Erythritol alters gene transcriptome signatures, cell growth, and biofilm formation in Staphylococcus pseudintermedius

Abstract

Background Erythritol was found to inhibit the growth of microorganisms. The present study aimed to demonstrate the growth inhibition of Staphylococcus pseudintermedius by erythritol and to define the changes in gene transcription signatures induced by erythritol. Changes in the gene transcription profiles were analysed by RNA sequencing and quantitative reverse transcription PCR. Gene ontology analysis was performed to assign functional descriptions to the genes. Results Erythritol inhibited S. pseudintermedius growth in a dose-dependent manner. We then performed a transcriptome analysis of S. pseudintermedius with and without 5% (w/w) erythritol exposure to validate the mechanism of growth inhibition. We revealed that erythritol induced up-regulation of three genes (ptsG, ppdK, and ppdkR) that are related to the phosphoenolpyruvate-dependent sugar phosphotransferase system (PTS). Glucose supplementation restored the up-regulation of the PTS-related genes in response to erythritol. In addition, erythritol down-regulated eleven genes that are located in a single pur-operon and inhibited biofilm formation of S. pseudintermedius. Conclusions These findings indicated that erythritol antagonistically inhibits PTS-mediated glucose uptake, thereby exerting a growth inhibitory effect on S. pseudintermedius. Moreover, erythritol inhibits the ‘de novo’ IMP biosynthetic pathway that may contribute to biofilm synthesis in S. pseudintermedius.

Erythritol alters phosphotransferase gene expression and inhibits the in vitro growth of Staphylococcus coagulans isolated from canines with pyoderma

“…GO analysis predicted that the transcripts of these two genes function as glycoside- and glucose-specific enzyme II components EIICBA, respectively. The upregulation of ptsG gene expression in response to erythritol in SC was in agreement with the data obtained in our recent study using an SP strain ( 26 ). The EIICBA are membrane permeases that play significant roles in the uptake of carbohydrates into the bacterial cytoplasm ( 32 , 33 ).…”

“…In the SP strain, erythritol upregulated ppdK and ppdkR , which are other PTS-related genes predicted to encode pyruvate phosphate dikinase (PPDK) and PPDK regulatory protein (PPDKR), respectively ( 26 ). The same study also revealed that erythritol downregulated the expression of pur operon genes involved in the synthesis of inosinic acid (IMP) leading to purine biosynthesis in the SP strain.…”

“…The following experiments were performed in accordance with the methodology used in a previous study ( 26 ). A single colony of JCM7470 was inoculated into 3 ml of National Institute of Technology and Evaluation Biological Resource Center (NRBC) #802 medium containing 1% hipolypepton (Fujifilm Wako, Osaka, Japan), 0.2% yeast extract (Nacalai Tesque Inc., Kyoto, Japan), and 0.1% MgSO 4 ·7H 2 O (Fujifilm Wako, pH 7.0), and incubated with rotation at 210 rpm until the optical density at 600 nm (OD 600 ) reached 3.4.…”

“…Very recently, we revealed that erythritol suppressed in vitro growth of the SP and SC strains JCM17571 and JCM7470, respectively ( 25 ). Furthermore, erythritol upregulated PEP-PTS-related genes ( ptsG, ppdK , and ppdkR ) in SP JCM17571 ( 26 ). However, the exact molecular mechanism by which erythritol suppresses the growth of SC has not been elucidated.…”

In Vitro/Vivo Mechanisms of Antibacterial Peptide NZ2114 against Staphylococcus pseudintermedius and Its Biofilms

“…When bacterial cells attach to the surface and expand to the biofilms, they are embedded in an extracellular slimy polymeric substance like a shield, therefore becoming more resistant to antimicrobials than those of single or multiple bacteria [38,39]. In the clinical treatment of canines, S. pseudintermedius isolated from the skin of diseased dogs usually has a strong biofilm-forming ability [3,40,41]. AMPs typically exert their effects on biofilms through the following ways: (1) inhibiting biofilm formation in the aggregation and adhesion stages of microorganisms, such as the inhibition effect of HBD2 on Pseudomonas aeruginosa's biofilm formation [42], and (2) the suppression of quorum sensing, such as the inhibitory effect of octopomycin on quorum sensing in Acinetobacter baumannii [43].…”