Ciencias Exactas y Ciencias de la Salud

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Design of droplet-based microfluidic devices and its application for protein fractionation
(Instituto Tecnológico y de Estudios Superiores de Monterrey, 2020-12-04) Hernández Cid, David; Mata Gómez, Marco Arnulfo; qro /|bqrotbecerra/tolmquevedo; Pérez González, Victor Hugo; Cervantes Avilés, Pabel Antonio; School of Engineering and Science; Campus Monterrey; González Valdéz, José Guillermo
Microfluidics technology offers new possibilities due to the nature of small scale, such as the high surface area to volume ratio. Droplet based microfluidics is an emerging field that has found its application in different areas, like material science, chemical reactions, and biochemical analysis. On the other hand, downstream processing operations like separation and purification still has some drawbacks like big reagents consumption, huge waste generation, extra steps like concentration or salts removal. Here is where microfluidics can be implemented to overcome some of the issues presented in this field. In this work, we designed and developed a microfluidic platform able to generate droplet-like aqueous two- phases system (ATPS) in a continuous manner. We started this project by performing a study of how different forces like surface tension, inertial forces, viscous forces, and geometry affect droplet formation. We found that all the previously mentioned parameters have an impact in different ways on the process of droplet formation. Afterwards, this knowledge was used to implement a microfluidic flow-focusing on the generation of droplet-based micro ATPS with the used of salts and PEG as the two liquid phases of the ATPS. To see that this ATPS droplet microfluidic system really worked, we tested its capacity to fractionate ribonuclease A (RNase A) on its native and PEGylated forms, which have affinity for the salt and PEG phases, respectively. Thus, we expected that proteins would get inside or outside droplets or streamlines according to their affinity for each of the two phases. In doing so, native, and PEGylated proteins were labeled with FITC and their movement from one to another phase or their retention in the phase where protein was placed were recorded. It was confirmed that proteins move or keep in their phase for which they have more affinity. This result opens the possibility for the development of microfluidic separators based on ATPS droplet formation.
Metabolic flux analysis of Xanthophyllomyces dendrorhous metabolism to understand the production of astaxanthin
(Instituto Tecnológico y de Estudios Superiores de Monterrey, 2020-06) Martínez Castro, Victor Ignacio; MATA GOMEZ, MARCO ARNULFO; 207149; Mata Gómez, Marco Arnulfo; lagdtorre, emipsanchez; Benavides Lozano, Jorge Alejandro; Licona Cassani, Cuauhtémoc; School of Engineering and Sciences; Campus Toluca; Goméz Sánchez, Carlos Eduardo
Carotenoid production by microorganisms, contrary to chemical synthesis, could fulfill the in-creasing demand for human consumption. The yeastX. drendrorhousis one of the most promis-ing and economically attractive microorganisms for industrial production of astaxanthin. Themetabolic pathway through which this yeast synthesize this valuable carotenoid is known. How-ever, the complex mechanisms that are involved in the process, the distribution of the metabolicfluxes and the rates at which the pathways work, remain unknown. Several studies have provideddifferent approaches to manipulate and improve carotenoid production inX. dendrorhousfromclassical mutagenesis to genetic engineering of the complete pathway covering improved precur-sor supply for carotenogenesis, enhanced metabolite flow into the pathway, and manipulating therelation C/N in the culture medium. However, it has not been reported quantitatively how nutri-ents, from the central metabolism and other pathways, converge in the carotenoids biosynthesis.In this study, the metabolism ofX. dendrorhousgrowing in a continuous culture, under two am-monia conditions, Limited (L) and Non-limited (NL). The metabolic flux analysis (MFA) allowedto understand the distribution the intracellular fluxes along the different metabolic pathways eval-uated, but most important, it elucidated that by limiting the ammonia assimilation flux(L= 0.002±1.1 E-05; NL= 0.004±4.3 E-05; g/gcellh), a promotion of the astaxanthin formation flux wasobtained (L= 86.4±0.6; NL= 0; ug/gcellh). This first approach will help to set a deeper studyin order to understand the metabolic pathways that regulate the flux towards the astanxanthinbiosynthesis

Ciencias Exactas y Ciencias de la Salud

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