Semillas criollas de maíz de Uruguay y contaminación con transgenes
Editores: Viviane Camejo Pereira e Fábio Dal Soglio. Universidad Federal do Rio Grande do Sul. Serie Ensino, Aprendizagem e Technologias, 2020. ISBN 978-65-5725-007-5. pp 135-159.
Galeano, P., Beltrán,M., Machado, S., Fossatti, M., González, T., Arleo, M., Porta, B., Vidal, R., Martinez, C., Franco Fraguas,L., Galván, G.
Capítulo V en el Libro “A conservação das sementes crioulas: uma visão interdisciplinar da agrobiodiversidade".
Nuevas metodologías para el análisis de microcistinas en peces. Estudio de Astraloheros facetus expuestos in vitro.
INNOTEC (2020) v.: 20, 10 - 29.
( link al artículo
Badagian, N., Letarmendia, M., Pirez, M., Carnevia, D., Brena, B.M.
La alta incidencia de floraciones de cianobacterias productoras de microcistinas en el país y la región representa un riesgo muy elevado para humanos y animales. A fin de estudiar el impacto y la presencia de las microcistinas (MCs) en ani-males, es importante disponer de métodos simples de bajo costo. Como primera aproximación a estos objetivos en peces, se estudiaron Astraloheros facetus (Cas-tañetas) expuestas a una floración de Microcystis spp (MCs 60 y 600 μgMCs/L) en un bioensayo subcrónico (18 días). Si bien no hubo mortalidad, la histopato-logía mostró infiltración grasa en el hígado, más relevante en los peces expuestos a la mayor concentración. Para analizar MCs en pescados se optimizaron dos mé-todos inmunoquímicos sensibles basados en un anticuerpo recombinante de llama (nanobody) de alta especificidad: ELISA y MALDI-TOF cuantitativo, utilizando partículas magnéticas funcionalizadas. Los métodos fueron recientemente desa-rrollados localmente. La excelente correlación ELISA/MALDI-TOF (rSpearman = 0,988, p< 10-7) resalta el potencial de este ELISA como herramienta simple y costo-efectiva para minimizar las muestras a analizar por métodos de referencia. Las concentraciones de MCs en las Castañetas fueron relevantes, acordes con bioensayos en otras especies y peces de la naturaleza. Esto destaca la importancia de analizar MCs en pescado para consumo.
Characterization of the cellulase-secretome produced by the Antarctic bacterium Flavobacterium sp. AUG42
Microbiological Research, vol 223-225, 13-21
Herrera,L.M., Braña, V., Franco Fraguas, L., Castro-Sowinski, S.
Flavobacterium sp. AUG42 is a cellulase-producing bacterium isolated from the Antarctic oligochaete Grania sp. (Annelida). In this work, we report that AUG42 produces a glycoside hydrolase cocktail with CMCase, PASCase and cellobiase activities (optimum pHs and temperatures ranging from 5.5 to 6.5 and 40 to 50 °C, respectively). The time-course analyses of the bacterial growth and cellulase production showed that the cocktail has maximal activity at the stationary phase when growing at 16 °C with filter paper as a cellulosic carbon source, among the tested substrates. The analyses of the CAZome and the identification of secreted proteins by shotgun Mass Spectrometry analysis showed that five glycoside hydrolyses are present in the bacterial secretome, which probably cooperate in the degradation of the cellulosic substrates. Two of these glycoside hydrolyses may harbor putative carbohydrate binding modules, both with a cleft-like active site. The cellulolytic cocktail was assayed in saccharification experiments using carboxymethylcellulose as a substrate and results showed the release of glucose (a fermentable sugar) and other reducing-sugars, after 24 h incubation. The ecological relevance of producing cellulases in the Antarctic environment, as well as their potential use in the bio-refinery industry, are discussed.
Enzymatic synthesis of non-natural trisaccharides and galactosides; Insights of their interaction with galectins as a function of their structure
Carbohydrate Research, v.: 472 p.:1 - 15
Porciúncula Gonzalez, C. Cagnoni, A.J., Mariño, K.V., Fontana, C., Saenz Mendez, P., Irazoqui, G., Giacomini, C.
Galectins are a family of carbohydrate-recognizing proteins that by interacting with specific glycoepitopes can mediate important biological processes, including immune cell homeostasis and activation of tolerogenic circuits. Among the different members of this family, Galectin 1 and 3 have shown pro-tumorigenic effects, being overexpressed in numerous neoplasic diseases, proving to be relevant in tumor immune escape, tumor progression and resistance to drug-induced apoptosis. Thus, generation of specific glycosides that could inhibit their pro-tumorigenic ability by blocking their carbohydrate recognition domain is one of the current major challenges in the field. Considering that galectin-ligand binding strength is closely related to the ligand structure, analysis of this relationship provides valuable information for rational design of high-affinity ligands that could work as effective galectin inhibitors. Taking profit of the ability of glycosidases to catalyze transglycosylation reactions we achieved the enzymatic synthesis of β-d-Galp-(1 → 6)-β-d-Galp-(1 → 4)-d-Glcp (2), a mixture of β-d-Galp-(1 → 6)-β-d-Glcp-(1 → 4)-d-Glcp (5) and β-d-Galp-(1 → 3)-β-d-Glcp-(1 → 4)-d-Glcp (6), and finally benzyl β-d-galactopyranoside (9), with reaction yields between 16 and 27%. All the galactosides were purified, and characterized using 1H and 13C nuclear magnetic resonance spectroscopy. Docking results performed between the synthesized compounds and human Galectin 1 (hGal-1) and human Galectin 3 (hGal-3) showed that the replacement of a glucose moiety linked to the terminal galactose with a galactose moiety, decreases the affinity for these galectins. Moreover, regarding the interglycosidic bond the most favorable β-Gal linkage seems to be β(1 → 4) followed by β(1 → 3) and β(1 → 6) for hGal-1, and β(1 → 4) followed by β(1 → 6) and β(1 → 3) for hGal-3. These results were in accordance with the IC50 values obtained with in vitro solid phase inhibition assays. Therefore, docking results obtained in this work proved to be a very good approximation for predicting binding affinity of novel galactosides.
Pseudozyma sp. isolation from Eucalyptus leaves and its hydrolytic activity over xylan
Biocatalysis and Agricultural Biotechnology, v. 21, p.10128
( DOI: 10.1016/j.bcab.2019.101282
Botto, E., Gioia, L., Menéndez, P., Rodríguez, P.
Eucalyptus leaves were investigated as a source for the isolation of xylanase producing microorganisms. A total of 37 isolates were obtained after a series of enrichment steps. Seven of the isolates were xylanase positive in an agar screening experiment and were further analyzed in liquid media with beechwood xylan as carbon source. A yeast identified as Pseudozyma sp. showed the highest xylanase activity in tested conditions. Afterwards, different lignocellulosic residues were studied as substrates for xylanase production by this strain and the best results were obtained with corncob. Yeast's xylanase with a molecular weight of 19.9 kDa showed the maximum activity at pH 4.8 and 50 °C. Thermostability was observed at 30 °C with a 60% activity retention after 10 days. By the hydrolytic activity of the enzyme was characterized as an endoxylanase, similar as the ones found in family GH 10, from the products obtained by beechwood xylan hydrolysis.
Survery of b-galactosidases properties: applications to transgalactosylation process
En: Beta Galactosidase Properties, Structure and Functions, 65-115, 2019, ed. Eloy Kras, Nova Science Publishers, Inc , New York,ISSN/ISBN: 978-1-53615-605-8
Porciúncula Gonzalez, C., Giacomini, C., Irazoqui, G.,
b-galactosidases (b-D-galactohydrolase, EC 188.8.131.52) are enzymes that hydrolyzes terminal b 1-4 galactosides. They belong to the GH 1, GH 2, GH 35 and GH 42 of the GH-A superfamily of glycoside hydrolases. They are widely distributed in nature and can be find in animals, plants and several microorganisms (yeast, fungi and bacteria). In nature, they function as hydrolases. In plants, they remove terminal b 1-4 galactose from polymers containing galactose, in animals and microorganisms they catalyzes the hydrolysis of lactose in galactose and glucose. The most studied b-galactosidases are those from microorganism, such as Escherichia coli, Aspergillus oryzae, Bacillus circulans, Streptococcus thermophilus, Kluyveromyces lactis. This allows a collection of b-galactosidases with different properties (as optimum pH and temperature, thermal stability, stability against pH, denaturing agents, etc.), which offer a great versatility of conditions to use these enzymes for different applications. One of the first biotechnological application of these enzymes was for lactose hydrolysis in order to produce lactose reduced milk and dairy products for consumption of lactose intolerant people. Besides, a reduced lactose content in dairy product avoids lactose crystallization improving their organoleptic properties. Nevertheless, under adequate conditions they are able to catalyze the synthesis of oligosaccharides and galactosides. Two methods have been used for this purpose, reverse synthesis and transglycosylation. The first one involves the inversion of the hydrolysis reaction starting from a mixture of monosaccharides and nucleophilic hydroxylated molecules. This method requires high concentration of reactants as well as the presence of co-solvents in order to reduce water activity. On the other hand transglycosylation mechanisms is kinetically controlled and catalyzes the transfer of a galactosyl moiety from a galactosyl donor (lactose, ortho-nitrophenyl-b-D-galactopyranosyde, lactulose) to an hydroxylated nucleophile. Aliphatic alcohols, hydroxylated aminoacids such as serine and threonine, polyols, flavonoids, and monosaccharides among others have proved to be good glycosyl acceptors. The use of the transgalactosylation mechanism is an excellent alternative to the complex chemical synthesis as it allows the synthesis of b-galactosides and b-galactooligosaccharides in a single step preserving the anomeric center configuration. Even though glycosidases are stereospecific they are not always regiospecific and so the generation of isomers mixtures could take place, depending on the structure of the acceptor molecule as well as on the source of the b-galactosidase. Regarding synthetic applications, they have been used for enzymatic synthesis of: galactooligosaccharides with prebiotic properties as food additives; galactooligosaccharides as building blocks for synthesis of branched oligosaccharides; galactosides with potential activity as galectin inhibitors or antitumor agent; alkyl galactosides as non-ionic surfactants. b-galactosidases have also been used for galactosylation of drugs in order to improve their hydrophilicity and bioavailability. In this chapter we will make an update regarding b-galactosidases classification, mechanisms, characterization of their properties and applications.
Oriented Functionalization of magnetic beads with in vivo biotinylated nanobodies for rapid MALDI-TOF MS ultrasensitive quantitation of microcystins in biological samples
Analytical Chemistry, 91, 15, 9925–9931
Pírez, M., Brena, B.M., González-Sapienza, G.
Here we present a new analytical method where immunoconcentration of the analyte is coupled to quantitative matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF MS) analysis allowing in minutes the identification and highly sensitive quantitation of microcystins (MCs) as model targets. The key element is a site-specific in vivo biotinylated nanobody of broad cross-reactivity with microcystins. The single biotin moiety at the C-terminus and the small size of the nanobody (15 kDa) enable its oriented and tightly packed immobilization on magnetic beads, providing a highly efficient capture of the toxin. The binding capacity of the bioadsorbent is partially loaded with an easily synthesized internal standard for MS quantitation. After capture, the beads are directly dispensed on the MALDI-TOF MS target enabling the identification and sensitive quantitation of the microcystin (MC) congeners. Since salts and contaminants are removed during the concentration step, no cleanup or other sample treatments are needed. The method was validated with a large number of water and serum samples with excellent precision and recovery at quantitation limits of 0.025 μg/L of MC.
Assessing multimetric trophic state variability during an ENSO event in a large estuary (Rio de la Plata, South America).
Regional Studies in Marine Sciences, v 28, 100565
Brugnoli, E., Muniz, P., Venturini, N., Brena, B.M., Rodríguez, A., Garcia Rodriguez, F.
During the “El Niño”- Southern Oscillation (ENSO) event 2009–2010, trophic state uni (TSI’s) and multimetric (TRIX’s) indexes were determined in the north coast of the Río de la Plata (RdlP). The relationships of such indexes with the hydrological, physical and chemical parameters were explored to identify climatic variability or anthropogenic effects on the eutrophication process. Eleven oceanographic sampling campaigns at 25 stations were conducted along the north coast of RdlP from 2009 to 2011. Temperature, salinity and oxygen saturation, water transparency (Secchi depth), chlorophyll (Chl a) content and total nutrient concentrations (total nitrogen and phosphorous) were determined. Trophic indexes (TSI Chl a, TSI TP, TRIX NP-Voll. and TRIX NP-Mvdeo.) were quantified and the RdlP flow was measured. RdlP flow showed significant differences between ENSO phases, with higher values during “El Niño” (EN), negative and significant correlations with salinity, Secchi depth and positive with total nutrients. The trophic status indexes showed spatial and temporal variability and associations with flow, physical and chemical parameters. Trophic state indexes showed spatial and temporal heterogeneity and were strongly associated to environmental conditions (hydrological and seasonal oscillations) during the study period but also to the anthropogenic factors affecting the north coast of RdlP. The system has been classified as mesotrophic to hypertrophic, depending on the selected indicator. According to these results, we suggest the use of a multimetric trophic state indicator, i.e., the TRIX index, as explains better the causes and effects of the eutrophication processes. To implement the TRIX index, (TRIX NP-Mvdeo.) we suggest researchers to consider a large number of local records (Chl a, nutrients, oxygen) at seasonal scales. This index is adapted to the environmental conditions of the study zone and complemented with other community structure indexes, and should be used for eutrophication assessment programs in the north coast of RdlP.
Combined Danio rerio embryo morbidity, mortality and photomotor response assay: A tool for developmental risk assessment from chronic cyanoHAB exposure.
Science of the Total Environment, v.: 697, 134210
Roegner, A., Truong, L., Weirich, C., Pirez, M., Brena, B.M.,Miller, T.M., Tanguay, R.
Freshwater harmful algal blooms produce a broad array of bioactive compounds, with variable polarity. Acute exposure to cyanotoxins can impact the liver, nervous system, gastrointestinal tract, skin, and immune function. Increasing evidence suggests chronic effects from low-level exposures of cyanotoxins and other associated bioactive metabolites of cyanobacterial origin. These sundry compounds persist in drinking and recreational waters and challenge resource managers in detection and removal. A systematic approach to assess the developmental toxicity of cyanobacterial metabolite standards was employed utilizing a robust and high throughput developmental Danio rerio embryo platform that incorporated a neurobehavioral endpoint, photomotor response. Subsequently, we applied the platform to cyanobacterial bloom surface water samples taken from temperate recreational beaches and tropical lake subsistence drinking water sources as a model approach. Dechorionated Danio rerio embryos were statically immersed beginning at four to six hours post fertilization at environmentally relevant concentrations, and then assessed at 24 h and 5 days for morbidity, morphological changes, and photomotor response. At least one assessed endpoint deviated significantly for exposed embryos for 22 out of 25 metabolites examined. Notably, the alkaloid lyngbyatoxin–a resulted in profound, dose-dependent morbidity and mortality beginning at 5 μg/L. In addition, hydrophobic components of extracts from beach monitoring resulted in potent morbidity and mortality despite only trace cyanotoxins detected. The hydrophilic extracts with several order of magnitude higher concentrations of microcystins resulted in no morbidity or mortality. Developmental photomotor response was consistently altered in environmental bloom samples, independent of the presence or concentration of toxins detected in extracts. While limited with respect to more polar compounds, this novel screening approach complements specific fingerprinting of acutely toxic metabolites with robust assessment of developmental toxicity, critical for chronic exposure scenarios.
Biodegradation of acid dyes by an immobilized laccase: an ecotoxicological approach
Environmental Science: water research & technology, 2018, 4 (12) , 2125 – 2135.
Gioia,L., Ovsejevi, K. , Manta,C. , Menéndez, P.
Synthetic dyes in watercourses resulting from industrial effluent discharges cause serious ecological impacts, besides carcinogenic and mutagenic effects on human health. Thus, it is important to develop effective methods to remove the dyes from industrial wastewaters, and also to carry out adequate toxicity studies to establish their safety. Azo dyes are the main class of industrial dyes and important environmental contaminants. We have examined the decolorization of two azo dyes (Acid Red 88 and Acid Black 172) by a native Trametes villosa laccase immobilized on thiolsulfinate-agarose as well as the effect of different redox mediators in the reactions. The method was effective for the decolorization of both dyes. The immobilization method did not affect the capacity of the biocatalyst for dye degradation. Therefore, the insoluble enzyme removed 97% of the color of AR88 and 92% of AB172 in 24 hours at 22 °C in the presence of the selected redox mediators, vanillin (0.1 mM) and violuric acid (1.0 mM), respectively. In addition, the immobilized enzyme kept 78% of its initial capacity for decolorization of AR88 after three cycles of use. The ecotoxicological evaluation of the solutions showed a great variation depending on the biological systems used. In the phytotoxicity test, the decolorization products were not toxic to plants, whereas in Daphnia magna and Microtox® bioassays an acute residual toxicity was found. The last outcome shows the importance of using a battery of bioassays to determine the remaining ecotoxicity of the treated effluents before their discharge into the aquatic environment.
Thiol-cyclodextrin: a new agent for controlling the catalytic activity of polyphenol oxidase from red delicious apple.
Journal of Food Science & Technology, 3(2)1-11, 2018.
Peralta-Altier, G., Manta, C., Ovsejevi, K.
Polyphenol oxidase (PPO, EC 184.108.40.206) is the main enzyme responsible for enzymatic browning, a natural process which produces deterioration of fruits and vegetables. One alternative to prevent this undesirable process is to inhibit the catalytic activity of PPO by encapsulating enzyme’s substrates in cyclodextrins (CD). In this article the effect of a Thiol-CD on PPO from Red Delicious apple was studied, demonstrating that this compound is a powerful tool for controlling oxidative processes in food. Thiol-CD could encapsulate the polyphenols, natural substrates of the enzyme, by means of the cyclodextrin hydrophobic internal cavity. Simultaneously, through the thiol group, it could inactivate the PPO by reducing the copper ions from the active site of the enzyme. Moreover, thiol moieties could decrease the browning by reducing the quinones generated by oxidative processes. The isolation and purification of PPO from apple was performed in order to study those effects, different polyphenols, chlorogenic acid (CA) and 4-methylcatechol (4-MC), were assayed as enzymatic substrates. Both β -CD and Thiol-CD exhibited better enzyme inhibition value for CA than for 4-MC. Moreover, Thiol-CD showed an extraordinary performance compared with β-CD. When CA 10 mM was used, 5 mM β-CD gave 11 % PPO inhibition, meanwhile nearly 100 times less concentration of Thiol-CD (45 μM) gave 100 % of enzyme inhibition.
Immobilization of b-galactosidase and a-mannosidase onto magnetic nanoparticles: A strategy for increasing the potentiality of valuable glycomic tools for glycosylation analysis and biological role determination of glycoconjugates
Enzyme and Microbial Technology, v. 117 p.45 - 55
Rodriguez, E., Francia, K., Brossard, N., García Vallejo, J.J., Kalay, H., van Kooyk, Y., Freire, T., Giacomini, C.
Glycans present in biological glycoconjugates have several structural and functional roles. Elucidation of glycan structure and biological function is critical to understand their role in physiological and pathogenic process, enabling the development of diagnostic methods and disease treatment. Immobilized glycosidases are powerful tools for glycan analysis, as they are able to remove specific carbohydrates without altering the protein structure. Here we describe the individual immobilization of Aspergillus oryzae β-galactosidase and Canavalia ensiformis α-mannosidase onto agarose and silica magnetic nanoparticles activated with cyanate ester groups. High immobilization yields (70-90%) were achieved, keeping above 60% of its original activity. Immobilized glycosidases were effective in the selective deglycosylation of model glycoproteins and a Fasciola hepatica lysate, evidenced by a decrease in specific lectin recognition of 40-50% after enzymatic deglycosylation. Immobilized glycosidases were reused for several deglycosylation cycles without loss of effectiveness. Their use was extended to the elucidation of the glycan role of native glycoconjugates. A decrease in the recognition of lactoferrin treated with α-mannosidase by a C-type lectin receptor, DC-SIGN was found. Also the specific deglycosylation of a F. hepatica lysate demonstrated the relevance of mannosylated glycans in the induction of Th2/Treg immune responses during the infection. Our results show successful immobilization of specific glycosidases in nano-supports and validate their utility to identify glycans biological functions.
Production and characterization of a beta-glucosidase from Issatchenkia terricola and its use for hydrolysis of aromatic precursors in Cabernet Sauvignon wine LWT-Food
Science and Technology v. 87, 515 – 522
De Ovalle, S., Cavello, I., Brena, B.M., Caballito, S., González-Pombo, P.
New enzymes isolated from the biodiversity of native wine ecosystems could contribute to increase the varietal character of regional wines. This study reports on the production and characterization of Issatchenkia terricola, beta-glucosidase and its potential to release red-wine aromatic compounds. The enzyme, a monomer of 48 kDa with an isoelectric point of 3.5 is tolerant to glucose and ethanol, properties compatible with enological use. Although fed-batch is usually the most suitable system for enzyme production in submerged culture, in this case the yield was practically the same as in batch culture. Enzyme productivity was increased 2-fold in synthetic medium with glucose with respect to the YPG and 3–8-fold with respect to other media assayed. After enzymatic treatment, GC-MS analysis of the released aglycones demonstrated significant increases in the concentration of phenols (83%) and norisoprenoids (65%). According to the judges of the sensory panel, the treatment resulted in a wine with dried fruits and raisins notes, as compared to the control, which was found more sweet and fruity. This, together with the lack of activity on anthocyanin glycosides, highlights the potential of this enzyme in enology, since its high selectivity allowed the development of aroma without compromising wine color.
Effects of wind mixing in a stratified water column on toxic cyanobacteria and microcystin-LR distribution in a subtropical reservoir
Bulletin of Environmental Contamination and Toxicology, v.101 (5): 611-616
González-Piana, M., Piccardo, A., Ferrer, C., Brena, B.M., Pírez, M., Fabian, D., Chalar, G.
We analyzed the effects of stratification changes due to wind on the vertical cyanobacteria distribution and microcystin-LR concentrations in a reservoir and assessed the implications for water management. Under stratified conditions, the highest microcystin concentrations (up to 4.16 µg/L) and toxic cyanobacteria biovolume occurred in the epilimnion (~ 1 m). The lowest microcystin concentrations were between 0.02 and 1.28 µg/L and occurred in the hypolimnion (~ 20 m). A cold front passage associated with high wind velocities induced water column mixing, promoting the redistribution of microcystin-LR and cyanobacteria throughout the water column and increasing their concentrations in deeper zones. Microcystin-LR concentration was positively correlated with cyanobacteria biovolume (r = 0.747) and chlorophyll a concentration (r = 0.798). Changes in thermal profile due to wind would imply a greater challenge for drinking water treatment plants, since high cyanobacterial and microcystin concentrations could reach deep-water intakes.
Tesis de maestría
Biocatálisis aplicada a la síntesis de radiotrazadores: síntesis de S-adenosil metionina por metodologías biocatalíticas
Diego Umpiérrez. Tutoras: Gabriela Irazoqui, Sonia Rodríguez .
Finalizada en 2023.
Análisis de glicoproteínas de Trypanosoma cruzi como potenciales ligandos de receptores de tipo Toll de la inmunidad innata
Mercedes Landeira. Tutoras: Teresa Freire, Cecilia Giacomini, Florencia Festari.
Finalizada en 2021.
Link a la tesis
Explotación del genoma de Issatchenika terricola para identificación de glicosidasas con potencial aplicación en enología
Juliette Dourron. Tutoras: Paula Gonzalez Pombo, Ana Ramón.
Finalizada en 2020.
Link a la tesis
Celulasas sicrófilas, una innovación en la industria del bioetanol
Lorena Herrera Marrero. Tutoras: Laura Franco Fraguas, Susana Castro.
Finalizada en 2019.
Link a la tesis
Tesis de doctorado
Producción, caracterización bioquímica e inmovilización de lipasas microbianas y sus aplicaciones
Agustín Castilla. Tutoras: Gabriela Irazoqui, Sonia Rodríguez.
Finalizada en 2022.
Link a la tesis
β-glucosidasas de cepas nativas y su potencial en la liberación de aromas de vino. Desarrollo de nuevas estrategias para su uso en la industria enológica.
Stefanie de Ovalle. Tutoras: Paula Gonzalez Pombo, Beatriz Brena.
Finalizada en 2021.
Link a la tesis
Desarrollo y validación de métodos sencillos y rápidos para cianotoxinas en el monitoreo ambiental
Vania Macarena Pírez. Tutores: Beatriz Brena, Gualberto González-Sapienza.
Finalizada en 2019.
Link a la tesis
Diseño racional y síntesis enzimática de galactósidos con potencial actividad como inhibidores de galectinas
Cecilia Porciúncula. Tutoras: Cecilia Giacomini, Gabriela Irazoqui, Patricia Saenz Méndez.
Finalizada en 2019.
Link a la tesis
Purificación, caracterización funcional y estructural de cisteín proteinasas presentes en frutos maduros de Bromelia antíacantha. Evaluación de sus posibles aplicaciones.
Diego Vallés. Tutora: Ana María Cantera.
Finalizada en 2019.
Link a la tesis