Article

Antibacterial Activity of Soil Bacteria against Escherichia coli and GC-MS Analysis of their Organic Compounds

Antimicrobial agents encompass a wide range of compounds, including antibiotics, bacteriocins, and lipopeptides, which play a crucial role in combating infectious diseases. Antibiotics, in particular, are secondary metabolites of low molecular weight predominantly synthesized by soil dwelling microorganisms. These microbial metabolites have long served as a vital source of clinically important therapeutic agents. Members of the genus Bacillus and other rhizosphere-associated bacteria are especially known for producing diverse antimicrobial substances. In this study, ten rhizosphere soil samples were collected from different sites in Sana’a city, Yemen. From these samples, 50 antibiotic-producing soil bacteria were isolated. Bioactive metabolites were extracted using the solvent extraction method with chloroform and Ethanol. The crude extracts were analyzed by Gas Chromatography–Mass Spectrometry (GC–MS) to identify their organic composition. Among the 50 bacterial isolates, 14 showed antibacterial activity against resistant Escherichia coli using the agar well diffusion method. Further secondary screening revealed that the filtrates of four isolates Pseudomonas fluorescens, Bacillus subtilis, Acinetobacter baylyi, and Azotobacter vinelandii exhibited the strongest antibacterial effects. GC–MS analysis showed that each isolate produced more than eighty organic compounds; however, only a subset demonstrated antibacterial activity. The most notable bioactive compounds detected included Phenol, 4-(2-aminoethyl)- (CAS: tyramine), 2,4-di-tert-butylphenol, and n-hexadecanoic acid. This study highlights the potential of rhizosphere soil bacteria as promising sources of novel bioactive compounds. The identification of active metabolites and their antibacterial properties against resistant E. coli underscores their possible application in the development of alternative therapeutic strategiesto address antibiotic resistance.

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Miadah M. Mojahed
Department of Biology, Faculty of Sciences, Sana’a University, Sana’a, Yemen
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Abdulrahman A. Humaid
Department of Biology, Faculty of Sciences, Sana’a University, Sana’a, Yemen
8067
Aasfar A, Bargaz A, Yaakoubi K, Hilali A, Bennis I. Nitrogen‑fixing Azotobacter species as potential soil biological enhancers for crop nutrition and yield stability. Front Microbiol. 2021;12:628379. doi:10.3389/fmicb.2021.628379.
8068
Ahmad F, Ahmad I, Khan M. Screening of free‑living rhizospheric bacteria for their multiple plant growth promoting activities. Microbiol Res. 2008;163(2):173–181. doi:10.1016/j.micres.2006.04.001.
8069
Ahmed SS, Benhadj D, Lamari F. GC–MS analysis of bioactive metabolites from halotolerant bacteria isolated from the Sebkha of Oran (Algeria) and their antimicrobial properties. Microorganisms. 2021;9(10):2132. doi:10.3390/microorganisms9102132.
8070
Ajilogba CF, Babalola OO. GC–MS analysis of volatile organic compounds from Bambara groundnut rhizobacteria and their antibacterial properties. World J Microbiol Biotechnol. 2019;35:83. doi:10.1007/s11274-019-2660-7
8071
Al-Abdulsalam H, Almalki M, Khalifa A. Isolation and characterization of antimicrobial metabolites producing bacteria from soils in Al‑Ahsa, Saudi Arabia. J Microbiol Biotechnol Food Sci. 2025;e11779. doi:10.55251/jmbfs.11779.
8072
Anantha Padmanabhan S, Deventhiran M, Saravanan P, Anand D, Rajarajan S. A comparative GC‑MS analysis of bacterial secondary metabolites of Pseudomonas species. Pharma Innov J. 2016;5(4):84–89.
8073
Bahram M, Hildebrand F, Forslund SK, Anderson JL, Soudzilovskaia NA, Bodegom PM, et al. Structure and function of the global topsoil microbiome. Nature. 2018;560(7717):233–237. doi:10.1038/s41586-018-0386-6.
8074
Berdy J. Bioactive microbial metabolites. J Antibiot (Tokyo). 2005;58(1):1–26. doi:10.1038/ja.2005.1.
8075
Cappuccino JG, Sherman N. Microbiology: A Laboratory Manual. 10th ed. Pearson Education; 2014.
8076
Cappuccino JG, Welsh C. Microbiology: A Laboratory Manual. 11th ed. Pearson; 2017.
8077
Caulier S, Nannan C, Gillis A, Licciardi F, Bragard C, Mahillon J. Overview of the antimicrobial compounds produced by members of the Bacillus subtilis group. Front Microbiol. 2019;10:302. doi:10.3389/fmicb.2019.00302
8078
Centers for Disease Control and Prevention (CDC). Antibiotic Resistance Threats in the United States [Internet]. 2023 [cited 2025 Jun 7]. Available from: https://www.cdc.gov/antimicrobial-resistance/
8079
Chen W, Gong L, Guo Z, Wang W. Antibacterial properties of plant essential oils and their components against foodborne pathogens. J Agric Food Chem. 2017;65(4):720–728. doi:10.1021/acs.jafc.6b04250
8080
Desbois AP, Smith VJ. Antibacterial free fatty acids: activities, mechanisms of action and biotechnological potential. Prog Lipid Res. 2010;49(1):1–13. doi:10.1016/j.plipres.2009.08.003
8081
Friedman M. Antibacterial, antiviral, and antifungal properties of caffeine and related methylxanthines. Food Chem. 2017;221:453–464. doi:10.1016/j.foodchem.2016.10.109.
8082
González H, et al. Green extraction of secondary metabolites from plants: obstacles, current status, and trends. Sustain Chem Environ. 2024;8:100157. doi:10.1016/j.scenv.2024.100157.
8083
Javed H, Riaz A, Qureshi A, Javed K, Mujeeb F, Ijaz F, et al. Isolation,characterization and screening of PGPR capable of providing relief in salinity stress. Eurasian J Soil Sci. 2020;9(2):85–91. doi:10.18393/ejss.650546.
8084
Kaaria P, Matiru V, Ndungu M. Antimicrobial activities of secondary metabolites produced by endophytic bacteria from selected indigenous Kenyan plants. Afr J Microbiol Res. 2012;6(45):7253–7258.
8085
Kai M. Diversity and distribution of volatile secondary metabolites throughout Bacillus subtilis isolates. Front Microbiol. 2020;11:559. doi:10.3389/fmicb.2020.00559.
8086
Khan MA, Yasien S, Iqbal MM, Javed M. Comparative evaluation of various extraction techniques for secondary metabolites from Bombax ceiba L. flowering plants along with in vitro anti‑diabetic performance. Bioengineering. 2022;9(10):486. doi:10.3390/bioengineering9100486.
8087
Numan M, Shah M, Asaf S, Rehman NU, Al-Harrasi A. Bioactive compounds from endophytic bacteria Bacillus subtilis strain EP1 with their antibacterial activities. Metabolites. 2022;12(12):1228. doi:10.3390/metabo12121228.
8088
Kovács A, Nagy R. Assessment of antibacterial potential using the agar well diffusion method. Eur J Microbiol Immunol. 2024;14(2):123–130. doi:10.1556/1886.2024.00015.
8089
Król E, Gąsiorowski K, Mucha P. Antibacterial and antifungal activity of cyclic siloxanes. Microbiol Biotechnol. 2017;101(11):4419–4433. doi:10.1007/s00253-017-8232-5.
8090
Kumar S, Sharma A, Verma A. Identification of bioactive secondary metabolites from Bacillus species by GC–MS profiling. J Appl Microbiol. 2022;134(2):453–468. doi:10.1111/jam.14999.
8091
Kurrey NK, Singh S, Yadav AN. Azotobacter: a potential bio‑fertilizer for soil and plant health management. Saudi J Biol Sci. 2020;27(12):3634–3640. doi:10.1016/j.sjbs.2020.09.054.
8092
Li Y, Xiong W, Wang Y, Sun Y, Zeng Q. Metabolic capabilities of Acinetobacter baylyi in the degradation of pollutants: implications for bioremediation. Front Microbiol. 2020;11:615. doi:10.3389/fmicb.2020.00615.
8093
Mahfouz N, Ferreira I, Beisken S, von Haeseler A, Heringa J. Antimicrobial resistance surveillance using machine learning methods. Nat Commun. 2024;15:1512. doi:10.1038/s41467-024-42021-7.
8094
Manga BS, Oyeleke SB. Essentials of M. Higton, Industrial Microbiology: a laboratory practical’s in microbiology. 1st ed. London: Blackwell Publisher/Tobes; 2008. p.56–76.
8095
Musliu A, Salawudeen W. Screening and isolation of the soil bacteria for ability to produce antibiotics. Eur J Appl Sci. 2012;4(5):211–215. doi:10.5829/idosi.ejas.2012.4.5.2011.
8096
Nakatsuji T. Antimicrobial property of lauric acid against Propionibacterium acnes: its therapeutic potential for inflammatory acne vulgaris. J Investig Dermatol. 2009;129(10):2480–2488. doi:10.1038/jid.2009.93.
8097
Qin S, Xiao W, Zhou C, Pu Q, Deng X, Lan L, et al. Pseudomonas aeruginosa: pathogenesis, virulence factors, antibiotic resistance, interaction with host, technology advances and emerging therapeutics. Signal Transduct Target Ther. 2022;7(1):199. doi:10.1038/s41392-022-01084-9.
8098
Qu Q, Zhang ZY, Peijnenburg WJGM, Liu WY, Lu T, Hu BL, et al. Rhizosphere microbiome assembly and its impact on plant growth. J Agric Food Chem. 2020;68(18):5024–5038. doi:10.1021/acs.jafc.0c00073.
8099
Santala S, Santala V. Acinetobacter baylyi ADP1—naturally competent for synthetic biology. Essays Biochem. 2021;65(2):309–318. doi:10.1042/EBC20200168.
8100
Shaikh S, Fatima J, Shakil S, Rizvi SMD, Kamal MA. Antibiotic resistance and extended-spectrum beta-lactamases: Types, epidemiology and treatment. Saudi J Biol Sci. 2023;30(1):103448. doi:10.1016/j.sjbs.2022.103448.
8101
Sorg O, Antille C, Kaya G, Saurat JH. Retinoids in cosmeceuticals. J Steroid Biochem Mol Biol. 2006;101(4–5):136–141. doi:10.1016/j.jsbmb.2006.09.033.
8102
Sumbul A, Tiwari A, Khare E. Volatile organic compounds from Pseudomonas fluorescens: Mechanisms of plant growth promotion and pathogen suppression. World J Microbiol Biotechnol. 2023;39:49. doi:10.1007/s11274-023-03873-0.
8103
Tiwari K, Gupta RK. Rare actinomycetes: A potential storehouse for novel antibiotics. Crit Rev Biotechnol. 2012;32(2):108–132. doi:10.3109/07388551.2011.562482.
8104
Todar K, Ubukata M, Hamada M. Microbiology: A human perspective. London: McGraw-Hill; 2005.
8105
Ventola CL. The antibiotic resistance crisis: part 1: causes and threats. Pharm Ther. 2023;48(2):71–77. Available from: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9876086/
8106
World Health Organization (WHO). Global Antimicrobial Resistance and Use Surveillance System (GLASS) Report 2024. Available from: https://www.who.int/publications/i/item/9789240070650
8107
Xun W, Yan R, Ren Y, Jin D, Xiong W, Zhang G, et al. Grazing-induced microbiome alterations drive soil organic carbon turnover and productivity in meadow steppe. Microbiome. 2018;6:170. doi:10.1186/s40168-018-0554-6
8108
Zhang Y, Wang Q, Yao H, Wang Y. Pyoverdine production in Pseudomonas fluorescens is influenced by carbon source availability. J Basic Microbiol. 2020;60(6):570–8. Available from: https://pubmed.ncbi.nlm.nih.gov/32500426
8109
Zhao J, Wang Y, Li L. GC–MS-based volatile organic compounds profile of Bacillus species and their pharmacological significance. Molecules. 2020;25(22):7556. doi:10.3390/molecules25227556
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Antibacterial Activity of Soil Bacteria against Escherichia coli and GC-MS Analysis of their Organic Compounds. (2026). Sana’a University Journal of Applied Sciences and Technology, 4(1), 1545-1551. https://doi.org/10.59628/jast.v4i1.1931

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