Involvement of AprD in regulating biofilm structure, matrix secretion, and cell metabolism of meat-borne Pseudomonas fragi during chilled storage

Abstract

Pseudomonas fragi is by far one of the most threatening species in the spoilage of chilled meat that is stored under aerobic conditions. The membrane protein AprD is a well-established regulator controlling protease secretion in Pseudomonas spp. However, its exact roles in modulating metabolic pathways and spoilage potential of P. fragi at the molecular level remain undefined. Here, an in-frame deletion mutation of aprD was used to explore the impacts on their biofilm structure, matrix secretion, and cell metabolism. The results showed that ΔaprD formed relatively disorganized loose aggregation in biofilm, resulting in a thinner structure and more dead cells. Meanwhile, marked changes in the content of extracellular carbohydrates and proteins were observed. Furthermore, intracellular metabolomic profiling revealed the involvement of aprD in several cellular metabolic pathways, mostly including the carbohydrate pathway, amino acid pathway, and nucleotide pathway, while the characterization of extracellular metabolism clarified the variations in the spoilage-related metabolites (e.g., creatine, IMP, spermine, fatty acids, amino acids, and oligopeptides) could be highly correlated with aprD deletion. In this finding, we indicated that aprD could be responsible for cell reproduction and in situ spoilage potential of P. fragi NMC25 during chilled storage by controlling related metabolism and nutrients utilization. Thus, our results will contribute to an improved understanding of the regulatory mechanism of aprD gene in meat spoilage contaminated with P. fragi, which can be valuable to ensure the quality and safety of meat.

Keywords: AprD; Biofilm structure; Extracellular polymeric substances; Meat spoilage; Metabolic alterations; Pseudomonas fragi.