Action mechanism of 6, 6′-dihydroxythiobinupharidine from Nuphar japonicum, which showed anti-MRSA and anti-VRE activities

Shinya Okamura, Eri Nishiyama, Tomohiro Yamazaki, Nao Otsuka, Shoko Taniguchi, Wakano Ogawa, Tsutomu Hatano, Tomofusa Tsuchiya, Teruo Kuroda

Research output: Contribution to journalArticlepeer-review

19 Citations (Scopus)


Background Multidrug-resistant bacteria, such as methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin resistant enterococci (VRE), cause serious infections at clinical sites, for which the development of new drugs is necessary. We screened candidates for new antibiotics and investigated its action mechanism. Methods An antimicrobial compound was isolated from an extract of Nuphar japonicum. Its chemical structure was determined by NMR, MS, and optical rotation. We measured its minimum inhibitory concentration (MIC) using the microdilution method. The effects of the compound on DNA gyrase and DNA topoisomerase IV were investigated with DNA supercoiling, decatenation, and cleavage assay. Results We isolated and identified 6,6′-dihydroxythiobinupharidine as the antimicrobial compound. The MIC of this compound was 1-4 μg/mL against various MRSA and VRE strains. We also demonstrated that this compound inhibited DNA topoisomerase IV (IC50 was 10-15 μM), but not DNA gyrase in S. aureus, both of which are known to be the targets of quinolone antibiotics and necessary for DNA replication. However, this compound only exhibited slight cross-resistance to norfloxacin-resistant S. aureus, which indicated that DTBN might inhibit other targets besides topoisomerase IV. These results suggest that 6,6′-dihydroxythiobinupharidine may be a potent candidate or seed for novel antibacterial agents. Conclusions DTBN from N. japonicum showed anti-MRSA and anti-VRE activities. DTBN might be involved in the inhibition of DNA topoisomerase IV. General significance DTBN might be useful as a seed compound. The information on the inhibition mechanism of DTBN will be useful for the modification of DTBN towards developing novel anti-MRSA and anti-VRE drug.

Original languageEnglish
Pages (from-to)1245-1252
Number of pages8
JournalBiochimica et Biophysica Acta - General Subjects
Issue number6
Publication statusPublished - Jun 2015


  • DNA topoisomerase IV
  • MRSA
  • Nuphar japonicum
  • VRE

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology


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