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Ginkgolic acids and Ginko biloba extract inhibit Escherichia coli O157:H7 biofilm formation

ABSTRACT: Contamination with enterohemorrhagic Escherichia coli O157:H7 (EHEC) is a worldwide problem but there is no effective therapy available for EHEC infection. Biofilm formation is closely related with EHEC infection and is one of the mechanisms of antimicrobial resistance. Antibiofilm screening of 560 plant secondary metabolites against EHEC shows that ginkgolic acids C15:1 and C17:1 at 5 μg/ml and Ginko biloba extract at 100 μg/ml significantly inhibited EHEC biofilm formation on the surface of polystyrene, nylon membrane, and glass. Importantly, the working concentration of ginkgolic acids and G. biloba extract did not affect bacterial growth and has been known to be non-toxic to human. Transcriptional analyses showed that ginkgolic acid C15:1 repressed curli genes and prophage genes in EHEC, which were corroborated by reduced fimbriae production and biofilm reduction in EHEC. Interestingly, ginkgolic acids and G. biloba extract did not inhibit the biofilm formation of commensal E. coli K-12 strain. The current study suggests that plant secondary metabolites are important resource of biofilm inhibitors, as well as other bioactive compounds. E. coli GeneChip Genome 2.0 Array (Affymetrix, P/N 900551, Santa Clara, USA) was used to study the differential gene expression of the E. coli O157:H7 cells after the treatment with ginkgolic acid C15:1 (0.005 mg/ml). Cells were inoculated into 25 ml LB medium in 250 mL shake flasks with a starting OD600 of 0.05, and cultured at 37oC for 8 h without shaking in the presence or absence of ginkgolic acid C15:1 (5 μg/ml). To prevent RNA degradation, RNase inhibitor (RNAlater, Ambion, TX, USA) was added, and the EHEC cells were collected by centrifugation at 10,000 rpm for 1 min. The cell pellets obtained were immediately frozen with dry ice and stored at -80°C. Total RNA was isolated using a Qiagen RNeasy mini Kit (Valencia, CA, USA).

SUBMITTER: Jintae Lee   Shi Y Ryu  Moo H Cho  Jin-Hyung Lee  Yong-Guy Kim 

PROVIDER: E-GEOD-49367 | ArrayExpress | 2015-07-29



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