Orientador: Ming Chih Chiu / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Engenharia de Alimentos / Made available in DSpace on 2018-08-22T11:15:33Z (GMT). No. of bitstreams: 1 Santos_CamiliaAoyagui_M.pdf: 3664368 bytes, checksum: 08f28439bf0d2c55027a15763d6a5b3e (MD5) Previous issue date: 2013 / Resumo: O óleo de palma possui lenta cristalização devido à alta energia livre de ativação de nucleação, atribuída às interações intermoleculares entre a estearina e a oleína e à presença dos diacilgliceróis. Aditivos podem ser adicionados ao óleo de palma na tentativa de modificar o seu comportamento de cristalização, adequando o seu uso aos processos industriais. Dentre esses aditivos se destacam os emulsificantes, que diferentemente da sua função usual, podem ser utilizados para alterar a estrutura cristalina e contribuir inclusive para a estabilidade de formas polimórficas desejadas nos óleos e gorduras onde são aplicados. Os ésteres de ácidos graxos de sorbitana têm demonstrado influência na morfologia dos cristais e na textura de misturas de óleos à base de palma. No entanto, estudos indicaram que os monoésteres de ácidos graxos de sorbitana disponíveis comercialmente possuíam diferentes composições em mono-, di- e triésteres de ácidos graxos de sorbitana. Esta variabilidade pode dificultar a comparação de resultados obtidos de cristalização de óleos e gorduras após adição destes emulsificantes entre os diversos estudos já realizados. O objetivo deste trabalho foi determinar a composição de vários ésteres de ácidos graxos comerciais, de diferentes fornecedores, e verificar se o comportamento de cristalização do óleo de palma refinado (RPO) adicionado dos emulsificantes foi influenciado pelas possíveis diferenças na composição deles. A análise de composição, realizada através da técnica de espectrometria de massas, EASI-MS, revelou diferenças na composição dos monolauratos de sorbitana (SMLa) dos fornecedores B, C e D e dos monoestearatos de sorbitana (SMS) dos fornecedores C, D e E. Entre os SMLa, o SMLa D foi o que apresentou o maior teor de trilaurato de sorbitana e de triésteres de modo geral, e dentre os SMS, o SMS E foi o que apresentou o maior teor de monoestearato de sorbitana, maior teor de monoésteres e menor teor de diésteres. Nas isotermas de cristalização realizadas a 25°C através da ressonância magnética nuclear (RMN), o SMLa D foi o que mais retardou o período de nucleação (em 128,6%) de RPO e o que possibilitou o menor teor máximo de gordura sólida (SFCMáx). Além disso, após análise dos eventos de cristalização por calorimetria diferencial de varredura (DSC), o SMLa D foi o único, dentre os SMLa, a retardar significativamente (p < 0,05) a nucleação e a cristalização dos TAGs de maior ponto de fusão de RPO. Em relação aos SMS, após análise das imagens da microestrutura dos cristais, notou-se que apenas a amostra com SMS E produziu cristais de morfologia e dimensões diferentes. Além disso, os resultados da análise de consistência mostraram que somente o SMS E aumentou significativamente (p < 0,05) a dureza do RPO nas temperaturas de 15°C, 20°C e 25°C. As diferenças encontradas na composição dos monolauratos e monoestearatos de sorbitana puderam ser associadas aos efeitos distintos no comportamento de cristalização do óleo de palma refinado / Abstract: Palm oil has a slow crystallization possibly due to the high free energy of activation of nucleation and additives can be added in an attempt to change its crystallization behavior adapting its use to industrial processes of margarine, shortenings, toppings and baking. Among these additives there are emulsifiers, which unlike its usual application can be used to alter the crystal structure and also contribute to the stability of desired polymorphic forms in the oil and fat where they are applied. The sorbitan fatty acid esters have been shown to influence the morphology, the distribution of crystals and the texture of blends of palm. However studies indicated that sorbitan monoesters commercially available had different compositions in mono-, di-and triesters of sorbitan fatty acids such variability may make it difficult to compare results obtained from crystallization of fats and oils after addition of emulsifiers among the many studies already conducted. The objective of this study was to determine the composition of several commercial sorbitan fatty acid esters from different suppliers and verify if the crystallization behaviour of refined palm oil (RPO) after addition of the emulsifiers was influenced by possible differences in their composition. The composition analysis using the technique of mass spectrometry, EASI-MS, revealed differences in the composition of sorbitan monolaurates (SMLa) from suppliers B, C and D and sorbitan monostearates (SMS) from suppliers C, D and E. Among SMLa, the SMLa D indicated the highest content of trilaurate and sorbitan triesters. Among SMS, SMS E presented the highest level of sorbitan monostearate, and higher content of monoesters and lower content of diesters. In the isothermal crystallization performed at 25°C by nuclear magnetic resonance (NMR), the SMLa D was the emulsifier the most retarded the nucleation period (at 128.6%) of the RPO and it resulted of the lowest maximum solid fat content (SFCMáx). Furthermore, after crystallization event analysis by differential scanning calorimetry (DSC), the SMLa D was the only one delayed significantly (p <0.05) the nucleation and crystallization of TAGs of higher melting point of the RPO. In relation to SMS, after examining the images of the microstructure of the crystals, it was noted that only the sample with SMS E produced crystals of different dimensions and morphology. Furthermore, the results of consistency analysis showed that only SMS E increased significantly (p <0.05) the hardness of the RPO at 15°C, 20°C and 25°C. The differences found in the composition of sorbitan monolaurates and monostearates could be associated with distinct effects on the crystallization behavior of refined palm oil / Mestrado / Tecnologia de Alimentos / Mestra em Ciência de Alimentos

Orientadores: Maria Regina Wolf Maciel, Cibelem Iribarrem Benites / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Engenharia Química / Made available in DSpace on 2018-08-18T15:37:18Z (GMT). No. of bitstreams: 1 Galucio_CleysondeSouza_M.pdf: 2671846 bytes, checksum: 073b9a1a95f494699d09f6ea38120b15 (MD5) Previous issue date: 2011 / Resumo: Monoacilgliceróis (MAG) são emulsificantes utilizados em produtos alimentícios, farmacêuticos e cosméticos. A obtenção de MAG a partir de óleos vegetais vem se destacando frente aos processos industriais devido aos benefícios que compostos naturais trazem à saúde humana. Industrialmente, são produzidos através da glicerólise química de triacilgliceróis (TAG), gerando cerca de 40 a 50% de MAG. Para algumas aplicações, é necessário concentrá-lo, sendo o processo de destilação molecular uma alternativa vantajosa, porque possibilita a concentração de compostos termicamente sensíveis e não utiliza solventes. Neste trabalho, foi realizado o estudo da glicerólise química do óleo de girassol (Helianthus annus), que se destaca por ser rico em ácido linoléico, pertencente à família ?-6. Após a reação, os MAG obtidos foram concentrados por destilação molecular. Foram utilizados dois destiladores moleculares de filme descendente agitado das marcas POPE e UIC-GmbH, salientando-se este último por apresentar as melhores condições de operação para o estudo de concentração de MAG. A técnica de planejamento de experimentos foi utilizada para o estudo dos fatores envolvidos na obtenção e concentração de MAG. Assim, nas condições otimizadas de glicerólise química, obteve-se 48,3% de MAG, 42,3% de diacilgliceróis (DAG), 6,9% de TAG e 2,8% de glicerol (GL). No processo destilação molecular foi realizada, primeiramente, a remoção de GL na corrente de destilado; a corrente de resíduo (rica em acilgliceróis) foi submetida a outra destilação molecular, visando aumentar a concentração de MAG. Na primeira destilação, sob temperatura do evaporador (TEV) de 90ºC, retirou-se 99,5% de GL. Na segunda destilação molecular concentraram-se os MAG em 82,6% na corrente de destilado, sob TEV de 170°C. Tanto o óleo vegetal quanto a mistura de acilgliceróis (oriunda da glicerólise) e o concentrado de MAG foram caracterizados por métodos físico-químicos, tomando como referência as características do óleo de girassol. Seguindo essas características de identidade e qualidade, os resultados obtidos apresentaram-se dentro dos padrões de legislação (ANVISA e CODEX). Ressalta-se que o conteúdo de ácidos graxos insaturados não foi alterado durante o processo de obtenção e concentração de MAG que apresentou características de qualidade aceitável para aplicação como emulsificante. A destilação molecular de filme descendente agitado (UIC-GmbH) mostrou-se uma ótima alternativa para concentração de MAG termicamente degradáveis. Portanto, nas condições estudadas, a glicerólise química e a destilação molecular mostraram-se processos viáveis para a obtenção de emulsificantes naturais concentrados / Abstract: Monoacylglycerols (MAG) are emulsifiers used in food, pharmaceutical and cosmetic products. MAG obtained from vegetable oils has been emphasizing to industrial processes due to the benefits that these natural compounds bring to human health. Industrially, these are produced by chemical glycerolysis of triacylglycerols (TAG), generating about 40 to 50% of MAG. For some applications, it is necessary to concentrate it and the process of molecular distillation is attractive because it enables the concentration of thermally sensitive compounds and does not use solvents. This work was carried out to study the chemical glycerolysis of sunflower (Helianthus annus), which is notable for being rich in linoleic acid (?-6 family). After reaction, the MAG obtained was concentrated by molecular distillation. Two falling film molecular distillers were used (POPE and UIC-GmbH), emphasizing this last one by better operating conditions for the study of concentration of MAG. The technique of experimental design was used to study the factors involved in MAG production and concentration. Thus, under optimized conditions, the chemistry glycerolysis obtain 48.3% of MAG, 42.3% diacylglycerols (DAG), 6.9% of TAG and 2.8% of glycerol (GL). Firstly, the molecular distillation process was conducted to remove the GL in the distillate stream, the residue stream (rich in acylglycerols) was feed to a second molecular distillation, to increase the MAG concentration. In the first distillation, at 90°C of evaporator temperature (TEV), 99.5% from GL was removed. In the second molecular distillation, MAG was concentrated at 82.6% in the distillate stream, under 170 °C of TEV. Both, the vegetable oil and the mixture of acylglycerols (from glycerolysis) and the MAG concentrate were characterized by physicochemical methods, with sunflower oil as reference. Following these characteristics of identity and quality, the results were inside the standards (ANVISA and CODEX). It is important to stand out that the unsaturated fatty acids were not changed during the process of obtaining and concentration of MAG showing acceptable quality characteristics for use it as an emulsifier. The agitated falling film molecular distillation (UIC-GmbH) was a great alternative to the concentration of thermally degradable MAG. Therefore, under these studied conditions, the chemical glycerolysis and the molecular distillation proved itself as a viable process for obtaining concentrated natural emulsifiers / Mestrado / Desenvolvimento de Processos Químicos / Mestre em Engenharia Química

A produção de compostos biologicamente ativos por micro-organismos marinhos tem atraído crescente interesse de biotecnólogos e microbiologistas nas últimas décadas. No presente trabalho 162 isolados de fungos filamentosos recuperados de amostras de água do mar e de dez diferentes macro-organismos marinhos foram avaliados quanto a produção de lipases pelo método de High Throughput Screening (HTS). Destes, 45 isolados tiveram a produção de lipases confirmada individualmente. A atividade lipolítica foi quantificada, e os resultados sugerem que fungos filamentosos oriundos de ambientes marinhos tem maior potencial para produção de lipases estáveis em pH alcalino. Entre estes, Fusarium sp. CBMAI 1227, Aspergillus parasiticus CBMAI 1228 e Trichoderma sp. CBMAI 1229 apresentaram atividade expressiva e de interesse biotecnológico em pH 8,0 (23,1, 12,7 e 12,2 U respectivamente). Os resultados deste trabalho demonstram o potencial dos fungos provenientes de ambiente marinho para aplicações biotecnológicas, e estimulam novos estudos envolvendo lipases. / The production of biologically active compounds by marine microorganisms has been drawing increasing interest of biotechnologists and microbiologists in the last decades. In the present work 162 filamentous fungi isolates recovered from water samples and ten different marine macroorganisms were evaluated for the production of lipases by the method of High Throughput Screening (HTS). Out of those, 45 had the lipase production individually confirmed. The lipolytic activity was quantified, and the results suggest that marine-derived filamentous fungi have a greater potential for the production of lipases stable under alkaline pH. Among these, Fusarium sp. CBMAI 1227, Aspergillus parasiticus CBMAI 1228 and Trichoderma sp. CBMAI 1229 showed expressive and biologically interesting activity in pH 8,0 (23,1, 12,7 and 12,2 U/min respectively). The results of the present work show the potential of marine-derived fungi in biotechnological applications, and stimulate new studies involving lipases.

Presented diploma thesis was focused on study of influence of stabilizers and other components on final quality of mixtures for ice-cream production. This thesis was worked out in cooperation with Frujo Co., Ltd., Tvrdonice to find out the most suitable stabilizer with positive influence on the quality of final product. In experimental part six mixtures of stabilizers, three mixtures for ice-cream production and particular ingredients for production of mixtures for ice-cream production were analysed. Saccharides and lipids were analysed only in mixtures of stabilizers. Content of saccharides was analyzed using spectrophotometry and HPLC/RI. Lipid components were analysed by TLC. Viscosity was measured by rotatory viscosimeter and the differences of flow curves of each mixture with different stabilizers were monitored. Based on viscosity measurements some ice-creams with different stabilizers were chosen for sensory analysis. Mixture of stabilizers “Stobicol” was analysed for active compounds and antioxidant stability. Microbiological stability was tested in all of the stabilizers and also in particular ingredients including final mixture for ice-cream production. In three ice-cream mixtures of different flavour protein composition was measured by microfluidic electrophoresis. The highest content of total and reducing saccharides was determined in hydrolyzed samples. The highest values of total saccharides were found in mixture of stabilizers “Meyprogen”, the highest values of reducing saccharides were detected in stabilizer “Swissgum”. Content of triglycerides was found out almost in all of the measured mixtures of stabilizers using TLC method even though according to formulas provided by producers there should not be any. Diglycerides were detected in “Meyprogen” too even though this mixture of stabilizers should contain only saccharides. Based on results of rheology measurements and in connection with sensory analysis “Meyprogen” stabilizer could be recommended as the best material for the industrial production.

京都大学 / 新制・課程博士 / 博士(農学) / 甲第23248号 / 農博第2455号 / 新制||農||1084(附属図書館) / 学位論文||R3||N5338(農学部図書室) / 京都大学大学院農学研究科農学専攻 / (主査)教授 松村 康生, 教授 丸山 伸之, 教授 谷 史人 / 学位規則第4条第1項該当 / Doctor of Agricultural Science / Kyoto University / DGAM

The lubricants that are applied during metal cutting acts on the interface between the tool and the nascent surfaces generated by the cutting process. Dispersions of oil in water made using suitable emulsifier(s) are used as metal cutting lubricants. The efficiency of the emulsion in rendering a low friction layer on the freshly cut surface will depend on the composition of the emulsion and on the speed, load and temperature characteristics in the tribological system. A unique tribometer which can perform friction testing on freshly cut surfaces has been designed and built for the experimental investigation. In this experimental facility experiments are conducted by performing cutting operation inside a pool of the lubricant and friction force is measured in-situ. Experiments at different loads and speeds were performed. The surfaces were subsequently subjected to spectroscopic analysis using X-ray Photoelectron spectroscopy and Fourier Transform Infrared Spectroscopy (FTIR). Lubricity of the base oils on nascent and preformed (oxidized) surfaces are compared by performing friction tests on surfaces which are cut and friction tested without exposing them to the environment, and on surfaces which were cut and exposed to the environment. While the freshly cut surfaces were seen to be sensitive to the structure of the base oil, the oxidized surfaces did not differentiate between the oil structures. Amongst the three base oils tested, aromatic oil was found to exhibit the least friction. This is attributed to tendency of the aromatic chains to react with the surface and form a film, due to the formation of radical anion-metal cation complexes. Results from spectroscopic investigations are presented to substantiate these arguments. The thesis then explores the differences in the tribological behavior promoted by an emulsion between, when it acts on a cut surface and is slid just once, and when it acts on a cut surface slid repeatedly. Due to repeated sliding, friction was found to decrease with sliding time (distance), and the transition from a freshly formed surface to a repeatedly slid one was found to follow a smooth transition. The improvement in lubricity is attributed to the formation of carboxylate type structures (C=O) which get generated due to the tribological action under repeated sliding conditions in the presence of water. Under repeated sliding conditions, the friction as a function of emulsifier concentration is found to exhibit a minimum at a value which is much below the critical micellar concentration of the emulsifier (CMC). However, the variation under continuous cutting followed a different pattern, with the friction undergoing a sharp decrease close to the CMC. The effect of speed on the tribological performance was investigated and friction was found to increase dramatically beyond a critical speed which is marked as the onset of starvation. The characteristic time required for a film to develop on a newly created surface, together with the contact pressure conditions dictated by the load and speed dictates starvation. The films formed at speeds corresponding to starvation conditions was found to have a significantly different chemical structure from that corresponding to a speed less than the starvation speed.. The effect of temperature was found to affect the lubricity adversely. At elevated temperature, the nature of the film was found to change to that to starved condition, even at a speed which does not register starvation when operating at a lower temperature. The effect of solubility of the emulsifier on the friction characteristics were explored by using emulsifiers of varying hydrophilic-lypophilic values (HLB). Lower HLB emulsifiers were found to exhibit lesser friction, than those corresponding to high HLB value. The variation in lubricity is examined in the light of the morphology of the micellar structures which evolve using these emulsifiers. The main conclusions of the thesis are: 1 Evaluation of lubricity of metal cutting fluids warrants a testing strategy which tests their lubricity on freshly cut surfaces. 2 The formation of carboxylate structures aids lubricity while using an emulsion; emulsions which can result in the formation of such structures exhibit better lubricity under cutting conditions. 3 Tribofilms which show characteristic peaks related to chemisorbed oxygen is found to exhibit good lubricity under the test conditions. 4 Emulsifiers which form lamellar micellar structures which aid easy shear give better lubricity in cutting than those which yield spherical micelles.

京都大学 / 0048 / 新制・論文博士 / 博士(農学) / 乙第12820号 / 論農博第2793号 / 新制||農||1025(附属図書館) / 学位論文||H26||N4815(農学部図書室) / 31307 / 京都大学農学研究科農学専攻 / (主査)教授 松村 康生, 教授 裏出 令子, 教授 安達 修二 / 学位規則第4条第2項該当 / Doctor of Agricultural Science / Kyoto University / DFAM

Τα γαλακτώματα είναι η κολλοειδής διασπορά δύο μη αναμίξιμων υγρών, τα οποία είναι κατά κανόνα θερμοδυναμικά ασταθή συστήματα. Οι πρωτεΐνες γάλακτος είναι γνωστές επιφανειοδραστικές ουσίες και ως εκ τούτου χρησιμοποιούνται ως συστατικά σε ένα ευρύ φάσμα γαλακτωμάτων τροφίμων. Σκοπός της παρούσας εργασίας είναι η εύρεση των κατάλληλων συνθηκών για την παρασκευή σταθερών γαλακτωμάτων πρωτεϊνών γάλακτος. Το μέγεθος των λιποσφαιριδίων διαδραματίζει τον κυρίαρχο ρόλο στη σταθερότητα του γαλακτώματος πρωτεϊνών γάλακτος. Η μέτρηση του μεγέθους των λιποσφαιριδίων έγινε με την τεχνική της Μονοφασικής Χρωματογραφίας Φυγοκεντρικού Πεδίου. Πιο συγκεκριμένα, μελετήθηκε η επίδραση της συγκέντρωσης (0,5 έως 3,0% w/w) και του τύπου (πρωτεΐνες ορού και καζεΐνες) των πρωτεϊνών γάλακτος, καθώς και των συνθηκών ομογενοποίησης (πίεση ομογενοποίησης 200 έως 600bar) του γαλακτώματος. Επίσης, μελετήθηκε η επίδραση της συγκέντρωσης γαλακτωματοποιητών εμπορίου (Tween 80) στη σταθερότητα των γαλακτωμάτων. Επιπρόσθετα, έγινε κινητική μελέτη συσσωμάτωσης των γαλακτωμάτων από πρωτεΐνες γάλακτος και στη συνέχεια μελετήθηκαν πιο συγκεκριμένα τα γαλακτώματα καζεϊνών, με σκοπό τον προσδιορισμό της σταθεράς ταχύτητας της συσσωμάτωσης των λιποσφαιριδίων σε θερμοκρασίες 30,5 και 80 ᵒC. Αυξάνοντας την πίεση ομογενοποίησης του γαλακτώματος παρατηρήθηκε μείωση της διαμέτρου των λιποσφαιριδίων. Τα γαλακτώματα που ομογενοποιήθηκαν σε πίεση μεγαλύτερη των 500 bar παρουσίασαν ευρύτερη κατανομή μεγέθους, λόγω της υψηλής θερμοκρασίας που αναπτύχθηκε κατά τη διάρκεια της ομογενοποίησης. Η πρωτεϊνική συγκέντρωση έχει σημαντικές επιπτώσεις στις φυσικοχημικές ιδιότητες του γαλακτώματος (λάδι σε νερό). Αυξανόμενης της συγκέντρωσης των πρωτεϊνών γάλακτος, μειώθηκε αισθητά η διάμετρος των λιποσφαιριδίων του γαλακτώματος και σε χαμηλές συγκεντρώσεις πρωτεϊνών (<1%κ.β.) παρατηρήθηκε σχηματισμός συσσωματωμάτων. Παρατηρήθηκε μικρή μεταβολή της διαμέτρου των λιποσφαιριδίων των γαλακτωμάτων που σχηματίστηκαν με διαφορετικές αναλογίες κλασμάτων πρωτεϊνών ορού/καζεϊνών. Οι δύο τύποι των πρωτεϊνών του γάλακτος παρουσίασαν πολύ καλή γαλακτωματοποιητική δράση και τα γαλακτώματα που σχηματίστηκαν ήταν πολύ σταθερά. Παρατηρήθηκε ότι, αυξάνοντας τη συγκέντρωση των πρωτεϊνών του ορού γάλακτος και ταυτόχρονα μειώνοντας τη συγκέντρωση των καζεϊνών, μειώθηκε η διάμετρος των λιποσφαιριδίων του γαλακτώματος. Η σταθερότητα των γαλακτωμάτων σε σχέση με τον χρόνο οφείλεται στη δομή των μορίων των πρωτεϊνών που σταθεροποιούν τα γαλακτώματα αυτά. Κατά συνέπεια, τα μόρια των καζεϊνών και των πρωτεϊνών ορού, προσδίδουν διαφορετικές ιδιότητες στα γαλακτώματα λόγω της διαφορετικής δομής τους. Τα γαλακτώματα που σχηματίστηκαν με καζεΐνες ήταν πιο ανθεκτικά στην θερμοκρασία και παρουσίασαν μακροπρόθεσμη σταθερότητα σε σχέση με τα γαλακτώματα που περιείχαν μόνο πρωτεΐνες ορού. Η υπολογισθείσα φαινόμενη σταθερά συσσωμάτωσης των γαλακτωμάτων καζεϊνών στη θερμοκρασία των 30,5 ᵒC βρέθηκε να είναι σχεδόν 14 φορές μικρότερη από αυτή των γαλακτωμάτων που συσσωματώθηκαν στη θερμοκρασία 80,0 ᵒC. Επομένως, η διαδικασία της συσσωμάτωσης συμβαίνει ταχύτερα σε πιο υψηλές θερμοκρασίες θέρμανσης για τα γαλακτώματα καζεϊνών, ωστόσο έφτασαν στο μέγιστο βαθμό συσσωμάτωσης σε ίδιο χρονικό διάστημα. / In the food industry, when referring to an oil-in-water emulsion, is usually described in which oil is dispersed in the form of small spherical droplets in the continuous phase. Food emulsions are thermodynamically unstable. Nevertheless, food scientists are able to slow down the above physicochemical mechanisms responsible for emulsion instability and thus, extend the self-life of such products by a relatively simple and well studied process, termed emulsification. Surface active materials termed emulsifiers, such as proteins help produce small droplets and contribute to the stability of the emulsion. Emulsifiers decrease the interfacial tension between the oil and water phases through rapid adsorption to the surface of the newly formed oil droplets. Milk proteins (caseins and whey proteins) are well known surfactants and hence are used as ingredients in a wide range of food emulsions. One of the important parameters affecting the quality, appearance and taste of the final food products is the particle size of the ingredients included. For example, particle size of fat globules plays predominant role in the stability of the milk-protein stabilized emulsion. Milk protein-stabilized model emulsions were formed using high-pressure homogenization and the effect of homogenization pressure during emulsification, protein concentration, type of milk proteins (casein and whey proteins) and the effect of the surfactant Tween-80 were studied. The kinetic of milk protein emulsions aggregation was also studied and moreover, the apparent rate constant was calculated for the aggregation of caseinate stabilized emulsions in different temperatures (30,5 and 80,0 ᵒC). Sedimentation field flow fractionation was employed for the size characterization of oil droplets and the results obtained are consistent with those of other studies. Increasing protein content results in significant reduction in emulsion particle size for the concentration range (0.5 – 3.0 % w/w) employed in this study. Low protein content (<1%) may be correlated with bridging flocculation leading to increased particle size, as indicated by optical microscopy. Similarly, increasing pressure during the homogenization process results in decreasing significantly the particle size of the oil-in-water emulsions, for the pressure range (200 – 600 bar) utilized in this study. Increased heating associated with high levels of pressure during the homogenization process, can result in changes in the oil or protein structure, which in turn may have an impact on the physicochemical properties of the oil-in-water emulsions on a long-term basis. The two types of milk proteins appeared to be both good emulsifiers and the formed emulsions were very stable. Increasing whey protein content and together decreasing the casein content, results in small reduction in emulsion particle size. Different proteins depending on their composition and structure posses’ properties, which render them, better emulsifiers than others. Caseinate stabilized emulsions were more resistant in heating time than whey stabilized emulsions. The calculated apparent rate constant for the aggregation of caseinate stabilized emulsions at the temperature of 30,5 ᵒC was found to be fourteen times smaller than the one at the temperature of 80,0 ᵒC. Therefore, the aggregation process is faster in high temperatures for caseinate stabilized emulsions, although the maximum of aggregation point is attained at the same time in both temperatures.

Esta memoria doctoral forma parte del proyecto financiado por el Ministerio de Ciencia e Innovación (CICYT, CTQ2010-21183-C02-01), el cual tiene entre sus objetivos la búsqueda de nuevas cepas con capacidad para producir emulsionantes naturales poliméricos. Existe un interés real en el aislamiento y caracterización estructural y funcional de nuevos exopolímeros (EPS) obtenidos a partir de microorganismos de ambientes extremos que pudieran ser utilizados con estos fines en diversas industrias. Para ello, se realizó en primer lugar el estudio de los EPS desde el punto de vista ultraestructural, de cepas antárticas adaptadas al frío, mediante técnicas de microscopía electrónica de transmisión (MET) después de procesar las muestras por criofijación a alta presión seguida de criosustitución (HPF-FS). Estas técnicas aportaron numerosos detalles sobre la ultraestructura de las distintas cepas y el material extracelular que producen, al lograr preservar el material biológico próximo a su estado natural. Se observó que este material polimérico extracelular es para la mayoría de las cepas complejo con la presencia de material capsular alrededor de las células y abundantes estructuras vesiculares dispersas en la matriz extracelular secretada por las bacterias. Es la primera vez que se describen y visualizan de manera tan clara y abundante las vesículas de membrana externa (VME) que producen bacterias no patógenas de ambientes naturales antárticos. De igual forma, es la primera vez que se reporta la presencia de estas estructuras en cepas de los géneros Marinobacter y Psychrobacter. Asimismo, se estudió el origen de las proteínas presentes en las VME mediante geles SDS-PAGE donde se mostraron los perfiles proteicos comparados con las proteínas de membrana externa. Esto evidenció el origen de estas VME formadas a partir de esta membrana, al presentar proteínas en la fracción de VME, que comigraron con proteínas que estaban presentes en la fracción de membrana externa, además en el perfil proteico de la membrana externa se detectaron proteínas adicionales que no estaban presentes en VME. Se estudió la influencia de la temperatura sobre la producción y morfología de VME producidas en la cepa S. livingstonensis NF22T Para esta bacteria psicrotolerante se demostró que la temperatura influye en la producción de VME. A bajas temperaturas la cantidad de VME que produce la cepa es mayor, su tamaño es menor y más regular y el perfil electroforético muestra la expresión diferencial de algunas proteínas, viéndose sobrexpresadas proteínas relacionadas con funciones de transporte a nivel de membrana. Se realizaron análisis proteómicos para identificar las proteínas presentes en las VME producidas a 4 y 16 ºC a partir de S. livingstonensis NF22T y S. vesiculosa M7T. Para ambas cepas se identificaron fundamentalmente proteínas de membrana externa y periplasmáticas con diferentes funciones fisiológicas, destacando por su abundancia las proteínas implicadas en el transporte y metabolismo de iones inorgánicos así como las relacionadas en la biogénesis de las envueltas celulares. El material extracelular (EPS) obtenido y purificado a partir de Shewanella vesiculosa M7T presentó mayor actividad emulsionante frente a aceites comestibles que los emulsionantes comercializados goma arábiga y xantano y su caracterización reveló que contiene abundantes VME y polímeros polisacarídicos, siendo sus componentes químicos mayoritarios azúcares neutros y aminados, lípidos de membrana y aminoácidos. / The present work is part of the research project (CICYT, CTQ2010-21183-C02-01), which has among its objectives the search for new strains capable to produce natural polymeric emulsifiers. There is a real interest in the isolation and structural and functional characterization of new exopolymers (EPS) derived from microorganisms of extreme environments that could be used for these purposes in various industries. Many Gram-negative, cold-adapted bacteria from the Antarctic environment produce large amounts of extracellular matter, which has potential biotechnology applications. We examined the ultrastructure of extracellular matter from Antarctic bacteria by transmission electronic microscopy after high pressure freezing and freeze substitution. All analyzed extracellular matter appeared as a netlike mesh composed of a capsular polymer around cells and large numbers of outer membrane vesicles (OMV), which have not been described for members of the genera Psychrobacter and Marinobacter so far. OMV showed the typical characteristics described for these structures, and seemed to be surrounded by the same capsular polymer as that found around cells. The analysis of OMV proteins from Antarctic strains by SDS-PAGE showed different banding profiles in OMV compared to the outer membrane, suggesting some kind of protein sorting during membrane vesicle formation. For the psychrotolerant bacterium, S. livingstonensis NF22T, the growth temperature seemed to influence the amount and morphology of OMV. In an initial attempt to elucidate the functions of OMV from S. livingstonensis NF22T and S. vesiculosa M7T we conducted a proteomic analysis on membrane vesicles obtained at 4 and 16°C. At both temperatures, OMV were highly enriched in outer membrane proteins and periplasmic proteins related to nutrient processing and transport in Gram-negative bacteria, suggesting that OMV could be related with nutrient sensing and bacterial survival. Differences were observed in the expression of some proteins depending on incubation temperature but further studies will be necessary to define their roles and implications in the survival of bacteria in the extreme Antarctic environment. The extracellular material (EPS) obtained and purified from Shewanella vesiculosa M7T had a higher emulsifying activity against edible oils than commercial emulsifiers like arabic and xanthan gums. Characterization of this EPS revealed that it contains abundant OMV and polysaccharides polymers, with neutral and amino sugars, membrane lipids and aminoacids as its major chemical components.

[ES] La presente Tesis doctoral plantea distintas estrategias para el diseño y desarrollo de oleogeles estables, con un perfil lipídico de alta calidad nutricional y la posterior incorporación de los oleogeles en la formulación de distintos alimentos. Se pretende diseñar alimentos mediante el reemplazo de grasas sólidas, ricas en ácidos grasos saturados y trans por oleogeles, que, por un lado, mantengan las propiedades texturales y organolépticas, y, por otro lado, presenten un perfil lipídico mejorado. En una primera etapa, se desarrollaron oleogeles elaborados con 1% de hidroxipropilmetilcelulosa y 0,6% de goma xantana empleando dos condiciones de secado diferentes: secado convencional en estufa a 80 °C durante 10 h 30 min y secado a vacío a 60 °C durante 24 h. Los aceites estructurados fueron aceite de oliva, lino, girasol y girasol alto oleico. La microestructura permitió apreciar oleogeles bien estructurados cuando se utilizaron los aceites de oliva, girasol y girasol alto oleico. La estabilidad física y las propiedades reológicas corroboraron la formación de oleogeles de alta estabilidad física, a lo largo de 35 días de almacenamiento, y con un comportamiento de gel sólido. Se observó que tanto el grado de insaturación del aceite como las condiciones de secado afectaron a la estabilidad física y química del oleogel. De esta manera, se obtuvieron oleogeles poco estructurados y no homogéneos al utilizar aceite con un alto grado de insaturación, como el aceite de lino, por secado convencional, mientras que no fue factible desarrollar oleogeles de lino con secado a vacío. Además, el secado en estufa convencional a 80 °C durante 10 h 30 min generó oleogeles de girasol y de girasol alto oleico con mayor estabilidad estructural y física que el secado a 60 °C durante 24 h. Los oleogeles de oliva y de girasol alto oleico producidos por secado convencional y los oleogeles de oliva y de girasol producidos por secado a vacío presentaron valores de estabilidad oxidativa primaria y secundaria dentro de los límites de aceptabilidad establecidos por el Codex Alimentarius. En una segunda etapa se desarrollaron cremas de cacao untables y panes dulces. En las cremas se estudió la microestructura, textura, reología y atributos sensoriales. La reformulación de las cremas con un reemplazo total (100%) y parcial (50%) de grasa de coco por oleogeles de oliva o girasol obtenidos por secado a vacío permitió mantener sus propiedades estructurales. Concretamente, la sustitución parcial de grasa de coco por oleogel de girasol permitió obtener cremas con atributos sensoriales como "apariencia cremosa", "textura cremosa" y "sabor a cacao", por lo que podría ser una alternativa viable para reformular cremas untables con un perfil nutricional más saludable. Los oleogeles de oliva o girasol alto oleico obtenidos por secado convencional y previamente caracterizados, se emplearon en la formulación de panes dulces elaborados al horno o al vapor. La reformulación con oleogeles permitió obtener panes con unas características estructurales y apariencia similar a la de los panes elaborados con margarina. La realización de una prueba triangular mostró diferencias en la apariencia de la miga y el sabor, mientras que no se observaron diferencias para el atributo textura entre los panes elaborados con oleogeles y con margarina. No se encontraron diferencias en la cantidad de ácidos grasos liberados tras la digestión in vitro entre los panes dulces elaborados con oleogeles y con margarina. Sin embargo, la velocidad inicial de la digestibilidad lipídica fue diferente dependiendo del tipo de procesado, horneado o vaporización, empleado en la elaboración de los panes. El desarrollo de oleogeles a base de hidrocoloides además de ofrecer a la industria alimentaria una alternativa como sustituto de grasas plásticas, podría investigarse como estrategia para modular la digestión de lípidos y brindar beneficios para la salud. Todas las estrategias abordadas en el transcurso de esta Tesis permitieron comprender y ahondar en los conocimientos que conducen a cómo reformular un alimento para mejorar su perfil lipídico sin comprometer sus características sensoriales y fisicoquímicas. / [CA] La present Tesi doctoral planteja diferents estratègies per al disseny i desenvolupament d'oleogels estables, amb un perfil lipídic d'alta qualitat nutricional, i la posterior incorporación dels oleogels en diferents aliments. Es pretén dissenyar aliments mitjançant el reemplaçament de greixos sòlids, rics en àcids grassos saturats i trans per oleogels, que d'una banda, mantinguen les propietats texturals i organolèptiques, i d'altra banda, presenten un perfil lipídic millorat. En una primera etapa, es van desenvolupar oleogels elaborats amb un 1% de hidroxipropilmetilcel·lulosa i un 0,6% de goma xantana emprant dues condicions d'assecat diferents: assecat convencional en estufa a 80 °C durant 10 h 30 min i assecat a buit a 60 °C durant 24 h. Els olis estructurats van ser d'oliva, de lli, de gira-sol i de gira-sol alt oleic. La microestructura va permetre apreciar oleogels ben estructurats quan es van utilitzar els olis d'oliva, gira-sol i gira-sol alt oleic. L'estabilitat física i les propietats reològiques van corroborar la formació d'oleogels d'alta estabilitat física, al llarg de 35 dies d'emmagatzematge, i amb un comportament de gel sòlid. Tant el grau d'insaturació de l'oli com les condicions d'assecat van afectar l'estabilitat física i química del oleogel. D'aquesta manera, es van obtindre oleogels poc estructurats i no homogenis en utilitzar oli amb un alt grau d'insaturació, com l'oli de lli, per assecat convencional, mentre que no va ser factible desenvolupar oleogels de lli amb assecat a buit. A més, l'assecat en estufa convencional a 80 °C durant 10 h 30 min va generar oleogels de gira-sol i de gira-sol alt oleic amb major estabilitat estructural i física que l'assecat a 60 °C durant 24 h. Els oleogels d'oliva i gira-sol alt oleic, produïts per assecat convencional i els oleogels d'oliva i gira-sol produïts per assecat a buit van presentar valors d'estabilitat oxidativa primària i secundària dins dels límits d'acceptabilitat establits pel Codex Alimentarius. En una segona etapa, es van desenvolupar cremes de cacau untables i pans dolços. En aquest treball, es va estudiar la microestructura, textura, reologia i atributs sensorials de les cremes untables. La reformulació de cremes amb un reemplaçament total (100%) i parcial (50%) de greix de coco per oleogels d'oliva o gira-sol obtinguts per assecat a buit, va permetre mantindre les propietats estructurals de les cremes untables. Concretament, la substitució parcial de greix de coco per oleogel de gira-sol va ser descrita amb atributs sensorials com a "aparença cremosa", "textura cremosa" i "sabor de cacau", la qual cosa podria ser una alternativa viable per a reformular cremes per a untar amb un perfil nutricional més saludable. Els oleogels d'oliva o gira-sol alt oleic obtinguts per assecat convencional i prèviament caracteritzats, es van empra en la formulació de pans dolços cuinats al forn o al vapor. Es van estudiar les propietats estructurals, i la digestibilitat lipídica in vitro del producte final. La reformulació amb oleogels va permetre obtindre pans amb unes característiques estructurals i aparença similar a la dels pans elaborats amb margarina. La realització d'una prova triangular va mostrar diferències en l'aparença de la molla i el sabor, mentre que no es van observar diferències per a l'atribut textura, entre els pans elaborats amb oleogels i amb margarina. No es van trobar diferències en la quantitat d'àcids grassos alliberats després de la digestió in vitro entre els pans dolços elaborats amb oleogels i amb margarina. No obstant això, la velocitat inicial de la digestibilitat lipídica va ser diferent depenent de la mena de processament, enfornat o vaporatge, emprat en l'elaboració dels pans. El desenvolupament d’oleogels a base de hidrocol∙loides, a més d ́oferir a la indústria alimentària una alternativa com a substitut de greixos plàstics, podria investigar-se com a estratègia per modular la digestió de lípids i brindar beneficis per la salut. Totes les estratègies abordades en el transcurs d'aquesta Tesi van permetre comprendre i aprofundir en els coneixements que condueixen a com reformular un aliment per a millorar el seu perfil lipídic sense comprometre les seues característiques sensorials i fisicoquímiques. / [EN] The research of this doctoral thesis proposes different strategies for the design and development of stable oleogels, with high nutritional lipid profile and the subsequent incorporation of the oleogels in the formulation of different . It aims to design foods by replacing solid fats, rich in saturated and trans fatty acids with oleogels, which, on the one hand, maintain the textural and organoleptic properties, and on the other hand, present an improved lipid profile. In a first stage, oleogels made with 1% hydroxypropylmethylcellulose and 0.6% xanthan gum were developed using two different drying conditions: conventional drying in an oven at 80 ° C for 10 h 30 'and vacuum drying at 60 ° C for 24 h. The structured oils were olive, flaxseed, sunflower and high oleic sunflower oil. The microstructure showed the oleogels structured, when olive, sunflower or high oleic sunflower oils were used. Physical stability and rheological properties corroborated the formation of oleogels with high physical stability, over 35 days of storage, and with a solid gel behaviour. Both the degree of unsaturation of the oil and the drying conditions affected the physical and chemical stability of the oleogel. In this way, unstructured non-homogeneous oleogels were obtained by using oil with a high degree of unsaturation, such as flaxseed oil, by conventional drying, while it was not feasible to develop flaxseed oleogels with vacuum drying. Furthermore, drying in a conventional oven at 80 ° C for 10 h 30 min generated sunflower and high oleic sunflower oleogels with more structural and physical stability than drying at 60 ° C for 24 h. Olive and high oleic sunflower oleogels, produced by conventional drying, and olive and sunflower oleogels produced by vacuum drying, presented primary and secondary oxidative values within the acceptability limits established by the Codex Alimentarius. In a second stage, spreadable cocoa creams and sweet breads were developed. The microstructure, texture, rheology and sensory attributes of spreadable creams were studied. The reformulation of cocoa creams with a total (100%) and partial (50%) replacement of coconut fat by olive or sunflower oleogels obtained by vacuum drying allowed to maintain the structural properties of spreadable creams. Specifically, the partial substitution of coconut fat for sunflower oleogel gave place to creams described with sensory attributes such as "creamy appearance", "creamy texture" and "cocoa flavor"; therefore, it could be a viable alternative to reformulate spreads with a healthier nutritional profile. The olive or sunflower oleogels obtained by conventional drying and previously characterized, were used in the formulation of sweet breads made in the oven or steamed. Replacement of margarine for oleogels produced breads with similar structural characteristics and appearance that those made with margarine. A triangular discriminatory test showed differences in the appearance of the crumb and the flavor, while no differences were observed for the texture attribute, between the breads made with oleogels or with margarine. No differences were found in the amount of fatty acids released after in vitro digestion between sweet breads made with oleogels and margarine. However, the initial rate of lipid digestibility was different depending on the type of processing, baking or steaming, used in the preparation of the breads. In addition to offering the food industry an alternative as a substitute for plastic fats, the debe lopment of hydrocolloid-based oleogels could be investigated as a strategy to modulate lipid digestion and provide health benefits. / Bascuas Véntola, SM. (2021). Estructuración de aceites mediante el uso de hidrocoloides para sustituir grasas plásticas en los alimentos [Tesis doctoral]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/174797