Proteomic Analysis Revealed Metabolic Inhibition and Elongation Factor Tu Deamidation by p-Coumaric Acid in Cronobacter sakazakii

Abstract

Screening drugs and compounds to fight against Cronobacter sakazakii (C. sakazakii), one of the most common pathogens that can cause fatal necrotizing enterocolitis, septicema and meningitis, is still needed. We found that p-coumaric acid (pCA) has an inhibitory effect on C. sakazakii in vitro and in vivo. Proteomic changes of C. sakazakii BAA-894 exposed to pCA were studied to reveal the antibacterial mechanisms involved. A total of 1,553 proteins were identified in C. sakazakii BAA-894 by label-free proteomics analysis. Fuzzy cluster analysis showed that 33 were up-regulated, and 110 were down-regulated with pCA treatment. Gene Ontology (GO) analysis concluded that pCA caused the change of metabolic state of bacteria and generally in the state of metabolic inhibition. KEGG Enrichment Analysis (KEGG) analysis showed that pCA inhibited energy metabolism and distorted the balance of amino acid metabolism. Posttranslational modification analysis showed that pCA affected the deamidation of three proteins, including Elongation factor Tu, one of the vital proteins in bacteria. Molecular docking suggested the hydrogen bond between the pCA carboxyl group and Elongation factor Tu Asn-64 might contribute to deamidation. Overall, we found that pCA interfered with cellular energy and amino acid metabolism and promoted elongation factor Tu deamidation, suggesting that pCA can be an effective natural substitute to control C. sakazakii.

EnvZ/OmpR Controls Protein Expression and Modifications in Cronobacter sakazakii for Virulence and Environmental Resilience

“…Our previous study found that the inhibition of C. sakazakii growth by p-coumaric acid is associated with the promotion of deamidation in bacterial elongation factors. 38 Additionally, the knockout of dnaK increased the deamidation of several proteins in C. sakazakii. 37 In this study, we discovered a connection between the envZ/ompR and protein deamidation.…”

“…An association was found between protein deamidation and growth disadvantage in C. sakazakii in our previous studies. 37,38 Proteomic analysis was employed to investigate potential protein deamidation resulting from the knockout of the envZ/ ompR gene. It was observed that 196 proteins exhibited deamidation modifications, among which 17 proteins (8.6%) showed a significant increase in deamidation after envZ/ompR knockout (Figure 5A).…”

Genetic determinants of thermal resistance in foodborne bacterial pathogens

“…Apart from the genomic islands and genes reported with a specific role in elevated thermal resistance, there has been evidence of a general stress tolerance mechanism mediated by the dnaK gene in C. sakazakii (Lu et al., 2023). A study that used gene deletion‐based assessment demonstrated that the dnaK gene in C. sakazakii is associated with critical attributes such as adhesion, infiltration, and resistance to acidity.…”

Proteomic Profiling of Burkholderia andropogonis

“…This is supported by the work reported by Wolska et al [38], that Eschericia coli dnaK and dnaJ mutants showed antibiotic susceptibility. DnaK is Hsp70, which is a molecular chaperone expressed in response to stress [45,46]. Hence, targeting molecular chaperones of such nature expressed by B. andropogonis, suppressing their expression and their resultant proteins thereof, might consequently result in finding an effective control method for this devastating plant pathogen.…”