Exploring clinically isolated Staphylococcus sp. bacteriocins revealed the production of amonabactin, micrococcin, and α-circulocin
Abstract
Background and Objectives: Bacteriocins are considered alternative non-conventional antimicrobials produced by certain bacteria with activity against closely related species. The present study focuses on screening, characterization, and partial purification of bacteriocins produced by Staphylococcus sp. isolated from different clinical sources such as pus and blood.
Materials and Methods: A total of 100 Staphylococcus isolates were screened for bacteriocin production using spot on lawn assay and agar diffusion method against five indicator bacteria. Bacteriocins from five selected highly active isolates were subjected to proteinase-K enzyme, different pH, and heating at different temperatures, and investigated the stabilities of their antimicrobials. Two selected isolates, MK65 and MK88, were molecularly identified by 16S rRNA gene sequencing, explored for the presence of 18 bacteriocin genes, and liquid chromatography-high resolution electrospray ionization mass spectrometry (LC-HRESIMS) was used to identify their different metabolites.
Results: Twenty isolates exhibited inhibitory effect against at least one indicator bacteria. Micrococcus luteus ATCC 4698 showed the highest sensitivity to such bacteriocins. Proteinase K, acidic pH, and heating at 100°C triggered marked activity inhibition. However, amylase enzyme, alkaline pH, and heating at 80°C caused trivial effects. Four out of eighteen bacteriocin genes were detected using PCR. Fermentation, partial purification, and LC-HRESIMS of total protein extracts of two selected isolates, MK65 and MK88, revealed the production of different antimicrobial peptides.
Conclusion: To the best of our knowledge, this is the first study to report the production of micrococcin and α-circulocin from Staphylococcus aureus MK65 and the production of amonabactin from Staphylococcus epidermidis MK88.
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11. Lotfy MM, Hassan HM, Mohammed R, Hetta M, El-Gendy AO, Rateb ME, et al. Chemical profiling and biological screening of some river Nile derived-microorganisms. Front Microbiol 2019; 10: 787.
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14. Naguib MM, Khairalla AS, El-Gendy AO, Elkhatib WF. Isolation and characterization of mercury-resistant bacteria from wastewater sources in Egypt. Can J Microbiol 2019; 65: 308-321.
15. El-Helw NO, El-Gendy AO, El-Gebaly E, Hassan HM, Rateb ME, El-Nesr KA. Characterization of natural bioactive compounds produced by isolated bacteria from compost of aromatic plants. J Appl Microbiol 2019; 126: 443-451.
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23. Daly KM, Upton M, Sandiford SK, Draper LA, Wescombe PA, Jack RW, et al. Production of the Bsa lantibiotic by community-acquired Staphylococcus aureus strains. J Bacteriol 2010; 192: 1131-1142.
24. Sashihara T, Kimura H, Higuchi T, Adachi A, Matsusaki H, Sonomoto K, et al. A novel lantibiotic, nukacin ISK-1, of Staphylococcus warneri ISK-1: cloning of the structural gene and identification of the structure. Biosci Biotechnol Biochem 2000; 64: 2420-2428.
25. Martínez B, Suárez JE, Rodríguez A. Lactococcin 972: a homodimeric lactococcal bacteriocin whose primary target is not the plasma membrane. Microbiology (Reading) 1996; 142: 2393-2398.
26. Carnio MC, Höltzel A, Rudolf M, Henle T, Jung G, Scherer S. The macrocyclic peptide antibiotic micrococcin P(1) is secreted by the food-borne bacterium Staphylococcus equorum WS 2733 and inhibits Listeria monocytogenes on soft cheese. Appl Environ Microbiol 2000; 66: 2378-2384.
27. Hsu C, Wiseman GM. The nature of epidermidins, new antibiotics from staphylococci. Can J Microbiol 1972; 18: 121-125.
28. He H, Shen B, Korshalla J, Carter GT. Circulocins, new antibacterial lipopeptides from Bacillus circulans, J2154. Tetrahedron 2001; 57: 1189-1195.
29. Konetschny-Rapp S, Jung G, Meiwes J, Zähner H. Staphyloferrin A: a structurally new siderophore from staphylococci. Eur J Biochem 1990; 191: 65-74.
30. Barghouthi S, Young R, Olson MO, Arceneaux JE, Clem LW, Byers BR. Amonabactin, a novel tryptophan- or phenylalanine-containing phenolate siderophore in Aeromonas hydrophila. J Bacteriol 1989; 171: 1811-1816.
31. Akeda Y, Shibata K, Ping X, Tanaka T, Taniguchi M. AKD-2A, B, C and D, new antibiotics from Streptomyces sp. OCU-42815. Taxonomy, fermentation, isolation, structure elucidation and biological activity. J Antibiot (Tokyo) 1995; 48: 363-368.
32. Saito M, Kawaguchi N, Hashimoto M, Kodama T, Higuchi N, Tanaka T, et al. Purification and structure of novel cysteine proteinase inhibitors, staccopins pi and p2, from staphylococcus tanabeensis. Agric Biol Chem 1987; 51: 861-868.
33. Zimmerman SB, Schwartz CD, Monaghan RL, Pelak BA, Weissberger B, Gilfillan EC, et al. Difficidin and oxydifficidin: novel broad spectrum antibacterial antibiotics produced by Bacillus subtilis. I. Production, taxonomy and antibacterial activity. J Antibiot (Tokyo) 1987; 40: 1677-1681.
34. Dairi T, Hasegawa M. Common biosynthetic feature of fortimicin-group antibiotics. J Antibiot (Tokyo) 1989; 42: 934-943.
35. Rahmdel S, Shekarforoush SS, Hosseinzadeh S, Torriani S, Gatto V. Antimicrobial spectrum activity of bacteriocinogenic Staphylococcus strains isolated from goat and sheep milk. J Dairy Sci 2019; 102: 2928-2940.
36. dos Santos Nascimento J, Fagundes PC, de Paiva Brito MA, dos Santos KR, do Carmo de Freire Bastos M. Production of bacteriocins by coagulase-negative staphylococci involved in bovine mastitis. Vet Microbiol 2005; 106: 61-71.
37. Ceotto H, Nascimento Jdos S, Brito MA, Bastos Mdo C. Bacteriocin production by Staphylococcus aureus involved in bovine mastitis in Brazil. Res Microbiol 2009; 160: 592-599.
38. Saitou N, Nei M. The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol 1987; 4: 406-425.
39. Kumar S, Stecher G, Tamura K. MEGA7: molecular evolutionary genetics analysis version 7.0 for bigger datasets. Mol Biol Evol 2016; 33: 1870-1874.
2. Nes IF, Diep DB, Holo H. Bacteriocin diversity in Streptococcus and Enterococcus. J Bacteriol 2007; 189: 1189-1198.
3. Chikindas ML, Weeks R, Drider D, Chistyakov VA, Dicks LM. Functions and emerging applications of bacteriocins. Curr Opin Biotechnol 2018; 49: 23-28.
4. O'Sullivan JN, Rea MC, O'Connor PM, Hill C, Ross RP. Human skin microbiota is a rich source of bacteriocin-producing staphylococci that kill human pathogens. FEMS Microbiol Ecol 2019; 95:fiy241.
5. Janek D, Zipperer A, Kulik A, Krismer B, Peschel A. High frequency and diversity of antimicrobial activities produced by nasal Staphylococcus strains against bacterial competitors. PLoS Pathog 2016; 12(8): e1005812.
6. Sandiford S, Upton M. Identification, characterization, and recombinant expression of epidermicin NI01, a novel unmodified bacteriocin produced by Staphylococcus epidermidis that displays potent activity against Staphylococci. Antimicrob Agents Chemother 2012; 56: 1539-1547.
7. Molham F, Khairalla AS, Azmy AF, El-Gebaly E, El-Gendy AO, AbdelGhani S. Anti-proliferative and anti-biofilm potentials of bacteriocins produced by non-pathogenic Enterococcus sp. Probiotics Antimicrob Proteins 2020: 10.1007/s12602-020-09711-1.
8. Dündar H, Atakay M, Çelikbıçak Ö, Salih B, Bozoğlu F. Comparison of two methods for purification of enterocin B, a bacteriocin produced by Enterococcus faecium W3. Prep Biochem Biotechnol 2015; 45: 796-809.
9. Ansari A, Zohra RR, Tarar OM, Qader SAU, Aman A. Screening, purification and characterization of thermostable, protease resistant Bacteriocin active against methicillin resistant Staphylococcus aureus (MRSA). BMC Microbiol 2018; 18: 192.
10. Lu Y, Yan H, Li X, Gu Y, Wang X, Yi Y, et al. Physicochemical properties and mode of action of a novel bacteriocin BM1122 with broad antibacterial spectrum produced by Lactobacillus crustorum MN047. J Food Sci 2020; 85: 1523-1535.
11. Lotfy MM, Hassan HM, Mohammed R, Hetta M, El-Gendy AO, Rateb ME, et al. Chemical profiling and biological screening of some river Nile derived-microorganisms. Front Microbiol 2019; 10: 787.
12. Ahmad MS, El-Gendy AO, Ahmed RR, Hassan HM, El-Kabbany HM, Merdash AG. Exploring the antimicrobial and antitumor potentials of Streptomyces sp. AGM12-1 isolated from Egyptian soil. Front Microbiol 2017; 8:438.
13. Tamura K, Dudley J, Nei M, Kumar S. MEGA4: molecular evolutionary genetics analysis (MEGA) software version 4.0. Mol Biol Evol 2007; 24: 1596-1599.
14. Naguib MM, Khairalla AS, El-Gendy AO, Elkhatib WF. Isolation and characterization of mercury-resistant bacteria from wastewater sources in Egypt. Can J Microbiol 2019; 65: 308-321.
15. El-Helw NO, El-Gendy AO, El-Gebaly E, Hassan HM, Rateb ME, El-Nesr KA. Characterization of natural bioactive compounds produced by isolated bacteria from compost of aromatic plants. J Appl Microbiol 2019; 126: 443-451.
16. Blinkova LP, Dorofeeva ES, Baturo AP, Romanenko EE, Katosova l K, Polikarpova SV, et al. [The detection of bacteriogenic causes of opportunistic infections]. Vestn Ross Akad Med Nauk 2008; (4): 14-18.
17. James R, Penfold CN, Moore GR, Kleanthous C. Killing of E. coli cells by E group nuclease colicins. Biochimie 2002; 84: 381-389.
18. Lynch D, O'Connor PM, Cotter PD, Hill C, Field D, Begley M. Identification and characterisation of capidermicin, a novel bacteriocin produced by Staphylococcus capitis. PLoS One 2019; 14(10): e0223541.
19. Padilla C, Lobos O, Brevis P, Abaca P, Hubert E. Effects of the bacteriocin PsVP-10 produced by Pseudomonas sp. on sensitive bacterial strains. Rev Latinoam Microbiol 2002; 44: 19-23.
20. Wladyka B, Wielebska K, Wloka M, Bochenska O, Dubin G, Dubin A, et al. Isolation, biochemical characterization, and cloning of a bacteriocin from the poultry-associated Staphylococcus aureus strain CH-91. Appl Microbiol Biotechnol 2013; 97: 7229-7239.
21. Kuipers OP, Rollema HS, de Vos WM, Siezen RJ. Biosynthesis and secretion of a precursor of nisin Z by Lactococcus lactis, directed by the leader peptide of the homologous lantibiotic subtilin from Bacillus subtilis. FEBS Lett 1993; 330: 23-27.
22. Bierbaum G, Götz F, Peschel A, Kupke T, van de Kamp M, Sahl HG. The biosynthesis of the lantibiotics epidermin, gallidermin, Pep5 and epilancin K7. Antonie Van Leeuwenhoek 1996; 69: 119-127.
23. Daly KM, Upton M, Sandiford SK, Draper LA, Wescombe PA, Jack RW, et al. Production of the Bsa lantibiotic by community-acquired Staphylococcus aureus strains. J Bacteriol 2010; 192: 1131-1142.
24. Sashihara T, Kimura H, Higuchi T, Adachi A, Matsusaki H, Sonomoto K, et al. A novel lantibiotic, nukacin ISK-1, of Staphylococcus warneri ISK-1: cloning of the structural gene and identification of the structure. Biosci Biotechnol Biochem 2000; 64: 2420-2428.
25. Martínez B, Suárez JE, Rodríguez A. Lactococcin 972: a homodimeric lactococcal bacteriocin whose primary target is not the plasma membrane. Microbiology (Reading) 1996; 142: 2393-2398.
26. Carnio MC, Höltzel A, Rudolf M, Henle T, Jung G, Scherer S. The macrocyclic peptide antibiotic micrococcin P(1) is secreted by the food-borne bacterium Staphylococcus equorum WS 2733 and inhibits Listeria monocytogenes on soft cheese. Appl Environ Microbiol 2000; 66: 2378-2384.
27. Hsu C, Wiseman GM. The nature of epidermidins, new antibiotics from staphylococci. Can J Microbiol 1972; 18: 121-125.
28. He H, Shen B, Korshalla J, Carter GT. Circulocins, new antibacterial lipopeptides from Bacillus circulans, J2154. Tetrahedron 2001; 57: 1189-1195.
29. Konetschny-Rapp S, Jung G, Meiwes J, Zähner H. Staphyloferrin A: a structurally new siderophore from staphylococci. Eur J Biochem 1990; 191: 65-74.
30. Barghouthi S, Young R, Olson MO, Arceneaux JE, Clem LW, Byers BR. Amonabactin, a novel tryptophan- or phenylalanine-containing phenolate siderophore in Aeromonas hydrophila. J Bacteriol 1989; 171: 1811-1816.
31. Akeda Y, Shibata K, Ping X, Tanaka T, Taniguchi M. AKD-2A, B, C and D, new antibiotics from Streptomyces sp. OCU-42815. Taxonomy, fermentation, isolation, structure elucidation and biological activity. J Antibiot (Tokyo) 1995; 48: 363-368.
32. Saito M, Kawaguchi N, Hashimoto M, Kodama T, Higuchi N, Tanaka T, et al. Purification and structure of novel cysteine proteinase inhibitors, staccopins pi and p2, from staphylococcus tanabeensis. Agric Biol Chem 1987; 51: 861-868.
33. Zimmerman SB, Schwartz CD, Monaghan RL, Pelak BA, Weissberger B, Gilfillan EC, et al. Difficidin and oxydifficidin: novel broad spectrum antibacterial antibiotics produced by Bacillus subtilis. I. Production, taxonomy and antibacterial activity. J Antibiot (Tokyo) 1987; 40: 1677-1681.
34. Dairi T, Hasegawa M. Common biosynthetic feature of fortimicin-group antibiotics. J Antibiot (Tokyo) 1989; 42: 934-943.
35. Rahmdel S, Shekarforoush SS, Hosseinzadeh S, Torriani S, Gatto V. Antimicrobial spectrum activity of bacteriocinogenic Staphylococcus strains isolated from goat and sheep milk. J Dairy Sci 2019; 102: 2928-2940.
36. dos Santos Nascimento J, Fagundes PC, de Paiva Brito MA, dos Santos KR, do Carmo de Freire Bastos M. Production of bacteriocins by coagulase-negative staphylococci involved in bovine mastitis. Vet Microbiol 2005; 106: 61-71.
37. Ceotto H, Nascimento Jdos S, Brito MA, Bastos Mdo C. Bacteriocin production by Staphylococcus aureus involved in bovine mastitis in Brazil. Res Microbiol 2009; 160: 592-599.
38. Saitou N, Nei M. The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol 1987; 4: 406-425.
39. Kumar S, Stecher G, Tamura K. MEGA7: molecular evolutionary genetics analysis version 7.0 for bigger datasets. Mol Biol Evol 2016; 33: 1870-1874.
| Files | ||
| Issue | Vol 13 No 2 (2021) | |
| Section | Original Article(s) | |
| DOI | https://doi.org/10.18502/ijm.v13i2.5983 | |
| Keywords | ||
| Staphylococcus sp.; Bacteriocins; Amonabactin; Micrococcin; Micrococcus luteus | ||
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How to Cite
1.
Kassem MA, Saafan AE, Bayomy F, Osama El-Gendy A. Exploring clinically isolated Staphylococcus sp. bacteriocins revealed the production of amonabactin, micrococcin, and α-circulocin. Iran J Microbiol. 2021;13(2):212-224.
