Cold set gelation of whey proteins : acidification rate, final pH and polysaccharide addition effects

Orientador: Rosiane Lopes da CunhaTese (doutorado) - Universidade Estadual de Campinas, Faculdade de Engenharia de AlimentosResumo: A gelificação a frio de soluções de isolado protéico de soro (WPI) foi realizada pela adição de diferentes quantidades de glucona-d-lactona (GDL) a soluções de WPI desnaturadas termicamente (80ºC/30 minutos). Foram analisados sistemas protéicos puros (WPI na concentração de 7% p/p) e mistos, após a adição de xantana ou guar (concentração WPI 5% fixa e de polissacarídeos, 0,1, 0,3 e 0,5% p/p). Estes sistemas exibiram diferentes taxas de acidificação e valores de pH final em torno do ponto isoelétrico das principais frações protéicas do soro (5,2 a 3,9). Nos sistemas protéicos puros foi possível distinguir dois estágios de desenvolvimento estrutural: 1-início da formação da rede até o ponto de gel, em que as propriedades reológicas não foram influenciadas pela taxa de acidificação, e 2- subseqüente desenvolvimento estrutural com a redução do pH devido a fortalecimento de ligações e rearranjos estruturais. O processo lento de acidificação levou a géis mais estruturados no pH final de 5,2 enquanto que a acidificação rápida produziu géis mais frágeis (pH 4,2), que mostraram rearranjo estrutural após a obtenção do pH final. No pH de equilíbrio em torno do pI, a capacidade de retenção de água foi menor, o que foi associado à menor solubilidade protéica (exceto da fração ß-Lg) nestas condições. A solubilidade protéica na presença ou ausência de condições desnaturantes indicou que interações eletrostáticas foram responsáveis pela manutenção da rede nos pHs 5,2 a 4,6, porém um maior caráter hidrofóbico foi visualizado no pH 4,2. A adição de polissacarídeos levou a uma descontinuidade da estrutura da rede do gel, com um aumento da quantidade de poros. A adição de xantana enfraqueceu a rede e diminuiu a sua deformabilidade. Em menores taxas de acidificação e concentração de xantana visualizou-se a formação de géis heterogêneos (separação de fases macroscópica), com maiores valores de lacunaridade (descontinuidade da microestrutura), e menores valores de dimensão fractal, Df, (agregados protéicos menores e menos complexos na rede). No entanto, o aumento da taxa de acidificação e diminuição do pH final levou à formação de géis homogêneos, com uma rede mais forte, um sistema menos poroso e com maiores valores de Df (agregados mais complexos na rede devido a maior interação eletrostática entre proteínas e xantana). Em elevadas concentrações de guar houve formação de géis não auto-sustentáveis com grande lacunaridade e menores Df. Em menores concentrações de guar observou-se a formação de uma rede contínua no gel (menos porosa), com menores valores de Df. Nos géis autosustentáveis foi visualizado que o aumento da taxa de acidificação levou a géis com maior valor de tensão de ruptura e módulo de elasticidade, maior Df e menor lacunaridade, o que foi associado ao aumento de interação proteína-proteína. A descontinuidade da microestrutura nos géis WPI-xantana foi resultado de fenômenos simultâneos de separação de fases (condição inicial dos sistemas no pH 6,7) e gelificação (fase de redução de pH), sendo, portanto influenciada pela concentração de xantana, taxa de acidificação e pH final do gel, enquanto que nos géis WPI-guar o efeito predominante do aumento da concentração de guar foi associado a efeitos de exclusão de volume entre as proteínas do soro e a guarAbstract: The cold set gelation of whey protein isolate (WPI) solutions was induced by addition of several amounts of glucone-d-lactone (GDL) to thermal denatured (80ºC/30min) WPI solutions. Pure whey protein systems (7% WPI w/w) and mixed WPI-xanthan or WPI-guar gels (5% WPI w/w with the addition of 0.1, 0.3 and 0.5% polysaccharide) were studied. The systems showed different acidification rates, which led to different final pH values near the isoelectric point (pI) of the main whey proteins (5.2 to 3.9). Two gelation stages were defined in pure whey protein systems: 1- until the gel point, in which the rheological properties were not influenced by the acidification rates; 2- gel structure development until the pH equilibrium, in which bonds strengthening and molecular rearrangements on gel network took place. Lower acidification rates led to stronger and structured gels at final pH 5.2. However, the increase in acidification condition caused the formation of weaker gels at final pH 4.2. In the latter case, molecular rearrangements on structure took place during a long time after the achievement of steady pH values. At this pH, the water retention capacity of the gels was also lower, which was attributed to more fragile network but mainly due to the lower protein solubility (except ß-Lg fractions) near the pI. The protein solubility in buffers also showed that electrostatic interactions were involved in gel structure stabilization at final pH range of 5.2-4.6 and a more hydrophobic contribution was observed at pH 4.2. The polysaccharide addition caused the formation of great discontinuity on gels structure, leading to an increment on gel porosity. The xanthan addition weakened the gels. Heterogeneous gels (macroscopic phase separation), with higher values of lacunarity (network discontinuity) and lower values of fractal dimension, Df, (smaller and less complex aggregates) were formed at lower acidification conditions and xanthan concentration. However, the increase in acidification rate led to homogeneous gels formation, with lower lacunarity. This could be explained by an increased electrostatic attraction between proteins and xanthan at pH<pI, leading to higher values of stress at rupture and elasticity modulus at final pHs 5.2 and 4.2, respectively. At higher guar concentration the WPI-guar gels were not self-supported, with greater lacunarity and lower Df values. More continuous network was observed for mixed gels at lower guar concentration. The increase in acidification rate led a reduction in lacunarity and an increase stress at rupture; elasticity modulus and Df values of self supported WPI-guar gels. This latter result was attributed to an increase in protein-protein interactions. The network discontinuity in mixed WPI-xanthan gels was attributed to simultaneous effects of phase separation and gelation. These effects were influenced by xanthan concentration, acidification rate and final pH. However, mixed WPI-guar gels showed a predominant effect of guar concentration on microstructure discontinuity, which was attributed to excluded volume effects between the whey proteins and guarDoutoradoDoutor em Engenharia de Alimento

Stability And In Vitro Digestibility Of Emulsions Containing Lecithin And Whey Proteins.

The effect of pH and high-pressure homogenization on the properties of oil-in-water (O/W) emulsions stabilized by lecithin and/or whey proteins (WPI) was evaluated. For this purpose, emulsions were characterized by visual analysis, droplet size distribution, zeta potential, electrophoresis, rheological measurements and their response to in vitro digestion. Lecithin emulsions were stable even after 7 days of storage and WPI emulsions were unstable only at pH values close to the isoelectric point (pI) of proteins. Systems containing the mixture of lecithin and WPI showed high kinetic instability at pH 3, which was attributed to the electrostatic interaction between the emulsifiers oppositely charged at this pH value. At pH 5.5 and 7, the mixture led to reduction of the droplet size with enhanced emulsion stability compared to the systems with WPI or lecithin. The stability of WPI emulsions after the addition of lecithin, especially at pH 5.5, was associated with the increase of droplet surface charge density. The in vitro digestion evaluation showed that WPI emulsion was more stable against gastrointestinal conditions.41322-3

Rheological characterization of rhea (Rhea americana) eggs subjected to different storage periods

Eggs are used as food industry ingredients due to their functional properties. Eggs from different bird species are used in industrial processing; therefore, rheological experiments were carried out at steady state to obtain the flow curves and also at a dynamic state to study the rheological properties of greater rhea eggs. The viscoelastic behavior of the fluids as a function of the coagulation temperatures for albumen and yolk and of the oscillatory shear frequency was studied. Fifteen rhea eggs stored for up to 28 days at 10 °C were analyzed at 1, 7, 14, 21, and 28 days. The albumen and yolk fractions showed pseudoplastic rheological behavior with small initial flow shear and dependence on shear time, with albumen demonstrating thixotropy and the yolk showing rheopexy. In low shear rheological tests, thermal gelation of albumen was observed, presenting a first change in the elastic modulus around 56 °C, and a second from 78 °C. In the yolk, a change was observed from 66 °C, with more structural variations in the development of the gel at higher temperatures. For rhea eggs, the gel structuring (gelation) temperature of albumen (80 °C) and egg yolk (69 °C) did not undergo variations during the storage periods. Compared to commercial eggs, the higher viscosity and gelation temperatures presented by the rhea eggs can help determine the size of the industrial processing system as they can be submitted to higher temperatures without modifying protein structures and hindering the process. Keywords: birds; denaturation temperature; flow; functional properties; protei

Protein-based structures for food applications: from macro to nanoscale

Novel food structures' development through handling of macroscopic and microscopic properties of bio-based materials (e.g., size, shape, and texture) is receiving a lot of attention since it allows controlling or changing structures' functionality. Proteins are among the most abundant and employed biomaterials in food technology. They are excellent candidates for creating novel food structures due to their nutritional value, biodegradability, biocompatibility, generally recognized as safe (GRAS) status and molecular characteristics. Additionally, the exploitation of proteins' gelation and aggregation properties can be used to encapsulate bioactive compounds inside their network and produce consistent delivery systems at macro-, micro-, and nanoscale. Consequently, bioactive compounds which are exposed to harsh storage and processing conditions and digestion environment may be protected and their bioavailability could be enhanced. In this review, a range of functional and structural properties of proteins which can be explored to develop macro-, micro-, and nanostructures with numerous promising food applications was discussed. Also, this review points out the relevance of scale on these structures' properties, allowing appropriate tailoring of protein-based systems such as hydrogels and micro- or nanocapsules to be used as bioactive compounds delivery systems. Finally, the behavior of these systems in the gastrointestinal tract (GIT) and the impact on bioactive compound bioavailability are thoroughly discussed.JM and AP acknowledge the Portuguese Foundation for Science and Technology (FCT) for their fellowships (SFRH/BPD/89992/2012 and SFRH/BPD/101181/2014). This work was supported by Portuguese FCT under the scope of the Project PTDC/AGR-TEC/5215/2014, of the strategic funding of UID/BIO/04469 unit and COMPETE 2020 (POCI-01-0145-FEDER-006684), and BioTecNorte operation (NORTE-01-0145-FEDER-000004) funded by the European Regional Development Fund under the scope of Norte2020—Programa Operacional Regional do Norte.info:eu-repo/semantics/publishedVersio

Influence of heat denaturation and the pH on rheological properties of whey protein and its thermodynamic compatibility with xanthan

Orientador: Rosiane Lopes da CunhaDissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Engenharia de AlimentosResumo: Proteínas e polissacarídeos correspondem a biopolímeros muito utilizados pela indústria de alimentos devido às suas características funcionais, porém apresentam uma incompatibilidade natural quando em soluções. Sistemas compostos por proteínas do soro e xantana foram avaliados neste trabalho em diferentes condições de pH, com as proteínas nativas e desnaturadas termicamente. Análises de eletroforese em gel de poliacrilamida foram realizadas para verificar o efeito do tratamento térmico nas proteínas do soro, mostrando que o aquecimento de soluções de proteínas do soro a 80oe por trinta minutos causou a formação de grandes agregados protéicos estabilizados principalmente por interações dissulfídicas. Estes agregados tiveram influência marcante no comportamento de fases destas proteínas com a xantana. Soluções nativas de isolado protéico de soro 5% (p/p) e xantana entre 0,01 % a 1 % se revelaram compatíveis, porém quando as proteínas foram termicamente desnaturadas, estes sistemas se mostraram termodinamicamente incompatíveis, separando em duas fases. Um diagrama de fases capaz de descrever a região de incompatibilidade termodinâmica desses sistemas foi determinado, porém limitações experimentais atribuídas principalmente à ineficiência da ultracentrifugação na separação das fases incompatíveis quando as soluções possuem elevada viscosidade impediu a determinação da binodal de equilíbrio. O principal mecanismo termodinâmico envolvido na incompatibilidade termodinâmica seria a baixa entropia quando as proteínas do soro estão agregadas, porém efeitos de volume excluído entre os agregados e as grandes moléculas de xantana também poderiam estar envolvidos nesta incompatibilidade, principalmente quando as soluções são diluídas. A técnica reológica conhecida como compressão biaxial imperfeita com lubrificação foi adequada na determinação de diferenças de consistência de géis ácidos de isolado protéico de soro 7% (p/p). As soluções (48h armazenamento/10°C) apresentaram estabilização de pH entre 3,36 e 5,63 e os índices de comportamento do fluido em regime extensional variaram entre 0,19 e 0,43, enquantoque..os. Indices .deconsistência . extensional. variaramentre.131.eA127 kPa.sn, indicando o caráter pseudoplástico destes géis. Os géis mais consistentes foram obtidos nos pHs 4,73 e 5,10. Interações dissulfídicas foram responsáveis pela estabilização dos agregados no tratamento térmico e podem estar envolvidas no processo de gelificação, enquanto que interações eletrostáticas seriam as principais envolvidas na agregação das proteínas durante a redução do pHAbstract: Proteins and polysaccharides are two biopolymers widely used in food industry for their functional characteristics, but these biopolymers show natural thermodynamic incompatibility in solution. Native and heat denatured whey proteins and xanthan systems were studied in this work in different pH conditions. Polyacrylamide gel electrophoresis (PAGE) analysis was used to verify the effect of heat treatment on whey proteins showing that the experimental conditions (80OC for 30 minutes) caused protein aggregation in large aggregates stabilized mainly by disulfíde interactions. The large protein aggregates had great influenceon phase behavior between such proteins and xanthan. Native whey protein (5% w/w) and xanthan (0,01-1% w/w) solutions are soluble, but such solutions beco me not soluble and show thermodynamic incompatibility, when the proteins are heat denatured. A phase diagram was established and it was able to describe the concentration region of ingredients incompatibility. The binodal line was not drawn due to experimental limitations, probably associated to the inefficiency of the uJtracentrifugation to separate the incompatible phases of high viscosity. Low mixture entropy due to protein aggregation is the main thermodynamic event leading to phase separation of this systems, but excluded volume effects between protein aggregates and great xanthan molecules could be involved in this incompatibility, mainly for diluted solutions. Imperfect lubricated squeezing flow viscosimetry analysis was able to distinguish consistency differences in acidified whey protein isolate (7% w/w) gels. The pH of whey protein solutions (48h storage/10°C) was stabilized between 3,36 and 5,63 and the flow index values under extensional fJow were between 0,19 and 0,43, while the consistency index values under extensíonal flow were between 131 and 4127 kPa.sn, indicating the shear thinning behavior of such gels. The strongest gels were obtaíned in the pH range between 4,73 and 5,10. Disulphide interactions were responsible for protein aggregation caused by the heat treatment, while eJectrostatic interactions could be the mainly factor determining protein aggregation duríng pH reductionMestradoMestre em Engenharia de Alimento

The Effect Of Transglutaminase From Streptomyces Sp. Cbmai 837 On The Gelation Of Acidified Sodium Caseinate

The influence of the transglutaminase (TG) from a newly isolated Brazilian Streptomyces sp. CBMAI 837 and sodium caseinate concentration (4% and 8%, w/w) on acid-gel properties was studied. Standard gels with and without commercial transglutaminase samples were tested in parallel. The mechanical properties of the samples (stress and strain at rupture and elasticity modulus) were evaluated using uniaxial compression measurements. Texture parameters showed that the commercial TG gels had greater values of elasticity modulus, while addition of the Streptomyces sp. CBMAI 837 TG to the gel led to the formation of less elastic, but more deformable gels. The electrophoresis results showed that the commercial TG enzyme in this system promoted higher molecular mass protein polymers than the enzyme from Streptomyces sp. CBMAI 837. Microscopy and water-holding capacity (WHC) observations showed that all the gel characteristics could be associated with specific interactions promoted by each TG tested. © 2010 Elsevier Ltd.2010673679Ando, H., Adachi, M., Umeda, K., Matsuura, A., Nonaka, M., Uchio, R., Purification and characteristics of a novel transglutaminase derived from microorganism (1989) Agriculture and Biological Chemistry, 53, pp. 2613-2617Barbut, S., Determining water and fat holding (1996) Methods of testing protein functionality, pp. 187-225. , Chapman & Hall, London, UK, G.M. 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Características físicas, composição químico-nutricional e dos óleos essenciais da polpa de Caryocar brasiliense nativo do estado de Mato Grosso

O fruto do pequizeiro (Caryocar brasiliense Camb.) apresenta considerável variabilidade em suas características físicas e químicas, segundo a região de origem, e pesquisas com frutos procedentes do Estado de Mato Grosso são escassas na literatura. Neste estudo, foram analisadas as características físicas, a composição químico-nutricional e os constituintes voláteis da polpa de pequi coletado em diferentes locais do Estado de Mato Grosso. Os frutos foram analisados quanto à cor (L*, a*, b*, hº e C*), à massa e ao rendimento do mesocarpo externo, pirênios e polpa. Foram determinados a composição centesimal, os teores de carotenoides totais, o perfil de ácidos graxos e de constituintes voláteis da polpa, e analisada a composição físico-química do solo dos locais de coleta.. Constatou-se que o fruto é composto por 80-85% de mesocarpo externo, 11-15% de pirênios e 4-7% de polpa, que é rica em lipídios (46-51g 100g-1), carotenoides totais (15-27mg 100g-1) - que foram associados à tonalidade e à intensidade da cor (hº e C*) da polpa - e fibra alimentar (8-15g 100g-1). Os principais constituintes voláteis da polpa de pequi foram o hexanoato de etila, (E)-&#946;-ocimeno e octanoato de etila. O isobutirato de alila, 3-hexanol, 4-metil-2-pentanol, mirceno, (Z)-di-hidroapofarnesol, &#946;-eudesmol e (E-E) geranilalinalool foram descritos pela primeira vez nos óleos essenciais da polpa de C. brasiliense. Grande parte da variância total dos voláteis foi explicada pelo conjunto de variáveis do solo, destacando-se o silte, o P e o Mg2+

Colloidal and Acid Gelling Properties of Mixed Milk and Pea Protein Suspensions

The present study aims to describe colloidal and acid gelling properties of mixed suspensions of pea and milk proteins. Mixed protein suspensions were prepared by adding pea protein isolate to rehydrated skimmed milk (3% w/w protein) to generate four mixed samples at 5, 7, 9, and 11% w/w total protein. Skimmed milk powder was also used to prepare four pure milk samples at the same protein concentrations. The samples were analyzed in regard to their pH, viscosity, color, percentage of sedimentable material, heat and ethanol stabilities, and acid gelling properties. Mixed suspensions were darker and presented higher pH, viscosity, and percentage of sedimentable material than milk samples. Heat and ethanol stabilities were similar for both systems and were reduced as a function of total protein concentration. Small oscillation rheology and induced syneresis data showed that the presence of pea proteins accelerated acid gel formation but weakened the final structure of the gels. In this context, the results found in the present work contributed to a better understanding of mixed dairy/plant protein functionalities and the development of new food products