorable characteristics have seen its use in industries such as cosmetics, personal care, textiles, and pharmaceuticals. The production of CGTase by wild-type Bacillus sp. is low, so recombinant Escherichia coli has been used for higher enzyme yields. Cell lysis and plasmid instability are among the challenges that emerge during recombinant enzyme excretion that hinder the production of recombinant CGTase in E. coli. In this study, a cell immobilization technique using pineapple peel was employed to overcome this problem. The effects of changing process parameters such as pH, contact time, and temperature on the immobilization of recombinant E. coli were studied, one parameter at a time. The optimal conditions for the production of cyclodextrin were pH 8 leading to a 55.95% immobilization yield, a contact time of 24 h for a 55.16% immobilization yield, and a temperature of 25 ℃ for 53.11% immobilization yield. In brief, pineapple peel was determined to be a suitable supporting matrix and optimized process parameters increased the immobilization of recombinant E. coli, improving CGTase production while maintaining low cell lysis.