Synergistic multi-enzyme system of Sphingobacterium sp. for enhanced penicillin G degradation: Pathways, proteomics, and binding dynamics analysis.

The persistent contamination of penicillin G sodium (PGNa) in pharmaceutical fermentation residues poses critical environmental and public health risks, demanding urgent global remediation strategies. While microbial degradation represents a promising solution, the enzymatic mechanisms governing PGNa detoxification remain poorly elucidated. This study demonstrates the exceptional PGNa degradation capacity of a multi-enzyme system derived from Sphingobacterium sp. SQW1. Notably, meropenem induction enhanced degradation enzyme activity by 13-fold compared to baseline levels. Furthermore, the effects of various mediators on the enzymatic catalysis of PGNa were systematically investigated. The degradation enzyme demonstrated robust thermal stability and broad pH adaptability, with particularly notable activity observed at 55 °C (1,585.32 U/mL). Through proteomics and binding analysis, AmpC was identified as the pivotal enzyme among 43 differentially expressed candidates, demonstrating strong PGNa affinity and catalytic stability. LC-MS-based pathway analysis identified three primary degradation routes: β-lactam ring hydrolysis, penicillin acylase-mediated side chain cleavage, and oxidative decarboxylation/demethylation cascades. Our findings provide the first molecular-level characterization of PGNa degradation by bacterial enzyme complexes, establishing a groundbreaking framework for enzymatic bioremediation of β-lactam antibiotic residues. This work advances both fundamental understanding and practical applications in waste fermentation residue management, offering an eco-efficient alternative to conventional antibiotic elimination technologies.