title: Unravelling the Structural and Molecular Basis Responsible for the Anti-Biofilm Activity of Zosteric Acid.
abstract: The natural compound zosteric acid, or p-(sulfoxy)cinnamic acid (ZA), is proposed as an alternative biocide-free agent suitable for preventive or integrative anti-biofilm approaches. Despite its potential, the lack of information concerning the structural and molecular mechanism of action involved in its anti-biofilm activity has limited efforts to generate more potent anti-biofilm strategies. In this study a 43-member library of small molecules based on ZA scaffold diversity was designed and screened against Escherichia coli to understand the structural requirements necessary for biofilm inhibition at sub-lethal concentrations. Considerations concerning the relationship between structure and anti-biofilm activity revealed that i) the para-sulfoxy ester group is not needed to exploit the anti-biofilm activity of the molecule, it is the cinnamic acid scaffold that is responsible for anti-biofilm performance; ii) the anti-biofilm activity of ZA derivatives depends on the presence of a carboxylate anion and, consequently, on its hydrogen-donating ability; iii) the conjugated aromatic system is instrumental to the anti-biofilm activities of ZA and its analogues. Using a protein pull-down approach, combined with mass spectrometry, the herein-defined active structure of ZA was matrix-immobilized, and was proved to interact with the E. coli NADH:quinone reductase, WrbA, suggesting a possible role of this protein in the biofilm formation process.
authors.affiliation: Department of Food Environmental and Nutritional Sciences, Università degli Studi di Milano, Milano, Italy.
authors.forename: Cristina
authors.initials: C
authors.lastname: Cattò
authors.affiliation: Department of Pharmaceutical Sciences, Università degli Studi di Milano, Milano, Italy.
authors.forename: Silvia
authors.initials: S
authors.lastname: Dell'Orto
authors.affiliation: Department of Food Environmental and Nutritional Sciences, Università degli Studi di Milano, Milano, Italy; Center for Biofilm Engineering, Montana State University, Bozeman, MT, United States of America.
authors.forename: Federica
authors.initials: F
authors.lastname: Villa
authors.affiliation: Department of Pharmaceutical Sciences, Università degli Studi di Milano, Milano, Italy.
authors.forename: Stefania
authors.initials: S
authors.lastname: Villa
authors.affiliation: Department of Pharmaceutical Sciences, Università degli Studi di Milano, Milano, Italy.
authors.forename: Arianna
authors.initials: A
authors.lastname: Gelain
authors.affiliation: Institute of Chemistry of Molecular Recognition-UOS Rome, CNR, Roma, Italy.
authors.forename: Alberto
authors.initials: A
authors.lastname: Vitali
authors.affiliation: Institute of Biochemistry and Clinical Biochemistry, Catholic University of Rome, Roma, Italy.
authors.forename: Valeria
authors.initials: V
authors.lastname: Marzano
authors.affiliation: Department of Food Environmental and Nutritional Sciences, Università degli Studi di Milano, Milano, Italy.
authors.forename: Sara
authors.initials: S
authors.lastname: Baroni
authors.affiliation: Department of Food Environmental and Nutritional Sciences, Università degli Studi di Milano, Milano, Italy.
authors.forename: Fabio
authors.initials: F
authors.lastname: Forlani
authors.affiliation: Department of Food Environmental and Nutritional Sciences, Università degli Studi di Milano, Milano, Italy.
authors.forename: Francesca
authors.initials: F
authors.lastname: Cappitelli
mesh_terms: D000838:Anions; D000900:Anti-Bacterial Agents / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology*; D018441:Biofilms / Q000187:drug effects* / Q000254:growth & development; D002264:Carboxylic Acids / Q000737:chemistry; D002934:Cinnamates / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology*; D004926:Escherichia coli / Q000187:drug effects* / Q000201:enzymology / Q000254:growth & development; D029968:Escherichia coli Proteins / Q000037:antagonists & inhibitors* / Q000737:chemistry; D006859:Hydrogen / Q000737:chemistry; D008826:Microbial Sensitivity Tests; D050296:Microbial Viability / Q000187:drug effects; D011485:Protein Binding; D012097:Repressor Proteins / Q000037:antagonists & inhibitors* / Q000737:chemistry; D054852:Small Molecule Libraries / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology*; D013329:Structure-Activity Relationship; D013463:Sulfuric Acid Esters / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology*
chemical_list: D000838:Anions; D000900:Anti-Bacterial Agents; D002264:Carboxylic Acids; D002934:Cinnamates; D029968:Escherichia coli Proteins; D012097:Repressor Proteins; D054852:Small Molecule Libraries; D013463:Sulfuric Acid Esters; C081759:WrbA protein, E coli; C503399:zosteric acid; D006859:Hydrogen