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 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 Taxonomía polifásica de actinomicetes útiles en biotecnología(Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos. Carrera de Ingeniería Bioquímica, 2018-12) Baquero Vayas, Diana Estefanía; Rodríguez Meza, Carlos AlbertoThirty strains of actinomycetes isolated from garden soil were characterized by polyphasic taxonomy and identified molecularly by sequencing the coding gene for 16S rRNA, the latter resulting in nine species closely related to the culture collection, these are Streptomyces griseiniger, Streptomyces cangkringensis, Streptomyces rhizosphaericus, Streptomyces sporoclivatus, Streptomyces malaysiensis, Streptomyces decoyicus, Streptomyces fildesensis, Streptomyces rhizosphaerihabitans, Streptomyces spiroverticillatus, thus obtaining a diversity percentage of 30 percent, this was corroborated by the variety of phenotypic and genotypic characteristics determined. The phenetic groups showed greater coherence and homogeneity compared to the genotypic ones, however, when analyzing them with the phylogenetic tree, relationship between the results was found. The collection of actinomycetes proved to be mostly, halophilic, neutrotolerant and mesophilic, in addition to expressing potential for the degradation of complex carbon sources and the use of amino acids as a carbon and nitrogen source. The importance of this work lies in the complementarity of these three tools for the characterization and identification of microorganisms, which continue from selective isolation, and the achievement of a profile to determine the biotechnological potential of the same, step that should be followed in this work. The option of taking the 16S phylogenetic analysis as a starting point for the characterization of microorganisms is also suggested, having as a basis the molecular identification to guide an adequate profiling of them with genotyping and finally phenotypic tests to determine their culture conditions and growth in order to identify its biotechnological utility.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 Determinación de la correlación entre métodos visuales ópticos y difusión en placa en el crecimiento de Escherischia coli(Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos. Carrera de Ingeniería Bioquímica, 2017-09) Fiallos Núñez, Johanna Elizabeth; Dugarte Jiménez, Nahir YerelyThe correlation between optical methods: Spectrophotometer and turbidimeter, visual method: McFarland scale and the reference method of plate diffusion were evaluated in order to determinate the concentration of Escherichia coli at 9 hours. The visual method doesn´t have a correlation with the reference method because is a qualitative method and the measurements are subjective. In the case of the optical methods, the spectrophotometer had an error of 91,4 percent in comparison of the turbidimeter with an error of 2,2 percent. The turbidimeter was the only method which has a correlation with the reference method and can be used as an alternative for the traditional plate diffusion method.Item Desarrollo de un método para recuento de Listeria monocytogenes utilizando turbidimetría(Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos. Carrera de Ingeniería Bioquímica, 2017-09) Bustos Cosios, Cristina Verónica; Arancibia Soria, Mirari YosuneThe present study was performed to develop a method for recounting Listeria monocytogenes using turbidimetry, the proposed method consists in a scale of four equidistant points, prepared with sulfate of sodium and chloride of barium that produce a precipitate of sulfate of barium, which resembles cellular concentration of 10 x 108 UFC/ml to 40 x108 UFC/ml, innovating the scale of turbidity proposed by McFarland in 1907. Using the proposed scale of precipitate of sulfate of barium, an interpolation equation was developed in which readings of the nephelometric turbidity units of microorganisms in culture medium are applied, predicting in a mathematical way the cell concentration in a liquid culture medium. The error rate of this method compared to the plate count method is 0,28 to 3,41 percent. Of the above is possible to use the turbidimeter to predict cell concentration effectively, simplifying microbiological management in investigative procedures, without the need for long preparation time to know the cellular concentration.Item Obtención de un hidrolizado de Spirulina (Arthrospira platensis) en polvo, mediante secado por aspersión, como ingrediente funcional.(Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos. Carrera de Ingeniería Bioquímica, 2017-04) Camacho Ayala, Tarsis Xavier; López Hernández, Orestes DaríoThe research project was based on an analysis of the Spirulina microalgae (Arthrospira platensis), which was studied by the application of several treatments with different specific conditions that allow a better assimilation of the alga, in case of being applied as active pharmaceutical ingredient or protein supplement. The best treatment was attained at the conditions: pH 5, a concentration of 5% (in relation to 1:20 of substrate: water) and at a hydrolysis temperature maintained in a range of 65 °C to 70 °C. The hydrolyzate was sprayed by spray drying which gave an optimum yield of 73.89% and a different coloration to Spirulina powder concentrate because of the structural modification of the phycobiliproteins in the hydrolytic process. In addition, the degree of hydrolysis of the powder was analyzed by a comparison of the amine nitrogen content of the hydrolyzate and Spirulina powder concentrate, SDS - PAGE electrophoresis and RP - HPLC chromatography techniques were also applied, which allowed to determine that the hydrolysis process was adequate and produced a complex mixture of free amino acids and low molecular weights peptides with with the necessary characteristics for the future application of Spirulina hydrolyzate as functional ingredient.