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Abstract Background An efficient biodegradation-strengthening approach was developed to improve penicillin G degradation from industrial bacterial residue in an expanded bed adsorption bioreactor (EBAB) is reported in this paper. Results Paracoccus sp. strain KDSPL-02 was isolated based on its ability to use penicillin G as the sole carbon and nitrogen source. Strain identification was based on analyses of morphology, physio-biochemical characteristics, and 16S rDNA sequences. The effects of temperature, pH, PVA-sodium alginate concentration, calcium chloride concentration and initial penicillin G concentration were investigated. Repeated operations of immobilized cells with EBAB, At initial penicillin concentrations below 2.0 g L− 1, the continuous mode could reach more than 20 times, and the degradation rate reached 100%. Conclusions The present study suggests that the EBAB system can be utilized for the simple and economical biodegradation of penicillin G from industrial bacterial residue.