Carrera de Biotecnología
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Item Formulación de un gel a base de extracto de polifenoles obtenidos de la cáscara de Theobroma cacao L. con actividad antioxidante(Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos y Biotecnología. Carrera de Biotecnología, 2026-07) Tuitise Varas, Dallyana Shirley; Fernández Rivero, DanaeThe husk of the Theobroma cacao L. pod is an agro-industrial residue containing phenolic compounds, which possess antioxidant activity and can be used in the development of products for the pharmaceutical and cosmetic sectors. Its use also contributes to the valorization of agricultural byproducts and the reduction of environmental impact. The extraction of phenolic compounds was carried out using an ultrasonic bath with a 2 cubed factorial design, where ethanol concentrations of 50 and 70 percent, solid-liquid ratios of 1:20 and 1:6, and temperatures of 30 and 40 degrees Celsius were evaluated. Subsequently, the selected extract was concentrated and incorporated into topical gel formulations at 3, 5, and 7 percent, corresponding to formulations A, B, and C. The results showed that the highest polyphenol recovery was obtained with 70 percent ethanol, a solid-to-liquid ratio of 1, at 20 and 30 degrees Celsius, reaching 19.03 mg of gallic acid per gram of dry sample. The extract exhibited antioxidant inhibition of 71.28 percent and an antioxidant capacity of 407,596 micromoles of Trolox per liter. All formulations met physicochemical criteria, and formulation A was selected as the most optimal, as it showed no significant differences compared to formulation B, achieving 54.028 percent inhibition and 305,759 micromoles of Trolox per liter. These results demonstrate the potential of cocoa husk as a source of bioactive compounds.Item Exploración de la capacidad de adsorción de las hojas de Jacinto de agua (Eichhornia crassipes) de diversos líquidos por ángulo de contacto y de su superficie adaxial mediante microscopía de fuerza atómica(Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos y Biotecnología. Carrera de Biotecnología, 2026-07) Tigselema Núñez, Jennifr Rubí; Casado Rojo, SantiagoThe foliar surface of aquatic plants constitutes a key interface in the interaction with substances present in the environment; however, there is limited information regarding the relationship between their nanoscopic structure and their behavior toward liquids of different natures. In this context, the present study aims to explore the adsorption capacity of Eichhornia crassipes leaves by determining contact angles and characterizing their adaxial surface. To this end, atomic force microscopy was used to analyze the surface topography at the nanoscopic scale and to determine roughness parameters in different leaf regions. Additionally, the interaction with water, mineral oil, and methanol was evaluated through contact angle measurements, and Fourier-transform infrared spectroscopy was performed to identify the main functional groups present on the leaf surface. The results revealed a heterogeneous topographical organization, characterized by the presence of epidermal structures and variations in roughness among regions, with high values in the central zone of the leaf. Likewise, a differential interaction was observed with the tested liquids, being greater for those with lower surface tension. Spectroscopic analysis confirmed the presence of structural polysaccharides, aromatic compounds, and lipid components associated with the plant cuticle. It is concluded that the adaxial surface of E. crassipes exhibits topographical and chemical characteristics that influence its interaction with different liquids. This study provides insights into the relationship between surface structure, chemical composition, and interfacial behavior in this aquatic species.Item Evaluación del potencial de los residuos agrícolas de maíz (Zea mays) y caña de azúcar (Saccharum officinarum) como sustratos para el cultivo del hongo cola de pavo (Trametes versicolor)(Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos y Biotecnología. Carrera de Biotecnología, 2026-07) Tenorio Quishpe, Martha Beatriz; Arancibia Soria, Mirari YosuneAgricultural residues from maize (Zea mays) and sugarcane (Saccharum officinarum) represent an abundant and underutilized lignocellulosic biomass in Ecuador, whose valorization through the cultivation of medicinal fungi constitutes a high-impact biotechnological alternative. Using a design with two factors: type of agricultural residue (maize and sugarcane) and proportion of structural material (25, 50 and 75 percent), six treatments (T1-T6) were generated with three replicates each, employing Trametes versicolor as the model organism, a basidiomycete capable of degrading lignin through an extracellular enzymatic system that includes laccase, manganese peroxidase (MnP) and lignin peroxidase (LiP); substrates were prepared by chopping, hydration to 85% moisture, pH adjustment between 4.5 and 6.5, and sterilization at 121 degrees Celsius for 90 minutes, before being inoculated with T. versicolor mycelium; response variables were mycelial colonization rate and percentage, analyzed by two-way ANOVA and Tukey HSD test (p less than 0.05). Results showed that both the type of agricultural residue and the proportion of structural material significantly influenced mycelial colonization of T. versicolor, with maize-based substrates containing a higher proportion of sawdust proving most suitable for fungal development under the evaluated conditions; these findings lay the groundwork for the development of sustainable production protocols for T. versicolor from local agroindustrial byproducts, with potential application in environmental biotechnology.Item Análisis cinético de la producción de ácido glucónico empleando residuos de cáscara de piña(Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos y Biotecnología. Carrera de Biotecnología, 2026-07) Solis Castro, Krystel Patricia; García Pazmiño, María EugeniaThe generation of agro-industrial residues derived from pineapple processing represents an environmental concern due to the large amount of discarded biomass. However, these residues contain fermentable sugars that can be utilized in biotechnological processes. The aim of this research was to analyze the kinetic behavior of gluconic acid production using pineapple peel as substrate. For this purpose, bacteria compatible with Gluconobacter oxydans were isolated from kombucha through cultivation on GYC medium supplemented with calcium carbonate, selecting those with acidifying capacity. Preliminary identification included Gram staining and biochemical tests. Subsequently, fermentations were carried out using pineapple peel extract as a carbon source. Microbial growth was evaluated by optical density, while gluconic acid production was determined by spectrophotometry at Five hundred forty nm using a colorimetric method based on hydroxylamine and ferric chloride. The results demonstrated the ability of the isolated strains to grow on the substrate and produce gluconic acid during fermentation. It was concluded that pineapple peel constitutes a viable raw material for the biotechnological production of gluconic acid and a sustainable alternative for the valorization of agro-industrial residues.Item Diseño de la variante mutante S188D de la enzima PETasa de Ideonella sakaiensis(Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos y Biotecnología. Carrera de Biotecnología, 2026-07) Robalino Sánchez, Génesis Aurora; García Solís, Mario DanielPolyethylene terephthalate PET accumulates in ecosystems at rates that conventional recycling methods cannot compensate for. The PETase from Ideonella sakaiensis, IsPETase, degrades PET, but its thermostability below 40 degrees Celsius limits its use in industrial applications. The structural comparison with the thermophilic cutinase LCC, whose D193 residue participates in salt bridge networks that sustain activity up to 70 degrees Celsius, motivated the S188D variation, which replaces the TCC codons with GAC, based on the ancestral mutant M8 architecture of IsPETase. The variant was cloned into the pET-30a(+) vector and expressed in E. coli Rosetta (DE3) under induction with IPTG 0.1 mM at 20 degrees Celsius. Purification by IMAC Ni NTA affinity chromatography produced 5,662 g of soluble protein, with fractions four and five showing the highest active protein concentration, 2,49 mg per mL, and the presence of the protein between the 26 to 34 kDa band was confirmed by SDS PAGE. The degradation assay showed that the activity of 1.5 mg of enzyme per g of micronized post consumer PET, without prior amorphization, at 50 degrees Celsius for 72 hours, reduced PET by an unspecified percentage compared with the control sample. This performance is acceptable, but it does not show significance compared with ancestral mutant variants. Therefore, it is necessary to integrate S188D with other cumulative mutations that strengthen the functional stability of IsPETase and consolidate it as a viable strategy for plastic biodegradation at an industrial scale.Item Evaluación de compuestos bioactivos a partir de microorganismos endófitos para el control de enfermedades por hongos fitopatógenos(Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos y Biotecnología. Carrera de Biotecnología, 2026-07) Quisnia Aldas, Angeles Monserrath; Garcés Moncayo, María DanielaThe present research addresses the problem of diseases caused by phytopathogenic fungi, which are responsible for significant losses in agricultural production and impacts on food security, generating growing interest in the development of sustainable biological strategies. Bioactive compounds produced by endophytic microorganisms isolated from medicinal plants from the Ecuadorian Amazon represent an alternative for the control of agriculturally relevant pathogens. A microbial bank of 104 endophytes was isolated and characterized, including 43 bacteria and 61 fungi. Among these, the isolates BEC-024 (Bacillus subtilis) and BEC-012 (Bacillus velezensis) showed the highest antagonistic potential against Alternaria spp., Botrytis spp., Fusarium spp., and Phoma spp., with larger inhibition halos and a broad spectrum of action. Metabolic expression under optimized conditions enabled the production of bioactive metabolites with high inhibitory activity, particularly in formula F3, composed of broad bean flour and glucose, which promoted mycelial inhibition against all evaluated pathogens. Subsequently, the compounds were concentrated and microencapsulated by spray drying using maltodextrin, obtaining an improved product. These findings demonstrate that the selected endophytes constitute an effective source of bioactive metabolites, whose microencapsulation enhances their stability and antifungal effect. The results suggest the application of these compounds as a biological alternative for the protection of agricultural crops, promoting sustainable, safe, and low-environmental-impact strategies against phytopathogenic fungi, thereby contributing to food security and the development of more responsible agricultural practices.Item Análisis de la capacidad antifúngica de polifenoles microencapsulados obtenidos a partir de orujo uva (Vitis vinífera l.) frente a Botrytis spp.. y Fusarium spp.(Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos y Biotecnología. Carrera de Biotecnología, 2026-07) Pilatasig Tibán, Myriam Rocío; López Hernández, Orestes DaríoThe generation of wine-related waste, such as grape pomace (Vitis vinifera L.), presents an environmental problem due to its high organic load and limited industrial valorization. Furthermore, the control of plant pathogens like Botrytis spp. and Fusarium spp. relies on synthetic fungicides, and their prolonged use has led to fungal resistance and adverse environmental effects. In this context, this project seeks to add value to grape pomace through the application of biotechnology, by utilizing bioactive compounds such as polyphenols, which possess antifungal, antioxidant, and anti-inflammatory properties, among others. Polyphenols were obtained using a solid-liquid extraction method with different concentrations of ethanol and water, and the total phenolic content was quantified using the Folin-Ciocalteu method. Ethanol at 96 percent showed the highest extraction efficiency. In microencapsulation, maltodextrin showed better performance and efficiency results compared to the maltodextrin-gum arabic combination. FTIR analysis confirmed the presence of functional groups characteristic of phenolic compounds and interactions between the extract and the encapsulating matrix. In vitro antifungal evaluation revealed the inhibitory activity of microencapsulated grape pomace polyphenols against Botrytis spp., with increasing halos observed as the polyphenol concentration increased, while Fusarium spp. showed no sensitivity under the tested conditions. In conclusion, microencapsulated grape pomace polyphenols represent a promising alternative for the biological control of Botrytis spp. This study could contribute to the development of environmentally friendly biofungicides and the biotechnological valorization of agro-industrial waste.Item Diseño de la variante mutante N233K de la enzima PETasa de Ideonella sakaiensis(Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos y Biotecnología. Carrera de Biotecnología, 2026-07) Orellana Cordova, Julio Jaker; García Solís, Mario DanielThe exponential increase in polyethylene terephthalate waste represents a severe global environmental crisis, demanding efficient biotechnological alternatives. This work evaluates the impact of the N233K point mutation on the M5 variant of the Ideonella sakaiensis PETase enzyme. This modification aims to rigidify a flexible surface loop through an intramolecular salt bridge to increase the thermal stability of the biocatalyst, offering a green and sustainable pathway aligned with the circular economy. Methodologically, the mutant gene was expressed in Escherichia coli Rosetta (DE3), yielding 2.09 grams of wet biomass in 550 milliliters of LB medium. After cell lysis by sonication, the recombinant protein was isolated using immobilized metal affinity chromatography (IMAC-FPLC) with a gross elution volume of 15 milliliters. Subsequently, the pool was purified and desalinated by gravity size-exclusion chromatography into a final volumen of 2 milliliters. Analytical quantification was performed on a spectrophotometer applying a corrected mass-specific extinction coefficient of 1.239 liters per gram centimeter. In conclusion, the process consolidated a real net mass of 5.745 milligrams of pure enzyme, which is equivalent to an overall yield of 7.74 percent and a final volumetric productivity of 11.490 milligrams per liter of culture. Gravimetric assays at 50 degrees Celsius for 72 hours confirmed an average mass loss of 10.87 percent in the plastic films, validating that the substitution autonomously confers the necessary thermostability to catalyze the hydrolysis of real PET.Item Análisis de variantes genéticas asociadas al cáncer de mama y su impacto en la prevalencia de la enfermedad estudiando diferentes poblaciones(Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos y Biotecnología. Carrera de Biotecnología, 2026-07) Mesias Barreno, Joel Esteban; Galarza Galarza, Cristian FernandoBreast cancer is a clinically relevant neoplasm due to its marked molecular heterogeneity, reflected in transcriptomic, regulatory, and genetic differences that influence tumor progression, therapeutic response, prognosis, and biological behavior of the disease. Therefore, its characterization using bioinformatics approaches allows for a more precise understanding of the variability among tumor subtypes, the recognition of mechanisms associated with aggressiveness, and provides useful information for the identification of potential biomarkers. Samples corresponding to the estrogen receptor-positive, progesterone receptor-positive, human epidermal growth factor receptor type 2- positive, and triple-negative breast cancer subtypes, obtained from NCBI, were analyzed using a bioinformatics workflow that included quality control, read preprocessing, alignment to the reference human genome, gene quantification, differential expression analysis, functional enrichment, detection of alternative splicing events, identification of genetic variants, functional annotation, and complementary review of prioritized candidate genes. The results showed that triple-negative breast cancer presented the most divergent transcriptomic profile, with a greater number of differentially expressed genes and processes associated with cell proliferation, DNA repair, RNA processing, extracellular matrix, tumor metabolism, and alternative splicing. The integration of differential expression, splicing, and variants allowed prioritizing genes such as QSOX1, MTCH2, and PTPRN2, providing relevant bioinformatics evidence for future research aimed at the experimental validation of biomarkers and the development of precision medicine strategies with possible application in the personalized management of breast cancer.Item Evaluación de la expresión génica de Hylocereus megalanthus (Pitahaya Amarilla) bajo condiciones de estrés por Metil Jasmonato (MeJa)(Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos y Biotecnología. Carrera de Biotecnología, 2026-07) Torres Merino, Joselyn Micaela; Lalaleo Córdova, Liliana PaulinaYellow pitaya (Hylocereus megalanthus) is a crop of increasing agricultural and commercial importance; however, the molecular information available on its defense mechanisms under stress conditions remains limited. This study evaluated the gene expression response of Hylocereus megalanthus using methyl jasmonate as an elicitor to simulate stress signals that the plant may experience in its environment in response to biotic factors. In vitro plants were treated with methyl jasmonate at a concentration of five hundred micromolar and analyzed at different exposure times (0, 4, 24, 48, 72) hours. The experimental approach included RNA extraction using TRI Reagent, complementary DNA (cDNA) synthesis, in silico identification of candidate genes associated with plant defense, preliminary primer validation through conventional PCR and electrophoresis, and relative expression analysis by RT qPCR using PowerUp SYBR Green Master Mix. During the treatment period, the plants did not show visible morphological alterations, which allowed the recovery of viable plant material for molecular analysis. RNA extraction and complementary DNA synthesis produced sufficient concentration for subsequent analyses, although variations among samples were observed. During preliminary validation, the reference genes Actin and EF1 alpha showed visible amplification, as did the candidate genes MYB, NAC, and APE. Subsequently, normalization with Actin allowed a clearer interpretation of relative expression, with MYB and APE showing the most consistent responses to the treatment. These findings provide useful information for understanding molecular defense responses in yellow pitaya and suggest MYB and APE as genes of interest for future biotechnological studies.