Abstract
Biofilm forming pathogens are among the major causes of hospital-acquired infections and are not much affected by antibiotic treatment. Consequently, novel agents and therapeutics are required urgently that possess antibacterial and antibiofilm activities. This study analyzed two bacteriocins from Lactobacillus plantarum subsp. argentoratensis SJ33 strain for their antibacterial and antibiofilm activity as well as cytotoxic properties. BacF1 and BacF2 showed broad spectrum activity against both Gram-positive (Listeria monocytogenes, Staphylococcus aureus) and Gram-negative (Pseudomonas aeruginosa, Escherichia coli) bacteria. Significant bactericidal action was also observed on S. aureus cells by pore formation. Additionally, bacteriocins disrupted biofilms formed by S. aureus and P. aeruginosa which were shown by crystal violet staining assay and visualized by fluorescence as well as scanning electron microscopy. Quantitative Real-Time PCR study revealed changes in gene expression of biofilm formation in S. aureus (ica) and P. aeruginosa (pelA, psl, rhlA). Cytotoxicity of bacteriocins was further analyzed on normal mammalian cells and Caenorhabditis elegans. Notably, bacteriocins showed no major effect on HEK-293 cell line and enhanced the survival of S. aureus infected HEK-293 cells. Similarly, no cytotoxic effect was visible on C. elegans even after treatment with higher concentration than MIC at different time intervals.
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All the data analyzed during this study are included in this article and its supplementary material.
References
Agaliya PJ, Jeevaratnam K (2013) Molecular characterization of lactobacilli isolated from fermented idli batter. Brazilian J Microbiol 44:1199–1206. https://doi.org/10.1590/S1517-83822013000400025
Ahn KB, Baik JE, Park OJ et al (2018) Lactobacillus plantarum lipoteichoic acid inhibits biofilm formation of Streptococcus mutans. PLoS ONE 13:1–16. https://doi.org/10.1371/journal.pone.0192694
Amortegui J, Rodríguez-López A, Rodríguez D et al (2014) Characterization of a new bacteriocin from Lactobacillus plantarum LE5 and LE27 isolated from ensiled corn. Appl Biochem Biotechnol 172:3374–3389. https://doi.org/10.1007/s12010-014-0757-x
Ansari A, Ibrahim F, Pervez S, Aman A (2020) Inhibitory mechanism of BAC-IB17 against β-lactamase mediated resistance in methicillin-resistant Staphylococcus aureus and application as an oncolytic agent. Microb Pathog 149:104499
Ansari A, Zohra RR, Tarar OM et al (2018) Screening, purification and characterization of thermostable, protease resistant Bacteriocin active against methicillin resistant Staphylococcus aureus (MRSA). BMC Microbiol 18:1–10. https://doi.org/10.1186/s12866-018-1337-y
Bakkiyaraj D, Pandian STK (2010) In vitro and in vivo antibiofilm activity of a coral associated actinomycete against drug resistant Staphylococcus aureus biofilms. Biofouling 26:711–717. https://doi.org/10.1080/08927014.2010.511200
Blanco AR, Sudano-Roccaro A, Spoto GC et al (2005) Epigallocatechin gallate inhibits biofilm formation by ocular Staphylococcal isolates. Antimicrob Agents Chemother 49:4339–4343. https://doi.org/10.1128/AAC.49.10.4339-4343.2005
Callewaert R, Holo H, Devreese B et al (1999) Characterization and production of amylovorin L471, a bacteriocin purified from Lactobacillus amylovorus DCE 471 by a novel three-step method. Microbiology 145:2559–2568. https://doi.org/10.1099/00221287-145-9-2559
Chopra L, Singh G, Kumar Jena K, Sahoo DK (2015) Sonorensin: a new bacteriocin with potential of an anti-biofilm agent and a food biopreservative. Sci Rep 5:1–13. https://doi.org/10.1038/srep13412
Chuah LO, Foo HL, Loh TC et al (2019) Postbiotic metabolites produced by Lactobacillus plantarum strains exert selective cytotoxicity effects on cancer cells. BMC Complement Altern Med 19:1–12. https://doi.org/10.1186/s12906-019-2528-2
Cirkovic I, Bozic DD, Draganic V et al (2016) Licheniocin 50.2 and bacteriocins from Lactococcus lactis subsp. lactis biovar. Diacetylactis BGBU1-4 inhibit biofilms of coagulase negative Staphylococci and Listeria monocytogenes clinical isolates. PLoS ONE 11:1–12. https://doi.org/10.1371/journal.pone.0167995
De Giani A, Bovio F, Forcella M et al (2019) Identification of a bacteriocin-like compound from Lactobacillus plantarum with antimicrobial activity and effects on normal and cancerogenic human intestinal cells. AMB Express 9(1):88
Dimitrov S, Wachsman M, Tomé E et al (2010) Characterisation of an antiviral pediocin-like bacteriocin produced by Enterococcus faecium. Food Microbiol 27:869–879. https://doi.org/10.1016/j.fm.2010.05.001
Fabian TJ, Johnson TE (1994) Production of age-synchronous mass cultures of Caenorhabditis elegans. J Gerontol 49:B145–B156. https://doi.org/10.1093/geronj/49.4.B145
Gerits E, Blommaert E, Lippell A et al (2016) Elucidation of the mode of action of a new antibacterial compound active against Staphylococcus aureus and Pseudomonas aeruginosa. PLoS ONE 11:1–17. https://doi.org/10.1371/journal.pone.0155139
Griffiths S, Maclean M, MacGregor SJ et al (2011) Decontamination of collagen biomatrices with combined pulsed electric field and nisin treatment. J Biomed Mater Res Part B Appl Biomater 96:287–293
Gusarov I, Gautier L, Smolentseva O et al (2013) Bacterial nitric oxide extends the lifespan of C. elegans. Cell 152:818–830. https://doi.org/10.1016/j.cell.2012.12.043
Hammami I, Triki MA, Rebai A (2011) Purification and characterization of the novel bacteriocin Bac IH7 with antifungal and antibacterial properties. J Plant Pathol 93:443–454
Hassan A, Usman J, Kaleem F et al (2011) Evaluation of different detection methods of biofilm formation in the clinical isolates. Brazilian J Infect Dis 15:305–311. https://doi.org/10.1590/S1413-86702011000400002
Héchard Y, Sahl HG (2002) Mode of action of modified and unmodified bacteriocins from Gram-positive bacteria. Biochimie 84:545–557
Hernández D, Cardell E, Zárate V (2005) Antimicrobial activity of lactic acid bacteria isolated from Tenerife cheese: Initial characterization of plantaricin TF711, a bacteriocin-like substance produced by Lactobacillus plantarum TF711. J Appl Microbiol 99:77–84. https://doi.org/10.1111/j.1365-2672.2005.02576.x
Ibrahim F, Siddiqui NN, Aman A et al (2020) Characterization, cytotoxic analysis and action mechanism of antilisterial bacteriocin produced by Lactobacillus plantarum isolated from cheddar cheese. Int J Pept Res Ther 26:1751–1764. https://doi.org/10.1007/s10989-019-09982-5
Ibrahim F, Zafar SB, Aman A et al (2019) Improvement of Lactobacillus plantarum for the enhanced production of bacteriocin like inhibitory substance using combinatorial approach. Biocatal Agric Biotechnol 22:101386. https://doi.org/10.1016/j.bcab.2019.101386
Jiang J, Shi B, Zhu D et al (2012) Characterization of a novel bacteriocin produced by Lactobacillus sakei LSJ618 isolated from traditional Chinese fermented radish. Food Control 23:338–344. https://doi.org/10.1016/j.foodcont.2011.07.027
Kaiser TDL, Pereira EM, dos Santos KRN et al (2013) Modification of the Congo red agar method to detect biofilm production by Staphylococcus epidermidis. Diagn Microbiol Infect Dis 75:235–239. https://doi.org/10.1016/j.diagmicrobio.2012.11.014
Kindoli S, Lee HA, Kim JH (2012) Properties of Bac W42, a bacteriocin produced by Bacillus subtilis W42 isolated from Cheonggukjang. J Microbiol Biotechnol 22:1092–1100. https://doi.org/10.1007/s10068-012-0232-9
Konai MM, Haldar J (2016) Lysine-based small molecules that disrupt biofilms and kill both actively growing planktonic and nondividing stationary phase bacteria. ACS Infect Dis 1:469–478. https://doi.org/10.1021/acsinfecdis.5b00056
Lakshmanan D, Harikrishnan A, Vishnupriya S, Jeevaratnam K (2019) Swarming inhibitory potential of cinnamtannin B1 from Cinnamomum tamala T. Nees and Eberm on Pseudomonas aeruginosa. ACS Omega 4:16994–16998. https://doi.org/10.1021/acsomega.9b02471
Leroy F, De Vuyst L (2010) Bacteriocins of lactic acid bacteria to combat undesirable bacteria in dairy products. Aust J Dairy Technol 65:143
Lin TH, Pan TM (2019) Characterization of an antimicrobial substance produced by Lactobacillus plantarum NTU 102. J Microbiol Immunol Infect 52:409–417. https://doi.org/10.1016/j.jmii.2017.08.003
Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2− ΔΔCT method. Methods 25:402–408
Lü X, Hu P, Dang Y, Liu B (2014) Purification and partial characterization of a novel bacteriocin produced by Lactobacillus casei TN-2 isolated from fermented camel milk (Shubat) of Xinjiang Uygur Autonomous region, China. Food Control 43:276–283. https://doi.org/10.1016/j.foodcont.2014.03.020
Mathur H, Field D, Rea MC et al (2018) Fighting biofilms with lantibiotics and other groups of bacteriocins. npj Biofilms Microbiomes. https://doi.org/10.1038/s41522-018-0053-6
Mathur T, Singhal S, Khan S et al (2006) Detection of biofilm formation among the clinical isolates of Staphylococci: an evaluation of three different screening methods. Indian J Med Microbiol 24:25–29. https://doi.org/10.4103/0255-0857.19890
Miao J, Guo H, Ou Y et al (2014) Purification and characterization of bacteriocin F1, a novel bacteriocin produced by Lactobacillus paracasei subsp. tolerans FX-6 from Tibetan kefir, a traditional fermented milk from Tibet, China. Food Control 42:48–53. https://doi.org/10.1016/j.foodcont.2014.01.041
Moh C, Engelhardt T, Albano H et al (2015) Antilisterial activity of bacteriocinogenic Pediococcus acidilactici HA6111-2 and Lactobacillus plantarum ESB 202 grown under pH and osmotic stress conditions. Food Microbiol 48:109–115. https://doi.org/10.1016/j.fm.2014.11.015
Moy TI, Ball AR, Anklesaria Z et al (2006) Identification of novel antimicrobials using a live-animal infection model. PNAS 103:10414–10419
Niu Q, Zhang L, Zhang K et al (2016) Changes in intestinal microflora of Caenorhabditis elegans following Bacillus nematocida B16 infection. Nat Publ Gr. https://doi.org/10.1038/srep20178
Okuda KI, Zendo T, Sugimoto S et al (2013) Effects of bacteriocins on methicillin-resistant Staphylococcus aureus biofilm. Antimicrob Agents Chemother 57:5572–5579. https://doi.org/10.1128/AAC.00888-13
Ong TH, Chitra E, Ramamurthy S et al (2019) Cationic chitosan-propolis nanoparticles alter the zeta potential of S. epidermidis, inhibit biofilm formation by modulating gene expression and exhibit synergism with antibiotics. PLoS ONE 14:1–13. https://doi.org/10.1371/journal.pone.0213079
Overhage J, Campisano A, Bains M et al (2008) Human host defense peptide LL-37 prevents bacterial biofilm formation. Infect Immun 76:4176–4182
Pal V, Pal A, Patil M et al (2010) Isolation, biochemical properties and application of bacteriocins from Pediococcus pentosaceous isolates. J Food Process Preserv 34:1064–1079
Rachid S, Ohlsen K, Witte W et al (2000) Effect of subinhibitory antibiotic concentrations on polysaccharide intercellular adhesin expression in biofilm-forming Staphylococcus epidermidis. Antimicrob Agents Chemother 44:3357–3363. https://doi.org/10.1128/AAC.44.12.3357-3363.2000
Ray Mohapatra A, Jeevaratnam K (2019) Inhibiting bacterial colonization on catheters: antibacterial and antibiofilm activities of bacteriocins from Lactobacillus plantarum SJ33. J Glob Antimicrob Resist 19:85–92. https://doi.org/10.1016/j.jgar.2019.02.021
Reenen CA Van, Dicks LMT, Chikindas ML (1998) Isolation, purification and partial characterization of plantaricin 423, a bacteriocin produced by Lactobacillus plantarum
Roe D, Karandikar B, Bonn-savage N et al (2018) Antimicrobial surface functionalization of plastic catheters by silver nanoparticles. J Antimicrob Chemother 61:869–876. https://doi.org/10.1093/jac/dkn034
Sadishkumar V, Jeevaratnam K (2018) Purification and partial characterization of antilisterial bacteriocin produced by Pediococcus pentosaceus KJBC11 from Idli batter fermented with Piper betle leaves. J Food Biochem 42:1–9
Sahoo TK, Jena PK, Patel AK, Seshadri S (2015) Purification and molecular characterization of the novel highly potent bacteriocin TSU4 produced by Lactobacillus animalis TSU4. Appl Biochem Biotechnol 177:90–104. https://doi.org/10.1007/s12010-015-1730-z
Sakurazawa T, Ohkusa T (2005) Cytotoxicity of organic acids produced by anaerobic intestinal bacteria on cultured epithelial cells. J Gastroenterol 40:600–609
Sand SL, Oppegård C, Ohara S et al (2010) Plantaricin A, a peptide pheromone produced by Lactobacillus plantarum, permeabilizes the cell membrane of both normal and cancerous lymphocytes and neuronal cells. Peptides 31:1237–1244. https://doi.org/10.1016/j.peptides.2010.04.010
Saranya S, Hemashenpagam N (2013) Purification and characterization of bacteriocin Produced by different Lactobacillus species isolated from fermented foods. Int J Microbiol Res 5:341–348. https://doi.org/10.9735/0975-5276.5.1.341-348
Sawa N, Koga S, Okamura K et al (2013) Identification and characterization of novel multiple bacteriocins produced by Lactobacillus sakei D98. J Appl Microbiol 115:61–69. https://doi.org/10.1111/jam.12226
Schägger H (2006) Tricine-SDS-PAGE. Nat Protoc 1:16–22. https://doi.org/10.1007/978-1-4939-8793-1_15
Son SJ, Park MR, Ryu SD et al (2016) Short communication: in vivo screening platform for bacteriocins using Caenorhabditis elegans to control mastitis-causing pathogens. J Dairy Sci 99:8614–8621. https://doi.org/10.3168/jds.2016-11330
Tan Y, Leonhard M, Ma S et al (2018) Efficacy of carboxymethyl chitosan against Candida tropicalis and Staphylococcus epidermidis monomicrobial and polymicrobial biofilms. Int J Biol Macromol 110:150–156. https://doi.org/10.1016/j.ijbiomac.2017.08.094
Tiwari SK, Srivastava S (2008) Purification and characterization of plantaricin LR14: a novel bacteriocin produced by Lactobacillus plantarum LR/14. Appl Microbiol Biotechnol 79:759–767. https://doi.org/10.1007/s00253-008-1482-6
Todorov SD, Dicks LMT (2005) Lactobacillus plantarum isolated from molasses produces bacteriocins active against Gram-negative bacteria. Enzyme Microb Technol 36:318–326. https://doi.org/10.1016/j.enzmictec.2004.09.009
Todorov SD, Dicks LMT (2009) Bacteriocin production by Pediococcus pentosaceus isolated from marula (Scerocarya birrea). Int J Food Microbiol 132:117–126
Vahedi Shahandashti R, Kasra Kermanshahi R, Ghadam P (2016) The inhibitory effect of bacteriocin produced by Lactobacillus acidophilus ATCC 4356 and Lactobacillus plantarum ATCC 8014 on planktonic cells and biofilms of Serratia marcescens. Turkish J Med Sci 46:1188–1196. https://doi.org/10.3906/sag-1505-51
Vaucher RA, Teixeira ML, Brandelli A (2010) Investigation of the cytotoxicity of antimicrobial peptide P40 on eukaryotic cells. Curr Microbiol 60:1–5. https://doi.org/10.1007/s00284-009-9490-z
Vidhyasagar V, Jeevaratnam K (2013) Bacteriocin activity against various pathogens produced by Pediococcus pentosaceus VJ13 isolated from Idly batter. Biomed Chromatogr 27:1497–1502. https://doi.org/10.1002/bmc.2948
Wen LS, Philip K, Ajam N (2016) Purification, characterization and mode of action of plantaricin K25 produced by Lactobacillus plantarum. Food Control 60:430–439. https://doi.org/10.1016/j.foodcont.2015.08.010
Wu S, Liu G, Jin W et al (2016) Antibiofilm and anti-infection of a marine bacterial exopolysaccharide against Pseudomonas aeruginosa. Front Microbiol 7:1–15. https://doi.org/10.3389/fmicb.2016.00102
Xu T, Wu Y, Lin Z et al (2017) Identification of genes controlled by the essential YycFG two-component system reveals a role for biofilm modulation in Staphylococcus epidermidis. Front Microbiol 8:1–17. https://doi.org/10.3389/fmicb.2017.00724
Yin H, Deng Y, Wang H et al (2015) Tea polyphenols as an antivirulence compound disrupt quorum-sensing regulated pathogenicity of Pseudomonas aeruginosa. Sci Rep 5:1–12. https://doi.org/10.1038/srep16158
Zhang J, Yang Y, Yang H et al (2018) Purification and partial characterization of Bacteriocin Lac-B23, a novel bacteriocin production by Lactobacillus plantarum J23, isolated from Chinese traditional fermented milk. Front Microbiol 9:1–7. https://doi.org/10.3389/fmicb.2018.02165
Zhao R, Duan G, Yang T et al (2015) Purification, characterization and antibacterial mechanism of bacteriocin from lactobacillus acidophilus XH1. Trop J Pharm Res 14:989–995
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Authors are thankful to DST-FIST, UGC-SAP for providing funds for the Department and IIT Madras, India for SEM analysis. Authors are also grateful to the University Grant Commission (UGC), New Delhi for the financial support.
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Ray Mohapatra, A., Lakshmanan, D., Mahesh, R. et al. Characterization and Cytotoxic Evaluation of Bacteriocins Possessing Antibiofilm Activity Produced by Lactobacillus plantarum SJ33. Int J Pept Res Ther 27, 1783–1797 (2021). https://doi.org/10.1007/s10989-021-10210-2
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DOI: https://doi.org/10.1007/s10989-021-10210-2