Identification of biosurfactant-producing fungi isolated from the Costa Rican mangrove
DOI:
https://doi.org/10.15359/ru.40-1.5Keywords:
haemolysis, drop-collapse, Trichoderma sp. , molecular identification, secondary metabolites, emulsification index, ITSAbstract
In Costa Rica, mangrove potential has not been studied from a biotechnological perspective, despite reports from other latitudes of promising fungi associated with these ecosystems, including biosurfactant potential. These molecules are primarily used in industry as emulsifiers; however, studies indicate their potential in healthcare, pharmaceutical, and agricultural applications. [Objective] This study aimed to screen and preliminarily identify fungal strains capable of producing biosurfactants by implementing a set of effective, low-cost, straightforward assays for future exploitation. [Methodology] Five fungal strains from the Manuel Antonio mangrove in Puntarenas, Costa Rica, were isolated, molecularly identified, and assessed for biosurfactant production utilizing the crude biosurfactant broth. The drop collapse test, oil displacement test, emulsification index, and hemolysis tests were assessed. [Results] The use of crude biosurfactant broth minimized the cost and time required for purifications, which enabled the reduction of screening time for all fermentations. Isolate MP4 displayed, overall and consistently, better performance than the other strains evaluated for growth and biosurfactant production assays, especially for the emulsification index and blood hemolysis test. ITS and LSU molecular markers were analyzed, and MP4 was taxonomically assigned as Trichoderma sp.; however, it was shown to be closely related to T. melanomagnum. [Conclusions] Results emphasize the unexplored potential of mangrove resources in Costa Rica for biosurfactant production, and although preliminary, they demonstrate the value of conducting future comprehensive studies to evaluate this fungal potential.
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References
Alberti, F., Foster, G. D., & Bailey, A. M. (2017). Natural products from filamentous fungi and production by heterologous expression. Applied microbiology and biotechnology, 101(2), 493–500. https://doi.org/10.1007/s00253-016-8034-2
Allam, Z., Bibri, S. E., & Sharpe, S. A. (2022). The Rising Impacts of the COVID-19 Pandemic and the Russia–Ukraine War: Energy Transition, Climate Justice, Global Inequality, and Supply Chain Disruption. Multidisciplinary Digital Publishing Institute 11(11), 99. https://doi.org/10.3390/resources11110099
Azmi, O.6 Seppelt, R. (1997). Fungi of the Windmill Islands, continental Antarctica. Effect of temperature, pH and culture media on the growth of selected microfungi. Polar Biol 18(2):128–134. https://doi.org/10.1007/s003000050167
Basu, S., Bose, C., Ojha, N., Das, N., Das, J., Pal, M., & Khurana, S. (2015). Evolution of bacterial and fungal growth media. Bioinformation, 11(4), 182–184. https://doi.org/10.6026/97320630011182
Bhardwaj, G., Cameotra, S. S., & Chopra, H. K. (2013). Utilization of oleo-chemical industry by-products for biosurfactant production. AMB Express, 3(1), 68. https://doi.org/10.1186/2191-0855-3-68
Bodour, A. A., & Miller-Maier, R. M. (1998). Application of a modified drop-collapse technique for surfactant quantitation and screening of biosurfactant-producing microorganisms. Journal of Microbiological Methods, 32(3), 273-280. https://doi.org/10.1016/S0167-7012(98)00031-1
Cooper, D. G., & Goldenberg, B. G. (1987). Surface-active agents from two bacillus species. Applied and environmental microbiology, 53(2), 224–229. https://doi.org/10.1128/aem.53.2.224-229.1987
Cortés, J., & Wehrtmann, I. S. (2009). Diversity of marine habitats of the Caribbean and Pacific of Costa Rica. In Marine Biodiversity of Costa Rica, Central America (pp. 1-45). Dordrecht: Springer Netherlands. https://doi.org/10.1007/978-1-4020-8278-8_1
da Silva, A. F., Banat, I. M., Giachini, A. J., & Robl, D. (2021). Fungal biosurfactants, from nature to biotechnological product: bioprospection, production and potential applications. Bioprocess and biosystems engineering, 44(10), 2003–2034. https://doi.org/10.1007/s00449-021-02597-5
da Silva, M. E. T., Nascimento, C. C., Junior, S. D., & Albuquerque, P. M. (2014). Biosurfactant production by Myrciaguianensis endophytic fungi. In BMC Proceedings 8(4), 213. https://doi.org/10.1186/1753-6561-8-S4-P213
de Souza Sebastianes, F. L., Romão-Dumaresq, A. S., Lacava, P. T., Harakava, R., Azevedo, J. L., de Melo, I. S., & Pizzirani-Kleiner, A. A. (2013). Species diversity of culturable endophytic fungi from Brazilian mangrove forests. Current genetics, 59(3), 153–166. https://doi.org/10.1007/s00294-013-0396-8
El-Enshasy, H. A. (2007). Filamentous fungal cultures–process characteristics, products, and applications. Bioprocessing for value-added products from renewable resources, 225-261. https://doi.org/10.1016/B978-044452114-9/50010-4
Fernandes, N. d. A. T., Simões, L. A., & Dias, D. R. (2023). Biosurfactants produced by yeasts: Fermentation, screening, recovery, purification, characterization, and applications. Fermentation. Multidisciplinary Digital Publishing Institute 9(3), 207. https://doi.org/10.3390/fermentation9030207
Ferreira, I. N. S., Rodríguez, D. M., Campos-Takaki, G. M., & da Silva Andrade, R. F. (2020). Biosurfactant and bioemulsifier as promising molecules produced by Mucor hiemalis isolated from Caatinga soil. Electronic Journal of Biotechnology, 47, 51-58. https://doi.org/10.1016/j.ejbt.2020.06.006
Gayathiri, E., Prakash, P., Karmegam, N., Varjani, S., Awasthi, M. K., & Ravindran, B. (2022). Biosurfactants: potential and eco-friendly material for sustainable agriculture and environmental safety—a review. Agronomy, 12(3), 662. https://doi.org/10.3390/agronomy12030662
Geiser, E., Wiebach, V., Wierckx, N., & Blank, L. M. (2014). Prospecting the biodiversity of the fungal family Ustilaginaceae for the production of value-added chemicals. Fungal biology and biotechnology, 1, 2. https://doi.org/10.1186/s40694-014-0002-y
Jia, S. L., Chi, Z., Liu, G. L., Hu, Z., & Chi, Z. M. (2020). Fungi in mangrove ecosystems and their potential applications. Critical reviews in biotechnology, 40(6), 852–864. https://doi.org/10.1080/07388551.2020.1789063
Johnson, P., Trybala, A., Starov, V., & Pinfield, V. J. (2021). Effect of synthetic surfactants on the environment and the potential for substitution by biosurfactants. Advances in colloid and interface science, 288, 102340. https://doi.org/10.1016/j.cis.2020.102340
Konishi, M., Hatada, Y., & Horiuchi, J. (2013). Draft Genome sequence of the Basidiomycetous yeast-like fungus Pseudozyma hubeiensis SY62, which produces an abundant amount of the biosurfactant Mannosylerythritol Lipids. Genome announcements, 1(4), e00409-13. https://doi.org/10.1128/genomeA.00409-13
Kosaric, N., & Sukan, F. V. (Eds.). (2014). Biosurfactants: production and utilization—processes, technologies, and economics (Vol. 159). CRC press. https://doi.org/10.1201/b17599
Laurie, J. D., Ali, S., Linning, R., Mannhaupt, G., Wong, P., Güldener, U., Münsterkötter, M., Moore, R., Kahmann, R., Bakkeren, G., & Schirawski, J. (2012). Genome comparison of barley and maize smut fungi reveals targeted loss of RNA silencing components and species-specific presence of transposable elements. The Plant cell, 24(5), 1733–1745. https://doi.org/10.1105/tpc.112.097261
Lorenz, S., Guenther, M., Grumaz, C., Rupp, S., Zibek, S., & Sohn, K. (2014). Genome Sequence of the basidiomycetous fungus Pseudozyma aphidis DSM70725, an efficient producer of biosurfactant Mannosylerythritol Lipids. Genome announcements, 2(1), e00053-14. https://doi.org/10.1128/genomeA.00053-14
Maheswari, N. U., & Parveen, L. F. (2012). Comparative study of biosurfactant by using Bacillus licheniformis and Trichoderma viride from paper waste contaminated soil. International Journal of Chemical Sciences. Sadguru Publications 10(3):1687–1697.
Marcelino, P. R. F., Peres, G. F. D., Terán-Hilares, R., Pagnocca, F. C., Rosa, C. A., Lacerda, T. M., & Da Silva, S. S. (2019). Biosurfactants production by yeasts using sugarcane bagasse hemicellulosic hydrolysate as new sustainable alternative for lignocellulosic biorefineries. Industrial Crops and Products, 129, 212-223. http://hdl.handle.net/11449/185383
Martinho, V., Dos Santos Lima, L. M., Barros, C. A., Ferrari, V. B., Passarini, M. R. Z., Santos, L. A., de Souza Sebastianes, F. L., Lacava, P. T., & de Vasconcellos, S. P. (2019). Enzymatic potential and biosurfactant production by endophytic fungi from mangrove forest in Southeastern Brazil. AMB Express, 9(1), 130. https://doi.org/10.1186/s13568-019-0850-1
McIntyre, M., Breum, J., Arnau, J., & Nielsen, J. (2002). Growth physiology and dimorphism of Mucor circinelloides (syn. racemosus) during submerged batch cultivation. Applied microbiology and biotechnology, 58(4), 495–502. https://doi.org/10.1007/s00253-001-0916-1
Morikawa, M., Hirata, Y., & Imanaka, T. (2000). A study on the structure-function relationship of lipopeptide biosurfactants. Biochimica et biophysica acta, 1488(3), 211–218. https://doi.org/10.1016/s1388-1981(00)00124-4
Morita, T., Konishi, M., Fukuoka, T., Imura, T., & Kitamoto, D. (2006). Discovery of Pseudozyma rugulosa NBRC 10877 as a novel producer of the glycolipid biosurfactants, mannosylerythritol lipids, based on rDNA sequence. Applied microbiology and biotechnology, 73(2), 305–313. https://doi.org/10.1007/s00253-006-0466-7
Mulligan, C. N., Cooper, D. G., & Neufeld, R. J. (1984). Selection of microbes producing biosurfactants in media without hydrocarbons. Journal of Fermentation Technology 62(4), 311-314.
Raddadi, N., Giacomucci, L., Marasco, R., Daffonchio, D., Cherif, A., & Fava, F. (2018). Bacterial polyextremotolerant bioemulsifiers from arid soils improve water retention capacity and humidity uptake in sandy soil. Microbial cell factories, 17(1), 83. https://doi.org/10.1186/s12934-018-0934-7
Rodríguez-Rodríguez, C. E., Zúñiga-Chacón, C., & Barboza-Solano, C. (2012). Evaluation of growth in diesel fuel and surfactants production ability by bacteria isolated from fuels in Costa Rica. Revista de la Sociedad Venezolana de Microbiología, 32(2), 116-120. https://www.redalyc.org/articulo.oa?id=199425417011
Saika, A., Koike, H., Hori, T., Fukuoka, T., Sato, S., Habe, H., Kitamoto, D., & Morita, T. (2014). Draft Genome Sequence of the yeast Pseudozyma antarctica type strain JCM10317, a producer of the glycolipid biosurfactants, Mannosylerythritol Lipids. Genome announcements, 2(5), e00878-14. https://doi.org/10.1128/genomeA.00878-14
Schirawski, J., Mannhaupt, G., Münch, K., Brefort, T., Schipper, K., Doehlemann, G., Di Stasio, M., Rössel, N., Mendoza-Mendoza, A., Pester, D., Müller, O., Winterberg, B., Meyer, E., Ghareeb, H., Wollenberg, T., Münsterkötter, M., Wong, P., Walter, M., Stukenbrock, E., Güldener, U., & Kahmann, R. (2010). Pathogenicity determinants in smut fungi revealed by genome comparison. Science (New York, N.Y.), 330(6010), 1546–1548. https://doi.org/10.1126/science.1195330
Schoch, C. L., Seifert, K. A., Huhndorf, S., Robert, V., Spouge, J. L., Levesque, C. A., Chen, W., Fungal Barcoding Consortium, & Fungal Barcoding Consortium Author List. (2012). Nuclear ribosomal internal transcribed spacer (ITS) region as a universal DNA barcode marker for Fungi. Proceedings of the National Academy of Sciences of the United States of America, 109(16), 6241–6246. https://doi.org/10.1073/pnas.1117018109
Sewalt, V., Shanahan, D., Gregg, L., La Marta, J., & Carrillo, R. (2016). The generally recognized as safe (GRAS) process for industrial microbial enzymes. Industrial Biotechnology, 12(5), 295-302. https://doi.org/10.1089/ind.2016.0011
Sharma G., & Pandey, R. (2010). Influence of culture media on growth, colony character and sporulation of fungi isolated from decaying vegetable wastes. Journal of yeast and fungal research, 1(8), 157–164. http://www.academicjournals.org/JYFR
Silva, A. C. S. D., Santos, P. N. D., Silva, T. A. L. E., Andrade, R. F. S., & Campos-Takaki, G. M. (2018). Biosurfactant production by fungi as a sustainable alternative. Arquivos do Instituto Biológico, 85, e0502017. https://doi.org/10.1590/1808-1657000502017
Singh, K., Nizam, S., Sinha, M., & Verma, P. K. (2012). Comparative transcriptome analysis of the necrotrophic fungus Ascochyta rabiei during oxidative stress: insight for fungal survival in the host plant. PloS one, 7(3), e33128. https://doi.org/10.1371/journal.pone.0033128
Solano-González, S., & Solano-Campos, F. (2022). Production of mannosylerythritol lipids: biosynthesis, multi-omics approaches, and commercial exploitation. Molecular omics, 18(8), 699–715. https://doi.org/10.1039/d2mo00150k
Solano-González, S., Darby, A. C., Cossar, D., & Caddick, M. X. (2019). High-quality draft Genome Sequence and annotation of the basidiomycete yeast Sporisorium graminicola CBS10092, a producer of Mannosylerythritol Lipids. Microbiology resource announcements, 8(42), e00479-19. https://doi.org/10.1128/MRA.00479-19
Stecher, G., Tamura, K., & Kumar, S. (2020). Molecular Evolutionary Genetics Analysis (MEGA) for macOS. Molecular biology and evolution, 37(4), 1237–1239. https://doi.org/10.1093/molbev/msz312
Syedd-León, R., Solano-Campos, F., Campos-Rodríguez, J., Pereira-Arce, D., Villegas-Peñaranda, L. R., & Sandoval-Barrantes, M. (2022). Fungal extracellular lipases from coffee plantation environments for the sustainable management of agro-industrial coffee biomass. Biomass, 2(2), 62-79. https://doi.org/10.3390/biomass2020005
Taniguti, L. M., Schaker, P. D., Benevenuto, J., Peters, L. P., Carvalho, G., Palhares, A., Quecine, M. C., Nunes, F. R., Kmit, M. C., Wai, A., Hausner, G., Aitken, K. S., Berkman, P. J., Fraser, J. A., Moolhuijzen, P. M., Coutinho, L. L., Creste, S., Vieira, M. L., Kitajima, J. P., & Monteiro-Vitorello, C. B. (2015). Complete Genome Sequence of Sporisorium scitamineum and Biotrophic Interaction Transcriptome with Sugarcane. PloS one, 10(6), e0129318. https://doi.org/10.1371/journal.pone.0129318
Thavasi, R., Sharma, S., & Jayalakshmi, S. (2011). Evaluation of screening methods for the isolation of biosurfactant producing marine bacteria. Journal of Petroleum and Environmental Biotechnology, 1(2), 1-7. https://doi.org/10.4172/2157-7463.S1-001
Villagrán, Z., Martínez-Reyes, M., Gómez-Rodríguez, H., Ríos-García, U., Montalvo-González, E., Ortiz-Basurto, R. I., Anaya-Esparza, L. M., & Pérez-Moreno, J. (2023). Huitlacoche (Ustilago maydis), an Iconic Mexican Fungal Resource: Biocultural Importance, Nutritional Content, Bioactive Compounds, and Potential Biotechnological Applications. Molecules (Basel, Switzerland), 28(11), 4415. https://doi.org/10.3390/molecules28114415
Vu, D., Groenewald, M., de Vries, M., Gehrmann, T., Stielow, B., Eberhardt, U., Al-Hatmi, A., Groenewald, J. Z., Cardinali, G., Houbraken, J., Boekhout, T., Crous, P. W., Robert, V., & Verkley, G. J. M. (2019). Large-scale generation and analysis of filamentous fungal DNA barcodes boosts coverage for kingdom fungi and reveals thresholds for fungal species and higher taxon delimitation. Studies in mycology, 92, 135–154. https://doi.org/10.1016/j.simyco.2018.05.001
Wada, K., Koike, H., & Morita, T. (2021). Draft Genome Sequence of a basidiomycetous yeast, Ustilago shanxiensis CBS 10075, which produces Mannosylerythritol Lipids. Microbiology Resource Announcements, 10(48), e0070621. https://doi.org/10.1128/MRA.00706-21
White, T.J. (1990) Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In: PCR Protocols, a guide to methods and applications, 315-322.
Wege, S. M., Gejer, K., Becker, F., Bölker, M., Freitag, J., & Sandrock, B. (2021). Versatile CRISPR/Cas9 systems for genome editing in Ustilago maydis. Journal of Fungi (Basel, Switzerland), 7(2), 149. https://doi.org/10.3390/jof7020149
Wehrtmann, I. S., Cortés, J., & Echeverría-Sáenz, S. (2009). Marine Biodiversity of Costa Rica: Perspectives and Conclusions. In: Wehrtmann, I.S., Cortés, J. (eds) Marine Biodiversity of Costa Rica, Central America. Monographiae Biologicae, (vol 86). Springer, Dordrecht. https://doi.org/10.1007/978-1-4020-8278-8_49
Youssef, N. H., Duncan, K. E., Nagle, D. P., Savage, K. N., Knapp, R. M., & McInerney, M. J. (2004). Comparison of methods to detect biosurfactant production by diverse microorganisms. Journal of Microbiological Methods, 56(3), 339–347. https://doi.org/10.1016/j.mimet.2003.11.001
Zamora-Trejos, P., & Cortés, J. (2009). Los manglares de Costa Rica: el Pacífico norte. Revista de Biología Tropical, 57(3), 473-488. https://doi.org/10.15517/rbt.v57i3.5469
Zhang, C. L., Liu, S. P., Lin, F. C., Kubicek, C. P., & Druzhinina, I. S. (2007). Trichoderma taxi sp. nov., an endophytic fungus from Chinese yew Taxus mairei. FEMS Microbiology Letters, 270(1), 90–96. https://doi.org/10.1111/j.1574-6968.2007.00659.x
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