Abstract
Chromobacterium violaceum, one free-living Gram-negative bacterium, is abundantly presented in tropics and sub-tropics soil and aquatic environment; it is also an opportunistic human pathogen. Here, two cinnamic acid derivatives, i.e., 4-dimethylaminocinnamic acid (DCA) and 4-methoxycinnamic acid (MCA), were identified as potential quorum sensing (QS) and biofilm inhibitors in C. violaceum ATCC12472. Both DCA (100 μg/mL) and MCA (200 μg/mL) inhibited the levels of N-decanoyl-homoserine lactone (C10-HSL) and reduced the production of certain virulence factors in C. violaceum, including violacein, hemolysin, and chitinase. Metabolomics analysis indicated that QS-related metabolites, such as ethanolamine and L-methionine, were down-regulated after treatment with DCA and MCA. Quantitative real-time polymerase chain reaction (qRT-PCR) demonstrated that DCA and MCA markedly suppressed the expression of two QS-related genes (cviI and cviR). In addition, DCA and MCA also inhibited biofilm formation and enhanced the susceptibility of biofilms to tobramycin, which was evidenced by scanning electron microscopy (SEM) and confocal laser scanning microscopy (CLSM). Our results indicated that DCA and MCA can serve as QS-based agent for controlling pathogens.
Key Points
• DCA and MCA inhibited QS and biofilm formation in C. violaceum.
• The combination of DCA or MCA and tobramycin removed the preformed biofilm of C. violaceum.
• DCA or MCA inhibited virulence factors and expressions of cviI and cviR of C. violaceum.
• DCA or MCA are potential antibiotic accelerants for treating C. violaceum infection.
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References
Adisakwattana S (2017) Cinnamic acid and its derivatives: mechanisms for prevention and management of diabetes and its complications. Nutrients 9(2):163. https://doi.org/10.3390/nu9020163
Ahmad A, Viljoen AM, Chenia HY (2015) The impact of plant volatiles on bacterial quorum sensing. Lett Appl Microbiol 60(1):8–19. https://doi.org/10.1111/lam.12343
Borges A, Serra S, Cristina Abreu A, Saavedra MJ, Salgado A, Simões M (2014) Evaluation of the effects of selected phytochemicals on quorum sensing inhibition and in vitro cytotoxicity. Biofouling 30:183–195. https://doi.org/10.1080/08927014.2013.852542
Brackman G, Celen S, Hillaert U, Van Calenbergh S, Cos P, Maes L, Nelis HJ, Coenye T (2011a) Structure-activity relationship of cinnamaldehyde analogs as inhibitors of AI-2 based quorum sensing and their effect on virulence of Vibrio spp. PLoS One 6(1):e16084. https://doi.org/10.1371/journal.pone.0016084
Brackman G, Cos P, Maes L, Nelis HJ, Coenye T (2011b) Quorum sensing inhibitors increase the susceptibility of bacterial biofilm to antibiotics in vitro and in vivo. Antimicrob Agents Chemother 55(6):2655–2661. https://doi.org/10.1128/AAC.00045-11
Burt SA, Ojo-Fakunle VT, Woertman J, Veldhuizen EJ (2014) The natural antimicrobial carvacrol inhibits quorum sensing in Chromobacterium violaceum and reduces bacterial biofilm formation at sub-lethal concentrations. PLoS One 9(4). https://doi.org/10.1371/journal.pone.0093414
Chen X, Wu H, Cao Y, Yao X, Zhao L, Wang T (2014) Ion-pairing chromatography on a porous graphitic carbon column coupled with time-of-flight mass spectrometry for targeted and untargeted profiling of amino acid biomarkers involved in Candida albicans biofilm formation. Mol BioSyst 10:74–85. https://doi.org/10.1039/c3mb70240e
Chen TT, Sheng J, Fu Y, Li M, Wang J, Jia AQ (2017) 1H NMR-based global metabolic studies of Pseudomonas aeruginosa upon exposure of the quorum sensing inhibitor resveratrol. J Proteome Res 16(2):824–830. https://doi.org/10.1021/acs.jproteome.6b00800
Clinical and Laboratory Standards Institute (CLSI) (2015) Methods for dilution antimicrobial susceptibility tests for bacteria that grow aerobically: Approved Standard 10th edn. M07-A10, CLSI, Wayne
D'Almeida RE, Molina RDI, Viola CM, Luciardi MC, Nieto Peñalver C, Bardón A, Arena ME (2017) Comparison of seven structurally related coumarins on the inhibition of quorum sensing of Pseudomonas aeruginosa and Chromobacterium violaceum. Bioorg Chem 73:37–42. https://doi.org/10.1016/j.bioorg.2017.05.011
Durán N, Justo GJ, Durán M, Brocchi M, Cordi L, Tasic L, Castro GR, Nakazato G (2016) Advances in Chromobacterium violaceum and properties of violacein—its main secondary metabolite: a review. Biotechnol Adv 34(5):1030–1045. https://doi.org/10.1016/j.biotechadv.2016.06.003
Evans KC, Benomar S, Camuy-Vélez LA, Nasseri EB, Wang X, Neuenswander B, Chandler JR (2018) Quorum-sensing control of antibiotic resistance stabilizes cooperation in Chromobacterium violaceum. ISME J 12(5):1263–1272. https://doi.org/10.1038/s41396-018-0047-7
Fukumoto A, Murakami C, Anzai Y, Kato F (2016) Maniwamycins: new quorum-sensing inhibitors against Chromobacterium violaceum CV026 were isolated from Streptomyces sp. TOHO-M025. J Antibiot 69(5): 395. https://doi.org/10.1038/ja.2015.126
Fuqua WC, Winans SC, Greenberg EP (1994) Quorum sensing in bacteria: the LuxR-LuxI family of cell density-responsive transcriptional regulators. J Bacteriol 176(2):269–275. https://doi.org/10.1128/jb.176.2.269-275.1994
Goo E, An JH, Kang Y, Hwang I (2015) Control of bacterial metabolism by quorum sensing. Trends Microbiol 23(9):567–576. https://doi.org/10.1016/j.tim.2015.05.007
Heurlier K, Dénervaud V, Haas D (2006) Impact of quorum sensing on fitness of Pseudomonas aeruginosa. Int J Med Microbiol 296(2–3):93–102. https://doi.org/10.1016/j.ijmm.2006.01.043
Iqbal MJ, Maqsood Y, Abdin ZU, Manzoor A, Hassan M, Jamil A (2016) SSR markers associated with proline in drought tolerant wheat germplasm. Appl Biochem Biotechnol 178(5):1042–1052. https://doi.org/10.1007/s12010-015-1927-1
Justo GZ, Duran N (2017) Action and function of Chromobacterium violaceum in health and disease: Violacein as a promising metabolite to counteract gastroenterological diseases. Best Pract Res Clin Gastroenterol 31(6):649–656. https://doi.org/10.1016/j.bpg.2017.10.002
Kumar R, Chhibber S, Harjai K (2009) Quorum sensing is necessary for the virulence of Pseudomonas aeruginosa during urinary tract infection. Kidney Int 76(3):286–292. https://doi.org/10.1038/ki.2009.183
Lee SY, Hwang IY, Jeong CS (2017) Protective effects of cinnamic acid derivatives on gastric lesion. Nat Prod Sci 23(4):299–305. https://doi.org/10.20307/nps.2017.23.4.299
Liu Z, Wang W, Zhu Y, Gong Q, Yu W, Lu X (2013) Antibiotics at subinhibitory concentrations improve the quorum sensing behavior of Chromobacterium violaceum. FEMS Microbiol Lett 341(1):37–44. https://doi.org/10.1111/1574-6968.12086
Malheiro JF, Maillard JY, Borges F, Simões M (2019) Evaluation of cinnamaldehyde and cinnamic acid derivatives in microbial growth control. Int Biodeterior Biodegrad 141:71–78. https://doi.org/10.1016/j.ibiod.2018.06.003
Morohoshi T, Kato M, Fukamachi K, Kato N, Ikeda T (2008) N-acylhomoserine lactone regulates violacein production in Chromobacterium violaceum type strain ATCC 12472. FEMS Microbiol Lett 279:124–130. https://doi.org/10.1111/j.1574-6968.2007.01016.x
Pan J, Ren D (2013) Structural effects on persister control by brominated furanones. Bioorg Med Chem Lett 23(24):6559–6562. https://doi.org/10.1016/j.bmcl.2013.10.070
Pomini AM, Paccola-Meirelles LD, Marsaioli AJ (2007) Acyl-homoserine lactones produced by Pantoea sp. isolated from the “maize white spot” foliar disease. J Agric Food Chem 55(4):1200–1204. https://doi.org/10.1021/jf063136a
Ravichandran V, Zhong L, Wang H, Yu G, Zhang Y, Li A (2018) Virtual screening and biomolecular interactions of CviR-based quorum sensing inhibitors against Chromobacterium violaceum. Front Cell Infect Microbiol 8:292. https://doi.org/10.3389/fcimb.2018.00292
Schmidley JW (1990) Free radicals in central nervous system ischemia. Stroke 21(7):1086–1090. https://doi.org/10.1161/01.str.21.7.1086
Sethupathy S, Ananthi S, Selvaraj A, Shanmuganathan B, Vigneshwari L, Balamurugan K, Mahalingam S, Pandian SK (2017) Vanillic acid from Actinidia deliciosa impedes virulence in Serratia marcescens by affecting S-layer, flagellin and fatty acid biosynthesis proteins. Sci Rep 7(1):16328. https://doi.org/10.1038/s41598-017-16507-x
Shepherd J, Swift T, Chang CY, Boyne JR, Rimmer S, Martin WHC (2019) Highly branched poly (N-isopropyl acrylamide) functionalized with an inducer molecule suppresses quorum sensing in Chromobacterium violaceum. Chem Commun 55(66):9765–9768. https://doi.org/10.1039/C9CC02524C
Shi Y, Chen QX, Wang Q, Song KK, Qiu L (2005) Inhibitory effects of cinnamic acid and its derivatives on the diphenolase activity of mushroom (Agaricus bisporus) tyrosinase. Food Chem 92(4):707–712. https://doi.org/10.1016/j.foodchem.2004.08.031
Singh VK, Mishra A, Jha B (2017) Anti-quorum sensing and anti-biofilm activity of Delftia tsuruhatensis extract by attenuating the quorum sensing-controlled virulence factor production in Pseudomonas aeruginosa. Front Cell Infect Microbiol 7:337. https://doi.org/10.3389/fcimb.2017.00337
Singh VK, Mishra A, Jha B (2019) 3-Benzyl-hexahydro-pyrrolo [1, 2-a] pyrazine-1, 4-dione extracted from Exiguobacterium indicum showed anti-biofilm activity against Pseudomonas aeruginosa by attenuating quorum sensing. Front Microbiol 10:1269. https://doi.org/10.3389/fmicb.2019.01269
Sirinavin S, Techasaensiri C, Benjaponpitak S, Pornkul R, Vorachit M (2005) Invasive Chromobacterium violaceum infection in children: case report and review. Pediatr Infect Dis J 24(6):559–561. https://doi.org/10.1097/01.inf.0000164761.81491.3f
Stauff DL, Bassler BL (2011) Quorum sensing in Chromobacterium violaceum: DNA recognition and gene regulation by the CviR receptor. J Bacteriol 193(15):3871–3878. https://doi.org/10.1128/JB.05125-11
Sun S, Tay QXM, Kjelleberg S, Rice SA, McDougald D (2015) Quorum sensing-regulated chitin metabolism provides grazing resistance to Vibrio cholerae biofilm. ISME J 9(8):1812–1820. https://doi.org/10.1038/ismej.2014.265
Truchado P, Giménez-Bastida JA, Larrosa M, Castro-Ibáñez I, Espín JC, Tomás-Barberán FA, García-Conesa MT, Allende A (2012) Inhibition of quorum sensing (QS) in Yersinia enterocolitica by an orange extract rich in glycosylated flavanones. J Agric Food Chem 60(36):8885–8894. https://doi.org/10.1021/jf301365a
Wu H, Moser C, Wang HZ, Høiby N, Song ZJ (2015) Strategies for combating bacterial biofilm infections. Int J Oral Sci 7(1):1–7. https://doi.org/10.1038/ijos.2014.65
Yarwood JM, Schlievert PM (2003) Quorum sensing in Staphylococcus infections. J Clin Invest 112(11):1620–1625. https://doi.org/10.1172/JCI20442
Yuan B, Wang X, Tang C, Li X, Yu G (2015) In situ observation of the growth of biofouling layer in osmotic membrane bioreactors by multiple fluorescence labeling and confocal laser scanning microscopy. Water Res 75:188–200. https://doi.org/10.1016/j.watres.2015.02.048
Zhou JW, Bi SY, Chen HJ, Chen T, Yang R, Li M, Fu Y, Jia AQ (2017) Anti-biofilm and antivirulence activities of metabolites from Plectosphaerella cucumerina against Pseudomonas aeruginosa. Front Microbiol 8:769. https://doi.org/10.3389/fmicb.2017.00769
Zhou JW, Luo HZ, Jiang H, Jian TK, Chen ZQ, Jia AQ (2018) Hordenine: a novel quorum sensing inhibitor and antibiofilm agent against Pseudomonas aeruginosa. J Agric Food Chem 66(7):1620–1628. https://doi.org/10.1021/acs.jafc.7b05035
Zhou JW, Ruan LY, Chen HJ, Luo HZ, Jiang H, Wang JS, Jia AQ (2019) Inhibition of quorum sensing and virulence in Serratia marcescens by Hordenine. J Agric Food Chem 67(3):784–795. https://doi.org/10.1021/acs.jafc.8b05922
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This study was funded by the National Key Research and Development Program of China (2017YFD0201401), National Natural Science Foundation of China (41766006), and Six Talent Peaks Project in Jiangsu Province.
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AQJ and JWZ conceived and designed research. AQJ and JWZ conducted experiments. WJC, PPZ, and HZL performed the experiments. WJC and AQJ wrote the paper. ST, JJL, and SMD provided technical supports. AQJ, WJC, and HZL analyzed data. All authors read and approved the paper.
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Cheng, WJ., Zhou, JW., Zhang, PP. et al. Quorum sensing inhibition and tobramycin acceleration in Chromobacterium violaceum by two natural cinnamic acid derivatives. Appl Microbiol Biotechnol 104, 5025–5037 (2020). https://doi.org/10.1007/s00253-020-10593-0
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DOI: https://doi.org/10.1007/s00253-020-10593-0