Carrera Ingeniería Bioquímica
Permanent URI for this collectionhttp://repositorio.uta.edu.ec/handle/123456789/809
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Item Diversidad biológica y metabólica de actinomicetes aislados de suelo y arena de ecosistemas glaciares de la Antártida y del norte del Chimborazo(Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos. Carrera de Ingeniería Bioquímica, 2015) Villacís Barrazueta, Juan José; Rodríguez Meza, Carlos AlbertoItem Caracterización de levaduras aisladas a partir de frutos de durazno (Prunus persica), fresa (Fragaria vesca) y manzana (Malus domestica)(Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos y Biotecnología. Carrera de Ingeniería Bioquímica, 2019-07) Escobar Reyes, Mercy Jeaneth; Rodríguez Meza, Carlos AlbertoThis paper shows the importance of the phenotypic and genotypic characterization for the identification of yeasts that were isolated from seasonal fruits peach, strawberry and apple from three different locations Montalvo-Palagua, Huachi Grande-San Alfonso, Alobamba – the triumph. The time, yeasts have generated a biotechnological interest due to their application in the industry, rapid growth and biomass production. The fermentation of the fruits made before the isolation allowed to obtain differences in the number of CFU for every ml according to the fruit and place of collection. A total of ninety four isolated and conserved strains were obtained. For the identification of these was taken different criteria as morphological where analyzed their characteristics macroscopic and microscopic, physiological criteria of growth in different ranges of pH, salinity, temperature and assimilation of Amino acids as a source of carbon and nitrogen. The characterization genotypic was performed by analyzing the digestion of the region ITS by means of RFLP, the results of numerical taxonomy obtained were analyzed in the statistical software NTSys which allowed to obtain a taxonomic determination with which it was possible to group and form sixteen species groups resulting from the analysis of numerical taxonomy of phenotypic data and fourteen species groups resulting from the numerical taxonomy of genotypic data. The final analysis of taxonomy Polifásica made for fifty-four strains of yeast where they grouped their characteristics resulting from the analysis phenotypic and Genotypic obtaining a final dendogram with a cut to 80 percent of similarity that allowed Obtain 18 species groups where it is observed its wide distribution for the fruits and places of where they were isolated the yeasts.Item Identificación de bacterias aisladas de ecosistemas glaciares, andinos y antárticos(Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos. Carrera de Ingeniería Bioquímica, 2019-05) Palacios Mazón, Diego Marcelo; Rodríguez Meza, Carlos AlbertoBacteria isolated from glacial, Andean and Antarctic ecosystems were identified. By rep-PCR using the BOX primer, 72 strains were genotypically characterized and a dendrogram was constructed by numerical taxonomy, using the Pearson coefficient and the UPGMA algorithm, obtaining the formation of 10 groups. From these, representatives were selected for the sequencing of the 16S rDNA gene and phenotypic characterization. The phylogenetic analysis of the 16S rDNA gene was performed by calculating the evolutionary distances of Jukes & Cantor and the Neighbor-Joining algorithm, allowing the identification of 11 different species. Of which, strains DG-222 and DG-203 probably belong to new species. However, further studies are necessary to demonstrate this. Through the phenotypic characterization, a dendrogram of numerical taxonomy of phenotypic data was constructed using the SSM coefficient and the UPGMA algorithm, obtaining the formation of 21 phenetic groups with 84,6 percent similarity. Most of the collection was determined as Gram negative, psychrotroph, neutrotolerant, weak halophilic or halophilic. In addition, new physiological characteristics of 4 different species were discovered. The analysis of polyphasic taxonomy with the strains that had fingerprinting patterns similar to the identified species, managed to locate 35 strains according to the molecular characterization. Forming 16 phenetic groups at 90 percent similarity and determining the polyphase taxonomic position of 48,6 percent of the collection. Probably, the diversity of the collection is underestimated and there could be species that have not been identified in this study. It is necessary to carry out a second analysis of the strains that were excluded in the analyzes.Item Determinación de la diversidad de actinomicetes aislados de ecosistemas glaciares de los Andes y la Antártida(Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos. Carrera de Ingeniería Bioquímica, 2019-05) Chamorro Sevilla, María Belén; Rodríguez Meza, Carlos AlbertoWere identified 152 strains of actinomycetes molecularly isolated from cold ecosystems of Antarctica, Cotopaxi and Chimborazo. Through the amplification of the fingerprinting patterns by rep-PCR, using the first BOX A1R1, I was able to construct the genotypic dendrogram generated by the Pearson coefficient and the UPGMA algorithm, obtaining 23 species-groups. The phylogenetic analysis of the 16S rDNA gene was constructed by calculating the evolutionary distances of Jukes & Cantor and the Neighbor - Joining algorithm. It was possible to identify 4 different species, which belonged to the genus Streptomyces, while the strain DE-067 was associated to the genus Nocardia. On the other hand, by phenotypic characterization it was determined that most of the strains of this study were mesophilic, weak halophilic and neutrotolerant. In addition, they have a great ability to metabolize amino acids as a source of carbon and nitrogen. With the obtained results, the phenotypic dendrogram was constructed by the SSM coefficient and the UPGMA algorithm, determining 23 species-groups at 89 percent similarity. Subsequently, the molecular identification of the collection was carried out using the 16S rDNA gene, with the strains that shared fingerprinting patterns like the identified species. Achieving taxonomically position 87 strains, in 14 groups-species at 80 percent similarity. As it is evident, it is necessary to complement the results obtained in this study, by means of the molecular identification of the strains that were not sequenced, to know with certainty the diversity of actinomycetes in the collection.Item Caracterización fenotípica de actinomicetes aislados de suelos contaminados con hidrocarburos(Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos. Carrera de Ingeniería Bioquímica, 2018-07) Chamorro Medina, Jessica Paola; Rodríguez Meza, Carlos AlbertoNinety-three strains of actinomycetes, isolated from soils contaminated with petroleum derivatives, from the Sacha field were used for the determination of phenotypic and metabolic diversity. Eighteen groups-species were identified, presenting a similarity greater than 90 percent, after performing numerical taxonomy of phenotypic data. Likewise, all crops were grown in Bushnell-Hass, enriched with gasoline, diesel and oil, as the only source of carbon and energy. It was determined that 89 percent of microorganisms can use different hydrocarbons in their metabolism and that only the representatives of two groups-species do not have this capacity. 92.2 percent of actinomycetes can use gasoline as a substrate, 83.9 percent diesel and 60 percent oil. The results show that this group of actinomycetes presents a great phenotypic and metabolic diversity. These characteristics could be studied to determine the potential use in in-situ bioremediation processes.Item Determinación preliminar de producción de compuestos bioactivos de actinomicetes y bacterias aislados de ecosistemas glaciares andinos y antárticos(Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos. Carrera de Ingeniería Bioquímica, 2018-03) Estrada Espín, María Cristina; Rodríguez Meza, Carlos AlbertoSeventy-nine actinomycete and thirty-three bacterial strains were isolated from soil and sand samples collected from glacial ecosystems from Chimborazo, Cotopaxi and Antarctica. Microorganisms were phenotypically characterized and their capacity to produce bioactive compounds against several microorganisms was determined. The results were analyzed by numerical taxonomy at 90 percent similarity. Actinomycetes were divided in forty phenetic groups, while bacteria were divided in twenty-nine. Fifty-five actinomycetes were isolated from Chimborazo, sixteen from Cotopaxi and four from Antarctica. In relation to bacteria seventeen strains came from Chimborazo, fourteen from Cotopaxi and one from Antarctica. The phenotypic data related to the growth depending on the temperatura, showed that all isolated microorganisms presented mesophilic characteristics. Respect to tolerance to NaCl, 61 percent bacteria presented growth characteristics over of salt levels up 15 percent w/v in medium , while actinomycetes only supported concentrations about 1 percent NaCl. Regarding production capacity of bioactive compounds, 84 percent of actinomycetes and 70 percent of bacteria isolated, showed antagonistic activity against the model microorganisms used. The actinomycetes DE002 and DE053 strains produced metabolites with bactericidal characteristics and DE006, DE031, DE081, DE083 and DE084 strains fungicidal metabolites. Bacteria were not able to produce compounds with bactericidal or fungicidal capacity and were only effective for certain species of microorganisms. Data obtained in the current investigation, demonstrated wide metabolic and phenotypic diversity of actinomycetes and bacteria present in glacial ecosystems. Further studies are necessary to explore its potential for the industrial production of bioactive compounds.Item Determinación preliminar de producción de compuestos bioactivos de actinomicetes y bacterias aislados de ecosistemas glaciares andinos y antárticos(Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos. Carrera de Ingeniería Bioquímica, 2018-03) Díaz Ortíz, Adriana Patricia; Rodríguez Meza, Carlos AlbertoSeventy-nine actinomycete and thirty-three bacterial strains were isolated from soil and sand samples collected from glacial ecosystems from Chimborazo, Cotopaxi and Antarctica. Microorganisms were phenotypically characterized and their capacity to produce bioactive compounds against several microorganisms was determined. The results were analyzed by numerical taxonomy at 90 percent similarity. Actinomycetes were divided in forty phenetic groups, while bacteria were divided in twenty-nine. Fifty-five actinomycetes were isolated from Chimborazo, sixteen from Cotopaxi and four from Antarctica. In relation to bacteria seventeen strains came from Chimborazo, fourteen from Cotopaxi and one from Antarctica. The phenotypic data related to the growth depending on the temperatura, showed that all isolated microorganisms presented mesophilic characteristics. Respect to tolerance to NaCl, 61 percent bacteria presented growth characteristics over of salt levels up 15 percent w/v in medium , while actinomycetes only supported concentrations about 1 percent NaCl. Regarding production capacity of bioactive compounds, 84 percent of actinomycetes and 70 percent of bacteria isolated, showed antagonistic activity against the model microorganisms used. The actinomycetes DE002 and DE053 strains produced metabolites with bactericidal characteristics and DE006, DE031, DE081, DE083 and DE084 strains fungicidal metabolites. Bacteria were not able to produce compounds with bactericidal or fungicidal capacity and were only effective for certain species of microorganisms. Data obtained in the current investigation, demonstrated wide metabolic and phenotypic diversity of actinomycetes and bacteria present in glacial ecosystems. Further studies are necessary to explore its potential for the industrial production of bioactive compounds.Item Primer estudio sobre la diversidad de levaduras asociada a un volcán: Tungurahua, Ecuador(Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos. Carrera de Ingeniería Bioquímica, 2015) Hidalgo Escobar, Karen Cristina; Serrano Bueno, GloriaEl volcán Tungurahua representa un ecosistema extremófilo en cuanto a sus características fisicoquímicas, las cuales deben ser soportadas por los organismos que lo habitan. Desde las faldas del volcán hasta zonas cercanas al cráter se establecieron trece puntos de muestreo, de los que se aislaron un total de doce levaduras. Estas fueron caracterizadas física y molecularmente, identificándose diez especies diferentes. El análisis filogenético las clasificó dentro de cinco órdenes distintos y, de forma equitativa, en dos familias (Basidiomicota y Ascomicota). El análisis comparativo de la diversidad de especies de levaduras asociadas a ecosistemas volcánicos, mostró que ésta, no fue influenciada por el tipo de hábitat y, posiblemente exista mayor relación entre poblaciones geográficamente próximas. Las especies Cryptococcus sp. y Rhodotorula mucilaginosa destacan por haber sido aisladas también de otros ecosistemas volcánicos. Asimismo, ésta última se considera ubicua, pues es capaz de colonizar casi todo tipo de ambiente. Los ensayos de temperatura y pH revelaron que estos factores influyentes en el crecimiento de las levaduras. Respecto a temperatura, se sugiere que el 90% pueden considerarse psicrófilas, cuya temperatura máxima de crecimiento no sobrepasa los 20 °C y, únicamente una especie, Lecythophora sp. fue capaz de crecer a 37 °C. En relación al crecimiento en medio acidificado, el 70% mostró un amplio rango de crecimiento en medio ácido (entre 2,5 y 6,5). Rhodotorula mucilaginosa, a pesar de presenta un amplio rango de crecimiento (entre 2,5 y 6,5), se puede identificar como acidófila, pues su pH óptimo de crecimiento está entre 2,5 y 4,5. Debido a las características extremófilas, en cuanto a la resistencia a bajas temperaturas y pH de las levaduras aisladas del volcán Tungurahua, estas se proponen como potencialmente aplicables en la industria biotecnológica.Item Determinación del potencial antimicrobiano de la microalga Chlorella extraída de las aguas empozadas de las acequias de Atocha, Tilulum y el Socavón del cantón Ambato(Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos. Carrera de Ingeniería Bioquímica, 2014) Barona Altamirano, Adriana Carolina; Valencia Silva, Alex FabiánLas microalgas son organismos unicelulares eucariotas fotosintéticos capaces de transformar la energía luminosa en energía química; las condiciones óptimas de crecimiento varían de una especie a otra, las investigaciones sobre el cultivo de microalgas son de gran importancia dada su amplia aplicación biotecnológica y comercial. Chlorella es una microalga verde unicelular de agua dulce, de forma esférica mide cerca de 2-10 micrones de diámetro, tiene alta concentración de clorofila, se reproduce rápidamente y de forma asexual, necesita carbono agua luz y pequeñas cantidades de minerales.Se recolecto diferentes muestras de aguas empozadas de Atocha, Tilulum y el Socavón de la ciudad de Ambato, los cultivos de la microalga Chlorella se aislaron utilizando el medio Basal de Bold debido a la cantidad de nutrientes que posee, las células tardaron 240 horas en alcanzar la fase exponencial para su posterior análisis. Se determinó el potencial antimicrobiano de la microalga en base a los resultados positivos en los ensayos en los que se utilizó el precipitado que resulta de la extracción del chlorellin con los solventes etanol y alcohol isopropílico. El análisis estadístico determinó que el mejor tratamiento fue Tilulum- Alcohol isopropílico al tener mayor el tamaño del halo de inhibición, cuya media fue de 28mm, ante la bacteria coco grampositiva, mientras que el menor tratamiento fue Socavón-etanol con una media de 0,11mm. En la última etapa de la investigación se realizó las pruebas de control positivo mediante la técnica del antibiograma utilizando la bacteria aislada y antibióticos como penicilina, demostrando que fue sensible con un halo de inhibición de 5mm ante la ampicilina y 1mm ante oxacilina, valores que están dentro del rango de sensibilidad de acuerdo a las tabla CIM (Concentración Mínima Inhibitoria).Item Determinación del efecto de las condiciones de almacenamiento sobre la viabilidad de esporas y micelio de actinomicetes aislados de la rizósfera de leguminosas(Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos. Carrera de Ingeniería Bioquímica, 2013) Andrade Proaño, Diana Carolina; Rodríguez Meza, Carlos AlbertoDurante la determinación del efecto de las condiciones de almacenamiento sobre la viabilidad de esporas y micelio de actinomicetes aislados de la rizósfera de leguminosas, se logró purificar, conservar y almacenar en cultivo puro, ochenta y seis cepas de actinomicetes, teniendo en cuenta el porcentaje de recuperación y pérdida de las mismas. De las doscientas setenta y siete cepas aisladas originalmente del proyecto Búsqueda de Actinomicetes Productores de Antibióticos en el año 2009, las cuales se entregaron conservadas en glicerol al 20% (w/v) y almacenadas en refrigeración entre 1°C y 3°C, únicamente se lograron purificar ochenta y seis cepas, 21 se purificaron a partir de la rizósfera de fréjol, 20 de alfalfa, 19 de haba, 18 de arveja y 8 de chocho. La purificación de cepas no fue como se esperaba, ya que se reportó únicamente un 31.0% de cepas purificadas, mientras que un 69.0% de cepas perdidas entre no crecidas y contaminadas que dan un total de 191 cepas. La taxonomía numérica de datos fenotípicos permitió obtener un número menor de grupos especie, que los obtenidos solo en base a la coloración de micelio aéreo, micelio del sustrato y pigmento difusible, mostrando que la adición de características fenotípicas permite delimitar con mayor precisión a los grupos- especie. El dendrograma obtenido al 90% de coeficiente de similaridad dividió a las 86 cepas de actinomicetes purificadas en 36 grupos-especie. Los datos obtenidos en la investigación demuestran que las condiciones de almacenamiento deficientes, influyen sobre la viabilidad de esporas y micelio de actinomicetes. Cabe recalcar que las técnicas de aislamiento y conservación inicial son la clave fundamental para poder establecer colecciones de cultivos por largos períodos de tiempos, los mismos que pueden ser utilizados en investigaciones futuras, permitiendo el descubrimiento de posibles nuevos compuestos bioactivos útiles en biotecnología.