Antimicrobial properties of bacillus licheniformis from agricultural lands in Tunceli

Abstract

This study investigated the antimicrobial potential of Bacillus licheniformis strains isolated from agricultural soils in Tunceli, Turkey. Soil samples were collected from minimally disturbed areas and bacterial strains were isolated using morphological, biochemical and molecular identification methods. The isolates were successfully confirmed as Bacillus licheniformis by 16S rDNA sequencing and phylogenetic analysis. The production of antimicrobial secondary metabolites was evaluated by culturing the isolates with different carbon sources (sucrose, glucose and fructose) and testing their activity against Escherichia coli (ATCC 25922) and Staphylococcus aureus (ATCC 25923) using the disk diffusion method. The results determined that sucrose was the most effective carbon source to enhance antimicrobial activity, especially against Escherichia coli. Antibacterial activity generally increased with longer incubation times. This research highlights the potential of Bacillus licheniformis as a source of antibacterial substances. It offers an environmentally friendly alternative for its use in agricultural applications for biological control of pathogens. In conclusion, the findings of this study contribute to knowledge about the antimicrobial properties of Bacillus licheniformis. Understanding the factors affecting carbon source availability and metabolite production is important to exploit the antimicrobial potential of Bacillus licheniformis.

Keywords

• soil bacteria
• secondary metabolites
• bacillus licheniformis
• 16S rDNA
• antibacterial

Tunceli tarım arazilerinden elde edilen bacillus licheniformis'in antimikrobiyal özellikleri

Abstract

Bu çalışmada, Türkiye Tunceli'deki tarım topraklarından izole edilen Bacillus licheniformis suşlarının antimikrobiyal potansiyelleri araştırılmıştır. Toprak örnekleri minimal olarak bozulmuş alanlardan toplanmış ve bakteri suşları morfolojik, biyokimyasal ve moleküler tanımlama yöntemleri kullanılarak izole edilmiştir. İzolatlar, 16S rDNA dizilemesi ve filogenetik analiz yoluyla Bacillus licheniformis oldukları başarılı bir şekilde doğrulanmıştır. Antimikrobiyal sekonder metabolitlerin üretimi, izolatların farklı karbon kaynaklarıyla (sükroz, glikoz ve fruktoz) kültürlenmesi ve disk difüzyon yöntemi kullanılarak Escherichia coli (ATCC 25922) ve Staphylococcus aureus'a (ATCC 25923) karşı aktivitelerinin test edilmesiyle değerlendirilmiştir. Sonuçlar, özellikle Escherichia coli'ye karşı antimikrobiyal aktiviteyi artırmak için en etkili karbon kaynağının sükroz olduğu belirlenmiştir. Antibakteriyel aktivite genellikle daha uzun inkübasyon süreleriyle artmıştır. Bu araştırma, Bacillus licheniformis'in antibakteriyel madde kaynağı olarak potansiyelini vurgulamaktadır. Patojenlerin biyolojik kontrolü için tarımdaki uygulamalarda kullanılması için çevre dostu bir alternatif sunmaktadır. Sonuç olarak, bu çalışmanın bulguları Bacillus licheniformis'in antimikrobiyal özellikleri hakkında bilgilere katkıda bulunmaktadır. Karbon kaynağındaki kullanılabilirliği ve metabolit üretimini etkileyen faktörleri anlamak, Bacillus licheniformis'in antimikrobiyal potansiyelinden yararlanmak için önemlidir.

Keywords

• toprak bakterileri
• ikincil metabolitler
• bacillus licheniformi
• 16S rDNA
• antibakteriyel

Supporting Institution

Bu çalışma, TÜBİTAK 2209/A - Üniversite Öğrencileri Yurt İçi Araştırma Projeleri Destek Programı (Proje no: 1919B012218280) kapsamında desteklenmektedir.

References

1. J. Jayaraj, S. Shibila, M. Ramaiah, T. A. Alahmadi, S. A. Alharbi, P. K. Mideen et al., “Isolation and identification of bacteria from the agricultural soil samples to tolerate pesticides dimethoate, thiamethoxam and imidacloprid,” Environmental Research Communications, vol. 5, no. 7, p. 075011, 2023.
2. M. Tudi, H. D. Ruan, L. Wang, J. Lyu, R. Sadler, D. Connell et al., “Agriculture development, pesticide application and its impact on the environment,” International Journal of Environmental Research and Public Health, vol. 18, no. 3, p. 1112, 2021.
3. B. Meza-Manzaneque, M. Pérez-Díaz, E. G. Biosca, and B. Álvarez, “Isolation and characterization of agricultural soil bacteria with biotechnological and biological control potential applications,” Biology and Life Sciences Forum, vol. 31, no. 1, p. 28, 2024.
4. G. A. Rook, C. A. Lowry, and C. L. Raison, “Microbial ‘old friends’, immunoregulation and stress resilience,” Evolution, Medicine, and Public Health, no. 1, pp. 46–64, 2013.
5. S. B. Peterson, S. K. Bertolli, and J. D. Mougous, “The central role of interbacterial antagonism in bacterial life,” Current Biology, vol. 30, no. 19, pp. R1203–R1214, 2020.
6. M. A. Eltokhy, B. T. Saad, W. N. Eltayeb, M. Y. Alshahrani, S. M. Radwan, and K. M. Aboshanab, “Metagenomic nanopore sequencing for exploring the nature of antimicrobial metabolites of Bacillus haynesii,” AMB Express, vol. 14, no. 1, p. 52, 2024.
7. S. Hallaj-Nezhadi, R. Hamdipour, M. Shahrvirani, R. Zare Tin, F. Chapeland-Leclerc, and G. Ruprich-Robert et al., “Antimicrobial activity of Bacillus sp. isolated strains of wild honey,” BMC Complementary Medicine and Therapies, vol. 22, p. 78, 2022.
8. M. Vehapi, B. İnan, S. Kayacan-Çakmakoğlu, O. Sağdıç, and D. Özçimen, “Production of Bacillus subtilis soil isolate as biocontrol agent under bioreactor conditions,” Archives of Microbiology, vol. 205, p. 52, 2023.

9. S. Caulier, C. Nannan, A. Gillis, F. Licciardi, C. Bragard, and J. Mahillon, “Overview of the antimicrobial compounds produced by members of the Bacillus subtilis group,” Frontiers in Microbiology, vol. 10, p. 302, 2019.
10. F. C. Yazdıç, A. Karaman, G. Torğut, and A. N. Karaaslan, “Antibacterial activity of novel synthesized chitosan graft poly N tertiary butylacrylamide neodymium composites for biomedical application,” Journal of Basic Microbiology, vol. 63, no. 9, pp. 1049–1056, 2023.
11. A. Saggese, Y. De Luca, L. Baccigalupi, and E. Ricca, “An antimicrobial peptide specifically active against Listeria monocytogenes is secreted by Bacillus pumilus SF214,” BMC Microbiology, vol. 22, p. 3, 2022.
12. M. O. Shleeva, D. A. Kondratieva, and A. S. Kaprelyants, “Bacillus licheniformis: A producer of antimicrobial substances, including antimycobacterials, which are feasible for medical applications,” Pharmaceutics, vol. 15, no. 7, p. 1893, 2023.
13. C. Harwood, J. Mouillon, S. Pohl, and J. Arnau, “Secondary metabolite production and the safety of industrially important members of the Bacillus subtilis group,” FEMS Microbiology Reviews, vol. 42, pp. 721–738, 2018.
14. Y. Wang, Y. Chen, J. Xin, X. Chen, T. Xu, and J. He, “Metabolomic profiles of the liquid state fermentation in co-culture of Eurotium amstelodami and Bacillus licheniformis,” Frontiers in Microbiology, vol. 14, 2023.
15. G. Singh, “Linking lichen metabolites to genes: Emerging concepts and lessons from molecular biology and metagenomics,” Journal of Fungi, vol. 9, 2023.
16. J. Pathma, G. R. Rahul, K. R. Kamaraj, R. Subashri, and N. Sakthivel, “Secondary metabolite production by bacterial antagonists,” Journal of Biological Control, pp. 165–181, 2011.
17. L. Liangzhi, H. Zheng, and Y. Yingjin, “Effects of propionate on streptolydigin production and carbon flux distribution in Streptomyces lydicus AS 4.2501,” Chinese Journal of Chemical Engineering, vol. 15, no. 2, pp. 143–149, 2007.
18. D. G. Cooper, C. R. Macdonald, S. J. B. Duff, and N. Kosaric, “Enhanced production of surfactin from Bacillus subtilis by continuous product removal and metal cation additions,” Applied and Environmental Microbiology, vol. 42, no. 3, pp. 408–412, 1981.
19. N. Iwase, M. S. Rahman, and T. Ano, “Production of iturin A homologues under different culture conditions,” Journal of Environmental Sciences, vol. 21, pp. S28–S32, 2009.
20. A. S. de Boer, F. Priest, and B. Diderichsen, “On the industrial use of Bacillus licheniformis: A review,” Applied Microbiology and Biotechnology, vol. 40, pp. 595–598, 1994.
21. A. Muras, M. Romero, C. Mayer, and A. Otero, “Biotechnological applications of Bacillus licheniformis,” Critical Reviews in Biotechnology, vol. 41, no. 4, pp. 609–627, 2021.
22. Z. Demirbağ and İ. Demir, Genel Mikrobiyoloji Laboratuvarı Uygulama Kitabı, Trabzon: KATÜ, 2005.
23. C. H. Collins and P. M. Lyne, Microbiological Methods, 3rd ed., 1970.
24. E. M. Mauti, G. O. Mauti, G. A. Ouno, B. M. Mabeya, and S. Kiprono, “Molecular identification of soil bacteria by 16S rDNA sequence,” Journal of Natural Sciences Research, vol. 3, no. 14, pp. 51–58, 2013.
25. T. Hall, “BioEdit: An important software for molecular biology,” GERF Bulletin of Biosciences, vol. 2, no. 1, pp. 60–61, 2011.
26. M. Z. Rahman, S. Uematsu, M. D. Coffey, S. Uzuhashi, H. Suga, and K. Kageyama, “Re-evaluation of Japanese Phytophthora isolates based on molecular phylogenetic analyses,” Mycoscience, vol. 55, pp. 314–327, 2014.
27. M. S. Wang, “Phylogenetic relationships between Bacillus species and related genera inferred from 16S rDNA sequences,” Brazilian Journal of Microbiology, vol. 40, pp. 505–521, 2009.
28. L. Lorenz, B. Lins, J. Barrett, A. Montgomery, S. Trapani, and A. Schindler, “Genomic characterization of six novel Bacillus pumilus bacteriophages,” Virology, vol. 444, pp. 374–383, 2013.
29. N. Taşar, F. C. Yazdıç, A. Karaman, and O. Gedik, “Foeniculum vulgare Mill ve Pimpinella anisum L türlerinin uçucu yağlarının antimikrobiyal aktivitesi,” International Journal of Innovative Engineering Applications, vol. 7, no. 1, pp. 119–122, 2023.
30. A. P. David and J. P. McCuen, Manual of BBL Products and Laboratory Procedures, 1988.
31. S. T. Williams, M. E. Sharpe, and J. G. Holt, Bergey’s Manual of Systematic Bacteriology, vol. 4, USA: Williams and Wilkins, 1993.
32. M. Park, C. Hong, B. Kim, D. Lee, and Y. Kim, “Integrated-omics study on the transcriptomic and metabolic changes of Bacillus licheniformis,” Journal of Agricultural and Food Chemistry, 2023.
33. N. Wang, J. Gao, L. Yuan, Y. Jin, and G. He, “Metabolomics profiling during biofilm development of Bacillus licheniformis,” International Journal of Food Microbiology, vol. 337, p. 108939, 2020.
34. C. Bang et al., “Metaorganisms in extreme environments,” Zoology, vol. 127, pp. 1–19, 2018.
35. A. Crits-Christoph et al., “Soil bacterial populations are shaped by recombination,” The ISME Journal, vol. 14, pp. 1834–1846, 2020.
36. F. C. Yazdıç, A. Karaman, and F. Yazdıç, “Sekonder metabolitlerin antimikrobiyal etkisi,” Turkish Journal of Agriculture-Food Science and Technology, vol. 6, pp. 1437–1443, 2018.
37. I. Gugel et al., “Fed-batch bioreactor cultivation of Bacillus subtilis,” Fermentation, vol. 10, p. 323, 2024.
38. Z. Liu, Y. Zhao, C. Huang, and Y. Luo, “Silent gene cluster activation in Streptomyces,” Frontiers in Bioengineering and Biotechnology, vol. 9, 2021.
39. G. Zong et al., “Elicitors to enhance antibiotic production,” Critical Reviews in Biotechnology, vol. 42, pp. 1260–1283, 2022.
40. S. Sánchez et al., “Carbon source regulation of antibiotic production,” The Journal of Antibiotics, vol. 63, pp. 442–459, 2010.
41. N. Suwannakul et al., “Targeting fructose metabolism in cholangiocarcinoma,” Genes & Diseases, vol. 9, pp. 1727–1741, 2021.