Microorganismos cultivables con actividad β-D-glucosidasa obtenidos en cápsulas de Vanilla planifolia, Vanilla pompona e híbrido de Costa Rica (V. planifolia x V. pompona)
Fecha
2022
Autores
Sánchez Vargas, Steven
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Editor
Universidad Nacional (Costa Rica)
Resumen
La acumulación de metabolitos secundarios en las especies comerciales del género Vanilla, siendo vainillina el más importante, la convierten en la segunda especia más cara a nivel mundial. Estos metabolitos se forman a partir de la actividad β-D-glucosidasa propia de sus cápsulas y de microorganismos presentes en el proceso de curado. El objetivo del presente trabajo fue determinar la presencia de microorganismos cultivables con actividad β-D-glucosidasa en cápsulas fisiológicamente maduras de V. planifolia, V. pompona e híbrido de Costa Rica ((V. planifolia × V. pompona) × V. planifolia), mediante su aislamiento, identificación y crecimiento en placas con sustratos, para un posible uso en el mejoramiento del curado en cápsulas de Vanilla comercial. El aislamiento de bacterias y levaduras se realizó previo al proceso de curado en caldos y agares de Tripticasa de Soya y Sabouraud Dextrosa, respectivamente. Se usó secuenciación de ADN del ARNr 16S para la identificación de los aislamientos bacterianos y del ITS para la identificación de los aislamientos fúngicos. Además, los sustratos arbutina, esculina y celobiosa se utilizaron para determinar la actividad β-D-glucosidasa. Los aislamientos a partir de las cápsulas de V. pompona tuvieron una mayor diversidad de micoorganismos. Con respecto a la actividad β-D-glucosidasa en bacterias, el filo Proteobacteria fué el más diverso y con un mayor número de aislamientos con actividad enzimática, principalmente en esculina y celobiosa. En levaduras, el filo con mayor número de aislamientos y más diverso fue Basidiomycota, en el cual, todos los aislamientos tuvieron una alta actividad sobre los sustratos arbutina y esculina. El principal aporte del trabajo se da en ser el primero en estudiar la actividad β-D-glucosidasa de microorganismos obtenidos a partir de cápsulas fisiológicamente maduras y previas a su curado de dos especies comerciales de Vanilla y el híbrido de Costa Rica. En conclusión, se obtuvo grupos de microorganismos no reportados previamente provenientes de cápsulas de V. planifolia, V. pompona e híbrido de Costa Rica con actividad β-D-glucosidasa, donde el grupo de las levaduras son las más ideales para futuras aplicaciones biotecnológicas en el curado de cápsulas de Vanilla con valor económico.
The accumulation of secondary metabolites in the commercial species of the Vanilla genus, vanillin being the most important, defined in the second most expensive spice worldwide. These metabolites are formed from the β-D-glucosidase activity of their capsules and from microorganisms present in the curing process. The objective of this study was to determine the presence of cultivable microorganisms with β-D-glucosidase activity in physiologically mature capsules of V. planifolia, V. pompona and hybrid from Costa Rica ((V. planifolia × V. pompona) × V. planifolia ), through its isolation, identification and growth in plates with substrates, for a possible use in the improvement of curing in commercial Vanilla capsules. The isolation of bacteria and yeasts was carried out prior to the curing process in Trypticase Soy and Sabouraud Dextrose broths and agars, respectively. DNA sequencing of 16S rRNA was used for identification of bacterial isolates and of ITS for identification of fungal isolates. In addition, the substrates arbutin, esculin, and cellobiose were used to determine β-D-glucosidase activity. Isolates from V. pompona capsules had a higher diversity of microorganisms. Regarding the β-D-glucosidase activity in bacteria, the Proteobacteria phylum was the most diverse and with a greater number of isolates with enzymatic activity, mainly in esculin and cellobiose. In yeasts, the phylum with the highest number of isolates and the most diverse was Basidiomycota, in which all isolates had a high activity on the substrates arbutin and esculin. The main contribution of this work is that it is the first to study the β-D-glucosidase activity of microorganisms obtained from physiologically mature capsules prior to curing of two commercial species of Vanilla and the hybrid from Costa Rica. In conclusion, groups of previously unreported discoveries were obtained from capsules of V. planifolia, V. pompona and hybrid from Costa Rica with β-D-glucosidase activity, where the group of yeasts are the most ideal for future biotechnological applications in the curing of Vanilla capsules with economic value.
The accumulation of secondary metabolites in the commercial species of the Vanilla genus, vanillin being the most important, defined in the second most expensive spice worldwide. These metabolites are formed from the β-D-glucosidase activity of their capsules and from microorganisms present in the curing process. The objective of this study was to determine the presence of cultivable microorganisms with β-D-glucosidase activity in physiologically mature capsules of V. planifolia, V. pompona and hybrid from Costa Rica ((V. planifolia × V. pompona) × V. planifolia ), through its isolation, identification and growth in plates with substrates, for a possible use in the improvement of curing in commercial Vanilla capsules. The isolation of bacteria and yeasts was carried out prior to the curing process in Trypticase Soy and Sabouraud Dextrose broths and agars, respectively. DNA sequencing of 16S rRNA was used for identification of bacterial isolates and of ITS for identification of fungal isolates. In addition, the substrates arbutin, esculin, and cellobiose were used to determine β-D-glucosidase activity. Isolates from V. pompona capsules had a higher diversity of microorganisms. Regarding the β-D-glucosidase activity in bacteria, the Proteobacteria phylum was the most diverse and with a greater number of isolates with enzymatic activity, mainly in esculin and cellobiose. In yeasts, the phylum with the highest number of isolates and the most diverse was Basidiomycota, in which all isolates had a high activity on the substrates arbutin and esculin. The main contribution of this work is that it is the first to study the β-D-glucosidase activity of microorganisms obtained from physiologically mature capsules prior to curing of two commercial species of Vanilla and the hybrid from Costa Rica. In conclusion, groups of previously unreported discoveries were obtained from capsules of V. planifolia, V. pompona and hybrid from Costa Rica with β-D-glucosidase activity, where the group of yeasts are the most ideal for future biotechnological applications in the curing of Vanilla capsules with economic value.
Descripción
Sánchez Vargas, Steven (2022). Microorganismos cultivables con actividad β-D-glucosidasa obtenidos en cápsulas de Vanilla planifolia, Vanilla pompona e híbrido de Costa Rica (V. planifolia x V. pompona). [Tesis de Licenciatura]. Universidad Nacional, Costa Rica.
Palabras clave
VAINILLA, MICROORGANISMOS, MICROORGANISMS, METABOLITOS SECUNDARIOS, BACTERIAS, SECONDARY METABOLITES