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Updated: Jul 5, 2025
Microtiter Dish Biofilm Formation Assay
Published on: January 30, 2011
Key biological processes and essential genes for Proteus mirabilis biofilm development inhibition by protocatechuic
Lu Tian1, Chang Gao1, Jiaxing Lu1
1School of Food Science and Engineering, Shaanxi University of Science and Technology, Xi'an, Shaanxi 710021, China.
Abstract:
Proteus mirabilis is an opportunistic pathogen linked to human urinary tract infections, and is potentially present as a foodborne pathogen within poultry products, including broiler chickens. This report outlines the inhibitory impacts of protocatechuic acid (PCA) on P. mirabilis isolated from a broiler slaughterhouse in China as well as its biofilm. This investigation encompasses assays related to motility and adhesion, bacterial metabolic activity, extracellular polymer (EPS) production, and scavenging capacity. The findings demonstrated that PCA reduced biofilm formation by 61 %. Transcriptomics findings identified that PCA limited the expression of genes like PstS that promote adhesin formation, rbsA and RcsB that alter bacterial chemotaxis, lipopolysaccharide synthesis genes LpxA and EptB, and cell wall synthesis genes MurF and MrdA, and affects the Regulator of Capsule Synthesis (RCS) two-component modulation system. Weighted gene co-expression network analysis (WGCNA) was conducted to identify the core genes. Furthermore, the binding sites of PCA to cytochrome oxidases cydA and cydB, two subunits of ATP synthase atpI and atpH, and ftsZ, which regulate bacterial division, were predicted via molecular docking. Metabolome analysis determined that PCA critically influenced coenzyme A biosynthesis, nucleotide metabolism, alanine, aspartic acid, and glutamate metabolic pathways of P. mirabilis. Therefore, PCA impacts metabolism within bacteria via various pathways, limiting the levels of extracellular polymer and bacterial viability to hinder biofilm formation. Additionally, we prepared an antibacterial plastic film containing protocatechuic acid using PVA as the monomer and CNC as the reinforcing agent. We examined the mechanical and antibacterial properties of this film. When used to wrap chicken, it reduced the total number of colonies, slowed the deterioration of chicken, and maintained the freshness of chicken. In conclusion, the information outlined in this study complements our comprehension of P. mirabilis inhibition by PCA and provides clues for the reduction of foodborne infections associated with P. mirabilis.
Insights
Protocatechuic acid (PCA) effectively inhibits Proteus mirabilis biofilm formation and bacterial viability. PCA also demonstrates potential in antibacterial films for preserving chicken products and reducing foodborne infections.
Area of Science:
- Microbiology
- Food Science
- Biochemistry
Background:
- Proteus mirabilis is an opportunistic pathogen found in poultry, posing a foodborne risk.
- Biofilm formation by P. mirabilis contributes to its pathogenicity and persistence.
- Understanding P. mirabilis inhibition mechanisms is crucial for food safety.
Purpose of the Study:
- To investigate the inhibitory effects of protocatechuic acid (PCA) on P. mirabilis and its biofilm.
- To explore the molecular mechanisms underlying PCA's antibacterial activity.
- To develop and evaluate an antibacterial film incorporating PCA for food preservation.
Main Methods:
- Assays for motility, adhesion, metabolic activity, and extracellular polymer (EPS) production.
- Transcriptomics and Weighted Gene Co-expression Network Analysis (WGCNA).
- Molecular docking simulations and metabolome analysis.
- Preparation and testing of PVA/CNC-based antibacterial films.
Main Results:
- PCA significantly reduced P. mirabilis biofilm formation by 61%.
- PCA downregulated genes involved in adhesion, chemotaxis, cell wall synthesis, and capsule formation.
- Molecular docking predicted PCA binding to key enzymes like cytochrome oxidase and FtsZ.
- Metabolome analysis revealed PCA's impact on central metabolic pathways.
- Antibacterial films containing PCA effectively preserved chicken products.
Conclusions:
- PCA effectively inhibits P. mirabilis through multiple molecular and metabolic pathways.
- PCA demonstrates potential as a natural antimicrobial agent for food safety applications.
- PCA-based films offer a promising strategy to reduce foodborne P. mirabilis contamination and extend shelf life.
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