ArticlePublished: 09 October 2025Zhenfeng Wang1,2 na1,Junhua Ma2 na1,Jintuan Lin2 na1,Congcong Li2,3 na1,Yong Xiang4,Zhijian Yu2,Bing Bai2,Guiqiu Li1,5 &…Ying Wei1,6 The Journal of Antibiotics (2025)Cite this articleSubjectsAntimicrobial resistanceClinical microbiologyProtein–protein interaction networksAbstractThe escalating prevalence of multidrug-resistant (MDR) gram-positive bacteria, including methicillin-resistant Staphylococcus aureus (MRSA) and linezolid-resistant Enterococcus faecalis, highlights the critical demand for new antibacterial agents that target resistance pathways. BB-Cl-Amidine is originally considered as a peptidyl arginine deiminase inhibitor and till now, its potential antimicrobial activity has not been explored. This study sought to evaluate the antibacterial effectiveness and underlying mechanisms of BB-Cl-Amidine against MDR gram-positive pathogens. The results showed that BB-Cl-Amidine exhibited potent antibacterial activity with minimum inhibitory concentration (MIC) values ranging from 25 μM to 50 μM against MRSA, E. faecalis and various of Gram-positive bacteria clinical isolates. At sub-MIC concentrations, BB-Cl-Amidine significantly reduced biofilm formation in both S. aureus and E. faecalis. Moreover, the increased permeability and depolarizing membrane potential of S. aureus was found by BB-Cl-Amidine. The antibacterial activity of BB-Cl-Amidine can be neutralized by cardiolipin (CL) and phosphatidylglycerol (PG). Furthermore, BB-Cl-Amidine exposure resulted in the abnormal expression of functional proteins correlated with the cell membranes and phospholipid metabolas. In summary, the potential antibacterial and anti-biofilm activities of BB-Cl-Amidine are demonstrated via membrane disruption, offering a promising scaffold for combating MDR Gram-positive infections.This is a preview of subscription content, access via your institutionAccess optionsSubscribe to this journalReceive 12 print issues and online access269,00 € per yearonly 22,42 € per issueLearn moreBuy this articlePurchase on SpringerLinkInstant access to full article PDFBuy nowPrices may be subject to local taxes which are calculated during checkoutFig. 1Fig. 2Fig. 3Fig. 4Fig. 5ReferencesAhmad-Mansour N, Loubet P, Pouget C, et al. Staphylococcus aureus toxins: an update on their pathogenic properties and potential treatments[J]. Toxins. 2021;13:677.Article CAS PubMed PubMed Central Google Scholar Piewngam P, Otto M. Staphylococcus aureus colonization and strategies for decolonization. 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BMC Microbiol. 2024;24:224.Article PubMed PubMed Central Google Scholar Download referencesAcknowledgementsThe authors express their gratitude to Wei Guang Pan and Jie Lian from the Department of Laboratory Medicine at Shenzhen Nanshan People’s Hospital for their assistance in identifying and preserving the bacterial isolates.FundingThis research was made possible through the following grants: National Natural Science Foundation of China (82172283); Guangdong Basic and Applied Basic Research Foundation (2022A1515110096, 2024A1515013276); Sanming Project of Medicine in Shenzhen (SMGC202305029); Shenzhen Key Medical Discipline Construction Fund (SZXK06162); and the Shenzhen Nanshan District Scientific Research Program of the People’s Republic of China (NS2024001; NS2023008; NSZD2024023; NSZD2024036; NS2022046; NS2024001Z; NS2024038).Author informationAuthor notesThese authors contributed equally: Zhenfeng Wang, Junhua Ma, Jintuan Lin, Congcong Li.Authors and AffiliationsClinical Medical College, Jiamusi University, Jiamusi, Heilongjiang, ChinaZhenfeng Wang, Guiqiu Li & Ying WeiDepartment of Infectious Diseases and Shenzhen Key Laboratory for Endogenous Infections, Shenzhen Nanshan People’s Hospital, Shenzhen, ChinaZhenfeng Wang, Junhua Ma, Jintuan Lin, Congcong Li, Zhijian Yu & Bing BaiSchool of Basic Medical Sciences, Shenzhen University Medical School, Shenzhen University, Shenzhen, ChinaCongcong LiMarketing Department ofGuangzhou branch ofVarian Medical Equipment Trading (Beijing) Corp, Guangzhou, ChinaYong XiangLaboratory Medicine Center, Nanshan People’s Hospital, Shenzhen, ChinaGuiqiu LiHeilongjiang Provincial Hospital, Harbin, Heilongjiang, ChinaYing WeiAuthorsZhenfeng WangView author publicationsSearch author on:PubMed Google ScholarJunhua MaView author publicationsSearch author on:PubMed Google ScholarJintuan LinView author publicationsSearch author on:PubMed Google ScholarCongcong LiView author publicationsSearch author on:PubMed Google ScholarYong XiangView author publicationsSearch author on:PubMed Google ScholarZhijian YuView author publicationsSearch author on:PubMed Google ScholarBing BaiView author publicationsSearch author on:PubMed Google ScholarGuiqiu LiView author publicationsSearch author on:PubMed Google ScholarYing WeiView author publicationsSearch author on:PubMed Google ScholarContributionsZ.W. taken part in an antimicrobial susceptibility test and growth curve experiments and drafted the manuscript. J.M. and J.L. performed the time-killing analysis. C.L. and Y.X. performed a biofilm assay and cytotoxicity assay. Y.W. taken part in the proteomics data analysis. B.B. participated in the collection and statistical analyses of the clinical isolates of S. aureus and E. faecalis coliform bacteria. Z.Y. and G.L. designed the study, took part in the analysis of the data, and provided important revisions of the manuscript for valuable intellectual content. valuable intellectual content provided important manuscript revisions. All authors have read and approved this manuscript.Corresponding authorsCorrespondence to Bing Bai, Guiqiu Li or Ying Wei.Ethics declarationsConflict of interestThe authors declare no competing interests.Ethical approvalApproval of ethics and permission for participation. All techniques were conducted in accordance with applicable norms and regulations, authorized by the Ethics Committee of Shenzhen Nanshan People’s Hospital, and in compliance with the Helsinki Declaration of 1964 and its later revisions or similar ethical standards. All experimental techniques involving human volunteers received approval from the Institutional Ethics Committee of Shenzhen Nanshan People’s Hospital. Bacterial strains were obtained in accordance with national standards for the regular clinical therapy of patients in China. Consequently, informed permission was not acquired, and a waiver of informed consent was sanctioned by the Institutional Ethics Committee of Shenzhen Nanshan People’s Hospital.Additional informationPublisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.Supplementary informationFigure S1 Evaluation of the effect of different concentrations of BB-Cl-Amidine on established biofilmsRights and permissionsSpringer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.Reprints and permissionsAbout this article