This study was conducted to develop and validate cleaning procedures for different processing equipment of varying complexity and to determine the efficacy of two different allergen tests. Following introduction of selected allergens to processing equipment, two treatments were applied - water wash or scrub/sanitize ? and a no clean was also evaluated. The equipment used consisted of a slicer, grinder, injector, vacuum tumbler, and plastic lugs. To introduce the allergen to the slicer, nine ready-to-eat hams were used. One hundred twenty-two kilograms of pork trim were ground, and a milk allergen was incorporated into the meat. The injector was contaminated with a food allergen by injecting boneless pork loins with a marinade containing soy flour. The slicer, grinder, injector, tumbler, and lugs were then subjected to randomized treatments. The results showed that the water wash and scrub/sanitize treatments did not differ significantly among the pieces of equipment tested. This study supported that both water wash and scrub/sanitize treatments can effectively removed allergens to a level below the industry threshold of 5 ppm.

Orientador: Flavia Maria Netto / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Engenharia de Alimentos / Made available in DSpace on 2018-08-24T21:18:27Z (GMT). No. of bitstreams: 1 Pessato_TassiaBatista_M.pdf: 1693373 bytes, checksum: c84093b37be90c1b31ce17c6722db6a1 (MD5) Previous issue date: 2014 / Resumo: O leite bovino é um dos alimentos mais alergênicos na primeira infância, sendo as caseínas e as duas principais proteínas do soro - ?-lactoalbumina (?-La) e ?-lactoglobulina (?-Lg) - os principais alérgenos. A hidrólise enzimática reduz a antigenicidade de proteínas e pode ser realizada com enzima livre ou imobilizada. A enzima imobilizada não requer inativação ao final da hidrólise, sendo retirada por filtração enquanto que a enzima livre precisa ser inativada, geralmente por aquecimento, para interromper a reação. Diferenças na atividade catalítica e de inativação podem acarretar em características diferentes dos hidrolisados formados. O objetivo desse trabalho foi avaliar os efeitos da hidrólise do isolado proteico de soro de leite bovino (IPS) com Alcalase livre (AL) e imobilizada em glioxil-agarose (AIm) nas características dos hidrolisados e na detecção de ?-La e ?-Lg por teste ELISA. As melhores condições de hidrólise do IPS com AL ou AIm foram estabelecidas por delineamento composto central rotacional (DCCR) 22. As variáveis independentes foram pH (7,0 a 9,0) e temperatura (45 a 65°C), a variável dependente foi grau de hidrólise (GH), determinado pelo método de pH-stat. Os hidrolisados foram caracterizados quanto ao perfil de hidrofilicidade por cromatografia líquida de alta eficiência de fase reversa (CLAE-FR) e, as análises posteriores, foram realizadas em hidrolisados tanto com AL (HAL) quanto com AIm (HAIm) obtidos em três condições de hidrólise. O perfil de massa molecular (MM) foi avaliado por cromatografia líquida de alta eficiência de exclusão molecular (CLAE-EM) e eletroforese (SDS-PAGE e SDS-PAGE/Tricina). A agregação dos hidrolisados foi estimada por turbidez e hidrofobicidade superficial (S0) e a detecção dos alérgenos foi realizada pelo método de Enzyme-Linked Immunosorbent Assay (ELISA) utilizando kits comerciais. Sob as mesmas condições de hidrólise, os valores de GH obtidos nos ensaios do DCCR com a AIm (9,5 a 22,2 %) foram, menores do que os obtidos com a AL (18 a 24 %), possivelmente em função de impedimentos estéricos resultantes da imobilização. As melhores condições de hidrólise do IPS com AIm, dentro das faixas de pH e temperatura estudadas, foram obtidas em temperaturas acima de 60°C. O DCCR com a AL não resultou em modelo devido à pequena variação dos resultados entre os ensaios. Os perfis cromatográficos (CLAE-FR) dos HAIm apresentaram mais picos na região de baixa hidrofilicidade do que os HAL. Os HAIm apresentaram peptídeos de MM maiores que os HAL, o que está relacionado aos menores GH produzidos com a AIm. Os HAL apresentaram menores valores de S0 que os HAIm, sugerindo a ocultação de sítios hidrofóbicos no interior dos agregados. As análises de turbidez em diferentes solventes mostraram que interações hidrofóbicas, pontes de hidrogênio e pontes dissulfeto são as principais forças envolvidas na formação dos agregados. A hidrólise com AL e AIm diminuiu significativamente a detecção das proteínas alergênicas. A detecção dessas proteínas foi maior nos HAIm devido, possivelmente, aos menores GH desses hidrolisados. Porém, os resultados sugerem que as características dos hidrolisados resultantes das condições de hidrólise e a formação de agregados estabilizados por diferentes interações também tiveram efeito na detecção dos alérgenos / Abstract: Cow's milk is one of the most allergenic foods in infancy, with the two major caseins and whey proteins - ?-lactalbumin (?-La) and ?-lactoglobulin (?-Lg) - the main allergens. Enzymatic hydrolysis considerably reduces the antigenicity of proteins, and can be performed with free or immobilized enzyme. The immobilized enzyme does not require inactivation at the end of hydrolysis and can be removed by filtration, while the free enzyme must be inactivated, usually by heating, to stop the reaction. Differences in catalytic activity and inactivation may result in hydrolysates with distinct characteristics. The objective of this study was to evaluate the effects of hydrolysis of whey protein isolate (WPI) with free Alcalase (AL) and Alcalase immobilized on glyoxyl agarose (Alm) on the characteristics of hydrolysates and detection of ?-La e ?-Lg by ELISA test. The best conditions for the hydrolysis of WPI using AL or Alm were established by 22 central composite rotational design (CCRD). The independent variables were pH (7.0 and 9.0) and temperature (45 to 65 °C), whereas the dependent variable was the degree of hydrolysis (DH) determined by the pH-stat method. All hydrolysates were characterized for the hydrophilicity profile by reverse phase high performance liquid chromatography (RP-HPLC), and the subsequent analyses were performed in both hydrolysates by AL (HAL) and Alm (HAlm) from three conditions of hydrolysis. The molecular weight profile (MW) was assessed by size exclusion high performance liquid chromatography (SE-HPLC) and electrophoresis (SDS-PAGE and SDS-PAGE/Tricine). The aggregation of hydrolysates was estimated by turbidity and surface hydrophobicity (S0) and the allergens ?-La and ?-Lg in the hydrolysates were detected by the Enzyme-Linked Immunosorbent Assay (ELISA) method using commercial kits. Under the same hydrolysis conditions, the DH values obtained with the Alm as defined by CCRD (9.5 to 22.2%) were lower than the values found for AL (18 to 24%), possibly due to the steric hindrance resulting from immobilization. The best conditions for hydrolysis of WPI by Alm within the pH and temperature ranges of this study were found at temperatures above 60 °C. The CCRD with the AL did not provide a model due to the little variation between trials. The chromatographic profiles (RP-HPLC) of HAlm exhibited more peaks in the hydrophilic low-complexity region than HAL. HAlm showed peptides with higher MW than HAL, which is related to the lower DH produced with Alm. The HAL had lower S0 values than HAlm, suggesting the hiding of hydrophobic sites in the interior part of the aggregates. The turbidity analyses showed that hydrophobic interactions, hydrogen bonds, and disulfide bonds were the main forces involved in the formation of aggregates. The hydrolysis with AL and Alm significantly decreased the detection of the allergenic proteins, which was higher in HAlm probably due to the lower DH of these hydrolysates. However, the results suggested that the characteristics of the hydrolysates resulting from the conditions of hydrolysis, and the formation of aggregates stabilized by different interactions also had an effect on the detection of allergens / Mestrado / Nutrição Experimental e de Alimentos / Mestra em Alimentos e Nutrição