Abstract
Screening for producers of potent antimicrobial peptides, resulted in the isolation of Bacillus cereus BGNM1 with strong antimicrobial activity against Listeria monocytogenes. Genome sequence analysis revealed that BGNM1 contains the gene cluster associated with the production of the lantibiotic, thusin, previously identified in B. thuringiensis. Purification of the antimicrobial activity confirmed that strain BGMN1 produces thusin. Both thusin sensitive and resistant strains were detected among clinical isolates of Streptococcus agalactiae. Random mutagenesis of a thusin sensitive strain, S. agalactiae B782, was performed in an attempt to identify the receptor protein for thusin. Three independent thusin resistant mutants were selected and their complete genomes sequenced. Comparative sequence analysis of these mutants with the WT strain revealed that duplication of a region encoding a 79 amino acids repeat in a C-protein α-antigen was a common difference, suggesting it to be responsible for increased resistance to thusin. Since induced thusin resistant mutants showed higher level of resistance than the naturally resistant B761 strain, complete genome sequencing of strain B761 was performed to check the integrity of the C-protein α-antigen-encoding gene. This analysis revealed that this gene is deleted in B761, providing further evidence that this protein promotes interaction of the thusin with receptor.
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References
Abriouel H, Franz CM, Ben Omar N, Gálvez A (2011) Diversity and applications of Bacillus bacteriocins. FEMS Microbiol Rev 35:201–232. https://doi.org/10.1111/j.1574-6976.2010.00244.x
Acedo JZ, Chiorean S, Vederas JC, van Belkum MJ (2018) The expanding structural variety among bacteriocins from Gram-positive bacteria. FEMS Microbiol Rev 42:805–828. https://doi.org/10.1093/femsre/fuy033
Ahmed Sheikh HM (2010) Antimicrobial activity of certain bacteria and fungi isolated from soil mixed with human saliva against pathogenic microbes causing dermatological diseases. Saudi J Biol Sci 17:331–339. https://doi.org/10.1016/j.sjbs.2010.06.003
Alvarez-Sieiro P, Montalbán-López M, Mu D, Kuipers OP (2016) Bacteriocins of lactic acid bacteria: extending the family. Appl Microbiol Biotechnol 100:2939–2951. https://doi.org/10.1007/s00253-016-7343-9
Asaduzzaman SM, Nagao J, Iida H, Zendo T, Nakayama J, Sonomoto K (2009) Nukacin ISK-1, a bacteriostatic lantibiotic. Antimicrob Agents Chemother 53:3595–3598. https://doi.org/10.1128/AAC.01623-08
Beric T, Kojic M, Stankovic S, Topisirović L, Degrassi G, Myers M et al (2012) Antimicrobial activity of Bacillus sp. natural isolates and their potential use in the biocontrol of phytopathogenic bacteria. Food Technol Biotechnol 50:25–31
Bierbaum G, Sahl HG (2009) Lantibiotics: mode of action, biosynthesis and bioengineering. Curr Pharm Biotechnol 10:2–18. https://doi.org/10.2174/138920109787048616
Collins FWJ, O’Connor PM, O’Sullivan O, Rea MC, Hill C, Ross RP (2016) Formicin—a novel broad-spectrum two-component lantibiotic produced by Bacillus paralicheniformis APC 1576. Microbiology 9:1662–1671. https://doi.org/10.1099/mic.0.000340
Cotter PD, Ross RP, Hill C (2013) Bacteriocins—a viable alternative to antibiotics? Nat Rev Microbiol 11:95–105. https://doi.org/10.1038/nrmicro2937
Darling AC, Mau B, Blattner FR, Perna NT (2004) Mauve: multiple alignment of conserved genomic sequence with rearrangements. Gen Res 14:1394–1403. https://doi.org/10.1101/gr.2289704
Dischinger J, Basi Chipalu S, Bierbaum G (2014) Lantibiotics: promising candidates for future applications in health care. Int J Med Microbiol 304:51–62. https://doi.org/10.1016/j.ijmm.2013.09.003
Dischinger J, Josten M, Szekat C, Sahl H-G, Bierbaum G (2009) Production of the novel two-peptide lantibiotic lichenicidin by Bacillus licheniformis DSM13. PLoS ONE 4:e6788. https://doi.org/10.1371/journal.pone.0006788
Draper LA, Cotter PD, Hill C, Ross RP (2015) Lantibiotic resistance. . Microbiol Molec Biol Rev 79:171–191. https://doi.org/10.1128/MMBR.00051-14
Eraso JM, Kachroo P, Olsen RJ, Beres SB, Zhu L, Badu T et al (2020) Genetic heterogeneity of the Spy1336/R28-Spy1337 virulence axis in Streptococcus pyogenes and effect on gene transcript levels and pathogenesis. PLoS ONE 15:e0229064. https://doi.org/10.1371/journal.pone.0229064
Fischbach MA, Walsh CT (2006) Assembly-line enzymology for polyketide and nonribosomal peptide antibiotics: logic, machinery, and mechanisms. Chem Rev 106:3468–3496. https://doi.org/10.1021/cr0503097
Gravekamp C, Horensky DS, Michel JL, Madoff LC (1996) Variation in repeat number within the alpha C protein of group B streptococci alters antigenicity and protective epitopes. Infect Immun 64:3576–3583. https://doi.org/10.1128/IAI.64.9.3576-3583.1996
Hasper HE, Kramer NE, Smith JL, Hillman JD, Zachariah C, Kuipers OP et al (2006) An alternative bactericidal mechanism of action for lantibiotic peptides that target lipid II. Science 313:1636–1637. https://doi.org/10.1126/science.1129818
Héchard Y, Sahl HG (2002) Mode of action of modified and unmodified bacteriocins from Gram-positive bacteria. Biochimie 84:545–557. https://doi.org/10.1016/s0300-9084(02)01417-7
Hopwood DA, Bibb MJ, Chater KF, Kieser T, Bruton CJ, Kieser HM et al (1985) Genetic manipulation of streptomyces—a laboratory manual. The John Innes Foundation, Norwich
Islam MR, Nagao J, Zendo T, Sonomoto K (2012) Antimicrobial mechanism of lantibiotics. Biochem Soc Trans 40:1528–1533. https://doi.org/10.1042/BST20120190
Jovcic B, Lepsanovic Z, Suljagic V, Rackov G, Begovic J, Topisirovic L et al (2011) Emergence of NDM-1 metallo-β-lactamase in Pseudomonas aeruginosa clinical isolates from Serbia. Antimicrob Agents Chemother 55:3929–3931. https://doi.org/10.1128/AAC.00226-11
Kiss A, Baliko G, Csorba A, Chuluunbaatar T, Medzihradszky KF, Alfoldi L (2008) Cloning and characterization of the DNA region responsible for megacin A-216 production in Bacillus megaterium 216. J Bacteriol 190:6448–6457. https://doi.org/10.1128/JB.00557-08
Kojic M, Strahinic I, Fira D, Jovcic B, Topisirovic L (2006) Plasmid content and bacteriocin production by five strains of Lactococcus lactis isolated from semi-hard homemade cheese. Can J Microbiol 52:1110–1120. https://doi.org/10.1139/w06-072
Lancefield RC (1934) A serological differentiation of specific types of bo- vine haemolytic streptococci (group B). J Exp Med 59:441–458. https://doi.org/10.1084/jem.59.4.441
Le Marrec C, Hyronimus B, Bressollier P, Verneuil B, Urdaci MC (2000) Biochemical and genetic characterization of coagulin, a new antilisterial bacteriocin in the pediocin family of bacteriocins, produced by Bacillus coagulans I4. Appl Environ Microb 66:5213–5220. https://doi.org/10.1128/aem.66.12.5213-5220.2000
Lindahl G, Stålhammar-Carlemalm M, Areschoug T (2005) Surface proteins of Streptococcus agalactiae and related proteins in other bacterial pathogens. Clin Microbiol Rev 18:102–127. https://doi.org/10.1128/CMR.18.1.102-127.2005
Maeland JA, Afset JE, Lyng RV, Radtke A (2015) Survey of immunological features of the alpha-like proteins of Streptococcus agalactiae. Clin Vaccine Immunol 22:153–159. https://doi.org/10.1128/CVI.00643-14
Marahiel MA, Essen LO (2009) Nonribosomal peptide synthetases: mechanistic and structural aspects of essential domains. Method Enzymol 458:337–351. https://doi.org/10.1016/S0076-6879(09)04813-7
Michel JL, Madoff LC, Olson K, Kling DE, Kasper DL, Ausubel FM (1992) Large, identical, tandem repeating units in the C protein alpha antigen gene, bca, of group B streptococci. Proc Natl Acad Sci USA 89:10060–10064. https://doi.org/10.1073/pnas.89.21.10060
Michel JL, Madoff LC, Kling DE, Kasper DL, Ausubel FM (1991) Cloned alpha and beta C-protein antigens of group B streptococci elicit protective immunity. Infect Immun 59:2023–2028. https://doi.org/10.1128/IAI.59.6.2023-2028.1991
Medema MH, Blin K, Cimermancic P, de Jager V, Zakrzewski P, Fischbach MA et al (2011) AntiSMASH: rapid identification, annotation and analysis of secondary metabolite biosynthesis gene clusters in bacterial and fungal genome sequences. Nucl Ac Res 39(Web Server issue):339–346. https://doi.org/10.1093/nar/gkr466
Morgan SM, O’connor PM, Cotter PD, Ross RP, Hill C (2005) Sequential actions of the two component peptides of the lantibiotic lacticin 3147 explain its antimicrobial activity at nanomolar concentrations. Antimicrob Agents Chemother 49:2606–2611. https://doi.org/10.1128/AAC.49.7.2606-2611.2005
Motta SA, Cladera-Olivera F, Brandelli A (2004) Screening for antimicrobial activity among bacteria isolated from the Amazon Basin. Braz J Microbiol. https://doi.org/10.1590/S1517-83822004000300007
Novovic K, Mihajlovic S, Vasiljevic Z, Filipic B, Begovic J, Jovcic B (2015) Carbapenem-resistant Acinetobacter baumanii from Serbia: revision of CarO classification. PLoS ONE 10:e0122793. https://doi.org/10.1371/journal.pone.0122793
Novovic K, Trudic A, Brkic S, Vasiljevic Z, Kojic M, Cirkovic I et al (2017) Molecular epidemiology of colistinresistant, carbapenemase-producing Klebsiella pneumoniae, Serbia, 2013–2016. Antimicrob Agents Chemother 61:e02550-e2616. https://doi.org/10.1128/AAC.02550-16
Oman TJ, van der Donk WA (2009) Insights into the mode of action of the two-peptide lantibiotic haloduracin. ACS Chem Biol 4:865–874. https://doi.org/10.1021/cb900194x
Rea MC, Ross RP, Cotter PD, Hill C (2011) Classification of bacteriocins from Gram-positive bacteria. In: Drider D, Rebuffat S (eds) Prokaryotic antimicrobial peptides. Springer, New York, pp 29–53
Sandiford SK (2014) Advances in the arsenal of tools available enabling the discovery of novel lantibiotics with therapeutic potential. Expert Opin Drug Discov 9:283–297. https://doi.org/10.1517/17460441.2014.877882
Saikia L, Gogoi N, Das PP, Sarmah A, Punam K, Mahanta B et al (2019) Bacillus cereus-attributable primary cutaneous anthrax-like infection in newborn infants, India. Emerg Infect Dis 25:1261–1270. https://doi.org/10.3201/eid2507.181493
Shenkarev ZO, Finkina EI, Nurmukhamedova EK, Balandin SV, Mineev KS, Nadezhdin KD et al (2010) Isolation, structure elucidation, and synergistic antibacterial activity of a novel two-component lantibiotic lichenicidin from Bacillus licheniformis VK21. Biochemistry 49:6462–6472. https://doi.org/10.1021/bi100871b
Singh M, Sareen D (2014) Novel LanT associated lantibiotic clusters identified by genome database mining. PLoS ONE 9:e91352. https://doi.org/10.1371/journal.pone.0091352
Steinberg DA, Hurst MA, Fujii CA, Kung AH, Ho JF, Cheng FC et al (1997) Protegrin-1: a broad-spectrum, rapidly microbicidal peptide with in vivo activity. Antimicrob Agents Chemother 41:1738–1742. https://doi.org/10.1128/AAC.41.8.1738
van Heel AJ, de Jong A, Song C, Viel JH, Kok J, Kuipers OP (2018) BAGEL4: a user-friendly web server to thoroughly mine RiPPs and bacteriocins. Nucl Ac Res 46:278–281. https://doi.org/10.1093/nar/gky383
Vasiljevic ZV, Novovic K, Kojic M, Minic P, Sovtic A, Djukic S et al (2016) Burkholderia cepacia complex in Serbian patients with cystic fibrosis: prevalence and molecular epidemiology. Eur J Clin Microbiol Infect Dis 35:1277–1284. https://doi.org/10.1007/s10096-016-2662-4
Xin B, Zheng J, Liu H, Li J, Ruan L, Peng D et al (2016) Thusin, a novel two-component lantibiotic with potent antimicrobial activity against several Gram-positive pathogens. Front Microbiol 7:1115. https://doi.org/10.3389/fmicb.2016.01115
Weissman KJ, Leadlay PF (2005) Combinatorial biosynthesis of reduced polyketides. Nat Rev Microbiol 3:925–936. https://doi.org/10.1038/nrmicro1287
Whelan F, Lafita A, Griffiths S, Cooper R, Whittingham J, Turkenburg J et al (2019) Defining the remarkable structural malleability of a bacterial surface protein Rib domain implicated in infection. Proc Natl Acad Sci USA 116:26540–26548. https://doi.org/10.1073/pnas.1911776116
Wilkinson HW, Eagon RG (1971) Type-specific antigens of group B type Ic streptococci. Infect Immun 4:596–604. https://doi.org/10.1128/IAI.4.5.596-604.1971
Willey JM, van der Donk WA (2007) Lantibiotics: peptides of diverse structure and function. Annu Rev Microbiol 61:477–501. https://doi.org/10.1146/annurev.micro.61.080706.093501
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This work was partly funded by the Ministry of Education, Science and Technological Development of the Republic of Serbia, Republic of Serbia (Grant No.173019, awarded to MK).
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The study was conceptualized by MK and NM. Laboratory work was done by NM, MO, POC and MK. Whole-genome analysis was carried out by BF and MK. Supervision, Project administration and Funding were carried out by MK. The manuscript was drafted by NM, MO and POC, reviewed by BJ and edited by MK, POC and PC. All authors have read and approved the final version of the manuscript.
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Mirkovic, N., Obradovic, M., O’Connor, P.M. et al. C-protein α-antigen modulates the lantibiotic thusin resistance in Streptococcus agalactiae. Antonie van Leeuwenhoek 114, 1595–1607 (2021). https://doi.org/10.1007/s10482-021-01626-3
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DOI: https://doi.org/10.1007/s10482-021-01626-3