Ciencias Exactas y Ciencias de la Salud
Permanent URI for this collectionhttps://hdl.handle.net/11285/551039
Pertenecen a esta colección Tesis y Trabajos de grado de las Maestrías correspondientes a las Escuelas de Ingeniería y Ciencias así como a Medicina y Ciencias de la Salud.
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- Effect of SOx from simulated flue gas on growth and gene expression of S-compounds in the microalgae Desmodesmus abundans(Instituto Tecnológico y de Estudios Superiores de Monterrey, 2024-11-11) Hernández Barrientos, Bryan Zeth; Pacheco Moscoa, Adriana; emimmayorquin; Mora Godinez, Shirley María; School of Engineering and Sciences; Campus MonterreyDesmodesmus abundans high CO2 acclimated strain (HCA) is a microalga adapted for 13 years to high CO2 atmospheres. Previous studies have shown overexpression of genes associated with purines and folate biosynthesis under model flue gas (250 000 ppm CO2, 700 ppm NO, and 100 ppm SO2). Therefore, it is hypothesized that a high S concentration in the growth medium generates biomass with desirable concentrations of S-containing compounds reflected in the overexpression of key genes involved in the biosynthesis of purines and folate, and other associated pathways such as cysteine, methionine, and S metabolism. D. abundans was cultured with different S concentrations in Erlenmeyer flasks to ¾ log phase (4 d) under 25% v/v CO2/air (continuous flow at 0.05 vvm), 300 mL BG11 medium, 25 ± 2 °C, 100 rpm, and 85 µmol PAR-photons m-2 s-1 of continuous light). Three experimental conditions in triplicate, and duplicated biologically, were tested: i) low S (BG11-3/4S, 7 mg L-1 SO42-), ii) high S (BG11-S+SOx, 100 ppm SO2 continuous supply), and iii) control (BG11, 29.2 mg L-1 SO42-). Gene expression analysis of pathways of interest was done using RNA-Seq. Analysis of differential expression of genes (DEGs) compared S treatments to the control (|Log2FC > 1| padj < 0.05). DEGs were obtained using CDS information of D. abundans HCA and filtered genes were mapped against KEGG pathways. Also, free amino acids were analyzed using a Waters ACQUITY UPLC® System with a fluorescence detector. Neither culture was inhibited by the growth condition (no lag phase) and exponential growth was similar among cultures; however, final biomass production was highest in the control (0.8 ± 0.1 g L-1 d.w.), followed by the high S and low S conditions (0.6 ± 0.06 and 0.51 ± 0.06 g L-1 d.w. respectively). Therefore, the presence of SOx slightly reduced by 19% final biomass production. A similar trend was observed for total protein content with a greater reduction of 38% under high S (56.2 ± 1.6%, 34.6 ± 0.41%, and 26.5 ± 3.7% d.w. for the control, high S, and limited S, respectively). DEGs showed that S-limitation resulted in up-regulation of sulfate assimilation- related genes but did not correlate to a greater quantity of S-containing amino acids and only the serine O-acetyltransferase (SAT) gene was upregulated in the cysteine and methionine pathways under this condition. Although upregulation in the folate biosynthesis pathway could be observed in both conditions, these genes were not related to the interconversion of folate derivatives. Given that gene expression of reactions of interest was not clearly observed through transcriptomic analysis, neither synthesis of S-containing amino acids, implementation of direct approaches based on sequence homology of specific genes and quantification of other S related compounds as folate should be pursue.
- Informe de control de calidad(Instituto Tecnológico y de Estudios Superiores de Monterrey, 2024-02-02) Sebastian Campos, Brigitte; Lorenadnldnvlndlvd
- Determination of parameters for the synthesis of carbon nanomaterials by ultrasonic spray-chemical vapor deposition(Instituto Tecnológico y de Estudios Superiores de Monterrey, 2022-12) Hurtado Alvarez, Agustín; SUSTAITA NARVAEZ, ALAN OSIRIS; 226812; Sustaita Narváez, Alan Osiris; puelquio/mscuervo; Ulloa Castillo, Nicolás Antonio; Lozano Sánchez, Luis Marcelo; González González, Francisco José; School of Engineering and Sciences; Campus Monterrey; Iturbe Ek, JackelineNanotechnology is a field that keeps growing every year, especially carbon nanomaterials like graphene or carbon nanotubes, which are the main ones employed in developing patents [1]. These nanomaterials exhibit outstanding properties like young modules around 1 TPa [2], experimental surface area above 1,300 m2g−1 [3], among many other thermal and conductivity properties. Although a lot of research is carried out, this is not reflected in the number of products on the market for daily use. The main boundaries that delay the adoption of these materials are cost/price, production methods scaling and material quality/consistency [4]. Chemical vapor deposition (CVD) is a methodology capable of synthesizing different carbon allotropes by varying the processing parameters [5]. Also, it is a very versatile methodology because it can use precursors in different physical states and chemical natures [6]. When the carbon precursors are in a liquid state with the catalyst dissolved, an activation mode is necessary to nebulize the solution to facilitate pyrolysis [7]. The ultrasonic spray is an activation mode that assists CVD by isolating the precursor solution through the acoustic cavitation that expands and compresses the droplets until it implodes into micron-sized droplets that facilitate the pyrolysis of the reactants [8]. In collaboration with the author of this thesis, Ceron designed and manufactured a CVD system assisted by ultrasonic spray to later automate it with routines to synthesize specific carbon nanomaterials [9]. Unlike other systems, this one includes a polytetrafluoroethylene (PTFE) membrane that separates the precursor solution from the transducer to increase the dispositive’s lifetime while allowing the user to change the nebulization chamber (useful for post-processing steps). The first trials of the system provide exciting results because they synthesize different morphologies by changing the processing parameters. However, a structured design of experiments (DOE) is needed to improve the synthesized product. For this reason, this thesis aimed to determine the conditions to synthesize a specific morphology with fewer defects and maximize mass production. To do so, 23 DOE was performed by duplicating. The independent variables were the concentration (molarity) of the catalyst in the precursor solution, the Ar flow (L/min) and the temperature. Meanwhile, the dependent variables were ID/IG rate and mass production (grams). Each experiment was characterized by Raman spectroscopy and Scanning Electron Microscopy (SEM). Raman spectra and SEM micrographs validate that the system designed can synthesize different kinds of carbon allotropes like carbon spheres or CNT. Current results suggest that the system can synthesize in a homogeneous way CNT at 800ºC. Nevertheless, future TEM characterization will allow us to validate the obtaining of MWCNT (as suggested by Raman analysis). A 23 design of experiments was done to minimize the number of defects in the synthesized carbon nanostructures, where it was found that at 800ºC, 0.15M and 1.5 L/min, the smallest value of ID/IG ratio was obtained. A similar study was performed to maximize mass production, where the highest mass is obtained at 800ºC, 0.1M and 1.0 L/min. Since the conditions of each study were different, it was looked for the parameters that balanced the two properties better, being 800ºC-0.1M-1.0L/min the conditions that maximized the mass production without affecting too much the quality of the graphene sheet because this condition was the second condition with fewer defects.
- Synergistic interaction between lac and uspA promoters’ expression systems for recombinant protein production in Escherichia coli(Instituto Tecnológico y de Estudios Superiores de Monterrey, 2021-12-06) Méndez Chavero, Jessica Paola; Benavides Lozano, Jorge Alejandro; puemcuervo/tolmquevedo; Chávez Santoscoy, Rocío Alejandra; Vargas Cortez, Teresa; School of Engineering and Sciences; Campus Monterrey; Hernández Pérez, JesúsRecombinant proteins need to be produced on a large-scale, short time and at a low cost. Different elements need to be controlled to achieve these characteristics. One of the most relevant is the promoter sequence since it is responsible for initiating the transcription process. Several promoters are already commercially used, but none is suitable for high-scale protein production due to the cost of the induction method, the leaky expression, the limited media options, etc. Furthermore, most current expression systems rely on the single use of a promoter to control the gene transcription. At the same time, in nature, this is not always the case since there are tandem promoters’ arraignments in control of a single gene. Using tandem promoters is an attractive idea to enhance the recombinant protein production considering that this significate the incorporation of more sites where the RNAP-complex could bind, enhancing recombinant protein production this way. Based on this, a triple-promoter expression system was developed in Escherichia coli to improve the yields of the red fluorescent recombinant protein (mRFP1). Two copies of the well-known and established lac promoter were used, and the sequence of the uspA promoter, which is the promoter of the most abundant protein in Escherichia coli according to the Protein Abundance Database PAXdb4.1, were used in this system. Additionally, the uspA promoter can be induced by several stress conditions. Thus, it was tested under oxidative starvation. Results indicated a 0.82 FU µg-1 mL-1 and 0.8 FU µg-1 mL yields of the mRFP1 when using the triple-promoter expression systems compared to a 0.59 FU µg-1 mL-1 of the single-promoter expression system. Thus, indicating an enhancement in the production yields of the target protein. These results are promising and open an opportunity for further research in the multiple-promoter system.
- Saccharification of chickpea supernatants made of extruded chickpea flour and their lactic acid fermentation with five different Lactobacillus strains(Instituto Tecnológico y de Estudios Superiores de Monterrey, 2021-12-06) Mariscal Orduño, Mireya; Serna Saldívar, Sergio Román Othón; puemcuervo; del Pino Espinosa Ramírez, Johanan; Santacruz López, Yolanda Arlette; School of Engineering and Sciences; Campus Monterrey; Pérez Carrillo, EstherGlucose-rich chickpea-based supernatants were developed with extruded chickpea (EF) flour enzymatically-treated with amyloglucosidase EF (AMG), and raw chickpea flour (RF) treated with α-amylase prior to catalysis with amyloglucosidase RF (Amyl + AMG). The resulting supernatants were subjected to lactic acid fermentation with five different Lactobacillus strains as starters (Lactobacillus plantarum BG112, Lactobacillus plantarum 299v, Lactobacillus plantarum V3, Lactobacillus plantarum 29, and Lactobacillus acidophilus LA3). At the end of fermentation, all strains produced pH drops to about 4.1 and yielded between 0.7-1.1 % of lactic acid. The La LA3 strain yielded the highest lactic acid whereas the counterpart Lp 29 was the lowest. Moreover, an active metabolism over glucose and α-galacto-oligosaccharides was found in all the strains. Lp 29 presented the highest growth rate in both supernatants. Interestingly, at the end of the programmed fermentation, all strains reached cell counts of about 109 CFU/mL (9 Log CFU/mL) with log units stable throughout 14 days of cold storage.
- Study of chemical diversity in Streptomyces venezuelae isolated from contrasting mexican environments using genome mining and bioactivity analysis(Instituto Tecnológico y de Estudios Superiores de Monterrey, 2021-11-25) Pérez García, Viridiana; Licona Cassani, Cuauhtémoc; puemcuervo; De la Torre Zavala, Susana; Verdel Aranda, Karina; School of Engineering and Sciences; Campus Monterrey; Aguilar Yáñez, Jose ManuelMicrobial natural products (NPs) have historically been considered one of the most important sources of chemical diversity at an industrial and pharmaceutical level. Many of these molecules are generated through secondary metabolic pathways, and their biosynthesis is mediated by ecological interactions such as defense, competition, and interactions with the environment. The genus Streptomyces represents one of the main groups of producers of specialized metabolites. Previous research in this genus has revealed that even strains of the same species can differ dramatically in the genes that code for these metabolites, indicating a lot of chemical variety to be discovered. Therefore, this work aimed to compare the chemical diversity at the level of biosynthetic genes clusters (BGCs) present in strains of the same species, Streptomyces venezuelae, isolated from two contrasting habitats: a highly oligotrophic environment and a nutrient-rich environment. It is hypothesized that these factors may influence the distribution and content of clusters and the generation of chemicals linked with them. Furthermore, genome mining research revealed a large diversity of BGCs from varied chemical families. These findings open the door to further research into the chemical diversity of Actinobacteria isolated from different geographical locations in the quest for bioactive molecules.
- Rational design and engineering of functionalized carbon nano-onions reinforced polymer nanocomposites for biomedical applications(Instituto Tecnológico y de Estudios Superiores de Monterrey, 2021-11-24) Velasco Delgadillo, Ramiro Manuel; Mamidi, Narsimha; puemcuervo; Sustaita, Alan O.; Cholula Díaz, Jorge Luis; School of Engineering and Sciences; Campus MonterreyRecently, materials chemistry has become an extensively studied research area due to its opportunity of creating or modifying existing materials, intending to improve biological, chemical, and mechanical properties and to further incorporate or substitute traditional methods. Carbon nano-onions (CNOs) are carbonaceous nanostructures that pose excellent physicochemical properties and when its surface gets modified in conjugation with biomaterials, the complex gains improved capabilities as drug uptake efficacy, tensile strength, thermal stability, hydrophobicity, cytocompatibility, thermosensitivity, drug release profile, among others, with potential application in a broad range of areas compared with pristine biomaterials. In this thesis, CNOs were fabricated, functionalized, and reinforced with different biomaterials to create emerging nanostructures to study their influence in mechanical, biological, and chemical properties in orthopedics and drug delivery applications. Herein, three different CNOs, poly 4-mercaptophenyl methacrylate (PMPMA)-CNOs, poly 4-hydroxyphenyl methacrylate (PHPMA)-CNOs, and poly (N-(4-aminophenyl) methacrylamide)) (PAPMA)-CNOs were attached to polymer-based nanocomposites for their potential use in orthopedic and drug delivery applications. In all cases, physicochemical properties of nanocomposites were systematically studied, as well as cytocompatibility studies to evaluate cell viability and proliferation, showing an increase in cytocompatibility with osteoblast cells and augmented tensile strength, toughness, and Young's modulus. In conclusion, the addition of functionalized CNOs considerably influences the mechanical and biological properties which could be advantageously used in biomedical applications.
- Optimization in the isolation and purification procedure of pristimerin, an anticancerigen, by Aqueous Two-phase Systems(Instituto Tecnológico y de Estudios Superiores de Monterrey, 2021-06-11) González Félix, Martha Alicia; González Valdez, José Guillermo; puemcuervo; Ibarra Herrera, Celeste Concepción; González Torres, Maykel; School of Engineering and Sciences; Campus Monterrey; Mejía Manzano, Luis AlbertoThe natural triterpene pristimerin possesses interesting potential against several types of cancer. However, its production at industrial levels requires novel environmentally friendly and efficient extraction procedures. Here we analyze an ethanol-phosphate Aqueous Two-Phase Systems (ATPS) and its design parameters such as Tie-Line Length (TLL) and volume ratio (VR) on the direct extraction of pristimerin, concluding that high TLL 70 % (w/w) and a low VR (0.33) values provide the best extraction yield and purity. Partition experiments provided a compound KP value of 3.12 ± 1.21, reaffirming its preference for the ethanolic-rich phase. Besides, green assessment positioned the extraction in the classification A with 90.93 of 100 points. Furthermore, analysis at different laboratory scales (1X, 10X and 100X) demonstrated the scalability and robustness. Finally, a complete bioprocess based on the obtained results is suggested as a tool to address the pharmaceutical potential of this molecule in a sustainable and efficient manner.
- Effect of processing conditions on physical-chemical and structural characteristics of cacao (Theobroma cacao L.)(Instituto Tecnológico y de Estudios Superiores de Monterrey, 2021-05-14) Guillén Guerrero, Karla María; De la Rosa Millán, Julián; puemcuervo; Morales de la Peña, Mariana; Tejada Ortigoza, Viridiana Alejandra; School of Engineering and Sciences; Campus MonterreyThree cocoa bean (Theobroma cacao L.) fermentations were carried out with a duration of 6 days simulating the box technique, with the aim to observe the biochemical changes and their effect on chemical and physicochemical characteristics, the temperature at the last fermentation stage was modified 50 °C for control, 40 °C (F40), and 60 °C (F60). Therefore, there were changes in pH into the cotyledons with 5.3, 5.7, and 5.0 for control, F40 and F60, respectively, which developed variations in the chemistry and structure of the cocoa bean. A proximal analysis was carried out (moisture, ash, proteins, lipids, and carbohydrates) as well as the determination of Free Amino Nitrogen (FAN), Phenolic Compounds (PC), Antioxidant Activity, Enzyme Activity, and Image structural Analysis. According to a Pearson Correlation Analysis and a Principal Component Analysis (PCA), the most significant response variables on day 6 of fermentation are FAN, with 11.41 ± 0.31, 10.38 ± 0.23, 12.50 ± 0.65 g / kg obtained for control, F40 and F60; and phenolic compounds with 42.00 ± 0.96, 66.43 ± 0.07 and 22.41 ± 0.96 mg Gallic Acid / g cocoa for control, F40 and F60 respectively. Changes in the behavior of enzymatic activity related to FAN and PC were also observed since proteases and amylases decreased their activity in F60, while they increased in lipases, inverse to the F40 treatments, compared with control. The degree of fermentation was determined with the cracking of the cotyledons as a parameter where 39% is observed for control, 32% for F40, and 49% for F60, showing that low and over fermentation temperatures cause variations in enzyme behavior, metabolite production and structure of cacao bean.
- Recombinant expression and in silico evaluation of AreV1, an Arenin variant.(Instituto Tecnológico y de Estudios Superiores de Monterrey, 2020-12-18) Teruel Barandiarán, Estefanía; Benavides Lozano, Jorge; tolmquevedo; Aguilar Jiménez, Oscar Alejandro; Chávez Santoscoy, Rocío Alejandra; School of Engineering and Sciences; Campus Monterrey; Hernández Pérez, JesúsIn the last decades, plants and animals described in Traditional Medicine practices have gained importance as sources of bioactive molecules since a great number of compounds have been isolated and characterized to develop new drugs. Although most of the current ethnopharmacological studies have focused on research bioactive compounds from plants, members of the anuran family Hylidae have also demonstrated to possess a plethora of molecules in their skin secretions with diverse capabilities. Among peptides, proteins, and low molecular weight compounds secreted by amphibians, protease inhibitors (PIs) have been proposed to be used as antimicrobial and antitumoral drugs that may possess high efficacy and efficiency. Recently, Arenin a Kunitz-like PI was isolated from the skin secretions of the canyon tree frog Dryophytes arenicolor. The DNA sequence encoding Arenin was modified by site-directed mutagenesis via PCR and the resultant variant, AreV1, was cloned, expressed, and purified by IMAC. Its 3D-structure was analyzed in silico through I-TASSER and COFACTOR servers and the structure modelling platform UCSF Chimera to assess the impact of the mutagenesis on the structure and further functionality of AreV1. Dissimilarities between AreV1 and Arenin were found at the C-terminus of the DNA sequence, altering ten amino acids (Leu45, Pro47, Trp48, Lys49, Ile50, Val51, Arg52, Pro53, Pro54 and Ala55). Despite these changes, according to the in silico analysis, the structural stability of AreV1 remained almost identical to Arenin, although the size of the variant polypeptide is three amino acids smaller. The analysis on AreV1 suggests that it has a great potential as trypsin regulator with a Lys13 as the residue located at the centre of the reactive site (P1 site). A tyrosine triplet (Tyr21, Tyr22 and Tyr23) associated with analgesic properties was also observed in AreV1.This research work aims to expand the knowledge on the activity optimization of naturally occurring molecules by presenting the recombinant production, purification and structural analysis of a variant from Arenin.
