Made available in DSpace on 2014-08-13T14:50:40Z (GMT). No. of bitstreams: 0 Previous issue date: 2013-09-06Bitstream added on 2014-08-13T18:01:29Z : No. of bitstreams: 1 000747140.pdf: 3556860 bytes, checksum: 8b2649d7b40ff90af861460125883363 (MD5) / O trabalho consiste em sintetizar e estudar amostras de cobalto e zinco contendo hidrolisados proteicos do soro do leite. A motivação do trabalho está centrada no fato deste ser um material altamente nutritivo, podendo portanto ser aproveitado como suprimento alimentar. A literatura recente tem reportado o fato de que a biodisponibilidade de oligoelementos, quando administrados na forma quelatada, é aumentada em comparação com os sais inorgânicos, comumente utilizados como fontes de minerais. Como objetivo secundário, tem-se a responsabilidade ambiental de evitar o descarte inadequado do soro de leite pelas indústrias de laticínios, já que trata-se de um produto altamente poluente (DBO da ordem de 25.000 - 80.000 mg O2/L). A fase inicial do trabalho foi realizada no Laboratório de Enzimologia da Faculdade de Ciências Farmacêuticas da UNESP de Araraquara e consistiu nas etapas de obtenção, tratamento, hidrólise enzimática e liofilização do soro do leite. Os resultados obtidos da eletroforese SDS-PAGE e dos aminogramas permitiram concluir que a hidrólise conduziu à formação de aminoácidos. A segunda fase do trabalho, realizada no Departamento de Química Geral e Inorgânica deste Instituto, compreendeu a síntese e estudo das diversas amostras obtidas, contendo cobalto ou zinco e o hidrolisado proteico, dependendo da rota de síntese escolhida e/ou das condições reacionais utilizadas (natureza do material de partida, solvente, pH, temperatura, etc.). A idéia maior do projeto é que essas amostras de coordenação contendo ligantes de origem natural possam integrar uma lista multimineral a ser testada posteriormente como metalofármacos (Ag, Pd, Pt) ou substâncias nutracêuticas (Cu, Zn, Co, Fe). / This study consists of synthesis and evaluation of cobalt and zinc compounds containing proteic hydrolyzates from whey. The motivation of the study is focused on the fact that is a highly nutritive material, which can be used, therefore, as food supply. The recent literature has reported that the bioavailability of trace elements, when administrated in the chelated form, is increased in comparison with the inorganic salts, mostly used as minerals sources. As a secondary objective, it has environmental responsibility to avoid the inappropriate discard of the whey by dairy industries, since it is a highly pollutant product (iological Oxygen Demand of the order of 25,000 - 80,000 mg O2/L). The initial stage of the study was performed in the laboratory of enzymology, Faculdade de Ciências Farmacêuticas of UNESP - Araraquara and consisted in the steps of extraction, treatment, hydrolysis and freezedrying of the whey. The results from SDS-PAGE electrophoresis and aminograms revealed that the enzymatic hydrolysis was effective and led to the release of amino acids. The second stage of the study, performed on Departamento de Química Geral e Inorgânica of this Institute, comprised the synthesis and study of the several compounds obtained containing cobalt or zinc ions and the proteic hydrolyzate, by different synthetic routes and experimental conditions (such as material nature, solvent, pH, temperature, etc.). The major goal of the project is that different coordination compounds containing natural ligands could compose a multimineral list to be tested as metallodrugs (Ag, Pd, Pt) or nutraceutical substances (Cu, Zn, Co, Fe).

Orientador: Regina Célia Galvão Frem Di Nardo / Banca: Flavia Maria Netto / Banca: Eduardo Tonon de Almeida / Resumo: O trabalho consiste em sintetizar e estudar amostras de cobalto e zinco contendo hidrolisados proteicos do soro do leite. A motivação do trabalho está centrada no fato deste ser um material altamente nutritivo, podendo portanto ser aproveitado como suprimento alimentar. A literatura recente tem reportado o fato de que a biodisponibilidade de oligoelementos, quando administrados na forma quelatada, é aumentada em comparação com os sais inorgânicos, comumente utilizados como fontes de minerais. Como objetivo secundário, tem-se a responsabilidade ambiental de evitar o descarte inadequado do soro de leite pelas indústrias de laticínios, já que trata-se de um produto altamente poluente (DBO da ordem de 25.000 - 80.000 mg O2/L). A fase inicial do trabalho foi realizada no Laboratório de Enzimologia da Faculdade de Ciências Farmacêuticas da UNESP de Araraquara e consistiu nas etapas de obtenção, tratamento, hidrólise enzimática e liofilização do soro do leite. Os resultados obtidos da eletroforese SDS-PAGE e dos aminogramas permitiram concluir que a hidrólise conduziu à formação de aminoácidos. A segunda fase do trabalho, realizada no Departamento de Química Geral e Inorgânica deste Instituto, compreendeu a síntese e estudo das diversas amostras obtidas, contendo cobalto ou zinco e o hidrolisado proteico, dependendo da rota de síntese escolhida e/ou das condições reacionais utilizadas (natureza do material de partida, solvente, pH, temperatura, etc.). A idéia maior do projeto é que essas amostras de coordenação contendo ligantes de origem natural possam integrar uma lista multimineral a ser testada posteriormente como metalofármacos (Ag, Pd, Pt) ou substâncias nutracêuticas (Cu, Zn, Co, Fe). / Abstract: This study consists of synthesis and evaluation of cobalt and zinc compounds containing proteic hydrolyzates from whey. The motivation of the study is focused on the fact that is a highly nutritive material, which can be used, therefore, as food supply. The recent literature has reported that the bioavailability of trace elements, when administrated in the chelated form, is increased in comparison with the inorganic salts, mostly used as minerals sources. As a secondary objective, it has environmental responsibility to avoid the inappropriate discard of the whey by dairy industries, since it is a highly pollutant product (iological Oxygen Demand of the order of 25,000 - 80,000 mg O2/L). The initial stage of the study was performed in the laboratory of enzymology, Faculdade de Ciências Farmacêuticas of UNESP - Araraquara and consisted in the steps of extraction, treatment, hydrolysis and freezedrying of the whey. The results from SDS-PAGE electrophoresis and aminograms revealed that the enzymatic hydrolysis was effective and led to the release of amino acids. The second stage of the study, performed on Departamento de Química Geral e Inorgânica of this Institute, comprised the synthesis and study of the several compounds obtained containing cobalt or zinc ions and the proteic hydrolyzate, by different synthetic routes and experimental conditions (such as material nature, solvent, pH, temperature, etc.). The major goal of the project is that different coordination compounds containing natural ligands could compose a multimineral list to be tested as metallodrugs (Ag, Pd, Pt) or nutraceutical substances (Cu, Zn, Co, Fe). / Mestre

Orientador: Rubens Monti / Banca: Thais Borges Cesar / Banca: João Atílio Jorge / Banca: Rosa dos Prazeres Melo Furriel Inocentes / Banca: Reinaldo Marchetto / Resumo: Os hidrolisados de proteínas do soro de queijo vêm sendo amplamente utilizados para diversos fins. Dependendo das enzimas utilizadas e meio de reação, podemos obter hidrolisados com diferentes características. O intuito do presente trabalho foi hidrolisar as proteínas do soro de queijo bovino de diferentes maneiras, variando-se a temperatura de reação e modo de adição das enzimas, a fim de se obter hidrolisados com diferentes graus de hidrólise para posterior aplicação em ensaios animais. Os hidrolisados escolhidos para aplicação nos ensaios biológicos foram obtidos pela ação das enzimas pepsina, tripsina, quimotripsina e carboxipeptidase a 50ºC. Dois hidrolisados diferentes foram preparados: um com maior quantidade de pequenos peptídeos (oligo, di e tripeptídeos) e outro com maior quantidade de aminoácidos livres. O soro sem hidrólise também foi utilizado como suplementação para um outro grupo de animais. A intenção foi testar os dois diferentes hidrolisados e o soro sem hidrólise como suplemento para ratos idosos, visando a minimização da sarcopenia. Diversos estudos apontam que fórmulas contendo oligopeptídeos, especialmente di- e tripeptídeos, possuem maior valor nutricional do que uma mistura equivalente de aminoácidos livres ou proteínas intactas. Tomando-se como base essa informação, delineou-se a pesquisa esperando-se que, o grupo suplementado com as proteínas parcialmente hidrolisadas obtivesse um melhor desempenho na minimização da sarcopenia. Todos os 3 grupos suplementados obtiveram aumento na massa muscular e óssea diferindo significativamente em relação ao controle (não suplementado) porem, o que apresentou efeitos mais positivos na minimização da sarcopenia foi realmente o grupo suplementado por proteínas parcialmente hidrolisadas. Paralelamente a esse estudo... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: The cheese whey hydrolysates have been widely used for various purposes. Depending on the enzymes used and the reaction medium, we can obtain hydrolysates with different characteristics. The purpose of this study was to hydrolyze bovine cheese whey proteins in different ways, varying the temperature of reaction and enzyme addition, in order to obtain hydrolysates with different degrees of hydrolysis for application in animal tests. The hydrolysates chosen for application in biological assays were obtained by the action of enzymes pepsin, trypsin, chymotrypsin and carboxypeptidase at 50°C. Two different hydrolysates were prepared: one with a larger amount of small peptides (oligo-, di-and tripeptides) and another with larger amount of free amino acids. Serum without hydrolysis was also used as a supplement to another group of animals. The intention was to test the two different hydrolysates and whey without hydrolysis as a supplement to old age rats in order to minimize the sarcopenia. Several studies indicate that formulas containing oligopeptides, especially di-and tripeptides, have higher nutritional value than an equivalent mixture of free amino acids or intact proteins. Taking this information as a basis, the research was then outlined expecting that the group supplemented with partially hydrolyzed protein could obtain a better performance in reducing the sarcopenia. The all 3 supplemented groups showed increase in muscle mass and bone and differ significantly from control (not supplemented). However, the group who presented the most positive effects in reducing the sarcopenia was actually the group supplemented with partially hydrolyzed protein. In parallel with this study, in sandwich PhD, the attainment of immobilized pepsin and trypsin (derivatives) in solid support, stable was studied, for further application to obtain... (Complete abstract click electronic access below) / Doutor

This thesis deals with the possibilities of biodegradation of whey by the means of a mixed thermophilic aerobic bacterial culture of the Bacillus and Thermus genera. After protein had been removed, the whey was used as a medium for cultivation of the thermophilic microorganisms. The cultivations took their course in a fermentation unit and in the Erlenmayer flasks in a heated shaker in a whey medium. The bioengineering characteristics of the cultivation processes and the degree of biodegradation of the whey were evaluated. Scale of the whey biodegration was judged through the analytical characteristics – concentration of biomass, laktose and a chemical oxygen demand (COD). A decrease of CHSK was detected in all cultivations. Maximal reduction of CHSK was happend always in c. first half of each exponential phase growt, i.e. about 15 ± 3 % after first growth phase and sumarily about 62 ± 4 % after second growth phase.

Presented work is focused on study of polyhydroxybutyrate production in bacteria. In theoretical part short characterization of PHB was given and the most common representative of wide group of polyhydroxyalkanoates (PHA) were described. Then, production of PHB and copolymer P(HB-co-HV) in selected bacterial strains was experimentally proven. First, PHB production in Bacillus megaterium using synthetic medium was studied. The PHB content in cells was increased during cultivation under limiting conditions, despite low growth. Addition of ethanol into production media resulted in increased PHB synthesis as well as biomass production (21 % PHB of 1,8 g/l biomass). Further, BM medium containing 8 g/l glucose was tested. PHB production was more than 1 g/l at significant growth increase when compared with synthetic medium. The bacteria B.megaterium showed, except glucose, ability to utilize maltose and xylose. Another cultivations were tested with bacterial strain Azotobacter vinelandii, which is capable of copolymer P(HB-co-HV) synthesis. Maximal growth and copolymer content was reached on Burk's medium with 30 g/l of glucose. Addition of peroxide to growth medium influenced P(HB-co-HV) synthesis to 46 % of 2,6 g/l biomass. Bacteria A.vinelandii showed the best growth on maltose, even compared with glucose (54 % copolymer of biomass content). Finally, PHB production on industrial waste product – whey was monitored. Using Plackett-Burman design for statistical media optimization, the whey content was modified. B.megaterium grown on adjusted whey reached 0,5 g/l PHB, 32 % of cell's content.

The Western Dairy Center at Utah State University had recognized through traiing of cheesemaking that a challenge that can exist for farmstead and artisan cheese manufacturing operations is disposal or utilization of the whey that is produced when milk is converted into cheese. Land application of whey is limited and can cause odor problems which would be detrimental to a cheesemaking operation located at the rual-urban interface. The project provided information in support of a research grant from the Western Sustainable Agribulture Research and Education program that was investigating economics and feasiblity of using anaerobic digestion for treatment of whey and cow manure mixtures. We performed initial trials to determine the level of whey that could be mixed with manure and have satisfactory operation of the digester to produce methane and a biomass. A continuous digester was used to produce biomass material for microbiological and physical testing for its suitability for use as a renewable potting mix or soil conditioner. The benefits from this research are that we have shown how a value-added product can be made by converting whey and manure into a deodorized biomass that could replace the use of peat moss, which is a non-renewable resource. This can improve the economics of using a bioreactor for whey disposal.

Hydrolysis of casein using chymotrypsin results in the formation of polypeptides (CH) with a hydrophobic aromatic amino acid on one end of the chain because the enzyme selectively cleaves the adjacent peptide-bond. Due to resonance of the aromatic micro-domain, thiols become redox-sensitive and actively participate in electron transfer. These types of amphipathic peptides also tend to be membrane-lytic. The two prong approach of this investigation was to, (1) assess antibacterial effect of the CH in beef steak, and (2), to determine its antioxidative efficacy as a constituent of Cheddar whey based edible coating mix. The edible coating prevented coliform growth even at a minute concentration range of 0.15-0.2 % (w/v). Marked antioxidative efficacy of the CH, particularly at a concentration of 0.3% (w/v), was also evident from its remarkable free radical scavenging ability and extended resilience in an abusive model system saturated with peroxyl-radicals generated through controlled pyrolysis.

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Protein structure affects the bioactivity and functionality of whey protein ingredients in food systems. Bioactivity of whey proteins and their derivatives are highly dependent upon primary, secondary and tertiary structure. The degree of denaturation of whey proteins is an important factor for determining how whey protein ingredients will perform in a food system. Several analytical methods have been developed to quantify protein denaturation of whey proteins. The goal of this project was to use a variety of analytical methods to quantify whey protein denaturation and to evaluate the correlation of denaturation to the functionality of whey protein powders. The objective of the first series of experiments was to compare three different analytical methods to measure denaturation of whey proteins in liquid whey obtained by various methods of separation and with varying degrees of heat treatment. A split plot experimental design was used. Raw bovine milk was skimmed and liquid whey was separated from the skim milk at natural pH. Three separation methods: 1) centrifugation, 2) membrane filtration and 3) enzyme coagulation, made up the first split plot. Each sub-plot of liquid whey was then divided into three split plots to receive heat treatment. Heat treatments were no heat, 76°C for fifteen seconds and 85°C for three minutes. Each of the resulting nine treatment combinations was analyzed by 1) polyacrylamide gel electrophoresis, 2) bicinchoninic acid-soluble protein assay and 3) fluorescence spectroscopy to determine the amount of denatured protein in the liquid whey. Fluorescence spectroscopy was found to be the most sensitive and reliable method for detecting differences in structure due to denaturation, while native polyacrylamide gel electrophoresis was found to be the least sensitive method. The sample which received the centrifugal treatment of isolation with no heat was found to be the most undenatured in structure while the sample which received the enzyme treatment of isolation with high heat was found to be the most denatured in structure. The objective of the second series of experiments was to evaluate the effect of denaturation on whey protein solubility in dried whey protein powders. Solubility is one of the most important functional properties to consider when selecting a whey protein ingredient, especially for beverage systems. Processing parameters are often manipulated in efforts to improve solubility. The protein structures of whey are considered to have an effect on solubility. Specifically, the degree of denaturation of whey proteins is thought to play a role in solubility. In this experimental design, raw bovine milk was skimmed and pasteurized then enzyme-coagulated at natural pH to separate the whey. Liquid whey was then split into three aliquots and each received one of the following treatments: 1) mild heat/ freeze dry, 2) mild heat/spray dry and 3) high heat/spray dry. Heat treatment was applied to liquid whey prior to concentration. Heat treated whey was then concentrated and dried. Powders were reconstituted and analyzed for denaturation using 1) bicinchoninic acid assay for soluble protein and 2) fluorescence spectroscopy and for solubility using an insolubility index. pH 4.6 solubility and fluorescence spectroscopy for quantifying denaturation correlated well to one another. Both found that the low heat treated samples were less denatured in structure than the sample which received the high heat treatment, regardless of drying method. However, the drying method of the protein powders was correlated to solubility rather than heat treatment. A correlation of denaturation measured in whey protein powders and solubility was apparent for the low heat, freeze dried sample and the high heat, spray dried sample. Several conclusions were made in this research. 1) Centrifugal force causes less denaturation than membrane filtration and enzyme coagulation, thus unheated liquid whey obtained by centrifugal force can be used as a control in research on denaturation. 1) Fluorescence spectroscopy is a better method for quantifying denaturation in liquid and powdered whey compared to native PAGE and pH 4.6 solubility measured by BCA. 3) Functional solubility is dependent on denaturation and can be correlated to analytical methods of measuring denaturation.