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O estudo da digestibilidade e do aproveitamento biológico do feijão (Phaseolus vulgaris, L. ),variedade Carioca e de suas principais frações protéicas foi realizado em ensaios biológicos com ratos, após tratamento térmico adequado (cocção). À metodologia convencional, baseada no \"Ganho de Peso\" e na razão entre o N ingerido e excretado, seguiu-se uma técnica utilizando feijões e suas frações protéicas previamente marcadas com 15N e 35S. A ingestão das rações causou respostas diferentes nos animais, específicos para cada fonte protéica. Os animais alimentados com o feijão integral tiveram reduzido aproveitamento biológico e excretaram 34% do nitrogênio ingerido, sendo este valor 3-4 vezes superior ao causado pela ingestão da ração controle de caseína. As frações, albumínicas e glutelínicas, apresentaram valores de \"NPR\" e \"Ganhos de Peso\" superiores, apesar de sua relativa baixa digestibilidade (70-80%). A globulina G1, altamente digerível (92%) teve uma utilização biológica muito reduzida, não permitindo o crescimento dos animais. O ensaio biológico do feijão e de suas proteínas, marcados isotopicamente, permitiu diferenciar a origem do nitrogênio e do enxofre fecal, se endógena ou exógena. A concentração de 15N em excesso, determinado nas fazes 12 horas após o início da ingestão das rações marcadas, consideramos ser de origem exógena, sendo esses valores interpretados comparativamente. A ingestão de feijões cozidos causou nos animais uma excreção de nitrogênio proveniente do próprio feijão, 10 vezes superior à elevada perda de nitrogênio endógeno, totalizando o equivalente a 34% do nitrogênio ingerido. Ratos alimentados com a globulina G1 excretaram nas primeiras 12 horas apenas quantidades muito reduzidas de 15N, e mesmo na fase de equilíbrio (4º - 6º dia de ingestão da reação marcada ) quando o 15N excretado corresponde ao exógeno + endógeno já marcado, a perda de N total não ultrapassou 10% do ingerido. A ingestão da fração albumínica, por sua vez, causou uma eliminação de 20% do N total ingerido, com participação tanto do N exógeno como endógeno. A interpretação dos resultados baseados na eliminação do enxofre e de seu radioisótopo 35S, mostrou um perfil de excreção semelhante ao observado para o 15N, embora em níveis levemente superiores. / Studies with growing rats were carried out to determine the digestibility, and nutritive value of beans (Phaseolus vulgaris, L.) Carioca variety, and their major protein fractions, after proper heat treatment in order to inactivate the antiphysiological factors. The conventional method based on weight gain and the relation between the ingested and excreted nitrogen was followed by a balance technique, employing either whole beans or their protein fractions, previously labeled with 15N and 35S. The ingestion of the diets showed different nutritive responses, which were specific for each of the diets. Low biological value and digestibility were observed by rats fed on whole bean-containing diets, and as a consequence an excretion of 34% the ingested nitrogen was noted. This value was 3-4 times higher than the one excreted by the casein-control animals. Higher vakues of \"weight gain\" and NPR were obtained with the albumin and glutelin based diets, although their digestibilities were relatively low (70 - 80%). The globulin G1 was highly digestible (92%) nevertheless it appeared to have very low nutritional values and did not allow the growth of rats. The biological assay with isotopically labeled beans and proteins, allowed to differenciate the origin of fecal nitrogen and sulfur, as exogenous or endogenous. The concentration of 15 N in excess, which appeared in the feces 12 hours after the first ingestion of the labeled diets, was considered to be of exogenous origin; these values can be interpreted comparatively. The ingestion of the cooked whole beans promoted an excretion of exogenous nitrogen, 10 times higher than that excreted by the rats on the globulin G1 diet, which with the high excretion of endogenous nitrogen, totalized 34% of the ingested one. Rats fed on the globulin G1 excreted, in the first 12 hours, only small amounts of 15N, even in the steady - state (4º - 6ºth day on feeding the labeled diet), when the excreted 15N corresponded to both, exogenous and labeled endogenous excretion reached, the maximum loss (about 10% of the ingested total nitrogen). The feeding of albumin based diets stimulated an excretion of 20% of the ingested nitrogen, due to exogenous and endogenous origin. The interpretation of the results based on the balance of total sulfur and their radioactive isotope (35S) showed an excretion pattern, similar to that of the 15N, although in absolute values (of excretion) it was a little higher than that representing the nitrogen loss.

This diploma thesis deals with fractionation of protein from hemp (Cannabis sativa L.) meal according to their solubility of the hemp varieties - Fedora 17 and Uso 31. The content of nitrogenous substances and protein content in the hemp meal of these studied varieties ranged from 23,6 % (Uso 31) to 31,5 % (Fedora 17) and from 100,1 mg.g-1 (Uso 31) to 130,7 mg.g-1 respectively. The fractionation of hemp protein (Fedora 17, Uso 31) confirmed that the main hemp protein fraction is an albumin fraction, with its relative abundance - 20,5 % of protein. As a next step, the spectra of hemp proteins were determined. The use SDS-PAGE analysis revealed albumin proteins in area of 7,4 53,6 kDa. In the globulin fraction was revealed by SDS-PAGE basic subunits of the major protein edestin in the area 19-21 kDa and acidic subunits in the area 34 kDa.

Made available in DSpace on 2014-06-11T19:23:26Z (GMT). No. of bitstreams: 0 Previous issue date: 2010-06-17Bitstream added on 2014-06-13T18:50:30Z : No. of bitstreams: 1 molina_jp_me_arafcf.pdf: 1805522 bytes, checksum: adb34fd5733f36fbfa386d389f68be89 (MD5) / Universidade Estadual Paulista (UNESP) / No presente trabalho, objetivou-se a determinar a composição química da farinha da semente de grão-de-bico, cultivar Cícero e do tremoço branco; o estabelecimento de condições de extração das proteínas; a separação das diferentes frações protéicas e caracterizou o comportamento térmico das frações protéicas. A composição química da farinha do grão-de-bico revelou valores de 18,72% de proteína, 10,02 % de umidade, 2,96% de cinzas e 2,6% de extrato etéreo. Já a composição química do tremoço revelou valores de 49,88% de proteína, 8,1% de umidade, 1,54% de cinzas e 2,94% de extrato etéreo. As frações do o grão-de-bico, cultivar Cícero, de maior conteúdo protéico foram a globulina com 88,67%, seguida da glutelina 74,78% e albumina 61,69% e prolamina 7,75%. Para o tremoço branco as frações de maiores conteúdos protéicos foram a albumina 91,33%, seguida da glutelina 89,24% e globulina 41,37%. O valor de energia gasta no processo endotérmico para o evento de desnaturação da proteína das frações protéicas do grão-de-bico, cultivar Cícero, expressos em H/Jg-1 é 190,3 para albumina, 154,3 para globulina, 140,4 para glutelina e 35,97 para prolamina. Para as frações protéicas do tremoço branco os valores expressos em H/Jg-1 é 173,6 para albumina, 208,8 para globulina e 126,0 para glutelina. O H/Jg-1 para prolamina não foi determinado. / The present study aimed to determine the chemical composition of the chickpeas, grow cicero and white lupine seed flour and the establishment of conditions for protein extraction, separation of different protein fractions and characterized the behavior thermal of the protein fractions. Now chemical composition of chickpeas flour revealed values of 18.72% protein, 10.02% moisture, 2.96% ash and 2.6% ether extract. Now the chemical composition of lupine revealed values of 49.88% protein, 8.1% moisture, 1.54% ash and 2.94% ether extract. The fractions for the chickpeas, Cicero is growing, of larger protein content were is globulin with 88.67%, followed by glutelin and albumin 74.78% 61.69% and 7.75% prolamin. For the white lupine, the fraction of larger protein content were is albumin 91.33%, followed by glutelin and globulin 89.24% 41.37%. The amount of energy expended in the endothermic process in the event of protein denaturation of the protein fractions of chickpeas, grow Cicero expressed in ΔH/Jg-1 is 190.3 for albumin, globulin to 154.3, 140, 4 to 35.97 for glutelin and prolamin. For the protein fractions of white lupine values in ΔH/Jg-1 is 173.6 for albumin, globulin and 208.8 to 126.0 for glutelin. The ΔH/Jg-1 to prolamin not determined.

Orientador: José Paschoal Batistuti / Banca: Rubens Monti / Banca: Jonas Contiero / Resumo: No presente trabalho, objetivou-se a determinar a composição química da farinha da semente de grão-de-bico, cultivar Cícero e do tremoço branco; o estabelecimento de condições de extração das proteínas; a separação das diferentes frações protéicas e caracterizou o comportamento térmico das frações protéicas. A composição química da farinha do grão-de-bico revelou valores de 18,72% de proteína, 10,02 % de umidade, 2,96% de cinzas e 2,6% de extrato etéreo. Já a composição química do tremoço revelou valores de 49,88% de proteína, 8,1% de umidade, 1,54% de cinzas e 2,94% de extrato etéreo. As frações do o grão-de-bico, cultivar Cícero, de maior conteúdo protéico foram a globulina com 88,67%, seguida da glutelina 74,78% e albumina 61,69% e prolamina 7,75%. Para o tremoço branco as frações de maiores conteúdos protéicos foram a albumina 91,33%, seguida da glutelina 89,24% e globulina 41,37%. O valor de energia gasta no processo endotérmico para o evento de desnaturação da proteína das frações protéicas do grão-de-bico, cultivar Cícero, expressos em H/Jg-1 é 190,3 para albumina, 154,3 para globulina, 140,4 para glutelina e 35,97 para prolamina. Para as frações protéicas do tremoço branco os valores expressos em H/Jg-1 é 173,6 para albumina, 208,8 para globulina e 126,0 para glutelina. O H/Jg-1 para prolamina não foi determinado. / Abstract: The present study aimed to determine the chemical composition of the chickpeas, grow cicero and white lupine seed flour and the establishment of conditions for protein extraction, separation of different protein fractions and characterized the behavior thermal of the protein fractions. Now chemical composition of chickpeas flour revealed values of 18.72% protein, 10.02% moisture, 2.96% ash and 2.6% ether extract. Now the chemical composition of lupine revealed values of 49.88% protein, 8.1% moisture, 1.54% ash and 2.94% ether extract. The fractions for the chickpeas, Cicero is growing, of larger protein content were is globulin with 88.67%, followed by glutelin and albumin 74.78% 61.69% and 7.75% prolamin. For the white lupine, the fraction of larger protein content were is albumin 91.33%, followed by glutelin and globulin 89.24% 41.37%. The amount of energy expended in the endothermic process in the event of protein denaturation of the protein fractions of chickpeas, grow Cicero expressed in ΔH/Jg-1 is 190.3 for albumin, globulin to 154.3, 140, 4 to 35.97 for glutelin and prolamin. For the protein fractions of white lupine values in ΔH/Jg-1 is 173.6 for albumin, globulin and 208.8 to 126.0 for glutelin. The ΔH/Jg-1 to prolamin not determined. / Mestre

O estudo da digestibilidade e do aproveitamento biológico do feijão (Phaseolus vulgaris, L. ),variedade Carioca e de suas principais frações protéicas foi realizado em ensaios biológicos com ratos, após tratamento térmico adequado (cocção). À metodologia convencional, baseada no \"Ganho de Peso\" e na razão entre o N ingerido e excretado, seguiu-se uma técnica utilizando feijões e suas frações protéicas previamente marcadas com 15N e 35S. A ingestão das rações causou respostas diferentes nos animais, específicos para cada fonte protéica. Os animais alimentados com o feijão integral tiveram reduzido aproveitamento biológico e excretaram 34% do nitrogênio ingerido, sendo este valor 3-4 vezes superior ao causado pela ingestão da ração controle de caseína. As frações, albumínicas e glutelínicas, apresentaram valores de \"NPR\" e \"Ganhos de Peso\" superiores, apesar de sua relativa baixa digestibilidade (70-80%). A globulina G1, altamente digerível (92%) teve uma utilização biológica muito reduzida, não permitindo o crescimento dos animais. O ensaio biológico do feijão e de suas proteínas, marcados isotopicamente, permitiu diferenciar a origem do nitrogênio e do enxofre fecal, se endógena ou exógena. A concentração de 15N em excesso, determinado nas fazes 12 horas após o início da ingestão das rações marcadas, consideramos ser de origem exógena, sendo esses valores interpretados comparativamente. A ingestão de feijões cozidos causou nos animais uma excreção de nitrogênio proveniente do próprio feijão, 10 vezes superior à elevada perda de nitrogênio endógeno, totalizando o equivalente a 34% do nitrogênio ingerido. Ratos alimentados com a globulina G1 excretaram nas primeiras 12 horas apenas quantidades muito reduzidas de 15N, e mesmo na fase de equilíbrio (4º - 6º dia de ingestão da reação marcada ) quando o 15N excretado corresponde ao exógeno + endógeno já marcado, a perda de N total não ultrapassou 10% do ingerido. A ingestão da fração albumínica, por sua vez, causou uma eliminação de 20% do N total ingerido, com participação tanto do N exógeno como endógeno. A interpretação dos resultados baseados na eliminação do enxofre e de seu radioisótopo 35S, mostrou um perfil de excreção semelhante ao observado para o 15N, embora em níveis levemente superiores. / Studies with growing rats were carried out to determine the digestibility, and nutritive value of beans (Phaseolus vulgaris, L.) Carioca variety, and their major protein fractions, after proper heat treatment in order to inactivate the antiphysiological factors. The conventional method based on weight gain and the relation between the ingested and excreted nitrogen was followed by a balance technique, employing either whole beans or their protein fractions, previously labeled with 15N and 35S. The ingestion of the diets showed different nutritive responses, which were specific for each of the diets. Low biological value and digestibility were observed by rats fed on whole bean-containing diets, and as a consequence an excretion of 34% the ingested nitrogen was noted. This value was 3-4 times higher than the one excreted by the casein-control animals. Higher vakues of \"weight gain\" and NPR were obtained with the albumin and glutelin based diets, although their digestibilities were relatively low (70 - 80%). The globulin G1 was highly digestible (92%) nevertheless it appeared to have very low nutritional values and did not allow the growth of rats. The biological assay with isotopically labeled beans and proteins, allowed to differenciate the origin of fecal nitrogen and sulfur, as exogenous or endogenous. The concentration of 15 N in excess, which appeared in the feces 12 hours after the first ingestion of the labeled diets, was considered to be of exogenous origin; these values can be interpreted comparatively. The ingestion of the cooked whole beans promoted an excretion of exogenous nitrogen, 10 times higher than that excreted by the rats on the globulin G1 diet, which with the high excretion of endogenous nitrogen, totalized 34% of the ingested one. Rats fed on the globulin G1 excreted, in the first 12 hours, only small amounts of 15N, even in the steady - state (4º - 6ºth day on feeding the labeled diet), when the excreted 15N corresponded to both, exogenous and labeled endogenous excretion reached, the maximum loss (about 10% of the ingested total nitrogen). The feeding of albumin based diets stimulated an excretion of 20% of the ingested nitrogen, due to exogenous and endogenous origin. The interpretation of the results based on the balance of total sulfur and their radioactive isotope (35S) showed an excretion pattern, similar to that of the 15N, although in absolute values (of excretion) it was a little higher than that representing the nitrogen loss.

O objetivo do estudo foi avaliar os efeitos da contagem de células somáticas (CCS) sobre características físico-químicas (pH, acidez, composição, cor), microbiológicas, índices de proteólise e lipólise, formação de compostos de Maillard, viscosidade aparente e frações protéicas do leite integral obtido pelo processamento direto de temperatura ultra alta (UAT). O leite cru foi avaliado quanto ao nível de células somáticas (CS) e características físicoquímicas. As amostras de leite foram colhidas em usina de beneficiamento de leite UAT no Estado de São Paulo. O leite cru foi submetido à pasteurização rápida e foi efetuada a esterilização industrial do leite por processo UAT direto, seguido do envase do produto. As análises no leite UAT foram realizadas nos dias 8, 30, 60, 90 e 120 após a fabricação. Quinze lotes de leite UAT foram avaliados para verificar a correlação entre a CCS e as variáveis estudadas. Adicionalmente, os lotes foram categorizados em grupos de baixa, intermediária e alta CCS. No leite cru, houve correlação negativa (r= -0,54, P=0,04) entre CCS e níveis de αS2 e ß-caseína. No leite UAT, o aumento da CCS foi acompanhado de uma diminuição dos níveis de αs1-caseína (r= -0,56, P=0,03) e ß-caseína (r= -0,56, P=0,03) aos 8 dias de armazenamento. No 60º dia após a fabricação a CCS foi correlacionada à diminuição de αS2-caseína (r= -0,69, P=0,005). A CCS exerceu efeito significativo (P=0,02) sobre o componente a*, com maiores (P=0,02) valores para o grupo de alta CCS em relação ao de CCS intermediária, e sobre a ß-caseína (P=0,02), com menores (P=0,03) concentrações no grupo de alta CCS comparado ao de CCS baixa. Os valores de pH, L*, proteína verdadeira e caseína apresentaram diminuição significativa ao longo do armazenamento. Contrariamente, a acidez, o componente a*, os ácidos graxos livres, a viscosidade e a proteólise aumentaram significativamente. A quantidade de caseína em relação à proteína verdadeira foi negativamente correlacionada à CCS no 120º dia de armazenamento. Observou-se redução significativa nas concentrações de frações protéicas durante o armazenamento, com exceção da αS2-caseína. A CCS no leite cru utilizado para a fabricação de leite UAT pode prejudicar a qualidade do produto por possíveis alterações decorrentes da proteólise no final da vida de prateleira. / The aim of this study was to evaluate the effects of somatic cell counts (SCC) on physicochemical characteristics (pH, acidity, composition, color), microbiology, proteolysis and lipolysis indexes, Maillard compound formation, apparent viscosity and protein fractions of whole milk obtained by direct ultra high temperature (UHT) process. The raw milk was analyzed for somatic cell (SC) levels and physico-chemical characteristics. Milk samples were collected in a dairy plant in São Paulo State. Raw milk was submitted to HT-ST pasteurization and industrial sterilization was taken by direct UHT process, followed by the packing. UHT milk analyses were conducted on days 8, 30, 60, 90 and 120 of storage. Fifteen batches of UHT milk were evaluated for correlations between SCC and the parameters studied. Additionally, UHT milks were categorized in selected groups of low, intermediate and high SCC. Raw milk presented a negative correlation (r= -0.54, P=0.04) between CSS and levels of αS2 and ß-casein. In UHT milk, SCC increase was followed by a reduction of αS1-casein (r= -0.56, P=0.03) and ß-casein (r= -0.56, P=0.03) concentrations on the 8th day of storage. On 60th day of storage, SCC was correlated with the reduction (r= -0.69, P=0.005) on αS2-casein concentration. SCC had significant effect on component a* (P=0.02), which values were higher (P=0.02) on high SCC group compared to the intermediate SCC group, and on ß-casein (P=0.02), with lower (P=0.03) concentrations in high SCC group than in low SCC group. Values of pH, L*, true protein and casein presented significant reduction during storage. Conversely, acidity, component a*, free fatty acids, viscosity and proteolysis increased significantly. Casein in relation to true protein was negatively correlated with SCC on 120th day of storage. It was observed a significant reduction on protein fractions concentrations in UHT milk during storage, except for αS2-casein. SCC in raw milk used for UHT milk production may prejudice the quality of the product through alterations caused by proteolysis in the end of shelf life.

O objetivo deste estudo foi avaliar os efeitos dos níveis de contagem de células somáticas (CCS) sobre as características físico-químicas e microbiológicas do leite semidesnatado e do creme de leite pasteurizados, ao longo de 28 e 30 dias de estocagem sob refrigeração. As frações protéicas do leite semidesnatado e o perfil de ácidos graxos do creme de leite pasteurizados foram pesquisados nos dias 1, 7, 14, 21 e 28 e 1, 10, 20 e 30 dias após a produção de cada produto. Foram definidos três grupos experimentais com baixa (< 100.000 cél.mL-1), intermediária (400.000 a 500.000 cél.mL-1) e alta (>750.000 cél.mL-1) CCS. O leite utilizado para compor cada grupo, em quatro repetições, foi coletado na granja leiteira da Fazenda da Aeronáutica, na cidade de Pirassununga-SP. O leite in natura de cada grupo foi submetido a desnate para obtenção do creme e do leite semidesnatado, tratados termicamente por pasteurização lenta e rápida, nesta sequência, seguido do envase asséptico em garrafas de polietileno de alta densidade (PEAD). No leite in natura, o percentual de gordura foi menor (P<0,05) no grupo com baixa CCS em relação aos demais grupos. Observou-se, ainda para o leite in natura, maior teor de proteínas (3,42%) no grupo de alta CCS. Para as frações protéicas do leite semidesnatado, não foram observadas diferenças significativas (P>0,05), com exceção da &beta;-caseína que apresentou maior concentração nos grupos de baixa e alta CCS (P<0,05). No grupo de CCS intermediária, o leite semidesnatado apresentou 8,98, 10,17 e 6,14 mg.mL-1 das frações &alpha;-caseína, &beta;-caseína e &kappa;-caseína, respectivamente. A participação da &alpha;S2-caseína, fração protéica verificada em menores proporções neste estudo, foi de 9,89; 10,59 e 14,97% em relação ao total de caseína, nos grupos de baixa, intermediária e alta CCS. Houve predominância dos ácidos palmítico (C16:0), esteárico (C18:0), oléico (C18:1) e mirístico (C14:0), em todos os grupos de contagem de células somáticas e nos tempos utilizados para avaliação do creme de leite. No entanto, a concentração de ácido oléico (C18:1) foi menor nos cremes com CCS intermediária e alta (P<0,05), em relação ao grupo de baixa CCS. Os ácidos saturados apresentaram variação de 53,9 a 58,1% em relação aos insaturados. Os ácidos de cadeia curta estiveram presentes no creme de leite em aproximadamente 8,15%, enquanto os de cadeia média e longa participam com cerca de 47 e 44%, respectivamente. A CCS influenciou na concentração de &beta;-caseína do leite semidesnatado, ao passo que, no creme de leite, a diminuição do teor de ácido oléico associada ao aumento da CCS indicou maior atividade lipolítica sobre os ácidos graxos de cadeia longa durante o período de 30 dias de estocagem. / The aim of this study was to evaluate the effects of somatic cell counts (SCC) levels on the physic-chemical and microbiological characteristics of both pasteurized semi-skimmed milk and milk cream, over 28 and 30 days of refrigerated storage. The semi-skimmed milk protein fractions were checked on days 1, 7, 14, 21 and 28, whereas the pasteurized milk cream fatty acids profile was checked on days 1, 10, 20 and 30 after each production. Three experimental groups were defined with low (<100.000 cel.mL-1), intermediate (400.000 to 500.000 cel.mL-1) and high (>750.000 cel.mL-1) SCC. The milk used in each group, with four repetitions, was collected in the Air Force dairy farm in Pirassununga, São Paulo state, Brazil. The raw milk in each group was skimmed to obtain the cream, thermally treated by slow pasteurization, and the semi-skimmed milk (fast pasteurization), followed by aseptic bottling in high density polyethylene bottles. In the raw milk, significant differences were verified by the Tukey test (P<0,05) for the parameter fat in the level of low SCC in relation to the other groups. It was also observed for the raw milk a higher rate of proteins (3,42%) in the group of high SCC, while the low count group presented 2,83% protein. For the protein fractions in the pasteurized semi-skimmed milk, with an exception of the &beta;-casein which presented statistical difference for the 5% significance level between the low and high SCC groups, no other significant differences were observed. In the intermediate SCC group the pasteurized semi-skimmed milk presented 8,98 mg.mL-1 for &alpha;-casein, 10,17 mg.mL-1 for &beta;-casein and 6,14 mg.mL-1 for &kappa;-casein fractions. The &alpha;s2-casein was the protein fraction with the lowest proportions in this study, with 9,89% (low SCC), 10,59% (intermediate SCC) and 14,97% (high SCC) in relation to the casein total. There was predominance of palmitic acids (C16:0), estearic (C18:0), oleic (C18:1) and miristic (C14:0) in all somatic cell counts groups and the times used to evaluate the pasteurized milk cream. However, lower levels of oleic acid (C18:1) were observed in creams with high and intermediate SCC, when compared with the lower SCC cream. The saturated acids presented variation of 53,9 to 58,1% in relation to the unsaturated ones. The short chain acids were found in the pasteurized milk cream with about 8,15%, while the ones with medium and long chains were 47% and 44% respectively. The SCC influenced the concentration of &beta;-casein in semiskimmed milk, while the decrease in oleic acid as observed in the cream samples containing higher SCC indicated higher lipolytic activity on the long-chain fatty acid in the product during 30 days of storage.

Milk is provided to new borns as their first food source and it contains essential nutrients, vitamins and other beneficial components, such as enzymes and antibodies that are required for rapid growth and development of the new born and for sustained growth over time. Milk contains two main types of proteins; casein proteins and whey proteins. Although casein proteins account for up to 80% of the proteins found in bovine milk, it is the whey protein that has become of high interest because of its bioactive content. Whey, a very watery mixture of lactose, proteins, minerals and trace amounts of fat, is formed from milk when the milk is coagulated and/or the casein proteins are removed from the milk. Bovine whey protein, including both the acidic and basic fractions (low and high isoelectric point, respectively), has previously been studied in vitro (cell based) and in vivo (using rats) for its impact on bone to determine if it can help improve bone mineral density and help reduce the risk of developing bone diseases, such as osteoporosis. Bone is constantly undergoing a remodelling process of being dissolved and reformed and the two main cell types responsible for this bone remodelling process are mature osteoclasts, which dissolve (resorb) bone, and osteoblasts, which reform the bone. Prior work has shown that acidic protein fractions derived from different sources of whey protein concentrate (WPC) have both in vivo and in vitro activity on bone, particularly anti-resorptive properties. However, the component(s) which confer activity have not yet been identified. In this thesis, work was undertaken to better understand the analytical composition of three types of WPC (cheese, mineral acid and lactic acid) and their associated acidic protein fractions and relate this to bone activity in the hope of identifying where the activity lies. Bone activity was assessed using in vitro screening with osteoblast cells (MC3T3-E1) and osteoclast cells (RAW 264.7). Comparison of the cell-based bone activity of the parent WPCs and corresponding acidic fractions indicated that the acidic fractions derived from both mineral acid and lactic WPC were superior in their ability to inhibit osteoclast development. Although compositional data was complex and definitive correlations with both bone bioactivities could not be made, it appeared that elements common to both the acidic fractions were a higher proportion of GLYCAM-1 and bone sialoprotein-1 (osteopontin). Further studies to more closely investigate the bone bioactivity of the acidic fractions are warranted.

Milk is provided to new borns as their first food source and it contains essential nutrients, vitamins and other beneficial components, such as enzymes and antibodies that are required for rapid growth and development of the new born and for sustained growth over time. Milk contains two main types of proteins; casein proteins and whey proteins. Although casein proteins account for up to 80% of the proteins found in bovine milk, it is the whey protein that has become of high interest because of its bioactive content. Whey, a very watery mixture of lactose, proteins, minerals and trace amounts of fat, is formed from milk when the milk is coagulated and/or the casein proteins are removed from the milk. Bovine whey protein, including both the acidic and basic fractions (low and high isoelectric point, respectively), has previously been studied in vitro (cell based) and in vivo (using rats) for its impact on bone to determine if it can help improve bone mineral density and help reduce the risk of developing bone diseases, such as osteoporosis. Bone is constantly undergoing a remodelling process of being dissolved and reformed and the two main cell types responsible for this bone remodelling process are mature osteoclasts, which dissolve (resorb) bone, and osteoblasts, which reform the bone. Prior work has shown that acidic protein fractions derived from different sources of whey protein concentrate (WPC) have both in vivo and in vitro activity on bone, particularly anti-resorptive properties. However, the component(s) which confer activity have not yet been identified. In this thesis, work was undertaken to better understand the analytical composition of three types of WPC (cheese, mineral acid and lactic acid) and their associated acidic protein fractions and relate this to bone activity in the hope of identifying where the activity lies. Bone activity was assessed using in vitro screening with osteoblast cells (MC3T3-E1) and osteoclast cells (RAW 264.7). Comparison of the cell-based bone activity of the parent WPCs and corresponding acidic fractions indicated that the acidic fractions derived from both mineral acid and lactic WPC were superior in their ability to inhibit osteoclast development. Although compositional data was complex and definitive correlations with both bone bioactivities could not be made, it appeared that elements common to both the acidic fractions were a higher proportion of GLYCAM-1 and bone sialoprotein-1 (osteopontin). Further studies to more closely investigate the bone bioactivity of the acidic fractions are warranted.