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Caracterización De Los Ácidos Grasos Y Actividad Antimicrobiana Del Extracto En Metanol De Holothuria Princeps (Holothuriida: Holothuriidae)

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Caracterización De Los Ácidos Grasos Y Actividad Antimicrobiana Del Extracto En Metanol De Holothuria Princeps (Holothuriida: Holothuriidae) ISSN Impreso: 0034-7744 ISSN electrónico: 2215-2075 DOI 10.15517/rbt.v69i1.41518 Caracterización de los ácidos grasos y actividad antimicrobiana del extracto en metanol de Holothuria princeps (Holothuriida: Holothuriidae) Yeinner Quiroz Lobo*1, Gilmar Santafé Patiño1 & Jorge A. Quirós-Rodríguez1 1. Grupo de Química de los Productos Naturales (PRONAT), Universidad de Córdoba, Cra. 6 Nº 74-103 Montería, Colombia; [email protected], [email protected], [email protected] * Correspondencia Recibido 22-IV-2020. Corregido 19-VIII-2020. Aceptado 01-X-2020. ABSTRACT. Characterization of fatty acids and antimicrobial activity of the methanol extract of Holothuria princeps (Holothuriida: Holothuriidae). Introduction: In recent decades, studies related to the search and characterization of bioactive molecules in marine organisms have increased exponentially, demon- strating the enormous wealth of secondary metabolites of diverse structural composition that cannot be found in organisms present in the terrestrial environment. A significant number of the new marine natural compounds discovered have contributed to solving some of the problems of humanity, mainly those related to human health. Objective: The purpose of this research is to evaluate the bactericidal and fungicidal activities of the methano- lic extract of sea cucumber Holothuria princeps collect from the bay of Cispatá in the Colombian Caribbean, in addition to chemically identifying its fatty acids. Methods: A methanolic extraction was performed from the collected biological material, by the cold maceration method. The extract obtained was fractionated using chromatographic techniques and the fatty acids were obtained, which were derivatized and identified by means of gas chromatography in coupling with mass spectrometry. The antibacterial and antifungal activities of the methanolic extract of Holothuria princeps was performed through the microdilution method against reference strains and clinical isolates. Results: We found 16 fatty acids present in Holothuria princeps according to the analysis of their mass spectra. Antibacterial activity showed that Enterococcus faecalis was the most susceptible to the extract at low concentrations, while Pseudomonas aeruginosa was the highest at the higher concentra- tions. In antifungal treatment, the fungus with the highest inhibition was the clinical isolate of Candida albicans (blood sample). Conclusions: Taking into account previous studies in the genus Holothuria, it is considered that the environment plays a fundamental role in the presence and diversity of fatty acids. The evaluation of the antibacterial activity against reference strains of Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, Klebsiella pneumoniae and Enterococcus faecalis demonstrated the existence of a considerable effect in the reduction of bacterial growth by the extracts applied, mainly at low concentrations (less than 1 000 ppm). On the other hand, the antifungal activity against the reference strain of Candida albicans and the clinical isolates of Candida albicans (blood sample) and Candida krusei (catheter sample), the extract showed that the best results were presented at higher concentrations (above 1 500 ppm). Key words: antibacterial; antifungic; Echinodermata; lipids; chromatography; mass Spectrometry. Quiroz Lobo, Y., Santafé Patiño, G., & Quirós-Rodríguez, J.A. (2021). Caracterización de los ácidos grasos y actividad antimicrobiana del extracto en metanol de Holothuria princeps (Holothuriida: Holothuriidae). Revista de Biología Tropical, 69(1), 36-44. DOI 10.15517/rbt.v69i1.41518 Durante millones de años, la vida mari- & Regueira, 2015). Desde esta perspectiva y na ha producido una inmensa variedad de teniendo en cuenta que los mares cubren la compuestos químicos de exclusivas estruc- mayor parte del planeta, la bioprospección turas y sofisticación (Fernández, Balmaseda, marina en comparación con la terrestre, puede 36 Rev. Biol. Trop. (Int. J. Trop. Biol.) • Vol. 69(1): 36-44, March 2021 considerarse como la forma más efectiva y pro- nutracéuticos y medicinales debido a su amplio metedora en cuanto al posible descubrimiento espectro de bioactividades, como son las acti- de nuevos compuestos bioactivos (Rizzo & vidades antimicrobiana, antiviral, antitumoral, Lo Giudice, 2018). Los invertebrados marinos antioxidante, entre otras (Chi, Cheung, Ng, & registran una gran diversidad de metabolitos Wong, 2015). Por su parte, los ácidos grasos se secundarios, alrededor de 10 000 nuevos com- encuentran entre los principales componentes puestos químicos son nombrados, estudiados y de las membranas celulares y son fundamenta- descritos anualmente (Valdés, Pérez, & Colom, les en el mantenimiento de señales nerviosas a 2010; Birks, 2015; Carroll, Copp, Davis, Key- lo largo del sistema neuronal (Giménez, 1998). zers, & Prinsep, 2019). En las últimas déca- Estos compuestos están involucrados en varias das se evidenció un continuo desarrollo de vías bioquímicas y son una fuente importante fármacos derivados de sustancias aisladas de de energía, son componentes estructurales de organismos marinos (Sima & Vetvicka, 2011; las membranas celulares e influyen en la per- Piñón, García, Mendiola, Pérez, & Monzote, meabilidad y tráfico de compuestos celulares, 2015). Un gran número de invertebrados viven la actividad de las proteínas y las rutas de seña- en íntima asociación con otros animales o plan- lización bioquímica (Goncalves et al., 2016). tas o microorganismos, factor que contribuye al Antes del descubrimiento y la utilización incremento de una gran variedad de metaboli- de los antimicrobianos por parte de la sociedad, tos secundarios biológicamente activos (Rama- las enfermedades infecciosas eran la principal sami et al., 2016). causa de muerte del ser humano, y continúan Como parte de los invertebrados, los holo- siéndolo en países en vías de desarrollo (Aties, turoideos, son organismos que habitan todos Duret, Tabío, & Pérez, 2017). Los microorga- los ambientes marinos, algunas especies pue- nismos han desarrollado nuevas estrategias que den desplegar capacidades natatorias (perma- les proveen resistencia a la acción de fárma- nentes o facultativas) en el ambiente pelágico, cos, por lo que existe una prioridad de salud mientras que otras ocupan distintos sustratos pública para explorar y desarrollar agentes en hábitats bentónicos (Martins, 2012). Estos antimicrobianos naturales más efectivos, con organismos, mezclan y consumen el material menores efectos secundarios, buena biodispo- orgánico de los sedimentos, permitiendo así, la nibilidad y de toxicidad mínima (Pérez, Falqué, entrada del oxígeno, contribuyendo a la estruc- & Dominguez, 2016). Por lo anterior, muchos turación del hábitat y el reciclaje de la mate- organismos marinos constituyen una potencial ria orgánica (Schneider et al., 2011; Purcell, fuente de antimicrobianos capaces de combatir Conand, Uthicke, & Byrne, 2016). A lo largo infecciones (Cusimano et al., 2019), en ese de la historia, la pesca de holoturoideos ha sido sentido, los extractos de especies de pepinos extensiva, debido principalmente a su amplio de mar, han demostrado una importante bioac- uso en la medicina tradicional asiática dadas las tividad (Sellem, Brahmi, Mnasser, Rafrafi, & numerosas aplicaciones contra la hipertensión, Bouhaouala-Zahar, 2017). el asma, el reumatismo y otras lesiones (Wang A pesar de la gran cantidad de compues- et al., 2012). Estos beneficios biomedicinales tos químicos aislados en las últimas décadas se atribuyen a la presencia de componentes quí- a partir de pepinos de mar (Costa Leal, Puga, micos, como esteroles, triterpenos, sulfatos de Serôdio, Gomes, & Calado, 2012; Santafé, condroitina y glicosaminoglicanos (Baharara, Guzmán, & Torres, 2014; Boufridi & Quinn, Amini, Kerachian, & Soltani, 2014). 2018 ), en muchas especies como Holothuria Otros compuestos de interés sintetizados (Theelothuria) princeps Selenka, 1867, son por los pepinos de mar, son los péptidos y muy pocos los reportes acerca de su compo- los ácidos grasos. Los primeros, son impor- sición química y estudios de bioprospección. tantes compuestos naturales biológicamente Es por eso, que en este trabajo se identificaron activos que tienen un alto potencial de valores los ácidos grasos presentes en especímenes de Rev. Biol. Trop. (Int. J. Trop. Biol.) • Vol. 69(1): 36-44, March 2021 37 H. princeps recolectados en la bahía de Cispa- Preparación del material biológico: De tá, Colombia, mostrando los resultados encon- los especímenes recolectados de H. princeps, trados al cuantificar la actividad antimicrobiana 8 fueron cortados enteros en pequeños trozos de su extracto metanólico. (350 g peso vivo) y se sometieron a maceración en frío con Metanol en ausencia de luz por 72 horas, luego se filtró y concentró bajo el prin- MATERIALES Y MÉTODOS cipio de destilación a presión reducida a 48 °C, Área de estudio: Los especímenes de obteniendo el extracto metanólico (2.5 g). Holothuria princeps fueron recolectados en el sector Banco de Arena, bahía de Cispatá Fraccionamiento cromatográfico: El en el Caribe Colombiano (09°25’26.15” N extracto metanólico (1.5 g) fue sometido a & 75°47’33.6” W) a profundidades menores cromatografía de columna (CC) usando sistema a 3 m. El sector se caracteriza por presentar de elución Bencina: Acetato de Etilo (7:1), las subfracciones obtenidas se monitorearon por un sustrato arenoso y fango arenoso, lo cual cromatografía de capa delgada (CCD), usando brinda disponibilidad
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