Carrera Ingeniería Bioquímica

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    Análisis bioinformático de péptidos con actividad antimicrobiana
    (2022-03) Acosta Bustos, Daniela Estefanía; García Solís, Mario Daniel
    This project was made with the purpose of studying the great antimicrobial potential of different proteins like response to developing of resistance of pathogenic microorganisms to conventional antimicrobial compounds. To reach this goal, a total of 126 peptides was collected and analyzed from the PDB and UniProt. The analyzed proteins belong to various species and have been into 12 groups such as cecropins, magainins, bombinins, dermaseptins, cathelicidins, brevinins, esculentins, ranalexins, defensins, drosocins, PR- 39 and gaeugerins. Multiple sequence alignment showed that peptides have highly conserved regions, which generate folding patterns responsible for their activity. When ordering the 126 antimicrobial peptides into 4 groups according to their secondary structure, a high conservation was identified between peptides of different classes and origin, this explains the similarities that these peptides present when interacting with the phospholipid bilayer of microorganisms, proving that the most used mechanisms of action is the “barrel stave” mode, the amphipathicity of these molecules allows the hydrophobic end to interact with the lipid core of the membrane, while the hydrophilic end is directed inwards, producing an aqueous pore. Finally, a phylogenetic tree was constructed by the maximum likelihood method for each class of antimicrobial peptides. From this analysis it was concluded that although the analyzed sequences belong to different species, these are related because they maintain the same evolutionary line and mechanism of action. In summary, this study offers a starting pint for the identification of peptides of interest through the use of open databases, which potentially antimicrobial activity.
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    Mecanismo, estructura e inhibición de la enzima Acetohidroxiácido Sintasa
    (Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos y Biotecnología. Carrera de Ingeniería Bioquímica, 2021-03) Albán Ruiz, Arianna Gabriela; García Solís, Mario Daniel
    The Acetohydroxy acid ctase (AHAS, E.C. 2.2.1.6) or acetolactate withouttase (ALS), is an enzyme dependent on thiamine diphosphate (ThDP) which is present in plants and microorganism. It participates in branched-chain amino acid biosynthesis BCAAs. In the presence of ThDP, FAD and a divalent metallic, it comprises the condensation of two pyruvate molecules into 2-acetolatate or a pyruvate molecule or one of 2-cetobutirate into 2-acete-2-hydroxybutyrate to form the valine, leucine and isoleucine. Structurally, four catalytic subunits (CSUs) interact with four (plant) or eight (yeast) regulatory subunits RSUs forming the enzyme complex AHAS. CSU owns the active site, while RSU increases the CSU activity and provides enzymatic sensitivity to feedback inhibition. Its inhibition has a biocidal effect, which occurs through different reactions and molecular interactions, where the best known is produced by molecules with herbicidal activity, making the enzyme an important target for the development of more than 58 herbicides. In recent years, research has focused on using AHAS inhibitor herbicides as antimicrobials, taking advantage of the absence of the enzyme in humans. Also, mutations and new inhibitors are being studied with promising results. Therefore, this study proposes to consolidate the available literature about the structure, function, and inhibition of AHAS to contribute to future projects by defining research needs in this area and the prospects for the use of the enzyme or its inhibitors in medical, biotechnological, bioinformatics, other applications.
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    Efecto de la aplicación postcosecha de ozono gaseoso sobre la tasa respiratoria y la calidad microbiológica de mora de Castilla (Rubus glaucus Benth)
    (Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos. Carrera de Ingeniería Bioquímica, 2018-11) Chonata Jiménez, Erika Brigitte; Arancibia Soria, Mirari Yosune
    The fragile structure of Andean blackberries (Rubus glaucus Benth) influences in a relatively short shelf-life and limited by the microbial contamination. The aim of the present investigation was to study the effect of the postharvest application of gaseous ozone (0.5 and 0.7 ppm) on the respiratory rate and the activity of native microorganisms (aerobic mesophiles, psychotrophs, molds and yeasts) and inoculated pathogens (Botrytis cinerea, Escherichia coli, Salmonella enterica, Staphylococcus aureus). The results show that the treatment with ozone did not affect the respiratory rate of the berries. In addition, an antimicrobial effect of ozone was observed in aerobic mesophiles, psychotrophs and molds and yeasts with growth inhibition of up to 0,75; 1,9 y 2,25 logarithmic units respectively. Regarding to the inoculated microorganisms, the maximum growth inhibition of B. cinerea was 0.69 logarithmic units with the concentration of 0.5 ppm O3. In the case of E. coli and S. enterica, we worked with blackberry pulp, with adjusted pH at optimum values for the growth of each bacterium, and there was limited reduction of the microbial load due to the surface action of the ozone. Finally, there was no growth of S. aureus due to factors other than the ozone action.
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    Extracción de capsaicina y evaluación de su actividad antimicorbiana frente a: Aspergillus niger, Bacillus cereus, Clostridium perfringens, Escherichia coli, Pseudomonas aeruginosa
    (Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos. Carrera de Ingeniería Bioquímica, 2017-04) Chipantiza Quinatoa, Hernán Patricio; Dugarte Jiménez, Nahir Yerely
    The purpose of this research was to optimize the process of extracting capsaicin (Oleoresin) from two different chilli by the continuous extraction method using a soxhlet extraction equipment modifying the number of discharges and the concentration temperature in the rotavapor. The number of discharges and the optical temperature for the variety of capsicum peppers was 3 and 40 C ° respectively, while for the variety of chilli peppers (frutescems) it was 2 and 40 C °. The ability of inhibition of the extracts obtained was evaluated in the pure state as incorporated into a chitosan polymer matrix in the resolution, film and microencapsulated film to preserve and mask the pepper extract. The antimicrobial activity was higher in Gram - (Escherichia.coli, Pseudomonas aeruginosa) in both the filmogenic and microencapsulated solutions. The formation of smaller halos in Gram + (Bacillus cereus, Clostridium perfringens) was observed and in fungi no halos of inhibition were formed.
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    Actividad antimicrobiana de recubrimientos de Quitosano y Ɛ-polisina en moras inoculadas con Escherichia coli, Salmonella enterica y Botrytis cinerea
    (Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos. Carrera de Ingeniería Bioquímica, 2017-02) Cabezas Medina, María Samantha; Horvitz Szoichet, Sandra Susana
    The aim of this research was to study the antimicrobial activity of chitosan and ε-polylysine coatings on blackberries (Rubus glaucus Benth) inoculated with Escherichia coli, Salmonella enterica (103 cfu•g-1) and Botrytis cinerea (103 conidias•ml-1). The inoculated blackberries were stored at 6 ± 1 °C for 10 days and microbiological analyses were evaluated every two days. Maximum inhibition of E. coli growth with chitosan coating was 3,22 log cfu•g-1 and S. enterica was 3,24 log cfu•g-1 for ten days of evaluation while growth of B. cinerea was inhibited in day 8. Likewise, ε-polylysine coating inhibited S. enterica growth during ten days and, on days 8 and 10, it reduced also the proliferation of E. coli (2,88 and 3,22 log cfu•g-1, respectively) and B. cinerea. Blackberries’ acid pH enhanced both, pathogenic bacteria’s inhibition and growth of the mold B. cinerea. From this work, it was concluded that chitosan and ε-polylysine coatings have antibacterial and antifungal activity and that this activity was higher as the exposure time of the coating with the microorganisms was longer.
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    Investigación de la actividad antimicrobiana y fitoquímica de extractos de plantas medicinales frente a los microorganismos patógenos Escherichia coli y Candida albicans
    (Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos. Carrera de Ingeniería Bioquímica, 2014) Yánez Arcos, Gabriela Isabel; Velasteguí Sánchez, José Ramiro
    Para inhibir el crecimiento de los microorganismos Escherichia coli y Candida albicans se realizaron 4 metodologías de obtención de los extractos de 8 especies vegetales. Las metodologías de extracción emplearon solventes de diferente polaridad tales como: agua, etanol y hexano. Las plantas medicinales estudiadas fueron: albahaca (Ocimum basilicum), ambo (Nicandra physalodes),guayaba (Psidium guajava), hierba luisa (Cymbopogon citratus), matico (Aristeguietia glutinosa), ortiga negra (Urtica dioica), paico (Chenopodium ambrosioides), tomillo (Thymus vulgaris). Los extractos que demostraron los mejores porcentajes de inhibición del crecimiento de Escherichia coli fueron la maceración en etanol de tomillo, con una media del 100% de inhibición del crecimiento; el macerado en etanol de paico, con una media del 100% de inhibición del crecimiento; decocción de tomillo, con una media del 75% de inhibición del crecimiento.Y, para la inhibición de Candida albicans fueron la maceración en etanol de paico, con una media del 100% de inhibición del crecimiento; el macerado en etanol de tomillo, con una media del 100% de inhibición del crecimiento; maceración en etanol de matico, con una media del 75% de inhibición del crecimiento. En el caso del extracto etanólico de paico este demostró que inhibe en un 100% el crecimiento de Escherichia coli a una concentración de 2500 mg/l.En cambio, el extracto etanólico de tomillo inhibe el crecimiento de la bacteria a una concentración de 5000 mg/l. Para la levadura Candida albicans el extracto etanólico de paico y el extracto etanólico de tomillo demostraron que inhiben el crecimiento de la levadura a una concentración de 2500 mg/l. Finalmente el análisis fitoquímico determinó que las hojas de paico y tomillo contienen metabolitos secundarios tales como: aceites esenciales, taninos, flavonoides y triterpenos, esteroides, alcaloides (paico) y saponinas (tomillo).
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    Determinación del potencial antimicrobiano de la microalga Chlorella extraída de las aguas empozadas de las acequias de Atocha, Tilulum y el Socavón del cantón Ambato
    (Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos. Carrera de Ingeniería Bioquímica, 2014) Barona Altamirano, Adriana Carolina; Valencia Silva, Alex Fabián
    Las microalgas son organismos unicelulares eucariotas fotosintéticos capaces de transformar la energía luminosa en energía química; las condiciones óptimas de crecimiento varían de una especie a otra, las investigaciones sobre el cultivo de microalgas son de gran importancia dada su amplia aplicación biotecnológica y comercial. Chlorella es una microalga verde unicelular de agua dulce, de forma esférica mide cerca de 2-10 micrones de diámetro, tiene alta concentración de clorofila, se reproduce rápidamente y de forma asexual, necesita carbono agua luz y pequeñas cantidades de minerales.Se recolecto diferentes muestras de aguas empozadas de Atocha, Tilulum y el Socavón de la ciudad de Ambato, los cultivos de la microalga Chlorella se aislaron utilizando el medio Basal de Bold debido a la cantidad de nutrientes que posee, las células tardaron 240 horas en alcanzar la fase exponencial para su posterior análisis. Se determinó el potencial antimicrobiano de la microalga en base a los resultados positivos en los ensayos en los que se utilizó el precipitado que resulta de la extracción del chlorellin con los solventes etanol y alcohol isopropílico. El análisis estadístico determinó que el mejor tratamiento fue Tilulum- Alcohol isopropílico al tener mayor el tamaño del halo de inhibición, cuya media fue de 28mm, ante la bacteria coco grampositiva, mientras que el menor tratamiento fue Socavón-etanol con una media de 0,11mm. En la última etapa de la investigación se realizó las pruebas de control positivo mediante la técnica del antibiograma utilizando la bacteria aislada y antibióticos como penicilina, demostrando que fue sensible con un halo de inhibición de 5mm ante la ampicilina y 1mm ante oxacilina, valores que están dentro del rango de sensibilidad de acuerdo a las tabla CIM (Concentración Mínima Inhibitoria).