Ciencia e Ingeniería en Alimentos y Biotecnología
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Item Caracterización de la celulosa bacteriana producida a través de combinaciones de Komagataeibacter xylinus DSM2004 y los consorcios PDC21 y PDC25(Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos y Biotecnología. Carrera de Biotecnología, 2024-08) Tiban Mullo, Bibiana Liseth; Cerda Mejía, Liliana AlexandraBacterial cellulose is a biopolymer of great commercial interest due to its high purity, high degree of crystallinity and superior mechanical properties compared to cellulose derived from vegetable sources. However, its high production cost has limited its use on an industrial scale. To reduce these costs, it is essential to explore techniques that optimize cellulose biosynthesis, such as the use of co-culture methods that increase the robustness of the production system and enhance the yield through synergistic interactions. In the present study, bacterial cellulose produced by combinations of a standard strain and the microbial consortia PDC21 and PDC25 was characterized, evaluating the production in co-cultures versus monocultures. Cellulose morphology was analyzed using scanning electron microscopy (SEM), chemical composition by Fourier transforms infrared spectroscopy (FT-IR) and crystallinity was evaluated by X-ray diffraction (XRD). Additionally, the resulting fermentation broths were characterized to understand the differences between co-culture and monoculture productions. The co-culture treatments demonstrated higher cellulose yields compared to their monocultures. Although all treatments exhibited similar chemical characteristics, variability was observed in the morphology and crystallinity of the cellulose produced. Significant differences were also identified in the physical and chemical characteristics of the fermentation broths. This study evidenced the potential of bacterial co-cultures to improve cellulose production efficiency.Item Obtención y caracterización de celulosa bacteriana producida por cepas acéticas endémicas del Ecuador(Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos y Biotecnología. Carrera de Biotecnología, 2023-09) Álvarez Morales, Juan José; García Solís, Mario DanielThe research and development of new bacterial strains producing bacterial cellulose (BC) is a topic of great relevance today, especially in the context of biotechnology and industrial applications. This study focuses on native strains from Ecuador, with special attention to acetic bacteria PDC 21 and PDC 25, opening new possibilities for BC production. The methodology of this study involved the axenic culture of the strains and the observation of the formation of BC biofilm. Fermentative processes were carried out and the BC production yield was evaluated in comparison with a control of Komagataeibacter xylinus (DSMZ 2004). In addition, acid, thermal, and mechanical treatments were performed to obtain nanocellulose. The morphology of the samples was analyzed by scanning electron microscopy (SEM), while Fourier Transform Infrared Spectroscopy (FTIR-IR) was used to evaluate their chemical properties. The study concludes with the proposal that strain PDC 25 could be a viable alternative for BC production in industrial applications. This conclusion is based on its production yield, glucose consumption, and the properties of the BC it produces, which are similar to those of strain DSMZ 2004, commonly used as a study model and in the industry. This finding opens new possibilities for BC production and biotechnology in Ecuador.Item Análisis in silico de las rutas metabólicas necesarias para la síntesis de celulosa bacteriana en Komagataeibacter xylinus a partir de lactosa(Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos y Biotecnología. Carrera de Ingeniería Bioquímica, 2022-09) Zapata Berrones, Karina Elizabeth; García Solís, Mario DanielBacterial Cellulose symbolizes a multifaceted resource of exceptional capabilities and fascinatingproperties. Currently, the demand for this exopolysaccharide has progressively increasing, in function of the optimization of processes and the implementation of agro-industrial waste as alternative carbon sources. In such a context, the archetype productor (K. xylinus) represents one of the most valuable organisms in the biotechnological field, a scenario that has made it possible to demonstrate its enormous power to synthesize BC from a wide range of substrates. However, some authors have elucidated its inability to metabolize lactose, caused by the lack of genes responsible for the transport and catabolism of this substrate. Here, DSM 2004 and the analogous strain E25 (identified by phylogenetic analysis), were studied with the aim of identifying missing genes and minimal genetic equipment. A critical analysis of metabolic pathways in BAL and BAA was carried out to list the transporter and the enzymes involved in the process of interest. In this, the summarized proteins were evaluated according to kinetic parameters, where the best results report access code and both peptide and nucleotide sequence of the gene that encodes them. In this, the genes of Escherichia coli K-12 (Lactose permease), Alicyclobacillus acidocaldarius ATCC 27009 (β – galactosidase), Bifidobacterium longum subsp. Infantis ATCC 15697 (galactokinase), and Escherichia coli XL1-Blue (galactose-1-P-uridyltransferase) were selected. Finally, the modeling of the new biosynthetic route presented two related modules: i) Transport and hydrolysis of Lactose and ii) Biosynthesis of CB, after the total replacement of glucose by whey lactose.Item Aislamiento y caracterización de cepas nativas de Komagataeibacter xylinus y comparación de su crecimiento en diferentes sustratos(Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos y Biotecnología. Carrera de Ingeniería Bioquímica, 2022-03) Dávalos Cerrón, Patricia Yissel; Cerda Mejía, Liliana AlexandraKomagataeibacter xylinus is the most efficient species, being able to synthesize bacterial cellulose biofilms (CB) from a great variety of carbon sources. This biopolymer has received considerable attention on the last few years due to diverse commercial applications, especially in the food industry and biomedical due to its high purity and variety in form and texture. In the present study, native strains of K. xylinus were isolated and characterized from acetic sources such as wine, wine vinegar and kombucha SCOBY in the media H-S and GYC comparing its production with the strain from control K. xylinus DSM 2004 in the different carbon sources (glucose, whey and ethylene glycol) in static cultivation. Two strains were isolated from the kombucha SCOBY. The results revealed that both strains denominated PDC 21 and PDC 25, and the control strain synthesized CB in different amounts from the glucose, ethylene glycol and fermented milk serum. The strain PDC 25 produced almost 8 more times CB than the strain control in the standard H-S medium compared with PDC 21 that only synthesized 0.4 times the CB humid, meanwhile with ethylene glycol PDC 25 synthesized 1.4 and 3.46 more times CB in the media H-S ethylene glycol and K- ethylene glycol respectively compared by the control strain. However, K xylinus DSM 2004 produced larger amounts of CB in the media that contained fermented milk serum.Item Análisis filogenético de Komagataeibacter xylinus K2G30=UMCC2756 para la producción de biofilms a partir de suero lácteo(Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos y Biotecnología. Carrera de Ingeniería en Alimentos, 2021-01) Balseca Mora, Evelyn Karina; Cerda Mejía, Liliana AlexandraBacterial cellulose, being an organic, renewable, and biodegradable biopolymer has received considerable attention. Here, the production of bacterial cellulose by Komagataeibacter xylinus K2G30 (UMCC 2756) was studied in static culture using Hestrin and Schramm (HS) medium with total replacement of glucose by whey and lactic acid. Furthermore, phylogenetic trees were made from sequences of the 16S ribosomal RNA gene from the species K. xylinus, K. europaeus, K. hansenii and K. intermedius, which revealed that they lacked the lac operon responsible for lactose hydrolysis. Due to these characteristics, they can explain that the highest production of BC by the K. xylinus K2G30 strain was achieved with the use of lactic acid as the sole carbon source. Results provide important information for future studies on the synthesis of BC from different carbon sources, including raw materials from industrial waste such as whey, which must be previously fermented for better assimilation.Item Producción de biofilms a partir de diferentes matrices biodegradables y evaluación de su actividad antibacteriana(Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos y Biotecnología. Carrera de Ingeniería en Alimentos, 2019-12) Guaypatín Sánchez, Edisson Fabricio; Cerda Mejía, Liliana AlexandraBiodegradable polymers have few viscoelastic, rupture, barrier and antibacterial properties affecting their use in industry. The objective of this research is to produce films with different biodegradable matrices, to improve their properties and evaluate their antibacterial effect by impregnating metal nanoparticles. It was determined that films produced by Komagataeibacter xylinus with a volume of culture medium of 50 ml produced thick films, as in films obtained from filmogenic mixtures with the increase to 20 ml of the final mixture. Stress tests such as axial force and tension related to the strength of materials were performed when axial force is applied in opposite directions; obtaining 21-24 N of axial force and 1.9-3.8 por ciento of tension for the commercial grade glucose per carrageenan mixture being the most representative test. The antibacterial evaluation showed that only films impregnated with silver have an antibacterial effect against E. coli and S.aureus; giving inhibition halos of 1.2 - 1.4 cm in diameter for CB films and 1.3 - 2.5 cm in diameter for films of filmogenic solutions evaluated in E.coli. On the other hand, values between 1.33 - 2.01 cm in diameter were obtained for CB films and 1.09 - 1.29 cm in diameter for films of filmogenic solutions evaluated in S. aureus.Item Producción de biofilm a partir de Komagataeibacter xylinus, utilizando diferentes residuos agroindustriales(Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos. Carrera de Ingeniería en Alimentos, 2019-06) Palacios Duchicela, Ronald Hendry; Terán Mera, David AndrésBacterial biofilms are being an important field of study, due to their wide use in different fields. However, due to its high production costs, its use is limited. For this reason, this work aimed to obtain biofilms using different agroindustrial wastes (sugarcane bagasse, rice bran and cocoa husk), using an H-S medium and minimum medium in static and agitated culture. Demonstrating that agroindustrial wastes contributes to the medium the necessary carbon and nitrogen source, so that the bacterium Komagataeibacter xylinus can develop and generate the biofilms. The biofilms obtained from the cocoa husk showed the best yield in the H-S medium with 17,8338 and 22,8312 percent for the static and agitation culture respectively, while in the minimum medium a yield of 23,3169 and 16,1095 percent was obtained for the static and agitation culture correspondingly; it even showed that there is no statistically significant difference with the standard (glucose). Also, an impregnation with silver and zinc particles was made to the biofilms produced from the cocoa husk. Furthermore, by X-ray diffraction analysis (XRD), it was possible to determine that the biofilms obtained corresponded to Cellulose I, cellulose Iα being present in a higher proportion. The scanning electron microscopy (SEM) helped to observe the structure of the biofilms and by means of a Mapping the presence of the Ag and Zn nanoparticles impregnated in the structure of the biofilm was verified.Item Estimación y efecto sobre la vida útil de fresa (Fragaria x ananassa) con el uso de biofilm producido por Komagataeibacter xylinus(Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos. Carrera de Ingeniería en Alimentos., 2019-05) Acosta Telenchana, Jorge Luis; Terán Mera, David AndrésThe strawberry is susceptible to physiological changes, great vulnerability to microbial attack, the benefits of edible coatings are: slow breathing rate, prolonged periods of storage, retention of firmness and reduced microbial growth, so the objective of this research work is estimate the useful life of strawberry (Fragaria x ananassa) with the use of biofilm produced by Komagataeibacter xylinus. It was determined that the samples with bacterial cellulose coating have an average life time of 12, 6 and 3 days while samples without coating the time was 8, 3 and 2 days at temperatures of 4, 25 and 37 Celsius degrees. The ultra-freeze analysis showed changes of 1 percent; 46.2 percent; 12.6 percent and 2.41 percent in samples without coating, on the other hand the samples with coating changes were 0.3 percent; 15.9 percent; 4 percent and 1.49 percent in pH, Acidity, Brix degrees and weight loss respectively. The samples containing bacterial cellulose coating impregnated with silver nanoparticles showed no growth of microorganisms during the analysis. The container of bacterial cellulose impregnated with natural gelatin presented a 15.3 percent weight loss, 76.4 Brix degrees and a dark red final coloration in the sample, the bacterial cellulose container with end seals presented greater weight loss of 19 percent. , 8 percent; 83.6 Brix degrees and a medium dark red final color.Item Estimación de vida útil de mora de castilla (Rubus glaucus Benth) a partir del recubrimiento de celulosa bacteriana producida por Komagataeibacter xylinus(Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos. Carrera de Ingeniería en Alimentos., 2019-01) Toscano Avila, Jonatan Alfonso; Cerda Mejía, Liliana AlexandraThe Blackberry (Rubus glaucus Benth) is an Andean crop with nutritional and antioxidant properties, whose intake potentiates the immune system and reduces the risk of developing degenerative and cardiovascular diseases. However, it is one of the most perishable fruits due to its high respiration rate and the lack of peel that protects the quality of the fruit, which makes it susceptible to microbial attack and rapid deterioration, which is a problem for commercialization. The objective of this research was to estimate the useful life of Castile mulberry (Rubus glaucus Benth) from the coating of bacterial cellulose produced by Komagataeibacter xylinus, applied in the postharvest quality of the Mora de Castilla, with the purpose of improving both physicochemical characteristics as nutritional. The treatment with biofilm coating of bacterial cellulose at a temperature of 4 Celsius degree allowed to increase the shelf life of the fruit up to 9 days, in addition to maintaining its characteristic texture, color, smell and taste, that is to say that the biofilm generated from Komagataeibacter xylinus, can be used as an antimicrobial agent, in edible coatings for fruits, vegetables, meats, etc.