Submitted by Renata Lopes (renatasil82@gmail.com) on 2017-01-09T13:58:43Z No. of bitstreams: 1 alancamposdasilva.pdf: 1251147 bytes, checksum: aa102a0e377862594d75e5865529db3d (MD5) / Approved for entry into archive by Diamantino Mayra (mayra.diamantino@ufjf.edu.br) on 2017-01-31T11:22:38Z (GMT) No. of bitstreams: 1 alancamposdasilva.pdf: 1251147 bytes, checksum: aa102a0e377862594d75e5865529db3d (MD5) / Made available in DSpace on 2017-01-31T11:22:38Z (GMT). No. of bitstreams: 1 alancamposdasilva.pdf: 1251147 bytes, checksum: aa102a0e377862594d75e5865529db3d (MD5) Previous issue date: 2016-08-12 / Doce de leite (DL) é um dos produtos lácteos concentrados mais populares no Brasil. Entretanto, o elevado valor calórico, teor de açúcar e/ou a presença de lactose limitam sua difusão entre os consumidores. Portanto, o desenvolvimento de produtos lácteos para dietas restritivas, principalmente, intolerância à lactose e valor calórico reduzido, é um desafio para a indústria laticinista e uma oportunidade de mercado. O objetivo deste trabalho foi desenvolver e caracterizar DL sem adição de sacarose e sem lactose (SL) e sem adição de sacarose (SA), bem como avaliar sua aceitação sensorial e intenção de compra pelos consumidores. Produziram-se os três tipos de DL, tradicional (TR), SL e SA. O leite destinado à produção do DL SL foi hidrolisado pela enzima β-galactosidase e o grau de hidrólise acompanhado pelo índice crioscópico. A fabricação ocorreu em tacho aberto com concentração para um teor de sólidos solúveis de 61 – 68 °Brix. Os DL foram analisados, quanto à hidrólise da lactose, por meio de Eletroforese de Carboidratos Assistida por Fluoróforo (FACE) e caracterizados através da composição centesimal, cor, cristais e requisitos microbiológicos. Foi realizada análise sensorial utilizando teste de aceitação em escala hedônica (1 a 9) e análise da intenção de compra (1 a 5). Os resultados foram avaliados pela ANOVA e comparação a posteriori de médias pelo teste t de Student com intervalo de confiança de 95%. Os leites utilizados nas produções demonstraram composição centesimal, crioscopia e acidez dentro dos padrões legais vigentes. A hidrólise da lactose no leite, pela enzima β-galactosidase, foi atingida com 80 minutos a 40 °C, na concentração de 0,7 g/L e confirmada no DL SL pela FACE por meio das bandas eletroforéticas de glicose e galactose. Os DL produzidos apresentaram características composicionais em conformidade com o Regulamento Técnico de Identidade e Qualidade (RTIQ), embora diferentes entre si. Em relação à cor, o DL SA exibiu coloração mais clara (L = 44,09 ± 0,73). O número de cristais por grama de doce de leite nos DL TR e SA foram 2,63 x 104 ± 19,00 e 4,63 x 106 ± 298,22, e tamanho de cristais 29,78 μm ± 27,84 e 24,83 μm ± 1,04, respectivamente. Quanto à aceitação sensorial, o DL TR obteve as maiores notas na maioria dos atributos julgados e na análise de intenção de compra. Cristais de lactose não foram percebidos no DL SL, possivelmente em função da hidrólise da lactose e sua aceitação foi igual (p>0,05) ou superior (p<0,05) ao DL TR em relação aos atributos cor, consistência e textura. O DL SA foi o menos aceito pelos consumidores. / Doce de leite (DL) is the one most popular concentrated dairy product in Brazil. However, the high-calorie, sugar content and/or the presence of lactose limit its spread among consumers. Therefore, the development of dairy products to restrictive diets, especially lactose intolerance and the reduced-calorie is a great challenge for the dairy industry and a market opportunity. Thus, the aim of this work was the development and characterization of DL with no added sucrose and lactose (SL) and without the addition of sucrose (SA) as well as, to evaluate the sensory acceptability and the purchase intent by consumers. Three sort of DL were produced, the traditional one (TR), SL and SA samples. For the preparation of the DL SL, the milk was hydrolyzed by β-galactosidase enzyme and the hydrolysis degree was accompanied by the cryoscopic index. The production took place in open pan with soluble solids concentration of 61-68 °Brix. DL were analyzed regarding lactose hydrolysis by Fluorophore-Assisted Carbohydrate Electrophoresis (FACE), and characterized as regards chemical composition, color, crystals presence and microbiological requirements. Sensory analysis was performed using acceptance test in hedonic scale (1 to 9) and the purchase intent analysis (1 to 5). The results were evaluated by ANOVA and a posteriori comparison of average by the Student's t test with confidence interval of 95%. The milk samples used in the DL production showed centesimal composition, freezing point and acidity values within the current legal standards. Lactose hydrolysis was achieved with 80 minutes at 40 ° C, with concentration of 0.7 g/L, confirmed in the DL SL sample by FACE analysis through glucose and galactose electrophoretic bands. The prepared DL showed compositional features in accordance with the Technical Regulation of Identity and Quality (RTIQ), even though different from each other.. Regarding color, the DL SA sample exhibited lighter color (L = 44.09 ± 0.73). The number of crystals per gram of doce de leite in the DL TR and SA sample was 2.63 x 104 ± 19.00 and 4.63 x 106 ± 298.22, with crystal size of 29.78 ± 27.84 µm and 24.83 ± 1.04 µm, respectively. As for sensory acceptance, DL TR obtained the highest scores in the most judge attributes and in the purchase intent analysis. Lactose crystals were not perceived in the DL SL sample, most likely as a result of the lactose hydrolysis and their acceptance was equal (p>0.05) or greater (p<0.05) when compared to color, texture and consistency of the DL TR sample. The DL SA sample was the least accepted by consumers.

Lactase persistence (LP) is a genetically determined trate caused by the expression of lactase in adulthood. Lactase is the intestinal enzyme responsible for digestion of milk sugar, lactose. Its production in the small intestine decreases during the childhood, this physiological condition is called lactose intolerance. However, in some individuals production of this enzyme is not stopped. The persistence of lactase activity is a recent phenomenon, which arose independently in several parts of the world over the past roughly 10,000 years, in connection with the emergence of agriculture, specifically milk production and is (likely) still under strong selection pressure. LP was first observed in Europe, where it is associated with a mutation -13 910*T. Frequency of this mutation correlates with latitude. In Africa, the presence of LP is conversely associated with herding and falls under the hypothesis of genetic and cultural co-evolution associated with cattle and the use of secondary food sources. Pastoral populations living in different areas of Africa have different LP mutations that are linked to their origin. Although many investigation on LP have already been carried out, neither analysed the Tuareg populations. This study is focused on the analysis LP mutations in 93 samples of Tuaregs from...

No description available.

Many consumers are unable to enjoy the benefits of milk, due to lactose-intolerance. Lactose-free milk is available, but at about 2 times the cost of regular milk or greater, it may be difficult for consumers to afford. The high cost of lactose-free milk is in part due to the added cost of the lactose hydrolysis process. Hydrolysis at refrigerated temperatures, possibly in the bulk tank or package, could increase the flexibility of the process, and potentially reduce the cost. A rapid β-galactosidase assay was used to determine the relative activity of commercially available lactase samples at different temperatures. Four enzymes exhibited low-temperature activity and were added to refrigerated raw and pasteurized milk at various concentrations and allowed to react for various lengths of time. The degree of lactose hydrolysis by each of the enzymes as a function of time and enzyme concentration was determined by HPLC. The two most active enzymes, as determined by the β-galactosidase assay, hydrolyzed over 98% of the lactose in 24 hours at 2°C using the supplier recommended dosage. The other two enzymes hydrolyzed over 95% of the lactose in 24 hours at two times the supplier recommended dosage at 2°C. Results were consistent in all milk types tested. The results show that it is feasible to hydrolyze lactose during refrigerated storage of milk using currently available enzymes.

No description available.

At the last time, we can see increased interest of lactose-free dairy products. The aim of the thesis was to evaluate offer of such products at selected retail chains in the South Bohemian region, then to conclude a sensory assessment of selected products and to verify of public awareness about lactose intolerance with a questionnaire survey. The widest offer of lactose-free dairy products has been found in Globus and Tesco, in the opposite the lowest offer has been found in Billa. Location of these products within the store was most the transparent in Globus (compared other stores). Three samples of white lactose free yoghurts and two samples of milk (with lactose and lactose-free) were evaluated by a sensory assessment. The sample of yoghurt from the company Madeta, line Nature was rated the best. On the contrary sample of yoghurt from the company Tatranská mliekáreň, line Nature´s Promise was rated the worst. The preference between two milk samples among the evaluators was balanced, despite large to moderate differences of intensity were described between them. Most respondents (79 %) knew what the term lactose intolerance means, but substantial part (22 %) of respondents badly indicated this the issue as allergy to milk casein. Because of high number of lactose-intolerant people in population this issue is still very actual, and it needs to be addressed.

In this work was studied the simultaneous influence of sodium alginate, gelatin and glutaraldehyde concentrations in the immobilization process of Beta-galactosidase from Aspergillus oryzae and the kinetic of lactose hydrolysis by the enzyme in the soluble and immobilized forms. The free enzyme was studied at 35°C and in a pH of 4.5, showing that for substrate concentrations up to 90g/L, the Michaelis-Menten model fitted the experimental data, with a Km and Vm value of 17.83 g/L (52.13 mM) and 1032.07 gglicose/L.min.mg proteína respectively. Galactose acted as a competitive inhibitor on the free enzyme kinetic, presenting kinect constants Ki and Km values of a 1.015 and e 17.61 g/L respectively. The enzyme was stable at pH values ranging from 4.5 to 7.0 and a maximum temperature activity of 55°C, with an activation energy of 6.9 kcal/mol. The thermal stability of the enzyme was studied from 53 to 65°C, presenting a half-life of 7.7 hours at 53°C. The activation energy for the thermal deactivation process was 88.14 kcal/mol. Through a central composite design, the sodium alginate, gelatin and glutaraldehyde concentrations that maximized the -galactosidase from Aspergillus oryzae activity in the inhibition process were, respectively, 6.60%(w/v), 4.05%(w/v) and 3.64%(v/v). The immobilized enzyme presented a 20% drop in activity after 25 uses. The immobilization yield found was 30%. The enzymatic activity for the immobilized form was maximum at pH of 5.0 and 60°C, determined through a central composite design. The reaction activation energy for the immobilized enzyme was 7.74 kcal/mol. The immobilized biocatalyst was stable on pH values ranging from 4.5 to 7.0. The half-life time of the immobilized enzyme was 12.8 hours at 53°C, with a activation energy for the thermal deactivation process value of 72.03 kcal/mol . The substrate concentration influence was studied from 5 to 140 g/L of lactose and the Michaelis-Menten model fitted the experimental data, with Vm and Km values of 1428.14 glactose/min.m3catalyst and 20.62 g/L (60.3 mM), respectively. It was observed a small resistance to lactose mass transfer at the biocatalyst particles, in the immobilized enzyme, due to the high effectiveness factor values. The inhibition model fitted the experimental data and the adjusted Km and Ki values were 16.7 and 9.6g/L, respectively. / Neste trabalho estudou-se a influência conjunta das concentrações de alginato de sódio, gelatina e glutaraldeído no processo de imobilização de Beta-galactosidase de Aspergillus oryzae e a cinética de hidrólise de lactose pela enzima nas formas solúvel e imobilizada. A cinética da enzima na forma livre foi estudada a 35°C, a pH 4,5, verificando que para concentrações de substrato de até 90g/L o modelo de Michaelis- Menten se ajustou aos resultados experimentais, com valores de Km e Vm iguais a 17,83 g/L (52,13 mM) e 1032,07 gglicose/L.min.mg proteína respectivamente. A galactose atuou como um inibidor competitivo na cinética da enzima na forma livre, apresentando constantes cinéticas Ki e Km com valores iguais a 1,015 e 17,61 g/L respectivamente. A enzima apresentou-se estável na faixa de pH de 4,5 a 7 e temperatura de máxima atividade de 55°C, com energia de ativação de 6,9 kcal/mol. A estabilidade térmica da enzima foi estudada na faixa de 53 a 65°C, apresentando meia vida de 7,7 horas a 53°C. A energia de ativação do processo de desativação térmica foi de 88,14 kcal/mol. Por meio de um planejamento composto central as concentrações de alginato de sódio, gelatina e glutaraldeido que maximizaram a atividade de b-galactosidase de Aspergillus oryzae no processo de imobilização foram, respectivamente, 6,60% (p/v), 4,05% (p/v) e 3,64% (v/v). A enzima imobilizada apresentou queda de 20% na atividade após 25 usos. O rendimento de imobilização encontrado foi de 30%. A atividade enzimática para a forma imobilizada foi máxima a pH a 5,0, a 60ºC, determinados através de um planejamento composto central. A energia de ativação da reação usando a enzima imobilizada foi 7,74 kcal/mol. O biocatalisador imobilizado apresentou-se estável na faixa de pH de 4,5 a 7. O tempo de meia vida da enzima imobilizada foi 12,8 horas a 53°C apresentando energia de ativação do processo de desativação térmica de 72,03 kcal/mol. A influência da concentração de substrato foi estudada para uma faixa de 5 a 140g/L de lactose e o Modelo de Michaelis-Menten ajustou-se bem aos dados experimentais, com valores de Vm e Km de 1032,07 glactose/min.m3catalisador e 20,62 g/L (60,3 mM), respectivamente. Em relação à enzima na forma imobilizada observou-se pequena resistência à transferência de massa de lactose nas partículas do biocatalisador, em função dos altos valores do fator de efetividade. O modelo de inibição competitivo ajustou-se bem aos dados experimentais e os valores de Km e Ki calculados foram 16,7 e 9,6g/L, respectivamente. / Doutor em Engenharia Química

Amadori-Produkte werden spontan während der ersten Phase der Maillard-Reaktion aus reduzierenden Zuckern und Aminen wie Lysin gebildet. Sie entstehen während der Erhitzung von Lebensmitteln und in vivo. Der enzymatische Abbau solcher spontan gebildeten Produkte ist Thema dieser Arbeit. Ein Teil untersuchte die Rolle von Oligosaccharid-Amadori-Produkten während der Verdauung von Kohlenhydraten im Dünndarm. Aufgrund ihrer strukturellen Ähnlichkeit mit bekannten Glycosidase-Inhibitoren wurde eine hemmende Wirkung der Amadori-Produkte auf die Kohlenhydratverdauung vermutet. Der andere Teil beschäftigte sich mit Fructosamin-3-kinase (FN3K) und dessen verwandtem Enzym Fructosamin-3-kinase-related Protein (FN3K-RP) aus humanen Erythrocyten. Diese Ketosaminkinasen werden als Proteinreparaturenzyme betrachtet, sogar als enzymatische Verteidigung gegen Glykierung in vivo diskutiert. Durch ihre Reaktion entstehen jedoch auch hoch-reaktive 1,2-Dicarbonylverbindungen, die weitere Proteinschäden bewirken können. Noch ist nicht klar, ob die Ketosaminkinasen die pathophysiologischen Folgen der Glykierung verhindern oder fördern. In dieser Arbeit wurde die Substratspezifität von Ketosaminkinasen mit einer Reihe von Amadori-Produkten untersucht. Damit könnten Inhibitoren zur weiteren Enzymcharakterisierung oder sogar für pharmazeutische Anwendungen identifiziert werden. Außerdem wurde die Variabilität der Enzymaktivitäten von Mensch zu Mensch in einer Kohorte von 100 Probanden untersucht. Als Modell für die menschliche Kohlenhydratverdauung im Dünndarm wurden Caco-2-Zellen als Monolayer etabliert. Deren Sucrase-Isomaltase kann die alpha-glycosidische Bindung in Amadori-Produkten von Maltose und Maltotriose mit Lysin und auch in Maltulose hydrolysieren. Trotz der Aminogruppe hemmen diese Amadori-Produkte die Maltosehydrolyse nur schwach als konkurrierende Substrate. Lactulosyllysin konnte nicht durch die Lactase der Caco-2-Zellen hydrolysiert werden. Tagatosyllysin und die Heyns-Produkte Glucosyllysin und Mannosyllysin hemmten die Lactosehydrolyse schwach. Alle beobachteten Hemmeffekte sind wahrscheinlich zu schwach, um während der Verdauung in vivo bedeutsam zu sein. Für FN3K konnte Desoxypiperidinofructose als kompetitiver Inhibitor identifiziert werden (Kic 0,006 mM). FN3K zeigte nur geringe Selektivität gegenüber Amadori-Produkten verschiedener Amine, ausgenommen aromatischer Amine. FN3K-RP war in Erythrocyten wesentlich aktiver als FN3K, auch wenn die Aktivität nicht selektiv inhibiert werden konnte. Beide Enzymaktivitäten unterscheiden sich unter den 100 Probanden, mit einer Spannweite von 3 bis 12 mU/g Hämoglobin für FN3K und 60 bis 135 mU/g Hb für FN3K und FN3K-RP zusammen. Es scheint eine Verbindung zwischen der Ketosaminkinase-Aktivität in Erythrocyten und Nierenerkrankungen, familiär auftretendem Diabetes mellitus, sowie familiär aufgetretenen Herzinfarkten oder Schlaganfällen zu bestehen, wie orientierende Auswertungen zeigten. Deshalb ist eine genauere Untersuchung der physiologischen Bedeutung der Ketosaminkinasen nötig. / Amadori products are formed spontaneously from reducing sugars and amines, e.g. lysine, during the first phase of the Maillard reaction. They occur in heated food and in vivo. The thesis focuses on the enzymatic degradation of such spontaneously formed compounds. One part of this work investigated the faith and impact of oligosaccharide derived Amadori products during small intestinal carbohydrate digestion. Due to their structural similarity with known glycosidase inhibitors, an inhibitory action of Amadori products towards carbohydrate digestions was assumed. The other part dealt with fructosamine-3-kinase (FN3K) and its related protein (FN3K-RP) from human erythrocytes. Such ketosamine kinases are regarded as protein repair enzymes, maybe even an enzymatic defence against glycation in vivo. While deglycating protein bound Amadori products, however, they produce highly reactive 1,2-dicarbonyl compounds, which can lead to further protein damage. It is unclear, whether the ketosamine kinase action prevents or supports the pathophysiological effects of glycation. This work studied the substrate specifity of ketosamine kinases with a variety of Amadori products, which could result in inhibitors for further enzyme characterisation or even pharmaceutical uses. Further, the variability of both enzyme activities in a cohort of 100 subjects was examined. As a model for human small intestinal carbohydrate digestion, a Caco-2 cell monolayer was employed. Their sucrase-isomaltase is able to hydrolyse the alpha-glucosidic linkage in Amadori products of maltose and maltotriose with lysine, as well as in maltulose. Despite their amino group, those amadori products inhibited maltose hydrolysis merely weakly as competing substrates. Lactulosyl lysine on the other hand could not be hydrolysed by Caco-2 lactase. Tagatosyl lysine and the Heyns products glucosyl lysine and mannosyl lysine showed weak inhibition of lactose hydrolysis. All observed inhibitory effects are probably too weak to be of importance during carbohydrate digestion in vivo. Deoxypiperidinofructose was identified as a competitive inhibitor of FN3K (Kic 0,006 mM). FN3K acted rather non-specific towards Amadori products of different amines, except aromatic amines. FN3K-RP showed much higher activity in erythrocytes than FN3K, although its activity could not be inhibited selectively. Both enzyme activities vary among 100 subjects, with a range of 3 to 12 mU/g hemoglobin for FN3K and 60 to 135 mU/g hb for FN3K and FN3K-RP together. Relations of ketosamine kinase activity in erythrocytes with renal diseases, familial diabetes mellitus and familial cardiovascular events seem to exist. Thus, investigating the physiological impact of ketosamine kinases is necessary.

In addition to the interaction of evolutionary forces, the population history of the African Sahel and Arabia has been influenced by the spread of Neolithic cultural innovations. The reflection of these processes today is a very complex structured diversity of the current populations, which is presented here through the analysis of several genetic markers. The aim is to provide a comprehensive view of the history of demographic processes in the Sahel and Arabia, by combining genetic, linguistic, subsistence and geographical data obtained from local populations. A study of a large dataset of mtDNA sequences showed that Arabia was a major crossroads in gene flow, and although it was colonized by anatomically modern humans from East Africa, today's differentiation from Africa is greater than the differentiation between local populations in these regions. Even the Sahel was an important biocorridor in the past. Today, we encounter populations of various subsistence strategies (nomadic pastoralists and settled farmers), between which gene flow has been severely restricted. A comparison of uniparently inherited loci in both groups points to different migratory activity in the eastern and western parts of the Sahel. Analyzes of Alu elements, which indicated the inclination of West African herders (Fulbs)...

[ES] La presente tesis doctoral, titulada "Microportadores inteligentes para la liberación controlada de sustancias de interés nutracéutico", se centra en el diseño y evaluación de sistemas híbridos orgánico-inorgánicos para proteger y liberar controladamente compuestos bioactivos. Dichos sistemas están basados en (i) materiales de sílice, principalmente partículas mesoporosas, como soporte inorgánico para almacenar y proteger la carga bioactiva; y (ii) una capa externa de biomoléculas como puerta molecular, que regula la liberación de la carga ante ciertos estímulos. En el primer capítulo de la tesis se describe el ácido oleico como puerta molecular. Este capítulo se subdivide en tres artículos diferentes, con distintos objetivos. En el primer artículo se emplea por primera vez el ácido oleico como puerta molecular de un soporte mesoporoso, cargado con la molécula modelo rodamina B. El material preparado es capaz de proteger la carga en las condiciones presentes en la boca y en el estómago, e inducir su liberación en el intestino con la acción surfactante de las sales biliares. El sistema se ha empleado para la liberación de vitamina B2, demostrando así la utilidad del diseño para la protección y liberación controlada de nutracéuticos. El segundo artículo evalúa la efectividad de esta puerta molecular en diferentes tipos de partículas mesoporosas de sílice, con diversos tamaños y estructuras de poro (MCM-41, MCM-48, SBA-15 y UVM-7). En todos los sistemas estudiados, la puerta molecular es capaz de mantener protegidas las moléculas cargadas, y liberarlas ante la presencia de sales biliares. El sólido basado en la estructura de UVM-7 se validó in vivo, observándose un retraso en la absorción intestinal de la rodamina gracias a su encapsulación. Por último, el tercer artículo incluido en este capítulo ha estudiado la posibilidad de incorporar puertas moleculares en filosilicatos. Se ha conseguido la protección y liberación controlada de biomoléculas de gran tamaño implicadas en el metabolismo humano (vitamina B12 y hematina) empleando filosilicatos funcionalizados con ácido oleico como puerta molecular. El segundo capítulo describe por primera vez el uso de la proteína zeína (prolamina de maíz) como puerta molecular. La presencia de la prolamina de maíz inhibe la salida de los compuestos antimicrobianos encapsulados (timol, carvacrol y cinamaldehído) liberándolos en presencia de las enzimas proteolíticas excretadas durante el crecimiento bacteriano. De todos los materiales desarrollados, el sistema cargado con cinamaldehído ha demostrado una inhibición del crecimiento de E. coli superior a la del compuesto libre. Finalmente, el tercer capítulo estudia la efectividad de la lactosa como puerta molecular para proteger aceites esenciales y liberarlos solo en las condiciones presentes en el intestino. Se han preparado tres materiales diferentes basados en MCM-41, cargados con timol, eugenol y cinamaldehído, y funcionalizados con lactosa para inhibir la salida de los compuestos. La acción enzimática de la lactasa secretada en el intestino es capaz de hidrolizar la puerta molecular en los correspondientes monosacáridos, liberando la carga a lo largo del lumen intestinal. Los microdispositivos diseñados han sido validados in vitro con células Caco-2, donde se ha observado el aumento de la capacidad citotóxica del cinamaldehído y la disminución de la permeabilidad a través del modelo de membrana intestinal gracias a su encapsulación. Finalmente, el microdispositivo cargado con cinamaldehído se ha validado in vivo ratificándose la disminución de la permeabilidad del compuesto y su mayor permanencia en el lumen intestinal. Así, la presente tesis doctoral ha demostrado la posibilidad de emplear biomoléculas sencillas de grado alimentario como puertas moleculares sobre diversos materiales de sílice. Estos nuevos sistemas han permitido proteger y liberar control / [CA] La present tesi doctoral, titulada "Microportadors intel·ligents per a l'alliberament controlat de substàncies d'interès nutracèutic", se centra en el disseny i avaluació de sistemes híbrids orgànic-inorgànics per a la protecció i alliberament controlat de compostos bioactius. Aquests sistemes estan basats en (i) materials de sílice, principalment partícules mesoporoses, com a suport inorgànic per emmagatzemar i protegir la càrrega bioactiva; i (ii) una capa externa de biomolècules com a porta molecular, que regula l'alliberament d'aquesta càrrega davant de determinats estímuls. En el primer capítol de la tesi es descriu l'àcid oleic com a porta molecular. Aquest capítol se subdivideix en tres articles diferents, amb objectius diferents. En el primer article s'empra per primera vegada l'àcid oleic com a porta molecular d'un suport mesoporós, carregat amb la molècula model rodamina B. El material preparat és capaç de protegir la càrrega en les condicions presents a la boca i a l'estómac, i induir el seu alliberament a l'intestí amb l'acció surfactant de les sals biliars. El sistema s'ha emprat per a l'alliberament de vitamina B2, demostrant així la utilitat del disseny per a la protecció i alliberament controlat de nutracèutics. El segon article avalua l'efectivitat d'aquesta porta molecular en diferents tipus de partícules mesoporoses de sílice, amb diverses mides i estructures de porus (MCM-41, MCM-48, SBA-15 i UVM-7). En tots els sistemes estudiats, la porta molecular és capaç de mantindre protegides les molècules carregades, i alliberar-les davant la presència de sals biliars. El sòlid basat en l'estructura de UVM-7 es validà in vivo, observant-se un retard en l'absorció intestinal de la rodamina gràcies a la seua encapsulació. Finalment, en el tercer article inclòs en aquest capítol s'ha estudiat la possibilitat d'incorporar portes moleculars en fil·losilicats. S'ha aconseguit la protecció i alliberament controlat de biomolècules de grans dimensions implicades en el metabolisme humà (vitamina B12 i hematina) emprant fil·losilicats funcionalitzats amb àcid oleic com a porta molecular. El segon capítol descriu per primera vegada l'ús de la proteïna zeïna (prolamina de dacsa) com a porta molecular. La presència de la prolamina de dacsa inhibeix la sortida dels compostos antimicrobians encapsulats (timol, carvacrol i cinamaldèhid) alliberant-los en presència dels enzims proteolítics excretades durant el creixement bacterià. De tots els materials desenvolupats, el sistema carregat amb cinamaldèhid ha demostrat una inhibició de l'creixement d'E. coli superior a la del compost lliure. Finalment, el tercer capítol estudia l'efectivitat de la lactosa com a porta molecular per protegir olis essencials i alliberar-los només en les condicions presents a l'intestí. S'han preparat tres materials diferents basats en MCM-41, carregats amb timol, eugenol i cinamaldèhid, i funcionalitzats amb lactosa per inhibir l'eixida dels compostos. L'acció enzimàtica de la lactasa secretada a l'intestí és capaç d'hidrolitzar la porta molecular en els corresponents monosacàrids, alliberant la càrrega al llarg del lumen intestinal. Els microdispositius dissenyats s'han validat in vitro amb cèl·lules Caco-2, on s'observà l'augment de la capacitat citotòxica del cinamaldèhid i la disminució de la permeabilitat a través del model de membrana intestinal gràcies a la seua encapsulació. Finalment, el microdispositiu carregat amb cinamaldèhid s'ha validat in vivo ratificant la disminució de la permeabilitat del compost i la seua major permanència al lumen intestinal. Així, la present tesi doctoral ha demostrat la possibilitat d'emprar biomolècules senzilles de grau alimentari com portes moleculars sobre diversos materials de sílice. Aquests nous sistemes han permès protegir i alliberar controladament diferents nutracèutics, millorant així la seua biodisponibilitat. / [EN] This PhD thesis, entitled "Smart microdevices for nutraceutical-delivery", is focused on the design and evaluation of organic-inorganic hybrid systems for the protection and controlled release of bioactive molecules. These systems are based on (i) silica materials, mainly mesoporous particles, as inorganic support to store and protect the bioactive cargo; and (ii) an outer layer of biomolecules that regulate the payload release triggered by certain stimuli. In the first chapter of the thesis oleic acid is described as a molecular gate. This chapter is subdivided into three different articles, with different objectives. In the first article, oleic acid is used for the first time as molecular gate of a mesoporous support, loaded with the rhodamine B model molecule. The designed material is capable of protecting the cargo under the conditions present in the mouth and stomach, and inducing its release in the small intestine with the surfactant action of bile salts. The system has been used for the release of vitamin B2, thus demonstrating the validity of the design for the protection and controlled release of nutraceuticals. The second article evaluates the effectiveness of this molecular gate in different types of mesoporous silica particles, with different sizes and pore structures (MCM-41, MCM-48, SBA-15 and UVM-7). In all the systems studied, the molecular gate is capable of keeping cargo molecules protected and releasing them in the presence of bile salts. The solid based on the structure of UVM-7 was validated in vivo, observing a delay in the intestinal absorption of rhodamine thanks to its encapsulation. Lastly, the third article included in this chapter has studied the possibility of incorporating molecular gates onto phyllosilicates. The protection and controlled release of large biomolecules involved in human metabolism (vitamin B12 and hematin) have been achieved using phyllosilicates functionalized with oleic acid as molecular gate. The second chapter describes for the first time the use of the protein zein (corn prolamine) as a molecular gate. The presence of corn prolamine inhibits the release of encapsulated antimicrobial compounds (thymol, carvacrol and cinnamaldehyde) releasing them in the presence of the proteolytic enzymes excreted during bacterial growth. Among all the materials developed, the cinnamaldehyde-loaded system has shown greater inhibition of E. coli growth than the free compound. Finally, the third chapter studies the effectiveness of lactose as a molecular gate to protect essential oils and release them only under the conditions present in the intestine. Three different materials based on MCM-41 loaded with thymol, eugenol, and cinnamaldehyde, and functionalized with lactose to inhibit the release of the compounds have been prepared. The enzymatic action of the lactase secreted in the intestine is capable of hydrolyzing the molecular gate into the corresponding monosaccharides, thus releasing the cargo along the intestinal lumen. The designed microdevices have been validated in vitro with Caco-2 cells, where an increase in the cytotoxic capacity of cinnamaldehyde and a decrease in permeability through the intestinal membrane model have been observed thanks to its encapsulation. Finally, the cinnamaldehyde-loaded microdevice has been validated in vivo, confirming the decrease in the permeability of the compound and its greater permanence in the intestinal lumen. Thus, the present PhD thesis has demonstrated the possibility of using simple food-grade biomolecules as gatekeepers on various silica materials. These new systems have allowed the protection and controlled release of different nutraceuticals, thus improving their bioavailability. / The authors also thank the Electron Microscopy Service at the UPV for support. The authors also thank Prof. Pedro Amorós for his explanations and guidance on the knowledge of phyllosilicates. / Poyatos Racionero, E. (2020). Smart microdevices for nutraceutical-controlled delivery [Tesis doctoral]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/159247 / TESIS