title: Antimicrobial mechanism of resveratrol-trans-dihydrodimer produced from peroxidase-catalyzed oxidation of resveratrol.
abstract: Plant polyphenols are known to have varying antimicrobial potencies, including direct antibacterial activity, synergism with antibiotics and suppression of bacterial virulence. We performed the in vitro oligomerization of resveratrol catalyzed by soybean peroxidase, and the two isomers (resveratrol-trans-dihydrodimer and pallidol) produced were tested for antimicrobial activity. The resveratrol-trans-dihydrodimer displayed antimicrobial activity against the Gram-positive bacteria Bacillus cereus, Listeria monocytogenes, and Staphylococcus aureus (minimum inhibitory concentration (MIC) = 15.0, 125, and 62.0 μM, respectively) and against Gram-negative Escherichia coli (MIC = 123 μM, upon addition of the efflux pump inhibitor Phe-Arg-β-naphthylamide). In contrast, pallidol had no observable antimicrobial activity against all tested strains. Transcriptomic analysis implied downregulation of ABC transporters, genes involved in cell division and DNA binding proteins. Flow cytometric analysis of treated cells revealed a rapid collapse in membrane potential and a substantial decrease in total DNA content. The active dimer showed >90% inhibition of DNA gyrase activity, in vitro, by blocking the ATP binding site of the enzyme. We thus propose that the resveratrol-trans-dihydrodimer acts to: (1) disrupt membrane potential; and (2) inhibit DNA synthesis. In summary, we introduce the mechanisms of action and the initial evaluation of an active bactericide, and a platform for the development of polyphenolic antimicrobials.
authors.affiliation: Department of Chemical and Biological Engineering, Rensselaer Polytechnic Institute, Center for Biotechnology and Interdisciplinary Studies, Troy, New York, 12180-3590.
authors.forename: Mauricio
authors.initials: M
authors.lastname: Mora-Pale
authors.affiliation: Department of Chemical and Biological Engineering, Rensselaer Polytechnic Institute, Center for Biotechnology and Interdisciplinary Studies, Troy, New York, 12180-3590.
authors.forename: Namita
authors.initials: N
authors.lastname: Bhan
authors.affiliation: Department of Chemistry and Chemical Biology, Rensselaer Polytechnic Institute, Center for Biotechnology and Interdisciplinary Studies, Troy, New York.
authors.forename: Sayaka
authors.initials: S
authors.lastname: Masuko
authors.affiliation: Henry Wellcome Building for Biocatalysis, Biosciences, College of Life and Environmental Science, University of Exeter, Exeter, Devon.
authors.forename: Paul
authors.initials: P
authors.lastname: James
authors.affiliation: Department of Chemistry and Chemical Biology, Rensselaer Polytechnic Institute, Center for Biotechnology and Interdisciplinary Studies, Troy, New York.
authors.forename: Julia
authors.initials: J
authors.lastname: Wood
authors.affiliation: NMR Core Facility, Rensselaer Polytechnic Institute, Center for Biotechnology and Interdisciplinary Studies, Troy, New York.
authors.forename: Scott
authors.initials: S
authors.lastname: McCallum
authors.affiliation: Department of Chemical and Biological Engineering, Rensselaer Polytechnic Institute, Center for Biotechnology and Interdisciplinary Studies, Troy, New York, 12180-3590.|Department of Chemistry and Chemical Biology, Rensselaer Polytechnic Institute, Center for Biotechnology and Interdisciplinary Studies, Troy, New York.|Department of Biology, Rensselaer Polytechnic Institute, Center for Biotechnology and Interdisciplinary Studies, Troy, New York.|Department of Biomedical Engineering, Rensselaer Polytechnic Institute, Center for Biotechnology and Interdisciplinary Studies, Troy, New York.
authors.forename: Robert J
authors.initials: RJ
authors.lastname: Linhardt
authors.affiliation: Department of Chemical and Biological Engineering, Rensselaer Polytechnic Institute, Center for Biotechnology and Interdisciplinary Studies, Troy, New York, 12180-3590. dordick@rpi.edu.|Department of Biology, Rensselaer Polytechnic Institute, Center for Biotechnology and Interdisciplinary Studies, Troy, New York. dordick@rpi.edu.|Department of Biomedical Engineering, Rensselaer Polytechnic Institute, Center for Biotechnology and Interdisciplinary Studies, Troy, New York. dordick@rpi.edu.|Department of Material Science and Engineering, Rensselaer Polytechnic Institute, Center for Biotechnology and Interdisciplinary Studies, Troy, New York. dordick@rpi.edu.
authors.forename: Jonathan S
authors.initials: JS
authors.lastname: Dordick
authors.affiliation: Department of Chemical and Biological Engineering, Rensselaer Polytechnic Institute, Center for Biotechnology and Interdisciplinary Studies, Troy, New York, 12180-3590. koffam@rpi.edu.|Department of Biology, Rensselaer Polytechnic Institute, Center for Biotechnology and Interdisciplinary Studies, Troy, New York. koffam@rpi.edu.
authors.forename: Mattheos A G
authors.initials: MA
authors.lastname: Koffas
mesh_terms: D000890:Anti-Infective Agents / Q000378:metabolism*; D004269:DNA, Bacterial / Q000378:metabolism; D004926:Escherichia coli / Q000187:drug effects* / Q000502:physiology; D020869:Gene Expression Profiling; D006094:Gram-Positive Bacteria / Q000187:drug effects* / Q000502:physiology; D008564:Membrane Potentials / Q000187:drug effects; D008826:Microbial Sensitivity Tests; D010084:Oxidation-Reduction; D009195:Peroxidase / Q000378:metabolism*; D000077185:Resveratrol; D013025:Glycine max / Q000201:enzymology; D013267:Stilbenes / Q000378:metabolism*
chemical_list: D000890:Anti-Infective Agents; D004269:DNA, Bacterial; D013267:Stilbenes; D009195:Peroxidase; D000077185:Resveratrol
keywords: DNA gyrase inhibition; antimicrobial; disrupted membrane potential; peroxidase; resveratrol