Publication: Efecto del procesado térmico, de presión y enzimático sobre alérgenos de frutos secos y su detección por PCR en tiempo real
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2018-12-21
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Universidad Complutense de Madrid
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Abstract
Los frutos secos son uno de los alimentos más frecuentemente implicados en reacciones alérgicas. El procesado térmico y no térmico de los alimentos puede producir cambios estructurales en las proteínas alergénicas modulando su capacidad antigénica. En esta Tesis Doctoral, titulada “Efecto del procesado térmico, de presión y enzimático sobre alérgenos de frutos secos y su detección por PCR en tiempo real”, se ha analizado el impacto del procesado sobre la inmunoreactividad de pistacho y anacardo. Los tratamientos utilizados han sido térmicos, de cocción (30 y 60 min) o con aplicación de presión (autoclave a 121⁰C/1.18 atm o138⁰C/2.56 atm durante 15 y 30 min), así como la combinación de éstos con hidrólisis enzimáticacon proteasas y sonicación. Se han empleado técnicas de inmunodetección in vitro, LC-MS/MS,pruebas cutáneas en pacientes alérgicos y ensayos de liberación de mediadores de la respuesta alérgica.La cocción no reduce significativamente la capacidad de unir IgE de pistacho y anacardo con respecto al control sin tratar , aunque sí disminuye la capacidad de inducir degranulación de basófilos. Al aplicar calor y presión, especialmente a 138⁰C/2.56 atm 30 minutos, la capacidad de anacardo y pistacho de unir IgE tanto in vitro como en la superficie de células efectoras disminuye considerablemente. La digestión enzimática desencadena la proteólisis de alérgenos de anacardo y en especial de pistacho, reduciendo su reactividad frente a anticuerpos IgE. La combinación de tratamientos térmicos con hidrólisis enzimática representa una eficiente estrategia para reducir la inmunoreactividad de estos frutos. Se han identificado péptidos de algunos alérgenos de anacardo y pistacho que resisten incluso al tratamiento combinado de calor, presión y digestión enzimática...
Tree nuts are one of the ingredients most frequently involved in allergic reactions.Thermal and non-thermal food processing can produce structural changes in allergenic proteins,modifying their antigenic capacity. In this Doctoral Thesis, entitled "Effect of thermal, pressure and enzymatic processing on tree nut allergens and their detection by real-time PCR", the impact of processing on pistachio and cashew immunoreactivity has been analysed. Thermal treatments were used, boiling (30 and 60 min) or heat and pressure (autoclave at 121⁰C / 1.18 atm or 138⁰C / 2.56 atm for 15 and 30 min), as well as the combination of these with enzymatichydrolysis using proteases under sonication. In vitro immunodetection techniques, LC-MS / MS,skin prick tests in allergic patients and release assays of allergic response mediators have been used. Boiling does not significantly reduce the IgE-binding capacity of pistachio and cashew compared to untreated control although it decreases the ability to induce degranulation of basophils. When heat and pressure is applied, especially at 138⁰C / 2.56 atm 30 minutes, the ability of cashew and pistachio to bind IgE, both in vitro and on the surface of effector cells, decreases considerably. The enzymatic digestion triggers the proteolysis of cashew allergensand especially of pistachio, reducing its reactivity against IgE antibodies. The combination of thermal treatments with enzymatic hydrolysis represents an efficient strategy to reduce the immunoreactivity of these tree nuts. Some peptides from cashew and pistachio allergenic proteins have been identified as resistant even after the combined treatment of heat, pressureand enzymatic digestion...
Tree nuts are one of the ingredients most frequently involved in allergic reactions.Thermal and non-thermal food processing can produce structural changes in allergenic proteins,modifying their antigenic capacity. In this Doctoral Thesis, entitled "Effect of thermal, pressure and enzymatic processing on tree nut allergens and their detection by real-time PCR", the impact of processing on pistachio and cashew immunoreactivity has been analysed. Thermal treatments were used, boiling (30 and 60 min) or heat and pressure (autoclave at 121⁰C / 1.18 atm or 138⁰C / 2.56 atm for 15 and 30 min), as well as the combination of these with enzymatichydrolysis using proteases under sonication. In vitro immunodetection techniques, LC-MS / MS,skin prick tests in allergic patients and release assays of allergic response mediators have been used. Boiling does not significantly reduce the IgE-binding capacity of pistachio and cashew compared to untreated control although it decreases the ability to induce degranulation of basophils. When heat and pressure is applied, especially at 138⁰C / 2.56 atm 30 minutes, the ability of cashew and pistachio to bind IgE, both in vitro and on the surface of effector cells, decreases considerably. The enzymatic digestion triggers the proteolysis of cashew allergensand especially of pistachio, reducing its reactivity against IgE antibodies. The combination of thermal treatments with enzymatic hydrolysis represents an efficient strategy to reduce the immunoreactivity of these tree nuts. Some peptides from cashew and pistachio allergenic proteins have been identified as resistant even after the combined treatment of heat, pressureand enzymatic digestion...
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Tesis de la Universidad Complutense de Madrid, Facultad de Ciencias Biológicas, Departamento de Bioquímica y Biología Molecular, leída el 05/11/2018. Tesis formato europeo (compendio de artículos)