Ciencia e Ingeniería en Alimentos y Biotecnología
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Item Evaluación del efecto de la enzima Cel6D de Paenibacillus barcinonensis en la degradación de residuos agroindustriales(Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos y Biotecnología. Carrera de Alimentos, 2025-02) Zurita Gancino, Joaquín Fernando; Cerda Mejía, Liliana AlexandraThe accumulation of agro-industrial residues generated by agricultural activity in Ecuador represents a significant environmental challenge because inadequate management can cause contamination and affect public health. The use of specific enzymes for cellulose degradation has been widely studied. Thus, this study focused on evaluating the effect of the enzyme Cel6D from Paenibacillus barcinonensis on the degradation of these residues, seeking a biotechnological alternative for their use and reduction of environmental impact. The methodology employed included the construction of a vector for the expression of the enzyme in Escherichia coli, followed by purification by affinity chromatography. The enzymatic activity of Cel6D was evaluated on various swollen and non-swollen agro-industrial substrates. The results showed that the enzyme presented enhanced enzymatic activity on swollen substrates, highlighting a yield of 1.84 49 milliunits of cellulose activity per milligram on cocoa husk and 1.49 49 milliunits of cellulose activity per milligram on rice bran, suggesting that swelling facilitates the access of the enzyme to cellulose. It was concluded that Cel6D is effective in the degradation of agro-industrial wastes, especially when the substrates are treated by acid swelling to increase its accessibility. This research highlights the importance of optimizing the swelling and purification processes to maximize enzyme activity and its potential application in biorefineries, thus contributing to a circular and sustainable economy in waste management in Ecuador.Item Evaluación de la capacidad antioxidante y estabilidad de un extracto obtenido a partir de residuos de hojas de la cosecha del maíz (Zea mays)(Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos y Biotecnología. Carrera de Biotecnología, 2025-02) Puruncajas Paucar, Angel Santiago; Fernández Rivero, DanaeMaize (Zea mays) is a major crop in Ecuador, generating high volumes of underutilized agricultural waste which, if poorly managed, could pollute the environment or have non-economic value. Maize leaves, rich in biomolecules such as chlorophyll and flavonoids, hold potential for industrial applications in food and cosmetics. This research aims to evaluate the antioxidant capacity and stability of maize leaf extracts to promote their utilization. Two extracts were prepared using 96 percent ethanol and an acetone-ethanol mixture in a 3 to 1 ratio, which were analyzed through FTIR spectrophotometry. Their antioxidant capacity was studied using the DPPH assay. The extracts were stored at different temperatures for four weeks, and their FTIR spectra and antioxidant capacity were examined. The FTIR study indicates the presence of hydroxylated compounds in both extracts, acetone extract containing more diverse groups. The antioxidant capacity was 107.15 and 104.39 micromoles of Trolox per gram of maize leaves for ethanol and acetone-ethanol extracts, respectively. The stability study showed changes in the FTIR spectra of both extracts, with an increase in the presence of new functional groups. The acetone-ethanol extract exhibited greater stability in antioxidant capacity, whereas the ethanol extract showed a significant reduction. Refrigeration proved to be the best condition for preserving antioxidant activity in both extracts.Item Obtención de astaxantina a partir de exoesqueletos de cangrejo rojo (Ucides occidentalis)(Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos y Biotecnología. Carrera de Alimentos, 2025-02) Chiliquinga Angulo, Joel Alexander; Cadena Carrera, Santiago EsmiroAstaxanthin is a carotenoid with outstanding biological activity, and its antioxidant capacity surpasses those of other carotenoids. Since the use of synthetic astaxanthin is restricted, there has been growing interest in obtaining this compound from natural sources. Astaxanthin is responsible for the characteristic reddish hue of the red crab (Ucides occidentalis). In Ecuador, approximately 40 million red crabs are caught annually for consumption, which generates a sanitation problem due to the large amounts of resulting by-products, as 70 to 80 percent corresponds to the inedible fraction of crustaceans. Considering this, the present study evaluated the overall extraction yield and the amount of astaxanthin present in each extract using extraction techniques such as Soxhlet and maceration, considering the extraction time. The overall extraction yield and astaxanthin content were evaluated by gravimetry and spectrophotometry, respectively. The highest overall yield was obtained using the Soxhlet technique at 210 min (0.120 percent), and the highest astaxanthin content was obtained using the Soxhlet technique at 180 min (332.132 microgram per gram). The antioxidant capacity of the extract with the highest astaxanthin content was evaluated by the DPPH method, where 48.779 percent inhibition equivalent to 250.620 micromoles of Trolox per liter was obtained. Therefore, the present investigation demonstrated the production of astaxanthin, evaluating its yield as a function of the extraction time with ethanol and its antioxidant capacity.Item Optimización del proceso de extracción de compuestos bioactivos mediante superficie de respuesta a partir de residuos de camarón(Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos y Biotecnología. Carrera de Alimentos, 2024-08) Villafuerte Herrera, Ronaldo Julián; Cadena Carrera, Santiago EsmiroAstaxanthin is a carotenoid with significant biological activity, even greater antioxidant activity than other carotenoids. Synthetic astaxanthin has usage restrictions, which has led to interest in obtaining it from natural sources. Astaxanthin is responsible for the pinkish color of shrimp, and the cultivation of shrimp has increased globally, along with its negative effects, primarily the generation of waste that constitutes approximately 25 to 60 percent by weight. Considering this, the present study evaluated the extraction of astaxanthin using Soxhlet and modeled the extraction kinetics from shrimp shells (Litopenaeus vannamei). The raw material was characterized by proximal analysis, particle size, and shape. The overall extraction yield and astaxanthin content were evaluated by gravimetry and spectrophotometry, respectively. The highest overall yield was obtained with the ethanol plus water solvent (3.75 percent), and the highest astaxanthin concentration was for the ethanol and methanol solvents (292.2 and 296.0 micrograms per gram, respectively). The model that best fit to describe the extraction kinetics was the Peleg model, with a determination coefficient between 0.93 and 0.98. This research demonstrates the potential recovery of astaxanthin from shrimp waste using ethanol.Item Caracterización cinética de las variantes silvestre y mutantes I208V, N212A y S238Y de la enzima PETasa de Ideonella sakaiensis(Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos y Biotecnología. Carrera de Biotecnología, 2024-08) Sierra Mena, Joselyn Patricia; García Solís, Mario DanielThe accumulation of plastic waste has reached critical levels, with 8 million tons polluting the oceans annually. The resulting microplastics enter ecosystems and organisms, posing a serious threat to environmental health. In response, bioremediation has emerged as a sustainable alternative, employing microorganisms or enzymes to degrade polymers. The enzyme IsPETase from Ideonella sakaiensis is particularly more effective than other PET-degrading enzymes. Thus, mutations close to the active site (S238Y, N212A and I208V) have been recently designed to improve its activity and thermostability, evaluated in qualitative analyses. Therefore, in this research the kinetics of these mutants was characterized by evaluating the kinetic parameters of the reactions catalyzed by these enzymes, using p(NP)-acetate as substrate and varying the temperature (25, 35 and 45 degrees Celsius). The data were analyzed with GraphPad Prism. A reduction in KM was observed in the mutants, indicating that the mutations increased the affinity for the substrate, although the decrease in kcat, reflected a lower catalytic capacity, especially for the I208V variant. Overall, all three mutations are less efficient than the WT variant in hydrolyzing phenyl esters under saturated conditions. Regarding the effect of temperature, the N212A variant showed the highest activity at 45 degrees Celsius, standing out as the most thermostable. This study aims to deepen the catalytic efficiency of IsPETase mutants, contributing to the development of biotechnological recycling for plastic wastes.Item Elaboración de una apanadura a base de residuos de camarón (litopenaeus vannamei), y panko(Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos y Biotecnología. Carrera de Alimentos, 2024-08) Hidalgo Mosquera, Annie Catherine; Paredes Escobar, Mayra LilianaThe inadequate management of waste has a significant environmental impact. The incorrect disposal of organic matter generates contamination, encourages the emission of gases, and generates unpleasant odors, as in the case of shrimp waste. The amount of bread waste represents a waste of valuable resources. It is essential to manage these wastes properly to mitigate their environmental impact and take advantage of their potential by reusing them or giving value to these materials. In the present study, a breading was developed using shrimp (Litopenaeus vannamei) and panko waste, for which the waste was selected through a visual evaluation of its odor, integrity and appearance characteristics. A convective drying, milling and sieving process was applied to obtain a homogeneous mixture that would allow adequate breading, which was adjusted according to sensory and technical-functional parameters such as oil absorption capacity and dough properties. In addition, the proximal composition of the most acceptable breading was determined. The results achieved demonstrate a crunchy texture and an enhancement of the flavor of the breaded food, characteristics that satisfy the consumer. The percentage yield obtained for shrimp shell flour was 29.0 percent and 68.9 percent for panko. The most acceptable formulation was the one obtained by treatment 3, which presented a low oil absorption index of 1.59, an intermediate texture between panko and breadcrumbs of 6386 gram-strength and a raw peak of 81 cP, which shows a viscosity like that of a gum. Regarding proximate composition and microbiological load, the product complied with INEN 2945 and NTE INEN 2532.Item Evaluación del efecto de la enzima Xyn30D de Paenibacillus barcinonensis en la degradación de residuos agroindustriales(Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos y Biotecnología. Carrera de Alimentos, 2024-08) Burbano Palacios, David Fernando; Cerda Mejía, Liliana AlexandraThe extraction and processing of raw materials in the wood, paper, and agricultural industries are responsible for greenhouse gas emissions and the destruction of biodiversity. In response, the use of renewable and earth-abundant raw materials, such as lignocellulose—a xylan-rich component—has been sought. However, the complex lignocellulosic structure presents a challenge to physical and chemical methods of degradation, leading to the proposed use of enzymes for this purpose. The bacterium Paenibacillus barcinonensis is known for its ability to produce extracellular enzymes that can break down biomass, including xylanases of the GH30 family. Among these enzymes, xylanase Xyn30D has demonstrated efficiency in lignocellulose degradation. In this project, we analyzed the activity of xylanase Xyn30D, from plasmid construction to expression in E. coli BL21 (DE3) and purification by immobilized metal affinity chromatography (IMAC). Furthermore, we evaluated its activity in four different agro-industrial wastes with high xylan content, which have a significant impact on the agricultural industry in Ecuador and worldwide. The substrates sugarcane bagasse and corn stover showed positive enzyme activity, while cocoa husk and rice husk did not exhibit significant values in terms of xylose units per gram of enzyme. This lack of activity is attributed to the presence of inhibitors in the chemical composition of the latter substrates.Item Evaluación de la biodegradación y propiedades mecánicas de diferentes formulaciones de bioplástico obtenidas a partir del aprovechamiento de colágeno residual y almidón de papa (Solanum tuberosum)(Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos y Biotecnología. Carrera de Biotecnología, 2024-08) Miquinga Lasluisa, Cinthya Alejandra; Flores Tapia, Nelly EstherPlastic pollution is one of the most critical problems we face, as the population continues to prefer this material for its versatility in daily life. However, the accumulation of plastics causes serious problems in terrestrial and marine ecosystems, affecting all beings that inhabit them. In addition, poor waste management has similar repercussions. This study focused on evaluating the biodegradation and mechanical properties of a bioplastic made from biopolymers recovered from organic waste, such as starch and collagen. For the production of bioplastics, collagen and starch were extracted by thermal hydrolysis and the wet method. Different formulations were used, varying the concentrations of starch and collagen, and two plasticizers were used: polyvinyl alcohol and glycerin. The aim was to develop a rapidly degrading material with optimal mechanical strength characteristics. It was determined that the most efficient collagen extraction was achieved at 120 minutes, with the highest yield. The presence of glycerin increased the flexibility of the sheets, while polyvinyl alcohol gave them more rigidity. The sample with a 1:1 ratio of flesh collagen showed higher tensile strength compared to the others. In biodegradation tests, faster decay was observed in water than on land; On land, the sheets with flesh collagen degraded more quickly. Therefore, it can be deduced that sheets made with flesh collagen and wet blue have optimal characteristics for the manufacture of bioplastics.Item Evaluación de la capacidad de biodegradación de contenedores fabricados a partir de recursos agroindustriales infrautilizados de la industria bananera(Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos y Biotecnología. Carrera de Biotecnología, 2024-08) Méndez López, Tomás Sebastián; Arancibia Soria, Mirari YosuneAccording to the United Nations, approximately 430 million tons of plastic are produced each year. Consequently, this waste represents 85 percent of the total contamination in bodies of water, 36 percent of which are disposable containers. To combat this problem, resources have been focused on the research of more environmentally friendly materials. In Ecuador, banana, due to its high production and waste generation, emerges as one of the best alternatives for the development of bio containers to replace single-use plastics. For this reason, this research evaluated the biodegradation capacity of containers made from underutilized agro-industrial resources of the banana industry. For this purpose, a buried test was carried out with samples of biodegradable banana containers (CBB) in agricultural soil for 15, 30, 45 and 60 days, where values of up to 52.53 percent biodegradation based on dry weight were obtained. On the other hand, in vitro biodegradation capacity was evaluated by inoculation of Aspergillus niger, where it was demonstrated that this strain prefers CBB as a substrate, compared to filter paper and single-use plastics, due to the greater availability of nutrients for its growth. Finally, through microbiological analysis and bibliographic research, it was demonstrated that the shelf life of CBB is equal to or greater than 60 days, since their physicochemical and usability characteristics are not compromised during this period.Item Obtención de nano-celulosa bacteriana utilizando hidrolizados de bagazo de caña y cáscara de cacao(Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos y Biotecnología. Carrera de Biotecnología, 2024-02) Sailema Sailema, Erick Fabricio; Fernández Rivero, DanaeCellulose is one of the most abundant biopolymers on Earth, and various applications in medicine and industry are being investigated. Most of it is found in plants (vegetable cellulose). However, the production of vegetable cellulose faces deforestation and costly purification processes problems. On the other hand, there are bacterial cellulose which has several advantages due to its characteristics; however, its production cost at industrial scale still exhibits problems. Therefore, use agro-industrial waste from cocoa and sugarcane as a carbon source to obtain bacterial cellulose presents a possible solution to the problem. Consequently, it was proposed to evaluate hydrolysis treatments on agro-industrial wastes to obtain fermentable sugars, which will be the carbon source in minimal mediums. Subsequently, cellulose yield produced by two acetic strains (PDC21 and PDC 25) and Komagataeibacter xylinus DSM 2004 (DSM 2004) was evaluated in the same medium. Once the appropriate treatment to produce cellulose was determined, which consisted of the use of cocoa shell hydrolyzate and the DSM 2004 bacteria, two cellulose hydrolysis methods were compared to obtain one of its derivatives, nano-cellulose, which was characterized morphologically by FT-IR and SEM. The research demonstrated the suitability of use hydrolysates of cocoa shell waste and sugarcane bagasse as low-cost carbon sources to produce bacterial cellulose. In addition, it was determined that the hydrolysis treatments used to obtain nano-cellulose do not influence its morphology.