Carrera Ingeniería Bioquímica
Permanent URI for this collectionhttp://repositorio.uta.edu.ec/handle/123456789/809
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Item Aprovechamiento de residuos orgánicos obtenidos en invernaderos de la provincia de Tungurahua(Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos y Biotecnología. Carrera de Ingeniería Bioquímica, 2022-09) Villacis Chiliquinga, Saul Elías; Fernández Rivero, DanaeThe development of greenhouse crops has increased in Ecuador in recent years, the main cultivated species are S. lycopersicum L (tomato), Cucumis sativus L (cucumber) and Capsicum anmuum L (pepper), of which it is estimated that the production of organic waste is around 151705.5 tons in the country, for which, the present bibliographic review focused on identifying possible alternatives for the use of agricultural waste as a source of extraction of metabolites and biomass for the production of biocomposites of biotechnological interest. The results of the research show that tomato fruits have a high content of lycopene, which can be extracted from a solid-liquid extraction with a mixture of hexane-ethanol-acetone solvents and by maceration in vegetable oil, methodologies that They allow to extract more than 90 percent of the lycopene from the tomato fruit, in addition, the possibility of using the biomass residues of the three species as a source for the generation of compost enriched with Cry proteins produced by Bacillus thuringiensis bacteria through solid state fermentation was evaluated. (FES), which are widely recognized for their ability to act as a non-phytotoxic insecticide. In conclusion, the research determined that agricultural residues produced in greenhouses can be used as a source of biocomposites of biotechnological interest in the country's food and agricultural industry.Item Desarrollo de un estudio de estabilización de residuos peligrosos para los laboratorios de la Facultad de Ciencia en Ingeniería en Alimentos y Biotecnología (FCIAB)(Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos y Biotecnología. Carrera de Ingeniería Bioquímica, 2022-09) Tene Moyolema, Wilmer Edison; Peñafiel Ayala, Rodny DavidThe current generation of waste has increased proportionally to population growth. Of these, the university sector is one of which generates waste from the use of laboratories, which by not treating it represents an imminent danger to the environment. Therefore, this project aims to carry out a study of stabilization-solidification of hazardous waste from the laboratories of the Faculty of Science and Engineering in Food and Biotechnology. Belonging to the Technical University of Ambato. Where an inventory was generated classifying waste into three large groups such as: waste from LACONAL laboratories, research, and academics. Additionally, it was estimated that the annual production of waste was 948 kg. Said residues were treated by means of a stabilization-solidification procedure based on pozzolanic portland cement and lime. With which 4 different treatments were elaborated with different proportions of cement, lime, and residue. Obtaining the following favorable treatments. For LACONAL waste, the fourth treatment, for academic laboratory waste, the second treatment, and for academic laboratory waste, the fourth treatment. Additionally, an advanced oxidation treatment was carried out with one of the residues, where favorable results were obtained, reducing the COD by 91 percent, however, when trying to scale this process and try to treat all the residues, a removal percentage of 65 was obtained percent, recommending the use of an additional treatment.Item Prospección de la enzima PETasa de Ideonella sakaiensis(Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos y Biotecnología. Carrera de Ingeniería Bioquímica, 2022-09) Manzano Rivera, Ruth Abigail; García Solís, Mario DanielPlastic is one of the most used materials worldwide, its use has been around for a long time because its composition is very resistant and malleable to be able to develop many necessary tools and products. One of the best-known plastics is PET, which tends to be very resistant when wanting to degrade it. Many types of degradation have been used such as chemical, physicochemical, and biological. However, it has great limitations by resorting to polluting reagents, exaggerated temperatures and poor handling of the waste that causes degradation. One of the enzymes that degrade PET IsPETase from the bacterium Ideonella sakaiensis. The action of different mutants and variants that increased from 1.4 to 120 times the activity of the enzyme was described, the crystal structures of their mutants were also shown with an image resolution ranging from 1.4 to 2.00 angstrom, the crystal structures were found in the PDB, each residue presented a suitable polarity to the degradation of the PET surface. The double mutant with the highest Tm value was W159H, F229Y with a degradation rate equivalent to 23.4 mg of PET per hour, per milligram of enzyme. The adjuvants found were adequate to measure the kinetic rate of product formation and the binding of the substrate to the active site, the binding of SDS and dodecyl trimethylammonium helped the substrate to enter more easily into the active site of the enzyme.Item Actualidad de la reutilización y reciclaje de desechos provenientes de curtiembres, revisión en Ecuador y el mundo(Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos y Biotecnología. Carrera de Ingeniería Bioquímica, 2022-09) Anchatipán Bastidas, Dayana Mishel; Flores Tapia, Nelly EstherTanneries are industries whose main activity is leather tanning, to obtain products of commercial value such as: bags, shoes, wallets, jackets, belts, etc. The leather tanning process generates solid, gaseous, and liquid waste, with solid and liquid waste being the most abundant. These are discharged into water resources and landfills; due to toxic compounds they contaminate these ecosystems causing the death of their flora and fauna. Therefore, the application of recycling processes and reuse of solid waste and liquid effluents from tanneries for their use as raw material to manufacture value-added products and the reuse of the same wastewater for the tanning process. The basis of this research project consists in the collection of information using the Connected-Paper and RedCube-Paper software where it was found that the solid waste was used for the production of glues, hydrolyzed collagen, production of gelatin and amino acids, biofuels. , fatliquoring for leather and biogas, while chrome is recovered from liquid effluents for transformation into chrome salts reused as a tanning agent, which is why this project aims to make the feasibility of applying recycling and reuse process strategies that are used worldwide for solid waste and liquid effluents generated by Ecuadorian tanneries, promoting the reduction of these wastes and the reduction of the environmental impact caused by them in Ecuador.Item Diseño y validación in silico de primers para la construcción de las variantes mutantes S121E, S121D, D186H y R280A de la enzima PETasa de Ideonella sakaiensis mediante tres diferentes métodos de mutagénesis dirigida al sitio(Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos y Biotecnología. Carrera de Ingeniería Bioquímica, 2022-03) Herrera Aldaz, Bryan Alexander; Cerda Mejía, Liliana AlexandraThe PETase enzyme from Ideonella sakaiensis (IsPETase) can be used to degrade PET, however, although it has been reported to date to have the highest enzymatic activity under normal conditions of all PET degrading enzymes, its low thermal stability limits its analysis and application. Therefore, by targeted mutagenesis, specific mutations can be introduced into the DNA to significantly increase its enzymatic activity. The purpose of this study was the design and in silico validation of primers for the construction of S121E, S121D, D186H and R280A mutant variants of IsPETase using three site-directed mutagenesis techniques: QuikChange, Q5 Site-Directed Mutagenesis and Phusion Site-Directed Mutagenesis. For the design of the primers, the parameters contained in the design guidelines for targeted mutagenesis methods were considered. The results showed that the 24 primers obtained meet the general criteria such as length, melting temperature, percentage of GC content and the position of the mutation in the primer. For the validation of the proposed primers, hairpin, autodimer and heterodimer analysis was performed with bioinformatics tools, the calculated values are within acceptable ranges. Finally, the specificity of the primers design was carried out by means of alignments, whose results indicate a high specificity for their respective targets. Likewise, an in silico PCR assay confirmed the specificity of the primers, which represent the expected amplification product, thus demonstrating the correct selection of the designed primers and the execution of the mutagenic PCR process.Item Análisis estructural de la enzima PETasa de Ideonella sakaiensis y enzimas homólogas(Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos y Biotecnología. Carrera de Ingeniería Bioquímica, 2022-03) Aguirre Muñoz, Adriana Alexandra; García Solís, Mario DanielPolyethylene terephthalate (PET), a low-cost thermoplastic produced industrially from fossil raw materials, is massively used in industry, and has become a high-impact pollutant. Among the latest discoveries related to the enzymatic treatment of PET is PETase from I. sakaiensis (IsPETase), which is a hydrolase capable of degrading PET. The present work was elaborated to establish possible mutations and modifications that improve the catalytic activity and enzymatic stability of IsPETase, for which a structural analysis of the same was carried out. Using the UNIPROT and PDBeFOLD databases, the enzymes that share a high degree of homology with IsPETase were identified. From these results, an analysis was carried out based on their percentage of sequence identity and secondary structure identity (SEQ and SSE respectively), E-value, Score, RMSD, and Q. The enzymes of greatest interest were selected for this analysis, the main ones being PbLipaseT, RgPETase, BbHydrolase, and SvCUT190, establishing that the N212S mutation can be used to improve the enzymatic activity and stability, in addition, possible substitutions to be studied were described to evaluate the effect that these would cause in the enzyme (S188E, S188A, and S114H). In the analysis of mutant variants, the presence of different amino acids in each case was identified; therefore, different possible substitutions to be made in the IsPETase enzyme were proposed, among the most relevant ones are W159F and S238T.Item Modelación matemática para la predicción de biomasa residual húmeda por efecto de poda, en las plantaciones de Persea americana del cantón Patate(Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos y Biotecnología. Carrera de Ingeniería Bioquímica, 2022-03) Aguaguiña Paguay, Oscar Mateo; García Pazmiño, María EugeniaThe objective of this research was to obtain a mathematical model for the prediction of residual biomass due to pruning in Persea americana (avocado) plantations from easily measurable dendrométric variables in the field. This species that has been cultivated in Ecuador due to its high consumption, which leads to accelerated forest growth, where pruning is carried out to control productivity, generating a considerable amount of organic waste that causes poor disposal and burning of the same, producing waste and gases that affect the planet's biosphere. Mathematical models for the prediction of wet residual biomass were presented applying the multiple linear regression technique, based on three variable selection criteria, where the statistical descriptions were analyzed and thus a regression model was selected from the selection step by step backwards to describe the behavior of the experimental data, given that it met most of the acceptance assumptions except for multicollinearity. Specifically, the model provides satisfactory results for this field with a determination coefficient of 62.68 percent, from the equation that involves the use of variables such as crown height, total height, stem diameter, crown diameter. The mathematical model was validated using the Cross-Validation technique, concluding that the data estimated by the model presented a good correlation with the real data.Item Elaboración de cerveza en base de harina de residuos agroindustriales de cacao(Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos y Biotecnología. Carrera de Ingeniería Bioquímica, 2021-01) Torres Guncay, Franco Michael; Terán Mera, David AndrésIn brewing beer, can be used different raw materials that have carbohydrates in their structure, barley is the main one but flours from other vegetables such as cocoa can also be used, these mixtures allow adding new organoleptic characteristics and reducing the cost of production of the beers. In this work a beer based on barley and cocoa residues flour (1 to 3) was obtained through the protocol for the elaboration of craft beer, in addition, the ability to degrade carbohydrates and obtain fermentable sugars from P. barcinonensis and enzymes, to incorporate it into the process. After obtaining the product, an analysis of pH, density, acidity, alcoholic degree was carried out following the Ecuadorian standards for this type of alcoholic beverage, later a tasting was carried out to determine its characteristics of color, flavor, consistency foam, cocoa aroma, alcohol aroma, turbidity and other parameters and a survey to know the acceptance of the product. Finally, the calculation of the production and sale prices was established, which were established at 1.53 dollars and 2.50 dollars, respectively. The label, the technical sheet, the description, and the composition of the final product were made, which serves as a basis for positioning this product in the market, with this work it was observed that it is feasible to develop a beer with another raw material and obtain a product that complies with the regulations, that is profitable and highly acceptable by consumers.Item Extracción asistida por ultrasonido de licopenos provenientes de residuos agroindustriales de tomate de árbol (Solanum betaceum Cav.) con capacidad antioxidante(Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos y Biotecnología. Carrera de Ingeniería Bioquímica, 2021-01) Proaño Tapia, Jennifer Karolina; Fernández Rivero, DanaeThe objective of this research was the extraction of lycopene’s with antioxidant capacity by ultrasound, using the agro-industrial residues of tree tomato (Solanum betaceum Cav.). The extraction was carried out from previously dry and ground vegetable residue using ethanol as solvent and an experimental 3 design was applied to develop an ultrasound technology for the extraction of lycopene’s. The influence of the factors plant material/solvent volume ratio (1/2.5; 1/5; 1/10), extraction temperature (20, 30 and 40 degrees Celsius) and extraction time (15, 30 and 60 minutes) on the extraction efficiency percentages. The process was optimized with a plant material/solvent volume ratio of 1:10, extraction temperature of 40 Celsius degrees and extraction time of 60 minutes. To verify the antioxidant activity of the lycopene-rich extract, in vitro tests were performed using the DPPH (2,2-diphenyl-1- 1-picrylhydrazyl) technique and in vivo with Saccharomyces cerevisiae Cav. as model. The extract rich in lycopene’s presented in the in vitro test a percentage of inhibition of free radicals of 60.23 percent, which is equivalent to 693.81 micromoles per liter of Trolox, the antioxidant activity in vivo was corroborated showing that the cell managed to overcome the oxidative stress when exposed to a concentration of 3 millimoles of hydrogen peroxide presenting cell growth values very similar to those obtained with vitamin C.Item Análisis Filogenético de Estructura y Genoma de enzima PETasa de Ideonella sakaiensis(Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos y Biotecnología. Carrera de Ingeniería Bioquímica, 2021-01) Garcés Cifuentes, Andrés Benjamín; Terán Mera, David AndrésThe bioinformatic studies presented on this work consisted of phylogenetic analysis, structure modeling and molecular docking. On the phylogenetic analysis, more than a hundred identified sequences like PETase from Ideonella sakaiensis were aligned with it. Said information was used to identify high homology sequences that codify for enzymes with possible PET hydrolytic activity. Through this procedure three enzymes were found that meet similar features. The enzyme Pbs from Acidovorax delafieldii (PbsA), DHL from Rizhobacter gummiphilus and DHL from Polyangium brachysporum. Protein structure modelling was performed using SWISSPROT. This needed since the three enzymes did not possess a crystallographic model present on any database. The templates used for the modelling reported are solution higher than 1.8 Armstroms. The molecular docking procedure was performed through AutodockTools. A PET monomer was used as ligand. As result the fixation energies of each enzyme were determined through enzymatic kinetics calculations. This value is a good indicator of substrateenzyme affinity. Finally, plasmid extraction, cell transformation and qualitative analysis of enzymatic activity was carried out in the E. coli (rosetta) cell line. The importance of this study lies in the discovery of PETase activity in already known proteins, which offers more biotechnological options for the treatment of plastic waste, which is an environmental problem that has progressively become a priority worldwide.