Carrera de Biotecnología

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    Expresión y purificación de una enzima PETasa obtenida mediante reconstrucción ancestral
    (Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos y Biotecnología. Carrera de Biotecnología, 2023-03) Quispe Gudiño, Melanie Dayana; Cerda Mejía, Liliana Alexandra
    In the world, there are a wide variety of organic polymers that have come to represent a significant problem of environmental contamination. Currently, the scientific field began to design appropriate and efficient alternatives to facilitate the recycling of plastic materials ecologically. Biodegradation by enzymes has become highly relevant in recent years due to its good capacity for plastic degradation. The present investigation aimed to express and purify an PETase enzyme obtained from an ancestral reconstruction (ASR); the ancestral enzyme was expected to have superior characteristics to I. sakaiensis PETase respect to its activity against PET and other organic polymers. The ancestral PETase N1 enzyme was expressed in E.coli Ta cells under stable conditions of 1,0 mM IPTG at room temperature, over 16 hours. The enzyme purification was carried out through affinity chromatography (FPLC) complemented with an SDS-PAGE electrophoresis to verify the presence of the ancestral PETase N1 enzyme according to its molecular weight. Over more, the purification used an extra serine protease inhibitor (PMSF) together with EDTA metalloprotease inhibitor. The activity of the ancestral PETase N1 enzyme was evaluated through degradation assays in six high-value polymers by SEM. The images obtained showed degradation features in the HDPE polymer, while in the LDPE, PS, and PVC polymers there was low activity. On the other hand, PP and PET polymers experienced a null activity of the enzyme that could be influenced by various factors.
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    Microencapsulación de microorganismos mediante secado por aspersión con aplicación industrial
    (Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos y Biotecnología. Carrera de Biotecnología, 2022-03) Altamirano Sánchez, María Fernanda; López Hernández, Orestes Darío
    Functional products containing beneficial microorganisms are gaining popularity on the world market. The viability of microorganisms present in products can be affected during processing, storage and ingestion by the consumer. Microencapsulation by spray drying makes it possible to prolong the consumption time of said products and preserve cell viability. In spray drying, a fluid is atomized in a stream of hot air to obtain dry particles with low water content. Therefore, in the present job it is presented a review of the most recent research on the spray drying of efficient microorganisms with application in the human food, animal food, agricultural and environmental and healthcare industries, with specific reference to the best conditions that allow obtaining powders with high cell viability. The main topics covered are the types of microencapsulated microorganisms, the objective of the study, viable cell count, growth medium, encapsulating agents, operating parameters, equipment and storage conditions. Within the analysis carried out, it can be determined that bacteria, yeasts, cyanobacteria and fungi have been microencapsulated for different purposes. Especially a large number of studies on bacteria with application in the human food industry have been reported, while in the healthcare industry a minimal amount of research has been published. On the other hand, it has been determined that the genus and species, the wall materials, as well as the operating parameters significantly influence cell viability and microencapsulation efficiency.