Gram-positive bacteria are responsible for a wide range of infections in humans. In most Gram-positive bacteria, sortase A plays a significant role in attaching virulence factors to the bacteria's cell wall. These cell surface proteins play a significant role in virulence and pathogenesis. Even though antibiotics are available to treat these infections, there is a continuous search for an alternative strategy due to an increase in antibiotic resistance. Thus, using anti-sortase drugs to combat these bacterial infections may be a promising approach. Here, we describe a method for targeting Gram-positive bacterial infection by combining curcumin and trans-chalcone as sortase A inhibitors. We have used curcumin and trans-chalcone alone and in combination as a sortase A inhibitor. We have seen ~78%, ~43%, and ~94% inhibition when treated with curcumin, trans-chalcone, and a combination of both compounds, respectively. The compounds have also shown a significant effect on biofilm formation, IgG binding, protein A recruitment, and IgG deposition. We discovered that combining curcumin and trans-chalcone is more effective against Gram-positive bacteria than either compound alone. The present work demonstrated that a combination of these natural compounds could be used as an antivirulence therapy against Gram-positive bacterial infection.

Department of Biotechnology National Institute of Pharmaceutical Education and Research Guwahati 781101 India

Department of Medicinal Chemistry National Institute of Pharmaceutical Education and Research Kolkata 700054 India

Zobrazit více v PubMed

Alharthi S, Popat A, Ziora ZM, Moyle PM (2023) Sortase A inhibitor protein nanoparticle formulations demonstrate antibacterial synergy when combined with antimicrobial peptides. Molecules 28(5):2114. https://doi.org/10.3390/molecules28052114 PubMed DOI PMC

Atkins KL, Burman JD, Chamberlain ES et al (2008) S. aureus IgG-binding proteins SpA and Sbi: host specificity and mechanisms of immune complex formation. Mol Immunol 45(6):1600–1611. https://doi.org/10.1016/j.molimm.2007.10.021

Bae J, Jeon JE, Lee YJ, Lee HS, Sim CJ, Oh KB, Shin J (2011) Sesterterpenes from the tropical sponge Coscinoderma sp. J Nat Prod 74:1805–1811. https://doi.org/10.1021/np200492k PubMed DOI

Becker S, Frankel MB, Schneewind O, Missiakas D (2014) Release of protein A from the cell wall of Staphylococcus aureus. Proc Natl Acad Sci USA 111(4):1574–1579. https://doi.org/10.1128/mBio.00575-13 PubMed DOI PMC

Bekker H, Berendsen HJC, Dijkstra EJ et al (1993) Gromacs-a parallel computer for molecular-dynamics simulations. In: 4th International Conference on Computational Physics PC92. World Scientific Publishing, pp 252–256

Benov L (2019) Effect of growth media on the MTT colorimetric assay in bacteria. PLoS ONE 14(8):e0219713. https://doi.org/10.1371/journal.pone.0219713 PubMed DOI PMC

Cascioferro S, Totsika M, Schillaci D (2014) Sortase A: an ideal target for anti-virulence drug development. Microb Pathog 77:105–112. https://doi.org/10.1016/j.micpath.2014.10.007 PubMed DOI

Chen F, Liu B, Wang D et al (2014) Role of sortase A in the pathogenesis of Staphylococcus aureus-induced mastitis in mice. FEMS Microbiol Lett 351(1):95–103. https://doi.org/10.1111/1574-6968.12354 PubMed DOI

De Jong NWM, Van Kessel KPM, Van Strijp JAG (2019) Immune evasion by Staphylococcus aureus. Microbiol Spectr 7(2). https://doi.org/10.1128/microbiolspec.GPP3-0061-2019

Dong J, Zhang L, Xu N, Zhou S, Song Y, Yang Q, Liu Y, Yang Y, Ai X (2021) Rutin reduces the pathogenicity of Streptococcus agalactiae to tilapia by inhibiting the activity of sortase A. Aquaculture 530:735743. https://doi.org/10.1016/j.aquaculture.2020.735743 DOI

Donlan RM (2001) Biofilm formation: a clinically relevant microbiological process. Clin Infect Dis 33(8):1387–1392. https://doi.org/10.1086/322972 PubMed DOI

Falugi F, Kim HK, Missiakas DM, Schneewind O (2013) Role of protein A in the evasion of host adaptive immune responses by Staphylococcus aureus. MBio 4(5):e00575–13. https://doi.org/10.1128/mBio.00575-13

Foster TJ, McDevitt D (1994) Surface-associated proteins of Staphylococcus aureus: their possible roles in virulence. FEMS Microbiol Lett 118(3):199–205. https://doi.org/10.1111/j.1574-6968.1994.tb06828.x PubMed DOI

Gu K, Ding L, Wang Z et al (2023) Wogonin attenuates the pathogenicity of Streptococcus pneumoniae by double-target inhibition of Pneumolysin and Sortase A. J Cell Mol Med 27(4):563–575. https://doi.org/10.1111/jcmm.17684 PubMed DOI PMC

Gupta A, Mahajan S, Sharma R (2015) Evaluation of antimicrobial activity of Curcuma longa rhizome extract against Staphylococcus aureus. Biotechnol Rep (Amst) 6:51–55. https://doi.org/10.1016/j.btre.2015.02.001 PubMed DOI

Hu P, Huang P, Chen MW (2013) Curcumin reduces Streptococcus mutans biofilm formation by inhibiting sortase A activity. Arch Oral Biol 58(10):1343–1348. https://doi.org/10.1016/j.archoralbio.2013.05.004

Jang KH, Chung SC, Shin J, Lee SH, Kim TI, Lee HS, Oh KB (2007) Aaptamines as sortase A inhibitors from the tropical sponge Aaptos aaptos. Bioorg Med Chem Lett 17:5366–5369. https://doi.org/10.1016/j.bmcl.2007.08.007 PubMed DOI

Jeon JE, Na Z, Jung M, Lee HS, Sim CJ, Nahm K, Oh KB, Shin J (2010) Discorhabdins from the Korean marine sponge Sceptrella sp. J Nat Prod 73:258–262. https://doi.org/10.1021/np9005629 PubMed DOI

Kang SS, Kim JG, Lee TH, Oh KB (2006) Flavonols inhibit sortases and sortase-mediated Staphylococcus aureus clumping to fibrinogen. Biol Pharm Bull 29:1751–1755. https://doi.org/10.1248/bpb.29.1751 PubMed DOI

Khatoon Z, McTiernan CD, Suuronen EJ, Mah TF, Alarcon EI (2018) Bacterial biofilm formation on implantable devices and approaches to its treatment and prevention. Heliyon 4(12):e01067. https://doi.org/10.1016/j.heliyon.2018.e01067 PubMed DOI PMC

Kim SH, Shin DS, Oh MN, Chung SC, Lee JS, Chang IM, Oh KB (2003) Inhibition of sortase, a bacterial surface protein anchoring transpeptidase, by β-sitosterol-3-O-glucopyranoside from Fritillaria verticillata. Biosci Biotechnol Biochem 67(11):2477–2479. https://doi.org/10.1271/bbb.67.2477 PubMed DOI

Kim SH, Shin DS, Oh MN, Chung SC, Lee JS, Oh KB (2004) Inhibition of the bacterial surface protein anchoring transpeptidase sortase by isoquinoline alkaloids. Biosci Biotechnol Biochem 68(2):421–424. https://doi.org/10.1271/bbb.68.421 PubMed DOI

Lee S, Song IH, Lee JH, Yang WY, Oh KB, Shin J (2014) Sortase A inhibitory metabolites from the roots of Pulsatilla koreana. Bioorg Med Chem Lett 24(1):44–48. https://doi.org/10.1016/j.bmcl.2013.12.006 PubMed DOI

Lee YJ, Han YR, Park W, Nam SH, Oh KB, Lee HS (2010) Synthetic analogs of indole-containing natural products as inhibitors of sortase A and isocitrate lyase. Bioorg Med Chem Lett 20(23):6882–6885. https://doi.org/10.1016/j.bmcl.2010.10.029 PubMed DOI

Li H, Chen Y, Zhang B et al (2016) Inhibition of sortase A by chalcone prevents Listeria monocytogenes infection. Biochem Pharmacol 106:19–29. https://doi.org/10.1016/j.bcp.2023.115447 PubMed DOI

Liu M, Lv Q, Xu J et al (2023) Isoflavone glucoside genistin, an inhibitor targeting Sortase A and Listeriolysin O, attenuates the virulence of Listeria monocytogenes in vivo and in vitro. Biochem Pharmacology 209:115447. https://doi.org/10.1016/j.bcp.2023.115447 DOI

Maggio B, Raffa D, Raimondi MV et al (2016) Discovery of a new class of sortase a transpeptidase inhibitors to tackle gram-positive pathogens: 2-(2-phenylhydrazinylidene) alkanoic acids and related derivatives. Molecules 21(2):241. https://doi.org/10.3390/molecules21020241 PubMed DOI PMC

Marraffini LA, DeDent AC, Schneewind O (2006) Sortases and the art of anchoring proteins to the envelopes of gram-positive bacteria. Microbiol Mol Biol Rev 70(1):192–221. https://doi.org/10.1128/MMBR.70.1.192-221.2006 PubMed DOI PMC

Mazmanian SK, Liu G, Ton-That H, Schneewind O (1999) Staphylococcus aureus sortase, an enzyme that anchors surface proteins to the cell wall. Science 285(5428):760–763. https://doi.org/10.1126/science.285.5428.760 PubMed DOI

Menichetti F (2005) Current and emerging serious Gram-positive infections. Clin Microbiol Infect 11:22–28. https://doi.org/10.1111/j.1469-0691.2005.01138.x PubMed DOI

Morris GM, Huey R, Lindstrom W, Sanner MF, Belew RK, Goodsell DS, Olson AJ (2009) AutoDock4 and AutoDockTools4: automated docking with selective receptor flexibility. J Comput Chem 30(16):2785–2791. https://doi.org/10.1002/jcc.21256 PubMed DOI PMC

Mulcahy ME, McLoughlin RM (2016) Host–bacterial crosstalk determines Staphylococcus aureus nasal colonization. Trends Microbiol 24(11):872–886. https://doi.org/10.1016/j.tim.2016.06.012 PubMed DOI

Nitulescu G, Margina D, Zanfirescu A, Olaru OT, Nitulescu GM (2021) Targeting bacterial sortases in search of anti-virulence therapies with low risk of resistance development. Pharmaceuticals (Basel) 14(5):415. https://doi.org/10.3390/ph14050415

Nitulescu G, Mihai DP, Nicorescu IM, et al (2019) Discovery of natural naphthoquinones as sortase A inhibitors and potential anti-infective solutions against Staphylococcus aureus. Drug Dev Res 80(8):1136–1145. https://doi.org/10.1002/ddr.21599

Oh I, Yang WY, Chung SC, Kim TY, Oh KB, Shin J (2011) In vitro sortase A inhibitory and antimicrobial activity of flavonoids isolated from the roots of Sophora flavescens. Arch Pharm Res 34(2):217–222. https://doi.org/10.1007/s12272-011-0206-0 PubMed DOI

Oh KB, Kim SH, Lee J, Cho WJ, Lee T, Kim S (2004) Discovery of diarylacrylonitriles as a novel series of small molecule Sortase A inhibitors. J Med Chem 47(10):2418–2421. https://doi.org/10.1021/jm0498708 PubMed DOI

Oh KB, Mar W, Kim S et al (2005) Bis(indole) alkaloids as sortase A inhibitors from the sponge Spongosorites sp. Bioorg Med Chem Lett 15(22):4927–4931. https://doi.org/10.1016/j.bmcl.2005.08.021 PubMed DOI

O’Toole GA (2011) Microtiter dish biofilm formation assay. J Vis Exp 47:e2437. https://doi.org/10.3791/2437

Park BS, Kim JG, Kim MR, Lee SE, Takeoka GR, Oh KB, Kim JH (2005) Curcuma longa L. constituents inhibit sortase A and Staphylococcus aureus cell adhesion to fibronectin. J Agric Food Chem 53(23):9005–9009. https://doi.org/10.1021/jf051765z

Paterson GK, Mitchell TJ (2004) The biology of Gram-positive sortase enzymes. Trends Microbiol 12(2):89–95. https://doi.org/10.1016/j.tim.2003.12.007 PubMed DOI

Rios JL, Recio MC (2005) Medicinal plants and antimicrobial activity. J Ethnopharmacol 100(1–2):80–84. https://doi.org/10.1016/j.jep.2005.04.025 PubMed DOI

Shukla SK, Rao TS (2017) An improved crystal violet assay for biofilm quantification in 96-well microtitre plate. Biorxiv 100214. https://doi.org/10.1101/100214

Si L, Li P, Liu X, Luo L (2016) Chinese herb medicine against Sortase A catalyzed transformations, a key role in gram-positive bacterial infection progress. J Enzyme Inhib Med Chem 31(1):184–196. https://doi.org/10.1080/14756366.2016.1178639 PubMed DOI

Smith EJ, Visai L, Kerrigan SW, Speziale P, Foster TJ (2011) The Sbi protein is a multifunctional immune evasion factor of Staphylococcus aureus. Infect Immun 79(9):3801–3809. https://doi.org/10.1128/IAI.05075-11 PubMed DOI PMC

Song W, Wang L, Jin M, Guo X, Wang X, Guan J, Zhao Y (2022) Punicalagin, an inhibitor of sortase A, is a promising therapeutic drug to combat methicillin-resistant Staphylococcus aureus infections. Antimicrob Agents Chemother 66(6):e00224–e322. https://doi.org/10.1128/aac.00224-22

Suree N, Yi SW, Thieu W et al (2019) Discovery and structure-activity relationship analysis of Staphylococcus aureus sortase A inhibitors. Bioorg Med Chem 17(20):7174–7185. https://doi.org/10.1016/j.bmc.2009.08.067 DOI

Thammavongsa V, Kim HK, Missiakas D, Schneewind O (2015) Staphylococcal manipulation of host immune responses. Nat Rev Microbiol 13(9):529–543. https://doi.org/10.1038/nrmicro3521 PubMed DOI PMC

Thappeta KRV, Zhao LN, Nge CE et al (2020) In-silico identified new natural sortase a inhibitors disrupt S. aureus biofilm formation. Int J Mol Sci 21(22):8601. https://doi.org/10.3390/ijms21228601

Ton-That H, Marraffini LA, Schneewind O (2004) Protein sorting to the cell wall envelope of Gram-positive bacteria. Biochim Biophys Acta 1694(1–3):269–278. https://doi.org/10.1016/j.bbamcr.2004.04.014 PubMed DOI

van der Maten E, de Jonge MI, de Groot R, van der Flier M, Langereis JD (2017) A versatile assay to determine bacterial and host factors contributing to opsonophagocytotic killing in hirudin-anticoagulated whole blood. Sci Rep 7:42137. https://doi.org/10.1038/srep42137 PubMed DOI PMC

Ventola CL (2015) The antibiotic resistance crisis: part 1: causes and threats. Pharm Ther 40(4):277–283

von Eiff C, Becker K, Machka K, Stammer H, Peters G (2001) Nasal carriage as a source of Staphylococcus aureus bacteremia. N Engl J Med 344(1):11–16. https://doi.org/10.1093/infdis/jis483 DOI

Wang J, Li H, Pan J, Dong J, Zhou X, Niu X, Deng X (2018) Oligopeptide targeting sortase a as potential anti-infective therapy for Staphylococcus aureus. Front Microbiol 9:245. https://doi.org/10.3389/fmicb.2018.00245 PubMed DOI PMC

Wang J, Shi Y, Jing S, Dong H, Wang D, Wang T (2019a) Astilbin inhibits the activity of sortase a from Streptococcus mutans. Molecules 24(3):465. https://doi.org/10.3390/molecules24030465 PubMed DOI PMC

Wang L, Bi C, Cai H et al (2015) The therapeutic effect of chlorogenic acid against Staphylococcus aureus infection through sortase A inhibition. Front Microbiol 6:1031. https://doi.org/10.3389/fmicb.2015.01031 PubMed DOI PMC

Wang W, Li Z, Wang J, Xu D, Shang Y (2019b) PSICA: a fast and accurate web service for protein model quality analysis. Nucleic Acids Res 47(1):443–450. https://doi.org/10.1093/nar/gkz402 DOI

Wang X, Luan Y, Hou J et al (2023) The protection effect of rhodionin against methicillin-resistant Staphylococcus aureus-induced pneumonia through sortase A inhibition. World J Microbiol Biotechnol 39:18. https://doi.org/10.1007/s11274-022-03457-4 DOI

Waterhouse A, Bertoni M, Bienert S et al (2018) SWISS-MODEL: homology modelling of protein structures and complexes. Nucleic Acids Res 46(1):296–303. https://doi.org/10.1093/nar/gky427 DOI

Wehrli PM, Uzelac I, Olsson T, Jacso T, Tietze D, Gottfries J (2019) Discovery and development of substituted thiadiazoles as inhibitors of Staphylococcus aureus Sortase A. Bioorg Med Chem 27(19):115043. https://doi.org/10.1016/j.bmc.2019.115043 PubMed DOI

Weiss WJ, Lenoy E, Murphy T et al (2004) Effect of srtA and srtB gene expression on the virulence of Staphylococcus aureus in animal models of infection. J Antimicrob Chemother 53(3):480–486. https://doi.org/10.1093/jac/dkh078 PubMed DOI

Won TH, Jeon JE, Kim SH et al (2012b) Brominated aromatic furanones and related esters from the ascidian Synoicum sp. J Nat Prod 75(12):2055–2061. https://doi.org/10.1021/np3005562 PubMed DOI

Won TH, Jeon JE, Lee SH, Rho BJ, Oh KB, Shin J (2012a) Beta-carboline alkaloids derived from the ascidian Synoicum sp. Bioorg Med Chem 20(13):4082–4087. https://doi.org/10.1016/j.bmc.2012.05.002 PubMed DOI

Woodford N, Livermore DM (2009) Infections caused by Gram-positive bacteria: a review of the global challenge. J Infect 59(1):4–16. https://doi.org/10.1016/S0163-4453(09)60003-7 DOI

Yang T, Zhang T, Guan XN, Dong Z, Lan L, Yang S, Yang CG (2020) Tideglusib and its analogues as inhibitors of Staphylococcus aureus SrtA. J Med Chem 63(15):8442–8457. https://doi.org/10.1021/acs.jmedchem.0c00803 PubMed DOI

Zhang B, Teng Z, Li X, Lu G, Deng X, Niu X, Wang J (2017) Chalcone attenuates Staphylococcus aureus virulence by targeting sortase A and alpha-hemolysin. Front Microbiol 8:1715. https://doi.org/10.3389/fmicb.2017.01715 PubMed DOI PMC

Zhang J, Liu H, Zhu K et al (2014) Antiinfective therapy with a small molecule inhibitor of Staphylococcus aureus sortase. Proc Natl Acad Sci USA 111(37):13517–13522. https://doi.org/10.1073/pnas.1408601111 PubMed DOI PMC

Zhulenkovs D, Rudevica Z, Jaudzems K, Turks M, Leonchiks A (2014) Discovery and structure-activity relationship studies of irreversible benzisothiazolinone-based inhibitors against Staphylococcus aureus sortase A transpeptidase. Bioorg Med Chem 22(21):5988–6003. https://doi.org/10.1016/j.bmc.2014.09.011 PubMed DOI