MICROORGANISMS AS CHEMICAL FACTORIES FOR ISOLATION OF META-BOLOMES FROM MESOPHILIC SOIL
Objective: To investigate the micro-flora of the mesophilic soil of Rajasthan for isolation of novel compounds having antibacterial potentials.
Methods: In present experiments, bacterial colonies were isolated from four different regions of Jaipur, Rajasthan and screened for antimicrobial efficacy against five selected pathogens Pseudomonas aeruginosa MTCC 7093, Staphylococcus aureus MTCC 7443, Escherichia coli MTCC 40, Klebsiella pneumoniae MTCC 530, and Bacillus subtillis MTCC 121. Antimicrobial efficacy against the selected strains was performed. The potential efficacy of the extract was also screened for gas chromatographyâ€“mass spectroscopy (GC-MS) analysis for novel metabolites screening. Further, potent bacterial strains were identified at the molecular level by 16S ribosomal deoxyribonucleic acid (DNA) sequencing method.
Results: After the primary screening, 29 microbial isolates were selected for the screening of bioactivity. Results displayed zones of inhibition ranging from 5 mm till maximum 13 mm. Soil testing indicated survival conditions for microbes isolated, and biochemical tests supported the identification of screened isolates. The potentially isolated strains S-III C, S-III D and S-IV D were identified at the molecular level using 16S ribosomal DNA sequencing as Bacillus shackletonii, Bacillus thuringiensis and Bacillus subtilis subsp. inaquosorum, respectively.
Conclusion: Extraction of active metabolites from soil microbiota, against five pathogenic bacteria, is far better, safe and economical method. This study will help in exploring new compounds against increasing number of resistant pathogenic strains with an aim to reduce demand of medicinal plants for extraction of effective antimicrobial compounds. Bacillus strains (S-IIIC, S-IIID and S-IVC) isolated from soil microflora possess antimicrobial activity and can be used for isolation of antibiotics at industrial levels.
2. Kamat U, Ferreira A, Savio R, Motghare D. Antimicrobial resistance among nosocomial isolates in a teaching hospital in Goa. Indian J Community Med 2008;33:89-92.
3. Sharma G, Dang S, Gupta S, Gabrani R. Identification and molecular characterization of bacteria having antimicrobial and antibiofilm activity. Int J Pharm Pharm Sci 2016;8:111.
4. Mathai E, Kaufmann ME, Richard VS, John G, Brahmadathan KN. Typing of Acinetobacter baumannii isolated from hospital-acquired respiratory infections in a tertiary care centre in southern India. J Hosp Infect 2001;47 Suppl 2:159-62.
5. Sahni V, Agarwal SK, Singh NP, Anuradha S, Sikdar S, Wadhwa A, et al. Candidemia-an under-recognized nosocomial infection in Indian hospitals. J Assoc Physicians India 2005;53:607-11.
6. Demain AL, Fang A. The natural functions of secondary metabolites. In History of Modern Biotechnology I: Springer Berlin Heidelberg; 2000. p. 1-39.
7. Berdy J. Bioactive microbial metabolites. J Anti biol 2005;58 Suppl 1:1-26.
8. Bibb MJ. Regulation of secondary metabolism in Streptomycetes. Curr Opin Microbiol 2005;8 Suppl 2:208-15.
9. Lamichhane B. Antibiotic resistance patterns of gram negative isolates in a tertiary care hospital of Nepal. Asian J Pharm Clin Res 2014;7 Suppl 3:30-3.
10. Siddhardha B. Studies on the isolation, characterization and bioevaluation of secondary metabolites from Aspergillus funiculosus, Aspergillus gorakhpurensis and Curvulariaoryzae. Ph. D dissertation, Jawaharlal Nehru technological University; 2010.
11. Davies, Davies D. Origins and evolution of antibiotic resistance. Microbiol Mol Biol Rev 2010;74 Suppl 3:417â€“33.
12. Kumari P, Kumar A, Somasundaram ST. Identification of genes/enzymes responsible for antibiotic resistance in Vibrio alginolyticus strain PTS 13. Int J Curr Pharm Res 2015;7:76-83.
13. Stein T. Bacillus subtilis antibiotics: structures, syntheses and specific functions. Mol Microbiol 2005;56:845â€“57.
14. Bizani D, Motta AS, Morrissy JA, Terra R, Souto AA, Brandelli A. Antibacterial activity of cerein 8A, a bacteriocin-like peptide produced by Bacillus cereus. Int Microbiol 2010;8:125-31.
15. Xie J, Zhang R, Shang C, Guo Y. Isolation and characterization of a bacteriocin produced by an isolated Bacillus subtilis LFB112 that exhibits antimicrobial activity against domestic animal pathogens. Afr J Biotechnol 2009;8:5611â€“9.
16. Teixeira ML, Dalla RA, Brandelli A. Characterization of an antimicrobial peptide produced by Bacillus subtilis subsp. spizezinii showing inhibitory activity towards Haemophilusparasuis. Microbiology 2013;159:980-8.
17. Magaldi S, Mata-Essayag S, De Capriles CH, Perez C, Colella MT, Olaizola C, et al. Well diffusion for antifungal susceptibility testing. Int J Infect Dis 2004;8 Suppl 1:39-45.
18. Saadoun I, Wahiby L, Ababneh Q, Jaradat Z, Massadeh M, Al-Momani F. Recovery of soil Streptomycetes from arid habitats in Jordan and their potential to inhibit multidrug resistant Pseudomonas aeruginosa pathogens. World J Microb Biot 2008;24:157-62.
19. Abussaud MJ, Alanagreh L, Elteen KA. Isolation, characterization and antimicrobial activity of Streptomyces strains from hot spring areas in the northern part of Jordan. Afr J Biotechnol 2013;12:7124-32.
20. Mandenius CF, BjÃ¶rkman M. Scale-up of cell bioreactors using biomechatronic design. Biotechnol J 2012;7:1026-39.
21. Pedersen-Bjergaard S, Rasmussen KE, Halvorsen TG. Liquidâ€“liquid extraction procedures for sample enrichment in capillary zone electrophoresis. J Chromatography 2000;902 Suppl 1:91-105.
22. Balouiri Mounyr, Moulay Sadiki, Saad Koraichi. IBN souda methods for in vitro evaluating antimicrobial activity: a review. J Pharm Anal 2016;6:71-9.
23. Sahin N, Ugur A. Investigation of the antimicrobial activity of some Streptomyces isolates. Turk J Biol 2003;27:73-8.
24. Verma B, Kumar P, Dhanasekaran D, Babalola OO, Banakar SP. Gas chromatography-mass spectrometry analysis and antibacterial activity of bluish green pigment from Pseudomonas sp. JJTBVK (KF836502). Braz Arch Biol Technol 2015;58 Suppl 4:628-35.
25. Kim OS, Cho YJ, Lee K, Yoon SH, Kim M, Na H, et al. Introducing EzTaxon-e: a prokaryotic 16S rRNA gene sequence database with phylotypes that represent uncultured species. Int J Syst Evol Microbiol 2012;62:716-21.
26. Frankland GC, Frankland PF. Studies on some new micro-organisms obtained from air. Philos Trans R Soc B 1887;178:257-87.
27. Claus DR. Genus bacillus cohn 1872. Bergey's manual of systematic bacteriology. 2nd ed.; 1986. p. 1105-39.
28. Berliner E. Ãœber die schlaffsucht der mehlmottenraupe (EphestiakÃ¼hniella Zell.) und ihren Erreger Bacillus thuringiensis n. sp. Z Angew Entomol 1915;2:29-56.
29. Olaya-Abril A, JimÃ©nez-MunguÃa I, GÃ³mez-GascÃ³n L, RodrÃguez-Ortega MJ. Surfomics: shaving live organisms for a fast proteomic identification of surface proteins. J Proteomics 2014;97:164-76.
30. Logan NA, Lebbe L, Verhelst A, Goris J, Forsyth G, RodrÃguez-DÃaz M, et al. Bacillus shackletonii sp. nov., from volcanic soil on Candlemas Island, South Sandwich archipelago. Int J Syst Evol Microbiol 2004;54 Suppl 2:373-6.
31. Palmisano MM, Nakamura LK, Duncan KE, Istock CA, Cohan FM. Bacillus sonorensis sp. nov., a close relative of Bacillus licheniformis, isolated from soil in the sonoran desert, Arizona. Int J Syst Evol Microbiol 2001;51:1671-9.
32. Shivaji S, Chaturvedi P, Suresh K, Reddy GS, Dutt CB, Wainwright M, et al. Bacillus aureus sp. nov., Bacillus aerophilus sp. nov., Bacillus stratosphericus sp. nov. andBacillus altitudinis sp. nov., isolated from cryogenic tubes used for collecting air samples from high altitudes. Int J Syst Evol Microbiol 2006;56:1465-73.
33. JimÃ©nez G, Urdiain M, Cifuentes A, LÃ³pez-LÃ³pez A, Blanch AR, Tamames J, et al. Description of Bacillus toyonensis sp. nov., a novel species of the Bacillus cereus group, and pairwise genome comparisons of the species of the group by means of ANI calculations. Syst Appl Microbiol 2013;36:383-91.
34. Rooney AP, Price NP, Ehrhardt C, Swezey JL, Bannan JD. Phylogeny and molecular taxonomy of the Bacillus subtilis species complex and description of Bacillus subtilis subsp. inaquosorum subsp. nov. Int J Syst Evol Microbiol 2009;59:2429-36.
35. Gatson JW, Benz BF, Chandrasekaran C, Satomi M, Venkateswaran K, Hart ME. Bacillus tequilensis sp. nov., isolated from a 2000-year-old Mexican shaft-tomb, is closely related to Bacillus subtilis. Int J Syst Evol Microbiol 2006;56:1475-84.
36. Nakamura LK, Roberts MS, Cohan FM. Relationship of Bacillus subtilis clades associated with strains 168 and W 23:a proposal for Bacillus subtilis subsp. subtilis subsp. nov. and Bacillus subtilis subsp. spizizenii subsp. nov. Int J Syst Bacteriol 1999;49:1211-5.
37. Ceylan O, Okmen G, Ugur A. Isolation of soil streptomyces as source antibiotics active against antibiotic resistant bacteria. Eur-Asia J Biol Sci 2008;2:73-82.
38. Edgar RC. MUSCLE: multiple sequence alignment with high accuracy and high throughput. Nucleic Acids Res 2004;32:1792-7.
39. Talavera G, Castresana J. Improvement of phylogenies after removing divergent and ambiguously aligned blocks from protein sequence alignments. Syst Biol 2007;56:564-77.
40. Dereeper A, Guignon V, Blanc G, Audic S, Buffet S, Chevenet F, et al. Phylogeny. fr: robust phylogenetic analysis for the non-specialist. Nucleic Acids Res 2008;36:465-9.
41. Eichorst SA, Breznak JA, Schmidt TM. Isolation and characterization of soil bacteria that define Terriglobus gen. nov., in the phylum Acidobacteria. Appl Environ Microbiol 2007;73:2708-17.
42. Kirk RE, Othmer DF. Encyclopedia of chemical technology. 2nd ed. New York: The Interscience Encyclopedia, Inc; 1953.
43. Kohama Y, T Teramoto, Y Kayamori, M Itoh, T Mimura, An Inada, et al. Nakanishi isolation and identification of hydroquinone in bakerâ€™s yeast. Agric Biol Chem 1990;54 Suppl 11:3051-2.
44. Jeyanthi V, Anbu P, Vairamani M, Velusamy P. Isolation of hydroquinone (benzene-1, 4-diol) metabolite from halotolerant Bacillus methylotrophicus MHC10 and its inhibitory activity towards bacterial pathogens. Bioprocess Biosyst Eng 2016;39 Suppl 3:429-39.
45. Bulama JS, Dangoggo SM, Halilu ME, Tsafe AI, Hassan SW. Isolation and characterization of palmitic acid from ethyl acetate extract of root bark of Terminalia glaucescens. J Chem Mater Res 2014;6 Suppl 12:140-4.