Suppression of predominant interfering bacteria in the purification process of myxobacteria
Abstract
Background and Objectives: Myxobacteria initially recognized by their complex life cycle and social behavior are progressively explored for their bioactive secondary metabolites. However, isolation of myxobacteria usually is accompanied by bacterial and fungal contaminations due to the direct cultivation of soil on isolation media, which results in severe challenges in the purification of myxobacteria. Therefore, it is necessary to improve their purification techniques from natural samples to the discovery of new biomolecules.
Materials and Methods: In the present study, six physichochemical methods were assessed for their efficacy in the purification of myxobacterial strains and specially from contaminants of Microvirga spp.
Results: Among the evaluated treatments, purification of fruiting bodies using a combination of ultrasonication and heat treatment was identified as the effective protocol with 80% success rate in the purification of myxobacterial strains and reducing up to 90% of the contaminating bacteria.
Conclusion: Concerning the problematic contamination of myxobacterial isolates, the introduced approach can retrieve the myxobacterial strains which are often suppressed by the over growth of contaminations especially root symbiotic bacteria namely Microvirga spp.
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10. Ardley JK, Parker MA, De Meyer SE, Trengove RD, O'Hara GW, Reeve WG, et al. Microvirga lupini sp. nov., Microvirga lotononidis sp. nov., and Microvirga zambiensis sp. nov. are Alphaproteobacterial root nodule bacteria that specifically nodulate and fix nitrogen with geographically and taxonomically separate legume hosts. Int J Syst Evol Microbiol 2012; 62: 2579-2588.
11. Parte AC. LPSN–List of Prokaryotic names with Standing in Nomenclature (bacterio. net), 20 years on. Int J Syst Evol Microbiol 2018; 68: 1825-1829.
12. Radl V, Simões-Araújo JL, Leite J, Passos SR, Martins LMV, Xavier GR, et al. Microvirga vignae sp. nov., a root nodule symbiotic bacterium isolated from cowpea grown in semi-arid Brazil. Int J Syst Evol Microbiol 2014; 64: 725-730.
13. Kanso S, Patel BKC. Microvirga subterranea gen. nov., sp. nov., a moderate thermophile from a deep subsurface Australian thermal aquifer. Int J Syst Evol Microbiol 2003; 53: 401-406.
14. Rosenberg, E (2012). Myxobacteria: development and cell interactions. Springer Sci Bus Med p. 45-48.
15. Rouhizohrab N, Mohammadipanah F. Thermostable alkaline serine protease production by the soil Myxobacterium of Archangium sp. UTMC 4504. Ind Biotechnol 2021; 17: 290-298.
16. Shimkets LJ, Dworkin M, Reichenbach H (2006). The Myxobacteria, In: Dworkin M, Falkow S, Rosenberg E, Schleifer KH, Stackebrandt E. (eds). The Prokaryotes. Springer New York, NY. p. 31-115.
17. Zhang H, Venkatesan S, Nan B. Myxococcus xanthus as a model organism for peptidoglycan assembly and bacterial morphogenesis. Microorganisms 2021; 9: 916.
18. Mohr KI, Stechling M, Wink J, Wilharm E, Stadler M. Comarison of myxobacterial diversity and evaluation of isolation success in two niches: Kiritimati Island and German compost. Microbiologyopen 2016; 5: 268-278.
19. Caputo A, Lagier J-C, Azza S, Robert C, Mouelhi D, Fournier P-E, et al. Microvirga massiliensis sp. nov., the human commensal with the largest genome. Microbiologyopen 2016; 5: 307-322.
20. Zhang L, Wang H, Fang X, Stackebrandt E, Ding Y. Improved methods of isolation and purification of myxobacteria and development of fruiting body formation of two strains. J Microbiol Methods 2003; 54: 21-27.
21. Liu B, Lin Y, LiFu F, LiuXing H. Isolation and purification of myxobacteria in Chengdu. Anim Husb Feed Sci 2011; 3: 25-27.
22. Serafini A, Riello E, Trojan D, Cogliati E, Palù G, Manganelli R, et al. Evaluation of new antibiotic cocktails against contaminating bacteria found in allograft tissues. Cell Tissue Bank 2016; 17: 619-628.
23. Herrmann J, Fayad AA, Müller R. Natural products from myxobacteria: novel metabolites and bioactivities. Nat Prod Rep 2017; 34: 135-160.
24. Chen L, Wang Z, Du S, Wang G. Antimicrobial activity and functional genes of actinobacteria from coastal wetland. Curr Microbiol 2021; 78: 3058-3067.
25. Wang DS, Xue QH, Zhu WJ, Zhao J, Duan JL, Shen GH. Microwave irradiation is a useful tool for improving isolation of actinomycetes from soil. Mikrobiologiia 2013; 82: 106-114.
26. Böllmann J, Rathsack K, Martienssen M. The precision of bacterial quantification techniques on different kinds of environmental samples and the effect of ultrasonic treatment. J Microbiol Methods 2016; 126: 42-47.
27. Kobayashi N, Bauer TW, Tuohy MJ, Fujishiro T, Procop GW. Brief ultrasonication improves detection of biofilm-formative bacteria around a metal implant. Clin Orthop Relat Res 2007; 457: 210-213.
28. JiangY, Cao Y, Zhao L, Wang Q, Jin R, He W, et al. Ultrasonic treatment of soil samples for actinomycete isolation. Wei Sheng Wu Xue Bao 2010; 50: 1094-1097.
29. Donald L, Pipite A, Subramani R, Owen J, Keyzers RA, Taufa T. Streptomyces: Still the Biggest Producer of New Natural Secondary Metabolites, a Current Perspective. Microbiol Res 2022; 13: 418-465.
30. TakahashiY, Omura S. Isolation of new actinomycete strains for the screening of new bioactive compounds. J Gen Appl Microbiol 2003; 49: 141-154.
31. Rana N, Khadka S, Marasini B, Joshi B, Poudel P, Khanal S, et al. Isolation and characterization of soil myxobacteria from Nepal. J Inst Sci Technol 2019; 24: 7-16.
32. Gaspari F, Paitan Y, Mainini M, Losi D, Ron EZ, Marinelli F. Myxobacteria isolated in Israel as potential source of new anti‐infectives. J Appl Microbiol 2005; 98: 429-439.
33. Zhang J, Song F, Xin YH, Zhang J, Fang C. Microvirga guangxiensis sp. nov., a novel alphaproteobacterium from soil, and emended description of the genus Microvirga. Int J Syst Evol Microbiol 2009; 59: 1997-2001.
34. Owusu EGA, MacRobert AJ, Naasani I, Parkin IP, Allan E, Yaghini E, et al. Photoactivable polymers embedded with cadmium-free quantum dots and crystal violet: efficient bactericidal activity against clinical strains of antibiotic-resistant bacteria. ACS Appl Mater Interfaces 2019; 11: 12367-12378.
35. Kim S-T, Yun S-C. Selection of KYC 3270, a cellulolytic myxobacteria of Sorangium cellulosum, against several phytopathogens and a potential biocontrol agent against gray mold in stored fruit. Plant Pathol J 2011; 27: 257-265.
36. Dworkin M, Falkow S, Rosenberg E, Schleifer K-H, Stackebrandt E (2006). The prokaryotes: volume 7: Proteobacteria: delta, epsilon subclass. Deeply Rooting Bacteria, Springer.
37. IizukaT, Jojima Y, Hayakawa A, Fujii T, Yamanaka S, Fudou R. Pseudenhygromyxa salsuginis gen. nov., sp. nov., a myxobacterium isolated from an estuarine marsh. Int J Syst Evol Microbiol 2013; 63: 1360-1369.
2. Wrótniak-Drzewiecka W, Brzezińska AJ, Dahm H, Ingle AP, Rai M. Current trends in myxobacteria research. Ann Microbiol 2016; 66: 17-33.
3. Dos Santos DFK, Kyaw CM, De Campos TA, Miller RNG, Noronha EF, Da Cunha Bustamante MM, et al ( 2014). The family Cystobacteraceae. In: Rosenberg E, DeLong, EF, Lory S, Stackebrandt E, Thompson F. (eds). The Prokaryotes. Springer, Berlin, Heidelberg p. 19-40.
4. Garcia RO, Krug D, Müller R. Chapter 3 Discovering natural products from myxobacteria with emphasis on rare producer strains in combination with improved analytical methods. Methods Enzymol 2009; 458: 59-91.
5. Mohr KI. Diversity of myxobacteria—we only see the tip of the iceberg. Microorganisms 2018; 6: 84.
6. Saadatpour F, Mohammadipanah F. Bioprospecting of indigenous myxobacteria from Iran and potential of Cystobacter as a source of anti-MDR compounds. Folia Microbiol (Praha) 2020; 65: 639-648.
7. Babadi ZK, Garcia R, Ebrahimipour GH, Risdian C, Kampfer P, Jarek M, et al. Corallococcus soli sp. Nov., a Soil Myxobacterium Isolated from Subtropical Climate, Chalus County, Iran, and Its Potential to Produce Secondary Metabolites. Microorganisms 2022; 10: 1262.
8. Shimkets LJ, Dworkin M, Reichenbach H (2006). The myxobacteria. In: Dworkin M, Falkow S, Rosenberg E, Schleifer KH, Stackebrandt E. (eds) The Prokaryotes. Springer, New York, NY. p. 31-115.
9. Holzinger A, Lütz‐Meindl U. Chondramides, novel cyclodepsipeptides from myxobacteria, influence cell development and induce actin filament polymerization in the green alga Micrasterias. Cell Motil Cytoskeleton 2001; 48: 87-95.
10. Ardley JK, Parker MA, De Meyer SE, Trengove RD, O'Hara GW, Reeve WG, et al. Microvirga lupini sp. nov., Microvirga lotononidis sp. nov., and Microvirga zambiensis sp. nov. are Alphaproteobacterial root nodule bacteria that specifically nodulate and fix nitrogen with geographically and taxonomically separate legume hosts. Int J Syst Evol Microbiol 2012; 62: 2579-2588.
11. Parte AC. LPSN–List of Prokaryotic names with Standing in Nomenclature (bacterio. net), 20 years on. Int J Syst Evol Microbiol 2018; 68: 1825-1829.
12. Radl V, Simões-Araújo JL, Leite J, Passos SR, Martins LMV, Xavier GR, et al. Microvirga vignae sp. nov., a root nodule symbiotic bacterium isolated from cowpea grown in semi-arid Brazil. Int J Syst Evol Microbiol 2014; 64: 725-730.
13. Kanso S, Patel BKC. Microvirga subterranea gen. nov., sp. nov., a moderate thermophile from a deep subsurface Australian thermal aquifer. Int J Syst Evol Microbiol 2003; 53: 401-406.
14. Rosenberg, E (2012). Myxobacteria: development and cell interactions. Springer Sci Bus Med p. 45-48.
15. Rouhizohrab N, Mohammadipanah F. Thermostable alkaline serine protease production by the soil Myxobacterium of Archangium sp. UTMC 4504. Ind Biotechnol 2021; 17: 290-298.
16. Shimkets LJ, Dworkin M, Reichenbach H (2006). The Myxobacteria, In: Dworkin M, Falkow S, Rosenberg E, Schleifer KH, Stackebrandt E. (eds). The Prokaryotes. Springer New York, NY. p. 31-115.
17. Zhang H, Venkatesan S, Nan B. Myxococcus xanthus as a model organism for peptidoglycan assembly and bacterial morphogenesis. Microorganisms 2021; 9: 916.
18. Mohr KI, Stechling M, Wink J, Wilharm E, Stadler M. Comarison of myxobacterial diversity and evaluation of isolation success in two niches: Kiritimati Island and German compost. Microbiologyopen 2016; 5: 268-278.
19. Caputo A, Lagier J-C, Azza S, Robert C, Mouelhi D, Fournier P-E, et al. Microvirga massiliensis sp. nov., the human commensal with the largest genome. Microbiologyopen 2016; 5: 307-322.
20. Zhang L, Wang H, Fang X, Stackebrandt E, Ding Y. Improved methods of isolation and purification of myxobacteria and development of fruiting body formation of two strains. J Microbiol Methods 2003; 54: 21-27.
21. Liu B, Lin Y, LiFu F, LiuXing H. Isolation and purification of myxobacteria in Chengdu. Anim Husb Feed Sci 2011; 3: 25-27.
22. Serafini A, Riello E, Trojan D, Cogliati E, Palù G, Manganelli R, et al. Evaluation of new antibiotic cocktails against contaminating bacteria found in allograft tissues. Cell Tissue Bank 2016; 17: 619-628.
23. Herrmann J, Fayad AA, Müller R. Natural products from myxobacteria: novel metabolites and bioactivities. Nat Prod Rep 2017; 34: 135-160.
24. Chen L, Wang Z, Du S, Wang G. Antimicrobial activity and functional genes of actinobacteria from coastal wetland. Curr Microbiol 2021; 78: 3058-3067.
25. Wang DS, Xue QH, Zhu WJ, Zhao J, Duan JL, Shen GH. Microwave irradiation is a useful tool for improving isolation of actinomycetes from soil. Mikrobiologiia 2013; 82: 106-114.
26. Böllmann J, Rathsack K, Martienssen M. The precision of bacterial quantification techniques on different kinds of environmental samples and the effect of ultrasonic treatment. J Microbiol Methods 2016; 126: 42-47.
27. Kobayashi N, Bauer TW, Tuohy MJ, Fujishiro T, Procop GW. Brief ultrasonication improves detection of biofilm-formative bacteria around a metal implant. Clin Orthop Relat Res 2007; 457: 210-213.
28. JiangY, Cao Y, Zhao L, Wang Q, Jin R, He W, et al. Ultrasonic treatment of soil samples for actinomycete isolation. Wei Sheng Wu Xue Bao 2010; 50: 1094-1097.
29. Donald L, Pipite A, Subramani R, Owen J, Keyzers RA, Taufa T. Streptomyces: Still the Biggest Producer of New Natural Secondary Metabolites, a Current Perspective. Microbiol Res 2022; 13: 418-465.
30. TakahashiY, Omura S. Isolation of new actinomycete strains for the screening of new bioactive compounds. J Gen Appl Microbiol 2003; 49: 141-154.
31. Rana N, Khadka S, Marasini B, Joshi B, Poudel P, Khanal S, et al. Isolation and characterization of soil myxobacteria from Nepal. J Inst Sci Technol 2019; 24: 7-16.
32. Gaspari F, Paitan Y, Mainini M, Losi D, Ron EZ, Marinelli F. Myxobacteria isolated in Israel as potential source of new anti‐infectives. J Appl Microbiol 2005; 98: 429-439.
33. Zhang J, Song F, Xin YH, Zhang J, Fang C. Microvirga guangxiensis sp. nov., a novel alphaproteobacterium from soil, and emended description of the genus Microvirga. Int J Syst Evol Microbiol 2009; 59: 1997-2001.
34. Owusu EGA, MacRobert AJ, Naasani I, Parkin IP, Allan E, Yaghini E, et al. Photoactivable polymers embedded with cadmium-free quantum dots and crystal violet: efficient bactericidal activity against clinical strains of antibiotic-resistant bacteria. ACS Appl Mater Interfaces 2019; 11: 12367-12378.
35. Kim S-T, Yun S-C. Selection of KYC 3270, a cellulolytic myxobacteria of Sorangium cellulosum, against several phytopathogens and a potential biocontrol agent against gray mold in stored fruit. Plant Pathol J 2011; 27: 257-265.
36. Dworkin M, Falkow S, Rosenberg E, Schleifer K-H, Stackebrandt E (2006). The prokaryotes: volume 7: Proteobacteria: delta, epsilon subclass. Deeply Rooting Bacteria, Springer.
37. IizukaT, Jojima Y, Hayakawa A, Fujii T, Yamanaka S, Fudou R. Pseudenhygromyxa salsuginis gen. nov., sp. nov., a myxobacterium isolated from an estuarine marsh. Int J Syst Evol Microbiol 2013; 63: 1360-1369.
| Files | ||
| Issue | Vol 14 No 5 (2022) | |
| Section | Original Article(s) | |
| DOI | https://doi.org/10.18502/ijm.v14i5.10968 | |
| Keywords | ||
| Myxobacteria; Strain purification; Heat treatment; Ultrasonication | ||
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How to Cite
1.
Rouhizohrab N, Mohammadipanah F. Suppression of predominant interfering bacteria in the purification process of myxobacteria. Iran J Microbiol. 2022;14(5):721-729.
