Aislamiento y caracterización de bacterias endémicas colombianas con capacidad de degradar tolueno
Isolation and characterization of Colombian endemic bacteria capable of degrading toluene
DOI:
https://doi.org/10.15446/rev.colomb.biote.v24n1.98613Palabras clave:
biodegradación, biorremediación BTEX, compuesto xenobiótico, crecimiento, Pseudomonas (es)español, biodegradation, bioremediation, growth, Pseudomonas, xenobiotic compounds (en)
Los hidrocarburos aromáticos monocíclicos: benceno, tolueno, etilbenceno y xileno (BTEX), presentes en crudo y refinados de petróleo, hacen parte de los compuestos con más impacto en el medio ambiente y la salud humana, debido a su naturaleza cancerígena, mutagénica y altamente tóxica. Esta investigación tuvo como objetivo obtener y caracterizar bacterias capaces de degradar tolueno. Se realizaron tres muestreos de suelo contaminado con hidrocarburos del Valle del Cauca en tres condiciones: gasolinería, derrame accidental y taller mecánico. Se aislaron bacterias capaces de crecer en tolueno vapor como única fuente de carbono y se caracterizaron a nivel morfológico, bioquímico y molecular. Para la caracterización molecular se amplificó, secuenció y analizó con herramientas bioinformáticas el gen ribosomal 16S. Se evaluó la utilización de tolueno directo con concentración al 1% como única fuente de carbono. Se logró aislar 29 bacterias con capacidad de metabolizar tolueno. La caracterización bioquímica y molecular identificó a las bacterias aisladas de suelo contaminado como Pseudomonas y Stenotrophomonas. Las bacterias aisladas en el taller mecánico resultaron ser los microorganismos con mejor crecimiento en tolueno como fuente de carbono, poseen un gran potencial para ser utilizadas para fines de biorremediación de suelos y aguas contaminadas con BTEX.
Monocyclic aromatic hydrocarbons: benzene, toluene, ethylbenzene, and xylene (BTEX), present in crude oil and refined petroleum products, they are the compounds with a big impact on the environment and human health, due to their carcinogenic, mutagenic, and highly toxic. This research aimed to obtain and characterize bacteria capable of degrading toluene. Three samples of soil contaminated with hydrocarbons were carried out in three different conditions: oil station, accidental oil spill, and mechanical workshop. Bacteria capable of growing in toluene vapor as the unique carbon source were isolated and characterized at a morphological, biochemical, and molecular level. For molecular characterization, the 16S ribosomal gene was amplified, sequenced, and analyzed with bioinformatic tools. The use of direct toluene with 1% concentration as the sole carbon source was evaluated. It was possible to isolate 29 bacteria with the capacity to metabolize toluene. Biochemical and molecular characterization identified bacteria isolated from contaminated soil as Pseudomonas and Stenotrophomonas. The isolated strains in the mechanical workshop were the microorganisms with the best growth in toluene as a carbon source, they have great potential to be used for the bioremediation of soils and waters with toluene pollution.
Referencias
Ahmed, N., Sik Ok, Y., Jeon, B.-H., & Ra, J. (2019). Assessment of benzene, toluene, ethyl-benzene, and xylene (BTEX) toxicity in soil using sulfur-oxidizing bacterial (SOB) bioassay. Chemosphere, 220, 651-657. doi:doi.org/10.1016/j.chemosphere.2018.12.102
Anzai, Y., Kim, H., Park, J. Y., Wakabayashi, H., & Oyaizu, H. (2000). Phylogenetic affiliation of the pseudomonads based on 16S rRNA sequence. International Journal of Systematic and Evolutionary Microbiology, 1563–1589. DOI: https://doi.org/10.1099/00207713-50-4-1563
Ashengroph, M., Nahvi, I., Zarkesh-Esfahani, H., & Momenbeik, F. (2011). Pseudomonas resinovorans SPR1, a newly isolated strain with potential of transforming eugenol to vanillin and vanillic acid. New Biotechnology, 28(6), 656–664. doi:10.1016/j.nbt.2011.06.009
Baghania, A., Sorooshianbc, A., Heydaria, M., Sheikhia, R., Golbaza, S., Ashournejadd, Q., . . . Shahsavani, A. (2017). A case study of BTEX characteristics and health effects by major point sources of pollution during winter in Iran. Environmental Pollution, 247, 607-617. doi:10.1016/j.envpol.2019.01.070
Bashandy, S. R., Abd-Alla, M. H., & Dawood, M. F. (2020). Alleviation of the toxicity of oily wastewater to canola plants by the N2-fixing, aromatic hydrocarbon biodegrading bacterium Stenotrophomonas maltophilia-SR1. Applied Soil Ecology, 154, 103654. doi:10.1016/j.apsoil.2020.103654
Benka-Coker, M. O., & & Ekundayo, J. A. (1997). Applicability of evaluating the ability of microbes isolated from an oil spill site to degrade oil. Environmental Monitoring and Assessment, 45(3), 259–272. doi:10.1023/a:1005774602112
Chaerun, S. K., Tazaki, K., Asada, R., & Kogure, K. (2004). Bioremediation of coastal areas 5 years after the Nakhodka oil spill in the Sea of Japan: isolation and characterization of hydrocarbon-degrading bacteria. Environment International, 30(7), 911–922. doi:10.1016/j.envint.2004.02.007
Claro, E., Cruz, J., Montagnolli, R., Lopes, P., Júnior, J., & Bidoia, E. (2018). Microbial Degradation of Petroleum Hydrocarbons: Technology and Mechanism. En V. Kumar, M. Kumar, & R. Prasad, Microbial Action on Hydrocarbons (págs. 125-141). Singapore: Springer. DOI: https://doi.org/10.1007/978-981-13-1840-5_6
Delgadillo-Ordoñez, N. C., Posada-Suárez, L., Marcelo, E., Cepeda-Hernández, M. L., & Sánchez-Nieves, J. (2017). Aislamiento e identificación de levaduras degradadoras de hidrocarburos aromáticos, presentes en tanques de gasolina de vehículos urbanos. Revista Colombiana de Biotecnología, 19(1), 141 - 151. DOI: https://doi.org/10.15446/rev.colomb.biote.v19n2.70278
Dellagnezze, B. M., Gomes, M. B., & Maia de Oliveira, V. (2018). Microbes and Petroleum Bioremediation. En V. Kumar, M. Kumar, & R. Prasad, Microbial Action on Hydrocarbons (págs. 97-123). Gateway East, Singapore: Springer. DOI: https://doi.org/10.1007/978-981-13-1840-5_5
Di Martino, C., López, N., & Raiger Iustman, L. (2012). Isolation and characterization of benzene, toluene and xylene degrading Pseudomonas sp. selected as candidates for bioremediation. International Biodeterioration & Biodegradation, 67, 15-20. doi:10.1016/j.ibiod.2011.11.004
El-Naas, M., Acio, J., & El Telib, A. (2014). Aerobic biodegradation of BTEX: Progresses and Prospects. Journal of Environmental Chemical Engineering, 2(2), 1104-1122. doi:10.1016/j.jece.2014.04.009
Faizal, I., Dozen, K., Hong , C., Kuroda, A., Takiguchi, N., Ohtake, H., . . . Kato, J. (2005). Isolation and characterization of solvent-tolerant Pseudomonas putida strain T-57, and its application to biotransformation of toluene to cresol in a two-phase (organic-aqueous) system. Environmental Biotechnology, 32, 542–547. DOI: https://doi.org/10.1007/s10295-005-0253-y
Garrity, G., Bell, J., & Lilburn, T. (2005). Class II. Betaproteobacteria class. nov.. En D. Brenner, N. Krieg, & J. Staley, Bergey’s Manual® of Systematic Bacteriology. Boston: Springer. doi:10.1007/978-0-387-29298-4_2
Hassan, H. A., & Aly, A. (2018). Isolation and characterization of three novel catechol 2,3-dioxygenase from three novel haloalkaliphilic BTEX-degrading Pseudomonas strains. International Journal of Biological Macromolecules, 108, 1107-1114. DOI: https://doi.org/10.1016/j.ijbiomac.2017.08.113
Hazrati, S., Rostami, R., Fazlzadeh , M., & Pourfarzi, F. (2016). Benzene, Toluene, Ethylbenzene and Xylene Concentrations in Atmospheric Ambient Air of Gasoline and CNG Refueling Stations. Air Quality. Atmosphere and Health, 9, 403-409. DOI: https://doi.org/10.1007/s11869-015-0349-0
Hendrickx, B., Junca, H., Vosahlova, J., Lindner, A., Rüegg, I., Bucheli-Witschel, M., & pringael, D. (2006). Alternative primer sets for PCR detection of genotypes involved in bacterial aerobic BTEX degradation: Distribution of the genes in BTEX degrading isolates and in subsurface soils of a BTEX contaminated industrial site. Journal of Microbiological Methods, 64(2), 250–265. DOI: https://doi.org/10.1016/j.mimet.2005.04.018
Hong, J. H., Kim, J., Choi, O. K., Cho, K.-S., & Ryu, H. W. (2005). Characterization of a diesel-degrading bacterium, Pseudomonas aeruginosa IU5, isolated from oil-contaminated soil in Korea. World Journal of Microbiology and Biotechnology, 21(3), 381–384. doi:10.1007/s11274-004-3630-1
Khodaei, K., Nassery, H. R., Asadi, M. M., Mohammadzadeh, H., & Mahmoodlu, M. (2017). BTEX biodegradation in contaminated groundwater using a novel strain (Pseudomonas sp. BTEX-30). International Biodeterioration & Biodegradation, 116, 234-242. doi:10.1016/j.ibiod.2016.11.001
Köhler, K., Rückert, C., Schatschneider, S., Vorhölter, F., Szczepanowski, R., Blank, L., . . . Schmid, A. (2013). Complete genome sequence of Pseudomonas sp. strain VLB120 a solvent tolerant, styrene degrading bacterium, isolated from forest soil. Journal of Biotechnology, 168(4), 729-730. doi:10.1016/j.jbiotec.2013.10.016
Kuhad, R., & Gupta, R. (2009). Biological Remediation of Petroleum Contaminants. En R. Kuhad, & R. Gupta, Advances in Applied Bioremediation, Soil Biology (Vol. 17, págs. 173-187). Berlin, Heidelberg: Springer. doi:10.1007/978-3-540-89621-0_9
Kumar, S., Stecher, G., Tamura, K., & Dudley, J. (2016). MEGA7: Molecular Evolutionary Genetics Analysis Version 7.0 for Bigger Datasets Downloaded from. Mol. Biol. Evol, 33(7), 1870–1874. https://doi.org/10.1093/molbev/msw054
Lozano-A, L., Bautista, M. A., Dussan-G, J., & Vives-Flórez, M. J. (2008). Dna extraction from heavy oil contaminated microcosms and rpob gene pcr amplification. Actualidades Biológicas, 30(88), 7-14.
Martinez, J., Jimenez, M., Soto, D., Lomas, M., & Sanchez, S. (2015). Aislamiento y caracterización de bacterias hidrocarburolíticas degradadoras de gasolina y diésel. Revista de Ciencias Naturales y Agropecuarias, 2(2), 155-161.
Mukherjee, P., & Roy, P. (2013). Persistent Organic Pollutants Induced Protein Expression and Immunocrossreactivity by Stenotrophomonas maltophilia PM102: A Prospective Bioremediating Candidate. BioMed Research International, 714232. doi:10.1155/2013/714232
Na, K., Moon, K., & Kim, Y. (2005). Source contribution to aromatic VOC concentration and ozone formation potential in the atmosphere of Seoul. Atmospheric Environment, 39, 5517–5524. doi:10.1016/j.atmosenv.2005.06.005
Nagarajan, K., & Loh, K.-C. (2015). Formulation of microbial cocktails for BTEX biodegradation. Biodegradation, 26(1), 51–63. doi:10.1007/s10532-014-9715-0
Nakagawa, Y., Muramatsu, Y., Miyashita, M., Sugimoto, M., & Yoshino, M. (2013). https://www.ncbi.nlm.nih.gov/nuccore/NR_114226. Obtenido de https://www.ncbi.nlm.nih.gov/nuccore/NR_114226
Palleroni, N. (2015). Stenotrophomonas. En Palleroni, & Bradbury, Bergey’s Manual of Systematics of Archaea and Bacteria (págs. 1–20). doi:10.1002 / 9781118960608.gbm01237 DOI: https://doi.org/10.1002/9781118960608.gbm01237
Park, J.‐B., Bühler, B., Panke, S., Witholt, B., & Schmid, A. (2007). Carbon metabolism and product inhibition determine the epoxidation efficiency of solvent‐tolerant Pseudomonas sp. strain VLB120ΔC. Biotechnology and Bioengineering, 98, 1219-1229. doi:10.1002/bit.21496
Patel, K., & Patel, M. (2020). Improving bioremediation process of petroleum wastewater using biosurfactants producing Stenotrophomonas sp. S1VKR-26 and assessment of phytotoxicity. Bioresource Technology, 123861. doi:10.1016/j.biortech.2020.123861
Ramos, J., Duque, E., Gallegos, M., Segura, A., & Marqués, S. (2004). Chapter 12 Regulation of toluene catabolic pathways and toluene efflux pump expression in bacteria of the genus Pseudomonas. En R. Vazquez-Duhalt, & R. Quintero-Ramirez, Studies in Surface Science and Catalysis (Vol. 151, págs. 341-372). Amsterdam, Netherlands: Elsevier. DOI: https://doi.org/10.1016/S0167-2991(04)80153-8
Tanaka, N., Miyazaki, S., & Sugawara, H. (2011). Stenotrophomonas maltophilia strain IAM 12423 16S ribosomal RNA, partial sequence. Obtenido de https://www.ncbi.nlm.nih.gov/nuccore/NR_041577
Tan, H.-M., Tang, H.-Y., & Joannou, C. (1993). The Pseudomonas putida ML2 plasmid-encoded genes for benzene dioxygenase are unusual in codon usage and low in G + C content. Gene, 130(1), 33-39. doi:10.1016/0378-1119(93)90343-2
Ugochukwu, U., & Fialips, C. (2017). Crude oil polycyclic aromatic hydrocarbons removal via clay-microbeoil. Chemosphere, 178, 65-72. doi:10.1016/j.chemosphere.2017.03.035
Wang, L., Tai, C., Wu, Y., Chen, Y., Lee, F., & Wang, S. (2010). Pseudomonas taiwanensis sp. nov., isolated from soil. International Journal of Systematic and Evolutionary DOI: https://doi.org/10.1099/ijs.0.014779-0
Cómo citar
APA
ACM
ACS
ABNT
Chicago
Harvard
IEEE
MLA
Turabian
Vancouver
Descargar cita
CrossRef Cited-by
1. José Alberto Orocio-Carrillo, Consuelo Bautista-Muñoz, María del Carmen Rivera-Cruz, Xavier M. Boldo, Said Cadena-Villegas, Antonio Juárez-Maldonado. (2025). Population dynamics of Pseudomonas spp. from the rhizosphere of Leersia hexandra during soil phytoremediation with crude oil in southeastern Mexico . International Journal of Phytoremediation, , p.1. https://doi.org/10.1080/15226514.2025.2584646.
2. Rojas-Flores Segundo, Magaly De La Cruz-Noriega, Nancy Soto-Deza, Nélida Milly Otiniano, Cabanillas-Chirinos Luis, Anibal Alviz-Meza. (2025). Fungifuels: Polyethylene Decomposition and Electricity Generation with Aspergillus ochraceopetaliformis in Microbial Fuel Cell Systems. Fermentation, 11(9), p.527. https://doi.org/10.3390/fermentation11090527.
3. Andrea N. Arias, Rocío Girón-Navarro, Ivonne Linares-Hernández, Verónica Martínez-Miranda, Elia Alejandra Teutli-Sequeira, Justo Lobato, Manuel A. Rodrigo. (2023). Removal of VOCs using electro-Fenton assisted absorption process. Journal of Environmental Chemical Engineering, 11(3), p.110041. https://doi.org/10.1016/j.jece.2023.110041.
4. M. E. Maya-Yescas, M. Gutiérrez-Rojas†, M. García-Rivero. (2024). Enhancing biodegradation of aged hydrocarbon-contaminated soils through toluene addition: assessing effects on solid and slurry phase treatments. Biodegradation, 35(6), p.939. https://doi.org/10.1007/s10532-024-10089-y.
Dimensions
PlumX
Visitas a la página del resumen del artículo
Descargas
Licencia

Esta obra está bajo una licencia internacional Creative Commons Atribución 4.0.
Esta es una revista de acceso abierto distribuida bajo los términos de la Licencia Creative Commons Atribución 4.0 Internacional (CC BY). Se permite el uso, distribución o reproducción en otros medios, siempre que se citen el autor(es) original y la revista, de conformidad con la práctica académica aceptada. El uso, distribución o reproducción está permitido desde que cumpla con estos términos.
Todo artículo sometido a la Revista debe estar acompañado de la carta de originalidad. DESCARGAR AQUI (español) (inglés).





