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Carrion Zavaleta Terecita Elizabeth Biblioteca Digital - Dirección de Sistemas de Informática y Comunicación UNIVERSIDAD NACIONAL DE TRUJILLO FACULTAD DE FARMACIA Y BIOQUÍMICA ESCUELA ACADÉMICO PROFESIONAL DE FARMACIA Y BIOQUÍMICA BIOQUIMICA Tesis II Y Actividad antibiótica y antioxidante de fracciones del extracto crudo de Marivirga lumbricoides FARMACIA PARA OPTAR EL GRADO DE BACHILLER EN DE FARMACIA Y BIOQUIMICA AUTORA: CARRION ZAVALETA, Terecita Elizabeth BIBLIOTECA ASESOR: Mg. GANOZA YUPANQUI, Mayar Luis TRUJILLO – PERÚ 2019 Esta obra ha sido publicada bajo la licencia Creative Commons Reconocimiento-No Comercial-Compartir bajola misma licencia 2.5 Perú. Para ver una copia de dicha licencia, visite http://creativecommons.org/licences/by-nc-sa/2.5/pe/ Biblioteca Digital - Dirección de Sistemas de Informática y Comunicación DEDICATORIA A Dios, por ser la fuente de energía infinita y equilibrio en la naturaleza, aquella motivación en momentos de mayor dificultad. A mi madre la Sra. Eva Dorliza Zavaleta Rodríguez, por su amor, trabajo y sacrificio en todos estos años, gracias a ti he logrado llegar hasta aquí́ y convertirme en lo que soy. A toda mi familia en especial a mi abuelita Teresa porqueBIOQUIMICA con sus oraciones, consejos Y y palabras de aliento hicieron de mí una mejor persona y de una u otra forma me acompañan en todos mis sueños y metas. FARMACIA A todo el equipo de investigación DEde la Facultad de Farmacia y Bioquímica porque con ellos aprendí el trabajo en equipo y me ayudaron a desenvolverme más en el campo profesional de la carrera BIBLIOTECA A mis amigos de la universidad y al grandioso grupo humano de investigación que sea formado en el año 2018, gracias a su apoyo trabaje con empeño, dedicación y cariño, y a todos quienes contribuyeron con un granito de arena para culminar con éxito la meta propuesta. Terecita Elizabeth Carrión Zavaleta i Esta obra ha sido publicada bajo la licencia Creative Commons Reconocimiento-No Comercial-Compartir bajola misma licencia 2.5 Perú. Para ver una copia de dicha licencia, visite http://creativecommons.org/licences/by-nc-sa/2.5/pe/ Biblioteca Digital - Dirección de Sistemas de Informática y Comunicación AGRADECIMIENTO Quiero expresar mi más grande y sincero agradecimiento a mi asesor el profesor Mayer Luis Ganoza Yupanqui, principal colaborador durante todo este proceso, quien con su dirección, conocimiento, enseñanza y colaboración permitió́ el desarrollo de este proyecto de investigación. Agradezco al proyecto PIBA-2-P-247-14-FONDECYT: “Biodiversidad mixobacterial marina del Perú: potencial fuente de biomoléculas antibióticas contra patógenos multidrogo resistentes” financiado con fondos del FONDEYCT/CONCYTECBIOQUIMICA con convenio N°092-2014-FONDECYT, por el apoyo de losY materiales para la culminación de la presente tesis. Finalmente quiero agradecer a los miembros del proyecto PIBA-2-P-247-14- FONDECYT: “Biodiversidad mixobacterialFARMACIA marina del Perú: potencial fuente de biomoléculas antibióticas contra patógenosDE multidrogo resistentes” BIBLIOTECA ii Esta obra ha sido publicada bajo la licencia Creative Commons Reconocimiento-No Comercial-Compartir bajola misma licencia 2.5 Perú. Para ver una copia de dicha licencia, visite http://creativecommons.org/licences/by-nc-sa/2.5/pe/ Biblioteca Digital - Dirección de Sistemas de Informática y Comunicación PRESENTACIÓN Señores miembros del jurado dictaminador: Dando cumplimiento a lo establecido por el reglamento de trabajos de investigación de la Facultad de Farmacia y Bioquímica de la Universidad Nacional de Trujillo, someto a vuestra consideración y elevado criterio profesional el presente informe de Tesis II intitulado: Actividad antibiótica y antioxidante de fracciones del extracto crudo de Marivirga lumbricoides. Es propicia esta oportunidad para manifestar mi sincero reconocimientoBIOQUIMICA a mi alma máter Y y su plana docente; que con su capacidad y buena voluntad, contribuyen en una formación profesional sólida. Guardo a su criterio, señores miembros del jurado, la calificación del presente trabajo de investigación científica. FARMACIA Trujillo, marzo de 2019 DE CARRION ZAVALETA TERECITA ELIZABETH BIBLIOTECA iii Esta obra ha sido publicada bajo la licencia Creative Commons Reconocimiento-No Comercial-Compartir bajola misma licencia 2.5 Perú. Para ver una copia de dicha licencia, visite http://creativecommons.org/licences/by-nc-sa/2.5/pe/ Biblioteca Digital - Dirección de Sistemas de Informática y Comunicación JURADO DICTAMINADOR Dr. CAMPOS FLORIAN, JULIO VÍCTOR PRESIDENTE BIOQUIMICA Y _______________________________________ Mg. GANOZA YUPANQUI, MAYAR LUIS FARMACIA ASESOR DE ______________________________________ Dr. QUISPE DÍAZ, IVÁN MIGUEL BIBLIOTECA MIEMBRO iv Esta obra ha sido publicada bajo la licencia Creative Commons Reconocimiento-No Comercial-Compartir bajola misma licencia 2.5 Perú. Para ver una copia de dicha licencia, visite http://creativecommons.org/licences/by-nc-sa/2.5/pe/ Biblioteca Digital - Dirección de Sistemas de Informática y Comunicación RESUMEN El objetivo de este trabajo de investigación fue evaluar la actividad antibiótica y antioxidante de las fracciones del extracto crudo de Marivirga lumbricoides aislada de origen marino. La biomasa de la bacteria se obtuvo por incubación en agar marino a 30 °C por 7 días, se empleó acetona como solvente extractor, se llevó a sequedad para obtener el extracto crudo, se fraccionó por Sephadex LH-20, se obtuvieron fracciones que fueron concentradas hasta extracto seco (ES), se evaluó la actividad antioxidante mediante 2,2- difenil-1-picril-hidrazilo (DPPH) y el ensayo ácido 2,2´-azinobis-(3-etilbenzotiazolina-6- sulfónico) (ABTS) expresados en equivalentes de Trolox (ET) y la actividad antibiótica mediante el método de difusión en disco y microdilución frente a Staphyloccocus aureus BIOQUIMICA ATCC 25923, Staphyloccocus aureus resistente a meticilinaY ATCC 43300, Bacillus subtilis ATCC 6633 y Escherichia coli ATCC 25922. Se obtuvieron 3 fracciones (F1, F2 y F3) con colores diferenciados, con halos de inhibición de crecimiento bacteriano entre 7,50 y 19,25 mm, con concentracionesFARMACIA mínimas inhibitorias (CMI) y concentraciones mínimas bactericidas (CMB) entreDE 0,62 y 20,00 mg/mL .En la actividad antioxidante se obtuvieron valores entre 4,34 ± 0,38 y 16,21 ± 3,00 mg ET/g ES (DPPH), y entre 9,74 ± 0,39 y 18,23 ± 1,18 mg ET/g ES (ABTS). Se concluye que la F2 presenta mejor actividad antibiótica como antioxidante. BIBLIOTECA Palabras claves: Marivirga lumbricoides, cromatografía, antioxidante, antibiótico. v Esta obra ha sido publicada bajo la licencia Creative Commons Reconocimiento-No Comercial-Compartir bajola misma licencia 2.5 Perú. Para ver una copia de dicha licencia, visite http://creativecommons.org/licences/by-nc-sa/2.5/pe/ Biblioteca Digital - Dirección de Sistemas de Informática y Comunicación ABSTRACT The objective of this research was to evaluate the antibiotic and antioxidant activity of the fractions of the isolated Marivirga lumbricoides raw extract of marine origin. The biomass of the bacteria was obtained by incubation in marine agar at 30 °C for 7 days, acetone was used as extractor solvent, dried to obtain the crude extract, fractionated by Sephadex LH-20, fractions were obtained that were concentrated to dry extract (DE), antioxidant activity was evaluated by 2,2-diphenyl-1-picryl-hydrazyl (DPPH) and the 2,2'-azinobis-(3-ethylbenzthiazoline-6-sulfonic acid) assay (ABTS) expressed in Trolox equivalents (TE) and antibiotic activity by disc diffusion and microdilution method against Staphylococcus aureus ATCC 25923, methicillin-resistantBIOQUIMICA Staphylococcus Y aureus ATCC 43300, Bacillus subtilis ATCC 6633 and Escherichia coli ATCC 259223 fractions were obtained ( F1, F2 and F3) with differentiated colors, with halos of inhibition of bacterial growth between 7.50 and 19.25 mm, with minimum inhibitory concentrations (MIC) and minimum bactericidalFARMACIA concentrations (CMB) between 0.62 and 20.00 mg/mL. In the antioxidant activityDE values were obtained between 4.34 ± 0.38 and 16.21 ± 3.00 mg TE/g DE (DPPH), and between 9.74 ± 0.39 and 18.23 ± 1.18 mg TE/g DE (ABTS). It is concluded that F2 presents better antibiotic activity as an antioxidant. Key words: Marivirga lumbricoides, chromatography, antioxidant, antibiotic. BIBLIOTECA vi Esta obra ha sido publicada bajo la licencia Creative Commons Reconocimiento-No Comercial-Compartir bajola misma licencia 2.5 Perú. Para ver una copia de dicha licencia, visite http://creativecommons.org/licences/by-nc-sa/2.5/pe/ Biblioteca Digital - Dirección de Sistemas de Informática y Comunicación INDICE I. INTRODUCCIÓN…………………………………………………………..1 II. MATERIAL Y MÉTODO…………………………………………………..5 III. RESULTADOS………………………………………………………….…12 IV. DISCUSIÓN…………………………………………………………..…....17 V. CONCLUSIONES………………………………………………………….24 VI. REFERENCIAS BIBLIOGRÁFICAS……………………………………..25 VII. ANEXOS…………………………………………………………………...30 BIOQUIMICA Y FARMACIA DE BIBLIOTECA vii Esta obra ha sido publicada bajo la licencia Creative Commons Reconocimiento-No Comercial-Compartir bajola misma licencia 2.5 Perú. Para ver una copia de dicha licencia, visite http://creativecommons.org/licences/by-nc-sa/2.5/pe/ Biblioteca Digital - Dirección de Sistemas de Informática y Comunicación I. INTRODUCCIÓN Los productos naturales han desempeñado un papel fundamental en el descubrimiento de fármacos entre ellos los nichos ecológicos marinos que son un punto de muestreo importante donde se encuentra una amplia diversidad de microorganismos como bacterias, hongos y cianobacterias,
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