title: Selective and Efficient Elimination ofVibrio choleraewith a Chemical Modulator that Targets Glucose Metabolism.
abstract: Vibrio cholerae, a Gram-negative bacterium, is the causative agent of pandemic cholera. Previous studies have shown that the survival of theseventhpandemic El Tor biotypeV. choleraestrain N16961 requires production of acetoin in a glucose-rich environment. The production of acetoin, a neutral fermentation end-product, allowsV. choleraeto metabolize glucose without a pH drop, which is mediated by the production of organic acid. This finding suggests that inhibition of acetoin fermentation can result inV. choleraeelimination by causing a pH imbalance under glucose-rich conditions. Here, we developed a simple high-throughput screening method and identified an inducer of medium acidification (iMAC). Of 8364 compounds screened, we identified one chemical, 5-(4-chloro-2-nitrobenzoyl)-6-hydroxy-1,3-dimethylpyrimidine-2,4(1H,3H)-dione, that successfully killed glucose-metabolizing N16961 by inducing acidic stress. When N16961 was grown with abundant glucose in the presence of iMAC, acetoin production was completely suppressed and concomitant accumulation of lactate and acetate was observed. Using a beta-galactosidase activity assay with a single-copy palsD::lacZreporter fusion, we show that that iMAC likely inhibits acetoin production at the transcriptional level. Thin-layer chromatography revealed that iMAC causes a significantly reduced accumulation of intracellular (p)ppGpp, a bacterial stringent response alarmone known to positively regulate acetoin production.In vivobacterial colonization and fluid accumulation were also markedly decreased after iMAC treatment. Finally, we demonstrate iMAC-induced bacterial killing for 22 differentV. choleraestrains belonging to diverse serotypes. Together, our results suggest that iMAC, acting as a metabolic modulator, has strong potential as a novel antibacterial agent for treatment against cholera.
authors.affiliation: Department of Microbiology and Immunology, Yonsei University College of Medicine Seoul, South Korea.
authors.forename: Young Taek
authors.initials: YT
authors.lastname: Oh
authors.affiliation: Department of Microbiology and Immunology, Yonsei University College of MedicineSeoul, South Korea; Brain Korea 21 Project for Medical Science, Yonsei University College of MedicineSeoul, South Korea.
authors.forename: Hwa Young
authors.initials: HY
authors.lastname: Kim
authors.affiliation: Department of Pharmacy, College of Pharmacy, Hanyang University Ansan, South Korea.
authors.forename: Eun Jin
authors.initials: EJ
authors.lastname: Kim
authors.affiliation: Department of Microbiology and Immunology, Yonsei University College of MedicineSeoul, South Korea; Brain Korea 21 Project for Medical Science, Yonsei University College of MedicineSeoul, South Korea.
authors.forename: Junhyeok
authors.initials: J
authors.lastname: Go
authors.affiliation: Department of Microbiology and Immunology, Yonsei University College of MedicineSeoul, South Korea; Brain Korea 21 Project for Medical Science, Yonsei University College of MedicineSeoul, South Korea.
authors.forename: Wontae
authors.initials: W
authors.lastname: Hwang
authors.affiliation: Bio and Drug Discovery Division, Korea Research Institute of Chemical Technology Daejeon, South Korea.
authors.forename: Hyoung Rae
authors.initials: HR
authors.lastname: Kim
authors.affiliation: Department of Pharmacy, College of Pharmacy, Hanyang UniversityAnsan, South Korea; Institute of Pharmacological Research, Hanyang UniversityAnsan, South Korea.
authors.forename: Dong Wook
authors.initials: DW
authors.lastname: Kim
authors.affiliation: Department of Microbiology and Immunology, Yonsei University College of MedicineSeoul, South Korea; Brain Korea 21 Project for Medical Science, Yonsei University College of MedicineSeoul, South Korea; Institute for Immunology and Immunological Diseases, Yonsei University College of MedicineSeoul, South Korea.
authors.forename: Sang Sun
authors.initials: SS
authors.lastname: Yoon
mesh_terms: D000093:Acetoin / Q000378:metabolism; D000143:Acids / Q000378:metabolism; D000818:Animals; D000900:Anti-Bacterial Agents / Q000737:chemistry / Q000494:pharmacology*; D050260:Carbohydrate Metabolism / Q000187:drug effects* / Q000235:genetics; D002771:Cholera / Q000188:drug therapy / Q000382:microbiology; D002855:Chromatography, Thin Layer; D003601:Cytotoxicity Tests, Immunologic; D004195:Disease Models, Animal; D004926:Escherichia coli / Q000235:genetics; D005285:Fermentation; D015964:Gene Expression Regulation, Bacterial / Q000187:drug effects; D005947:Glucose / Q000378:metabolism*; D057166:High-Throughput Screening Assays / Q000379:methods; D006863:Hydrogen-Ion Concentration; D051379:Mice; D050296:Microbial Viability; D008855:Microscopy, Electron, Scanning; D014733:Vibrio / Q000145:classification; D014734:Vibrio cholerae / Q000187:drug effects* / Q000235:genetics / Q000254:growth & development* / Q000378:metabolism*; D001616:beta-Galactosidase / Q000032:analysis
chemical_list: D000143:Acids; D000900:Anti-Bacterial Agents; D000093:Acetoin; D001616:beta-Galactosidase; D005947:Glucose
keywords: Vibrio cholerae; acetoin; glucose metabolism; inducer of medium acidification (iMAC); mixed acid fermentation