Orientador: Celio Kenji Miyasaka / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Engenharia de Alimentos / Made available in DSpace on 2018-08-08T11:44:53Z (GMT). No. of bitstreams: 1 Lollo_PabloChristianoBarboza_M.pdf: 4184736 bytes, checksum: 3b13bac14d821be3f7d31a4ad595e6b2 (MD5) Previous issue date: 2007 / Resumo: O futebol assim como outras modalidades esportivas vem utilizando os conhecimentos científicos produzidos para a preparação dos atletas. Esse melhor preparo resulta em distâncias percorridas durante o jogo cada vez maiores além de um melhor desenvolvimento da musculatura para desempenhar as tarefas necessárias durante as partidas. É reconhecido que essa atividade física eleva as necessidades protéicas dos atletas, porém não se sabe exatamente qual é o requerimento protéico dos atletas de diferentes modalidades esportivas. Os principais motivos citados para esse aumento no requerimento protéico de atletas são: hipertrofia muscular (em determinadas fases do treinamento); oxidação de proteínas corporais durante atividades de longa duração para fornecimento de energia (via esqueleto carbônico dos aminoácidos de cadeia ramificada - BCAA); danos em proteínas musculares decorrentes de alterações fisiológicas causadas pelo exercício (queda de pH, elevação da temperatura intramuscular e tensões mecânicas nos músculos e demais estruturas do aparelho locomotor). As proteínas de soro de leite são consideradas excelente fonte de BCAA, e possuem alto valor biológico. Objetivo: verificar os efeitos da suplementação com proteínas de soro de leite na composição corporal, desempenho físico e parâmetros bioquímicos de futebolistas. Metodologia: quarenta e oito futebolistas juvenis (masculino), idade 16,7 + 0,6 anos, medindo 179,2 cm + 6,7 e pesando 74,42 kg + 6,44, foram realizados 2 experimentos (n=24 em cada experimento), o primeiro experimento submeteu os 24 atletas a suplementação protéica diária de 1g.kg-1.dia-1 mais 0.4 g.kg-1.dia-1 de carboidrato (sacarose) por 8 semanas com as diferentes proteínas: a) caseína (CAS, n=8); b) proteína do soro de leite isolada (PSLI1, n=8); c) proteína do soro de leite hidrolisada (PSLH2 n=8). O segundo experimento submeteu 24 atletas à suplementação protéica ou glicídica diária de 1g.kg-1.dia-1 por 12 semanas com: a) maltodextrina (MALTO, n=8); b) PSLI2 (n=8); c) PSLH2 (n=8). Foram realizados testes antropométricos (composição corporal), de desempenho físico ¿ ¿Yo-yo intermittent recovery level 2¿, saltos verticais, 4 minutos contra o relógio, 3000m e 3200m à 85% da freqüência cardíaca (FC) máxima e os parâmetros bioquímicos: ácido úrico, colesterol, HDL, creatinina, glicose plasmática e dosagem das enzimas: creatina quinase (CK) e lactato desidrogenase (LDH). Resultados: após a suplementação, os grupos CAS e PSLI2 aumentaram significativamente a massa muscular em 2,83% e 3,36% respectivamente. No desempenho físico, foi observado que os atletas dos grupos PSLI2 e PSLH2 aumentaram a distância percorrida em 4,44% e 3,41% respectivamente no teste 4 minutos contra o relógio. No teste de 3200m em 85% da FC máxima o tempo dos atletas dos grupos PSLI2 e PSLH aumentaram o tempo em 5,48% e 6,8%. Nos parâmetros bioquímicos analisados verificamos queda significativas nas enzimas indicadoras de lesão muscular nos atletas dos grupos PSLH2 e aumento significativo no ácido úrico nos atletas dos grupos, PSLI1, MALTO, PSLI2 e PSLH2 / Abstract: The soccer as well as other sporting modalities comes using the knowledge produced for training the athletes. These better training results in large distances covered during the game. So the better preparation of the muscle is need for the soccer players. The physical activity raises the proteins requirements, however, how much is not known accurately. The main reasons cited for this increase in the proteins requirements in athlete are: muscular hypertrophy (in determined phases of the training); body protein oxidation during exercise of long term for energy supply (main skeleton carbonic of the branched chain amino acids - BCAA); damages in muscular proteins decurrently of physiological alterations caused by the exercise (fall of pH and rise of the temperature intramuscular and mechanical tensions in the muscles and another structures of the locomotive device). The whey protein is excellent source of (BCAA), and protein of high biological value. Objective: to verify the effect of the supplementation with whey proteins in the body composition, physical performance and biochemistry parameters of soccer players. Methodology: Forty eight youthful soccer players (masculine), age 16.7 + 0.6 years, heighted 179.2 cm + 6.7 and weighted 74.42 kg + 6.44, in 2 groups (n=24 in each group), the first group was submitted to the daily protein supplementation of 1g.kg-1.dia-1 plus 0.4g.kg-1.dia-1 of carbohydrate (sucrose) for 8 weeks with different proteins: a) the casein (CAS, n=8); b) whey protein isolated (PSLI1, n=8); c) whey protein hydrolyzed (PSLH2 n=8). The group 2 was submitted to daily the protein supplementation of 1g.kg-1.dia-1 for 12 weeks with: a) maltodextrine (MALTO, n=8); b) PSLI2 (n=8); c) PSLH2 (n=8). Anthropometric tests had been carried through (body composition), of physical performance ¿ Yo-yo intermittent recovery level 2, jump tests, 4 minutes against the clock, 3000m and 3200m to 85% of the cardiac frequency (FC) maximum ¿ and biochemistry tests - acid uric, cholesterol, HDL, creatinine, plasmatic glucose and enzymes: lactate dehydrogenize (LDH) and creatina kinase (CK) and anthropometry. Results: After the supplementation groups CAS and PSLI2 had increased significantly the muscular mass in 2,83% and 3,36% respectively. In the physical performance, it was observed that groups PSLI2 and PSLH2 had increased in the distance covered in 4,44% and 3,41% for PSLI2 respectively in test 4 minutes against the clock. In the test of 3200m to 85% of the maximum FC the time of the groups PSLI2 and PSLH had increased the time in 5,48% and 6,8%. In the biochemistry parameters it was verified significantly reduction in enzymes of muscular injury in group PSLH2 and significant increase in the uric acid of groups PSLI1, MALTO, PSLI2 and PSLH2 / Mestrado / Nutrição Experimental e Aplicada à Tecnologia de Alimentos / Mestre em Alimentos e Nutrição

Orientadores: Rosiane Lopes da Cunha, Ângelo Luiz Fazani Cavallieri / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Engenharia de Alimentos / Made available in DSpace on 2018-08-20T01:10:51Z (GMT). No. of bitstreams: 1 Mantovani_RaphaeladeAraujo_M.pdf: 4371985 bytes, checksum: 490b60583a49fed7d2421e32dcf9160e (MD5) Previous issue date: 2012 / Resumo: As proteínas do soro do leite (WPI) e lecitina de soja são amplamente utilizadas em alimentos devido às suas excelentes propriedades emulsificantes. Este trabalho visou avaliar as propriedades emulsificantes da mistura de WPI e lecitina em diferentes condições de pH e concentração dos ingredientes. A primeira etapa consistiu no estudo das interações entre os emulsificantes em meio aquoso em diferentes razões WPI:lecitina e condições de pH. Os resultados mostraram que na razão 1:1 e em pH abaixo do pI da proteína, no qual os emulsificantes encontravam-se opostamente carregados, foi favorecida a formação de complexos eletrostáticos. Na segunda etapa, emulsões O/A contendo proteínas do soro e/ou lecitina foram avaliadas através da estabilidade à cremeação, microestrutura, distribuição do tamanho de gota, densidade de carga superficial, reologia, eletroforese em gel de poliacrilamida e digestão in vitro, verificando-se a influência do pH e da pressão de homogeneização. As emulsões estabilizadas somente por proteínas apresentaram separação de fases e comportamento não-Newtoniano somente em pH próximo ao pI. As emulsões contendo somente lecitina não separaram de fases e apresentaram comportamento Newtoniano. Os sistemas em pH abaixo do pI, contendo a mistura de emulsificantes apresentaram elevada instabilidade cinética devido à forte interação eletrostática entre os componentes. No entanto, em pH próximo ao pI da prote, a interação foi favorável e levou ao aumento da estabilidade das emulsões e a menores tamanhos de gota, resultando em fluidos pouco viscosos. O aumento da pressão de homogeneização, em emulsões com pH próximo e acima do pI favoreceu a formação de agregados de alta massa molecular, o que não foi observado em pH mais ácido. Também não foi notada a formação de agregados na presença de lecitina em nenhuma das condições de pH. Os ensaios de eletroforese deram indícios de que a interação entre os emulsificantes levou a modificações na estrutura das proteínas e revelaram maior afinidade da lecitina com a ?-lactoalbumina (?-la). Finalmente, na etapa de simulação do processo de digestão as proteínas mostraram-se mais resistentes, uma vez que a presença de lecitina promoveu a liberação do óleo da emulsão logo no início da etapa gástrica. De maneira geral, a utilização das proteínas do soro juntamente com a lecitina como emulsificante levou a resultados satisfatórios pois permitiu o desenvolvimento de sistemas estáveis à cremeação em diferentes valores de pH, inclusive em pH próximo ao pI, e consideravelmente resistentes às condições adversas do trato gastrointestinal / Abstract: Whey proteins (WPI) and soybean lecithin are widely used in food due to its excellent emulsifying properties. This study aimed to evaluate the emulsifying properties of the mixture of whey protein and lecithin at different conditions of pH and concentration of ingredients. Firstly, the interactions between emulsifiers in an aqueous medium at different ratios (WPI:lecithin) and pH conditions were studied. The results showed that at mixing ratio of 1:1 and at pH below the isoelectric point of the protein (pI) in which the emulsifiers were oppositely charged, was favored the formation of electrostatic complexes. Afterwards, O/W emulsions containing WPI and/or lecithin were prepared and the influence of pH and pressure homogenization were studied. Stability, microstructure, droplet size distribution, surface charge density, rheology, electrophoresis in polyacrylamide gel (SDS-PAGE) and digestion in vitro were evaluated. The emulsions stabilized by proteins showed phase separation and non-Newtonian behavior only in pH close to the pI. The emulsions containing only lecithin were stable and showed Newtonian behavior. Systems containing the mixture of emulsifiers at pH below the pI showed high kinetic instability due to the strong electrostatic interactions between the components. However, at pH close to the pI the interaction was favorable and led to an increase of the emulsion stability and to smaller droplet sizes, resulting in lower viscosities. When the pressure homogenization was increased, emulsions at a pH close to and above the pI favored the formation of high molecular weight protein aggregates which was not observed in emulsions at lower pH. In the presence of lecithin, protein aggregates were not formed in any of the pH conditions. The SDS-PAGE analysis showed that the interaction between emulsifiers led to modifications in the structure of proteins and indicated a higher affinity of lecithin to ?-lactalbumin (?-la). Finally, proteins were more resistant to digestion, since the presence of lecithin promoted release of oil emulsion at the beginning of gastric step. In general, the use of whey protein with lecithin as emulsifier led to satisfactory results because it allowed the development of stable emulsions to creaming at different pH values, even at pH close to the pI, and substantially resistant to the adverse conditions of the gastrointestinal tract / Mestrado / Engenharia de Alimentos / Mestre em Engenharia de Alimentos

Whey is a by-product of cheese manufacture, containing a mixture of proteins of commercial value, each having unique attributes for nutritional, biological and food ingredient applications. A tremendous amount of whey, normally treated as a waste product, is produced worldwide each year. This work describes the cation-exchange adsorption of the two major whey proteins, alpha-lactalbumin (ALA) and beta-lactoglobulin (BLG) with the purpose of optimising a process for isolating them from whey. Adsorption of pure BLG and ALA was studied onto SP Sepharose FF using 0.1M acetate buffer. Batch experiments were carried out at various pH values for ALA and BLG, and the relevant Langmuir isotherm parameters, dissociation constant, Kd, and maximum binding capacity, qm, were determined. The optimum pH for separation was chosen to be pH 3.7. At pH 3.7, both Kd and qm pertaining to ALA were found to have higher numerical values than those of BLG, implying different characteristics of adsorption of the two proteins on this adsorbent. The Kd for the former protein was almost four times larger than the latter, while qm was 1.3 times higher. Packed-bed column adsorption was performed using a 1-ml column at pH 3.7, flow rate 1 ml/min and initial concentration of 3 mg/ml for BLG and 1.5 mg/ml for ALA both in 0.1M sodium acetate buffer. The t1/2 for the resulting ALA breakthrough was 75% longer than its BLG counterpart. The above results suggest the possibility of the occurrence of competitive adsorption between the proteins when adsorbed simultaneously. In traditional batch uptake experiments, the kinetic rate constants of ALA and BLG in both the single- and two-component systems were determined using the simple kinetic model. The values so obtained implied that BLG was adsorbed faster than ALA. In the confocal laser scanning microscopy experiments, the different behaviour of ALA and BLG in the single-component system with regard to their penetration within the adsorbent beads suggested that the two proteins have different transport mechanisms governing their adsorption. The two-component system results showed that ALA was able to displace BLG in spite of the lower affinity of the former protein to the adsorbent. The packed-bed adsorption and elution of a mixture of ALA and BLG were then investigated under the above conditions but using a 5-ml column. BLG breakthrough occurred first, and its concentration in the outlet exceeded its feed value by 1.6 fold before declining to the feed value, followed by the breakthrough of ALA. ALA displaced and eluted all the BLG from the column in a pure form. Pure ALA could then be eluted with good recovery. The single- and two-component breakthrough curves for ALA and BLG were simulated by the simple kinetic model using the isotherm parameters, but the overshoot phenomenon could only be predicted after correcting these parameters. The evidence of the competitive nature of adsorption observed in binary mixtures was used to develop a facile separation procedure for the two proteins from aqueous solutions of whey concentrate powders. A novel consecutive two-stage separation process was developed to separate ALA and BLG from whey concentrate mixtures. Almost all the BLG in the feed was recovered, with 78% being recovered at 95% purity and a further 20% at 86% purity. In addition, 67% of ALA was recovered, 48% at 54% purity and 19% at 60% purity. The correction factors employed for the pure binary mixture were used to simulate the breakthrough curves of the two proteins in experiments conducted with whey concentrate in each of the two stages of the novel separation process, and there was agreement between the experimental and theoretical results.

Orientador: Flavia Maria Netto / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Engenharia de Alimentos / Made available in DSpace on 2018-08-14T13:49:46Z (GMT). No. of bitstreams: 1 Fernandes_MicheleAugusto_M.pdf: 2050778 bytes, checksum: aedc46d9b78bf413f7e34cfbe46a36a6 (MD5) Previous issue date: 2009 / Resumo: A ß-Lactoglubulina (ß-Lg) é uma das proteínas mais antigênicas presente no leite bovino. Tratamentos físicos, químicos ou enzimáticos podem alterar a antigenicidade desta proteína. Em trabalho anterior, verificou-se que o potencial antigênico da ß-Lg é reduzido quando polimerizada pela enzima transglutaminase (TG) na presença de cisteína (Cys). No entanto, o efeito da polimerização sobre o valor nutricional da ß-Lg ainda não é conhecido. O presente estudo teve como objetivo avaliar o efeito da reação de polimerização catalisada pela TG na digestibilidade in vitro e atividade antigênica da ß-Lg antes e após a ação das enzimas gastrintestinais. A ß-Lg polimerizada pela TG (0, 10 ou 25 U g-1), após tratamento térmico ou na presença de agentes redutores Cys (0, 0,1 e 0,25 mol L- 1) ou ditiotreitol (DTT 0,02 mol -1), foi avaliada quanto à digestibilidade in vitro, utilizando as enzimas pepsina e pancreatina. As amostras, antes e após a digestão in vitro, foram caracterizadas pelos métodos SDS-PAGE, SDSPAGE/ tricina e cromatografia líquida de alta eficiência de fase reversa (CLAE-FR). Posteriormente, as amostras foram avaliadas quanto à antigenicidade, exceto aquelas na presença de DTT, por meio do método de Imunoblote, utilizando soro de camundongos BALB/c sensibilizados com a ß-Lg na forma nativa (ß-Lg N) ou com ß-Lg polimerizada com 25 U de TG g-1, na presença de 0,25 mol L-1 de Cys (ß-Lg 0,25Cys 25TG). A adição de TG resultou na formação de polímeros com massa molar igual ou acima de 97,4 kDa, principalmente na presença de agentes redutores alcançando aproximadamente 96% de polimerização na presença de DTT e 91% na presença de Cys. A digestibilidade in vitro da ß-Lg N foi 53,6% e todos os tratamentos realizados aumentaram a digestibilidade da proteína em até 79%. Os maiores valores de digestibilidade foram obtidos quando a ß-Lg foi tratada com agentes redutores. O processo de polimerização também teve efeito positivo na digestibilidade, principalmente para as amostras polimerizadas na presença de Cys ou DTT, atingindo valores acima de 75%. A análise por Imunoblote mostrou que a polimerização da ß-Lg na presença de agente redutor Cys, na concentração de 0,25 mol L-1, reduziu o reconhecimento da ß-Lg pelas IgE específicas presente nos soros dos animais sensibilizados com ß-Lg N ou com ß- Lg 0,25Cys 25TG. Após a digestão com pepsina e pancreatina, as amostras polimerizadas pós-tratamento térmico ou na presença de Cys apresentaram redução da antigenicidade, como também os digeridos da ß-Lg tratada com Cys (não polimerizada com TG). A desnaturação pelo agente redutor Cys e a polimerização por TG em ambas as condições estudadas mostraram ser métodos efetivos no aumento da digestibilidade da ß-Lg. Por sua vez, a combinação destes métodos com a digestão por enzimas gastrintestinais levou à redução da antigenicidade da proteína, já que os peptídeos gerados apresentaram potencial antigênico baixo / Abstract: The ß-Lactoglubulin (ß-Lg) is one of the most antigenic proteins present in the bovine milk. Physical, chemical or enzymatic treatments can alter the antigenicity of this protein. In previous study, it was shown that the antigenic potential of ß-Lg is reduced when polymerized by transglutaminase (TG) in the presence of cysteine (Cys). However, the effect of polymerization on the nutritional properties of ß-Lg is still unknown. The present study aimed to evaluate the effect of the polymerization reaction catalyzed by TG on the in vitro digestibility and the antigenic activity of ß- Lg, before and after simulate digestion with gastrointestinal enzymes. The in vitro digestibility of the ß-Lg treated with TG (0, 10 or 25 U g-1), after heat treatment or in the presence of reducing agents Cys (0, 0.1 and 0.25 mol L-1) or dithiothreitol (DTT 0.02 mol L-1), was evaluated using the gastrointestinal enzymes, pepsin and pancreatin. The samples, before and after in vitro digestion, were characterized by SDS-PAGE, SDS-PAGE/tricine and reversed-phase high performance liquid chromatography (RP-HPLC). Subsequently, the samples were evaluated for antigenicity, except those prepared in the presence of DTT, by immunochemical methods (Immnoblotting), using sera from BALB/c mice sensitized with native ß-Lg (ß-Lg N) or ß-Lg polymerized with 25 U of TG g-1 in the presence of 0.25 mol L-1 of Cys (ß-Lg 0.25Cys 25TG). The formation of polymers with molar mass equal to or above 97.4 kDa was observed with the addition of TG, especially in the presence of reducing agents, reaching approximately 96% of polymerization in the presence of DTT and 91% in the presence of Cys. The in vitro digestibility of native ß-Lg was 53.6% and the all treatments performed increased the digestibility of protein up to 79%. The highest values of digestibility were obtained in the presence of reducing agents. The polymerization also had a positive effect on the digestibility, especially for those samples polymerized in the presence of Cys or DTT, with values above 75%. Analysis by immunoblotting showed that the polymerization of ß-Lg in the presence of 0.25 mol L-1 Cys, reduced the recognition of ß-Lg specific IgE present in the sera of animals sensitized with ß-Lg N or ß-Lg 0.25 Cys 25TG. After digestion with pepsin and pancreatin, the samples polymerized after heat treatment or in the presence of Cys showed reduced antigenicity. The digest of the samples treated with Cys (not treated with TG) was not recognized as antigens. The denaturation by Cys and polymerization by TG in both conditions were effective in increasing the digestibility of ß-Lg. In turn, the combination of these methods with digestion by gastrointestinal enzymes led to reduction of antigenicity of the protein and that peptides generated showed low antigenic potential / Mestrado / Nutrição Experimental e Aplicada à Tecnologia de Alimentos / Mestre em Alimentos e Nutrição

The proposal of this work is to evaluate a feasible alternative to minimize the environmental problem caused by the disposal of liquid waste from cheese production in the municipality of Nossa Senhora de Lourdes, in the state of Sergipe, Brazil. Moreover, it also proposes the recovering of whey proteins which have a high economic value for several industrial sectors. At present, the whey has been disposed of without any treatment, damaging the environment and resulting in the loss of whey proteins, although they constitute a profitable economic alternative as well as a rich feed resource. With the objective of evaluating the dimensions of the problem, surveys were carried out at the cheese manufacturers in that municipality in order to verify the economic, social and environmental conditions of the region. Whey samples were collected for laboratory tests and analyses. The survey results have shown that subsistence farming is one of the main economic activities in that municipality. The local community complains about the lack of alternatives to the promotion of socioeconomic development. The tests and analyses of the milk serum in cheese making reveal a high level of organic substances (49.000 mg/L), which can contaminate the ground and compromise the quality of water resources. This action can be minimized through the appropriate waste treatment by ultra-filtration. In this work, it was possible to concentrate the whey proteins in 36,85% and reduce the Biochemical Oxygen Demand (BOD) to 30,61%. With this result, it is possible to obtain lower pollutant emissions to environment and it values are added to an abundant municipal product. Therefore, is possible to promote industrial growth balanced with sustainable development, implementing alternatives that can minimize social, economic and environmental inconveniences at the semi-arid region of Sergipe, aiming at assuring better quality of life for the community of that region. / O presente trabalho de pesquisa propõe avaliar uma alternativa viável para minimizar a agressão ambiental provocada pelo descarte do rejeito líquido proveniente da produção de queijo do município de Nossa Senhora de Lourdes, além de recuperar as proteínas do soro que apresentam um elevado valor econômico para diversos setores industriais. Atualmente, a prática de descartar o soro é realizada sem nenhum tratamento, prejudicando o meio ambiente e perdendo-se uma fonte alternativa de renda e alimentação, as proteínas. Com a finalidade de avaliar a dimensão dessa problemática, foram aplicados questionários nas fábricas de queijo do município para verificar as condições sócio-econômicas e ambientais da região. Amostras do soro eliminado no processo produtivo foram coletadas para testes e análises posteriores em laboratório. Os resultados revelaram que o município estudado tem a agropecuária de subsistência como uma das principais atividades econômicas. Neste aspecto, a comunidade local reclama a falta de alternativas capazes de promover o desenvolvimento sócio-econômico. Os testes e análises do soro de leite eliminado na produção do queijo, apontam para uma elevada carga orgânica (49.000 mg/L), a qual pode contaminar o solo, bem como comprometer a qualidade dos recursos hídricos. Esta ação pode ser minimizada através do tratamento adequado do rejeito, por ultrafiltração. No presente trabalho, foi possível concentrar as proteínas do soro em 36,85%, e reduzir em 30,61% a DBO (Demanda Bioquímica de Oxigênio). Com este resultado, tem-se uma menor emissão de poluentes para o meio ambiente, além de agregar valores a um produto abundante no município. Portanto, é possível promover o crescimento industrial equilibrado com o desenvolvimento sustentável, implementando alternativas capazes de minimizar inconvenientes de ordem social, econômica e ambiental no semi-árido sergipano, na perspectiva de garantir uma melhor qualidade de vida para a comunidade daquela região.

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L’étuvage de poudre de protéines sériques (PS) de lait permet l’amélioration des propriétés techno-fonctionnelles des protéines : aptitude à la formation de gel, mousse, ou émulsion. L’objectif de cette thèse était d’étudier le mécanisme de formation et les propriétés physico-chimiques et rhéologiques d’assemblages de PS de taille micronique ou AMI produits par étuvage à 100 °C d’une poudre d’isolat de PS ajustée à pH 9.5 et à une activité d’eau proche de 0,2. Pour cela, la caractérisation morphologique des assemblages, la dénaturation des protéines, la diminution de lactose libre et la formation de produits de Maillard ont été suivies au cours du temps d’étuvageLe rôle de chacun des constituants de la poudre a aussi été étudié en faisant varier sa composition. Les résultats obtenus montrent que l’étuvage à pH 9.5 permet la formation d’assemblages de taille et de forme relatives à celle des particules de poudre étuvée. Dans un premier temps, des réactions de glycation des protéines s’opèrent, des agrégats solubles sont produits et les poudres brunissent, tandis que les produits terminaux de la réaction de Maillard se forment. Ensuite, la quantité d’agrégats solubles diminue avec la formation concomitante des AMI. Les AMI possèdent des propriétés de rétention d’eau très élevées ce qui leur confère des propriétés viscosifiantes. L’addition de lactose à la poudre d’isolat de PS permet d’accélérer la cinétique de formation des AMI et d’obtenir des AMI plus denses en protéines. Les caséines sont aussi intégrées dans les AMI, mais ont très peu d’effets sur leurs propriétés stru / The dry heating of whey protein powder increases the functional properties of proteins such as their ability to form gels, foams or emulsions. The aim of this PhD thesis was to study the mechanism of formation and the physicochemical and rheological properties of protein assemblies of micrometer sizes that were produced by dry heating at 100 °C a whey protein powder isolate adjusted at pH 9.5 at a water activity of 0.2. Thus, morphological properties, protein denaturation, lactose conjugation and products of the Maillard reaction were studied during the time course of the dry heating. The role of each compound of the powder has also been studied by modification of the powder composition. Dry heating at pH 9.5 was able to form large assemblies of size and shape related to the size of the dry heated powder particles.In the first step, the glycation of proteins increased. Soluble aggregates formed and the powder browning progressed, while advanced glycated end products of the Maillard reaction were found. Then, the content in soluble aggregates decreased and large protein assemblies of micrometer sizes formed. These large assemblies were highly hydrated, and had very high viscosity values. Addition of lactose to the whey protein isolate powder allowed increasing the speed of formation of these large assemblies, increasing their density, and reaching higher yields of production. Caseins were able to join to whey proteins in the large assemblies, and had very poor effects on their structural properties and ability to retain the aqueous phase.

L’objectif de ce travail est d’identifier les conditions et les mécanismes permettant la création d'aptitudes nouvelles ou améliorées des protéines du lactosérum à la stabilisation et texturation des mousses alimentaires.À cette fin, nous avons étudié la rhéologie interfaciale de protéines adsorbées à l’interface eau/air, les réarrangements topologiques à l’échelle de quelques films liquides, la stabilité et la rhéologie de mousses de protéines. L’étude a porté à la fois sur un mélange de protéines du lactosérum et sur sa protéine majoritaire purifiée, la ß-lactoglobuline. Pour identifier les liens avec leurs propriétés structurales et physico-chimiques, des modifications des protéines ont été générées par étuvage de poudres. Plusieurs paramètres d’étuvage ont été variés simultanément. Une large gamme de modifications structurales des protéines a été obtenue grâce au contrôle de ces paramètres. Nous avons mis en évidence que de petites modifications structurales des protéines ont des conséquences majeures sur la rhéologie interfaciale, la dynamique des réarrangements de films, la stabilité et la rhéologie des mousses.Les effets de l’étuvage des poudres sur les propriétés des mousses sont complexes, car ils dépendent étroitement de la combinaison des effets des paramètres d’étuvage, comme de la propriété de la mousse qui est mesurée. Parallèlement, l’examen de la variabilité des comportements à plusieurs échelles apporte un éclairage original sur la contribution de la rhéologie interfaciale aux propriétés de mousses de protéines. Il met notamment en évidence l’intérê / The objective of this work is to identify the conditions and mechanisms of the creation or improvement of the stability and rheology of whey proteins foams. To this aim, we studied the interfacial rheology of protein layers adsorbed at the air/water interface, the liquid films dynamics after a topological rearrangement, the stability and rheology of whey protein foams. Both a mixture of whey proteins and purified ß-lactoglobulin, used as a model protein, were studied. To study the relationships with protein structure, proteins were modified by dry-heating of whey protein powders. A wide variety of structural changes was obtained by varying simultaneously multiple dry-heating parameters.Interestingly, low-extent structural modifications have a dramatic impact on interfacial rheology, liquid film dynamics, foam stability and foam rheology. The effects of dry-heating parameters on the foam properties are complex and depend on their combination and the considered foam feature. Our original multiscale approach (interface, film dynamics and foam) sheds light on the contribution of the interfacial rheology to protein foam properties. In particular, foam dynamics have been shown to play a predominant role.

The phenolic compounds as food components represent the largest group of secondary metabolites in plant foods. The phenolic compounds, e.g. chlorogenic acid (CQA), are susceptible to oxidation by enzymes specially, polyphenol oxidase (PPO) and at alkaline conditions. Both enzymatic and non-enzymatic oxidations occur in the presence of oxygen and produce quinone, which normally further react with other quinone to produce colored compounds (dimers), as well as is capable of undergoing a nucleophilic addition to proteins. The interactions of proteins with the phenolic compounds have received considerable attention in the recent years where, plant phenolic compounds have drawn increasing attention due to their antioxidant properties and their noticeable effects in the prevention of various oxidative stress associated diseases. Green coffee beans are one of the richest sources of chlorogenic acids. Therefore, a green coffee extract would provide an eligible food relevant source for phenolic compounds for modification of proteins. The interaction between 5-CQA and amino acid lysine showed decrease in both free CQA and amino acid groups and only a slight effect on the antioxidative capacity depending on the reaction time was found. Furthermore, this interaction showed a large number of intermediary substances of low intensities. The reaction of lysine with 5-CQA in a model system initially leads to formation of 3-CQA and 4-CQA (both are isomers of 5-CQA), oxidation giving rise to the formation of a dimer which subsequently forms an adduct with lysine to finally result in a benzacridine derivative as reported and confirmed with the aid of HPLC coupled with ESI-MSn. The benzacridine derivative containing a trihydroxy structural element, was found to be yellow, being very reactive with oxygen yielding semiquinone and quinone type of products with characteristic green colors. Finally, the optimal conditions for this interaction as assessed by both the loss of CQA and free amino groups of lysine can be given at pH 7 and 25°C, the interaction increasing with incubation time and depending also on the amount of tyrosinase present. Green coffee bean has a higher diversity and content of phenolics, where besides the CQA isomers and their esters, other conjugates like feruloylquinic acids were also identified, thus documenting differences in phenolic profiles for the two coffee types (Coffea arabica and Coffea robusta). Coffee proteins are modified by interactions with phenolic compounds during the extraction, where those from C. arabica are more susceptible to these interactions compared to C. robusta, and the polyphenol oxidase activity seems to be a crucial factor for the formation of these addition products. Moreover, In-gel digestion combined with MALDI-TOF-MS revealed that the most reactive and susceptible protein fractions to covalent reactions are the α-chains of the 11S storage protein. Thus, based on these results and those supplied by other research groups, a tentative list of possible adduct structures was derived. The diversity of the different CQA derivatives present in green coffee beans complicates the series of reactions occurring, providing a broad palette of reaction products. These interactions influence the properties of protein, where they exposed changes in the solubility and hydrophobicity of proteins compared to faba bean proteins (as control). Modification of milk whey protein products (primarily b-lactoglobulin) with coffee specific phenolics and commercial CQA under enzymatic and alkaline conditions seems to be affecting their chemical, structural and functional properties, where both modifications led to reduced free amino-,thiol groups and tryptophan content. We propose that the disulfide-thiol exchange in the C-terminus of b-lactoglobulin may be initiated by the redox conditions provided in the presence of CQA. The protein structure b-lactoglobulin thereupon becomes more disordered as simulated by molecular dynamic calculation. This unfolding process may additionally be supported by the reaction of the CQA at the proposed sites of modification of -amino groups of lysine (K77, K91, K138, K47) and the thiol group of cysteine (C121). These covalent modifications also decreased the solubility and hydrophobicity of b-lactoglobulin, moreover they provide modified protein samples with a high antioxidative power, thermally more stable as reflected by a higher Td, require less amount of energy to unfold and when emulsified with lutein esters, exhibit their higher stability against UV light. The MALDI-TOF and SDS-PAGE results revealed that proteins treated at alkaline conditions were more strongly modified than those treated under enzymatic conditions. Finally, the results showed a slight change in emulsifying properties of modified proteins. / Für die Verbesserung von Nahrungsmitteleigenschaften können Modifikationen an verschiedenen Inhaltsstoffen vorgenommen werden. Beispielsweise werden bereits Proteine miteinander verknüpft und bilden sogenannte „Crosslinks“ oder vernetzte Biomoleküle. Diese werden für die Herstellung fester, viskoelastischer Produkte, die zum Verdicken als auch zum Stabilisieren von Emulsionen oder Schäumen eingesetzt werden, genutzt. Da die Verbraucher sich Zunehmens mit gesundheitsfördernden Lebensmitteln befassen, ist das Einbringen von gesundheitsfördernden Inhaltsstoffen wie z.B. phenolische Verbindungen, immer mehr in den Fokus der Forschung gerückt. Demnach ist das wissenschaftliche Bestreben phenolische Verbindungen in die Vernetzung von Proteinen mit einzubeziehen und deren positive Wirkungen (antioxidativ) auszunutzen, vorteilhaft. Als Phenole werden Verbindungen bezeichnet, die eine oder mehrere Hydroxygruppen am Benzolring aufweisen. Phenole liegen in der Enolform vor, da diese, bedingt durch den Erhalt des aromatischen Benzolringes, energetisch begünstigt ist. Kaffeesäure ist eine Hydroxyzimtsäure und in Kaffeebohnen zu finden. Der am häufigsten anzutreffende Ester besteht aus Kaffee- und Chinasäure. Der einfachste Vertreter ist die Chlorogensäure (5-Caffeoylchinasäure, 5-CQA), die in vielen Pflanzenteilen enthalten ist. Chlorogensäure und ihre Derivate besitzen ebenfalls antioxidative Eigenschaften. Zusätzlich wirken sie auf Enzyme, die an entzündlichen- oder allergischen Reaktion teilnehmen, inhibierend. Während Verarbeitungs- und Lagerungsprozessen können phenolische Komponenten pflanzlicher Lebensmittel mit den Aminosäuren der Proteine in Lebensmitteln reagieren. Solche Reaktionen können die physikalisch-chemischen Eigenschaften von Proteinen verändern und deren ernährungsphysiologische Wertigkeit vermindern. Proteine weisen verschiedene reaktive Seitengruppen (Sulfhydryl-, Hydroxyl-, Aminogruppen) auf, mit denen sie über kovalente und nicht-kovalente Wechselwirkungen mit Phenolen Verbindungen eingehen können. Zu den nicht-kovalenten Verbindungen gehören u. a. Wasserstoffbrückenbindungen, hydrophobe Wechselwirkungen und Ionenbindungen. Phenole (z.B. Chlorogensäuren) können bei Anwesenheit von Sauerstoff enzymatisch bzw. nichtenzymatisch oxidiert werden. Die Reaktionsprodukte (Chinone) bilden anschließend mit reaktiven Thiol- bzw. Aminogruppen von Proteinen Addukte. Die Erfassung dieser verschiedenen Facetten von Interaktionen stellt somit die primäre Forschungsaufgabe im Rahmen dieser Arbeit. Die primäre Aufgabe der vorliegenden Arbeit besteht demzufolge in der Etablierung der Analysen- und der Charakterisierungsmöglichkeiten solcher Wechselwirkungen (Bindung) pflanzlicher Verbindungen bzw. deren Reaktionsprodukten mit Proteinen u.a. über massenspektrometrische Methoden. Da die Wechselwirkung mit Proteinen auch zu Veränderungen der Proteinstruktur führt, können deren funktionelle Eigenschaften auch verändert sein. Dies soll anhand der Messung von isolierten Proteinen die an der Wechselwirkung beteiligt sind, nachgewiesen werden. Anschließend sollen über Docking-Untersuchungen die entsprechenden Bindungsstellen näher charakterisiert werden. Durch die vorliegenden Ergebnisse wurden mögliche Reaktionen von phenolischen Verbindungen mit Proteinen, näher charakterisiert. Es wurde festgestellt, dass die Apfelsorte Braeburn über die höchste PPO- Enzymaktivität beim gleichzeitigen niedrigen CQA Gehalt im Vergleich zu den anderen untersuchten Sorten verfügt. Die PPO/Tyrosinase modulierte Reaktionen zwischen CQA und Lysine wurden in Abhängigkeit der vorherrschenden Bedingungen optimiert und die Reaktionsprodukte analysiert. In dem zweiten Teil wurden solche Reaktionsmöglichkeiten in den Grünen Kaffeebohnen lokalisierte und modelliert. Dazu wurden die sortenabhängige CQA-Zusammensetzung ermittelt und die möglichen Reaktionen mit den Hauptspeicherproteinen des Kaffees dargestellt. Im letzten Teil wurden dann diese Reaktionen mit Molkenproteinen simuliert und Einflüsse auf die Struktur und die funktionellen Eigenschaften erfasst. Die Ergebnisse belegen eine umfangreiche und sehr heterogene Adduktbildung mit den Aminoseitenketten des Lysins und Cysteins. Ein Katalog der unterschiedlichen Reaktionsprodukte wurde erstellt und am Protein modelliert. Die entsprechende Veränderung an die Proteinstruktur wurde experimentell belegt und der Einfluss wurde in den technofunktionelle Eigenschaften (wie die Löslichkeit, Emulgierbarkeit usw.) wiederspiegelt. Ein Anstieg des antioxidativen Potentials der Proteine wurde erreicht und diese so modifizierten Proteine wurden weiter zur Stabilisierung und Produktentwicklung getestet. Die ersten Ergebnisse eröffnen Nutzungsmöglichkeiten der modifizierten Proteine zur Verkapselung von bioaktiven Sekundären Pflanzenstoffen.

Les émulsions eau/eau (W/W) ont suscité un grand intérêt en raison de leur potentiel d'application dans différentes industries telles que l'agroalimentaire, les produits pharmaceutiques, les cosmétiques et les soins personnels. Le caractère particulier des émulsions W/W est leur stabilisation par ajout de particules. L’objectif de ce travail de thèse est de comprendre cet aspect en étudiant une émulsion modèle W/W à base de dextran et du poly(oxyde d'éthylène) stabilisée par des particules à base de protéines du lactosérum. Dans un premier temps, nous avons étudié l'effet de la morphologie des particules protéiques et leur partitionnement sur la stabilité des émulsions W/W. En particulier, la stabilité s’est révélée dépendre de la structure des particules quand ses derniers étaient sous forme de microgels, d’agrégats fractals ou de fibrilles. Il a été montré que la stabilité s'améliorait lorsque les particules se localiser préférentiellement dans la phase continue. Deuxièmement, nous avons étudié la gélification, des microgels et des agrégats fractals, induite en réduisant le pH entre 6,5 et 3,5 ou en ajoutant 0,3 M NaCl à pH 7,0 aussi bien quand l’excès des particules se situe dans la phase continue ou dispersée. Dans le premier cas, un réseau se formé dans la phase continue de dextran, permettant d’inhiber le crémage des gouttelettes de PEO, les agrégats fractals étant plus efficaces que les microgels. Dans le second cas, des particules protéiques denses pourraient être formées par gélification des gouttelettes de dextran dispersées. Finalement, nous avons exploré l'adsorption des protéines natives sur les particules de latex et leur capacité à stabiliser les émulsions. / Water/water (W/W) emulsions have attracted great interest recently due to their high potential for applications in different industries such as food and beverages, pharmaceutical, cosmetics and personal care. An important issue is the stabilization of W/W emulsions by adding particles. The aim of the research for this thesis was to shed light on this issue by studying a model W/W emulsion formed by mixing dextran and poly(ethylene oxide) with particles based on whey proteins. Firstly, we studied the effect of the morphology of protein particles and their partitioning on the stability of W/W emulsions. The stability was different when microgels, fractal aggregates or fibrils were added. We showed that stability improved when the particles partitioned to the continuous phase. Secondly, we investigated gelation of the fractal aggregates and microgels induced by reducing the pH between 6.5 and 3.5 or by adding 0.3M NaCl at pH 7.0 with excess particles either in the continuous or he dispersed phase. In the first case, a network was formed in the continuous dextran phase, making it possible to arrest creaming of PEO droplets, fractal aggregates being more effective than microgels. In the second case, dense protein particles could be formed by gelation of the dispersed dextran droplets. Thirdly, we explored the effect of adsorbing native proteins unto latex particles on their capacity to stabilize W/W emulsions.