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Caracterización Tecnológica, Química, Bioquímica Y Sensorial Del Queso De Valdeón Con I.G.P

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Caracterización Tecnológica, Química, Bioquímica Y Sensorial Del Queso De Valdeón Con I.G.P Caracterización tecnológica, química, bioquímica y sensorial del queso de Valdeón con I.G.P. durante la maduración Technological, chemical, biochemical and sensory characterization of Valdeón cheese with P.G.I. during ripening Isabel Diezhandino Hernández 2015 La autora de esta Memoria ha sido beneficiaria de una ayuda destinada a financiar la contratación predoctoral de personal investigador de reciente titulación (beca PIRTU) concedida por la Consejería de Educación de la Junta de Castilla y León y cofinanciada por el Fondo Social Europeo (ORDEN EDU/1064/2009 de 14 de mayo (BOCYL nº 93 de 20 de agosto de 2009)). Las investigaciones de esta Tesis han sido desarrolladas en la Universidad de León dentro del proyecto L021A12-2 financiado por la Consejería de Educación de la Junta de Castilla y León (ORDEN EDU/671/2012 de 8 de Agosto (BOCYL nº 159 de 20 de agosto de 2012)). “La posibilidad de realizar un sueño es lo que hace que la vida sea interesante” Paulo Coehlo “Algunas personas quieren que algo ocurra, otras sueñan con que pasará, otras hacen que suceda” Michael Jordan Agradecimientos Esta Tesis ha sido posible gracias a la colaboración, ayuda y trabajo de muchas personas a las que estaré siempre agradecida. Al Dr. Fresno, Dr. Prieto y Dr. Arenas, directores de esta Tesis, por depositar su confianza en mí, por su dedicación, ayuda y paciencia. A la Quesería Picos de Europa S.L. por su colaboración en este trabajo realizando la fabricación de los quesos. Al resto de los profesores del Área de Tecnología de los Alimentos por sus enseñanzas y valiosos consejos. A Domingo por ser mi guía durante esta aventura. Por todas sus enseñanzas en el laboratorio, por las horas de trabajo juntos, por su compañía durante los viajes a la quesería, por su amistad. Gracias a ti todo ha sido más sencillo. A Leticia por su disposición. Por su ayuda en el laboratorio de microbiología. Por su inestimable apoyo. Por ser para mi muchísimo más que la Técnico de Laboratorio. A Avelino por acogerme durante mi estancia en Cork y formar una pequeña familia durante esos maravillosos tres meses. Al resto de compañeros del Área de Tecnología de los Alimentos, por hacer que ir a trabajar nunca supusiera un esfuerzo. Por los grandes momentos que hemos pasado, por el rato de café a media mañana, por esa pequeñita familia que hemos formado en torno a una mesa en la hora de la comida, por las cenas, por las risas, por todo. Y en especial a Noelia (por su ayuda y colaboración, por ser un apoyo importantísimo para mí durante la Tesis), a Erica (por su gran ayuda durante la etapa final de este trabajo), a Dolores (por las conversaciones juntas), a Ana (por esos momentos en los cursos de Doctorado) y a Patri (por esos primeros meses de laboratorio con nuestras amigas las aminas y por las horas de despacho juntas). To Dr. McSweeney for his supervision and advices and all the members of the Laboratory of School of Food and Nutritional Science of the University of Cork. A la Dra. Recio por admitirme en su equipo y formar parte de mi formación. Y a todo su equipo del Departamento de Bioactividad y Análisis de los Alimentos (Instituto de Investigación en Ciencias de la Alimentación, CSIC, Madrid). A la Dra. Costell por permitirme formar parte de su equipo en el Instituto de Agroquímica y Tecnología de los Alimentos (CSIC, Valencia) y a la Dra. Bayarri por su supervisión y consejos. A mis padres, Antonio y Mª Asunción, y a mi hermana Beatriz, por confiar siempre en mí y estar siempre a mi lado aún en la distancia. A mis amigas y amigos porque sin todos los momentos que he pasado a su lado esta Tesis tampoco hubiera sido posible. A Jose, por todo. Por su compresión, por saber entenderme (aún cuando ni yo misma era capaz de hacerlo), por su cariño, su apoyo y confianza. Gracias. ÍNDICE Resumen /Summary ……………………………………………………………………………. 1 1. Introducción ………………………………………………………………………………… 11 1.1. El queso azul ……………………………………………………………………… 13 1.1.1. Características químicas y fisico-químicas …………………………... 13 1.1.2. Microbiología del queso azul ………………………………………… 16 1.1.3. Metabolismo de la lactosa en los quesos azules ……………………... 19 1.1.4. Proteólisis en los quesos azules ……………………………………... 20 1.1.5. Péptidos bioactivos …………………………………………………... 26 1.1.6. Lipólisis en los quesos azules ………………………………………... 29 1.1.7. Compuestos volátiles en los quesos azules …………………………... 30 1.1.8. Propiedades sensoriales en los quesos azules ………………………... 35 1.2. El queso de Valdeón ……………………………………………………………… 39 1.3. Referencias ………………………………………………………………………... 41 2. Justificación y Objetivos …………………………………………………………………… 49 3. Material y Métodos …………………………………………………………………………. 53 3.1. Elaboración del queso …………………………………………………………….. 55 3.2. Análisis microbiológicos …………………………………………………………. 57 3.3. Análisis químicos y fisico-químicos ……………………………………………… 58 3.4. Determinación de los principales índices de maduración del queso ……………… 58 3.4.1. Estudio de la glucólisis ………………………………………………. 58 3.4.2. Estudio de la proteólisis ……………………………………………… 58 3.4.2.1. Fracciones nitrogenadas ………………………………….. 58 3.4.2.2. Electroforesis de las caseínas …………………………….. 59 3.4.2.3. Análisis del perfil de péptidos ……………………………. 59 3.4.2.4. Análisis de los aminoácidos ……………………………… 60 3.4.2.5. Análisis de la concentración de aminas biógenas ………... 61 3.4.2.6. Análisis de péptidos con actividad biológica …………….. 63 3.4.2.7. Análisis de la actividad de la plasmina …………………... 65 3.4.3. Estudio de la lipólisis ………………………………………………… 65 3.4.3.1. Análisis de ácidos grasos libres ………………………….. 65 3.5. Estudio de las propiedades sensoriales …………………………………………… 67 3.5.1. Análisis sensorial mediante panel de catadores ……………………… 67 3.5.2. Análisis del perfil de textura …………………………………………. 68 3.5.3. Estudio reológico del queso ………………………………………….. 68 3.5.4. Determinación del color ……………………………………………… 69 3.6. Análisis estadístico ………………………………………………………………... 69 3.7. Referencias …………………………………………………………………........... 70 4. Resultados …………………………………………………………………………………... 73 Microbiological, physic-chemical and proteolytic changes in a Spanich blue cheese during ripening (Valdeón cheese) …………………………………………………………………. 75 Rheological, textural, colour and sensory characteristics of a Spanish blue cheese (Valdeón cheese) ……………………………………………………………….................... 85 Microbiological, chemical, textural and sensory changes in a Spanish blue cheese (Valdeón cheese) made at different seasons. 95 Microbiological, chemical, textural and sensory changes in a Spanish blue cheese (Valdeón cheese) made from raw or pasteurized milk ……………………………………. 115 Changes in free fatty acid profile during ripening of a Spanish blue veined cheese (Valdeón) made in different seasons and from pasteurized and raw milk ………………. 135 Evolution of free amino acids and biogenic amines contents during ripening of a Spanish blue cheese (Valdeón cheese) ……………………………………………………………… 157 Peptidomic study of Spanish blue cheese (Valdeón) and changes after simulated gastrointestinal digestion ………………………………………...………………………… 179 5. Discusión General …………………………………………………………………………… 191 5.1. Influencia del tiempo de maduración en las características de la IGP queso de Valdeón ……………………………………………………………………………. 193 5.2. Influencia de la estación de elaboración en las características de la IGP queso de Valdeón ……………………………………………………………………………. 200 5.3. Influencia del tratamiento térmico aplicado a la leche en las características de la IGP queso de Valdeón …………………………………………………………….. 204 5.4. Presencia de péptidos bioactivos en la IGP queso de Valdeón antes y tras simulación de digestión gastrointestinal …………………………………………… 208 5.5. Referencias …………………………………………………………………………. 210 6. Conclusiones/Conclusions…………………………………………………………………… 217 RESUMEN / SUMMARY Resumen / Summary RESUMEN Durante los últimos años, en España se han llevado a cabo varios estudios de tipificación de quesos que abarcan la práctica totalidad de Denominaciones de Origen Protegidas (D.O.P.) e Indicaciones Geográficas Protegidas (I.G.P.) existentes. Sin embargo, existen algunas variedades de las que sólo se conocen algunos datos, siendo este el caso de la I.G.P. queso de Valdeón. Esta Memoria de Tesis Doctoral surge con el objetivo principal de caracterizar la I.G.P. Queso de Valdeón estudiándose la influencia que tiene el tiempo de maduración, la estación de elaboración y el tratamiento térmico aplicado a la leche en la calidad final de esta variedad. Para ello, se elaboraron un total de 12 lotes, 8 de ellos a partir de leche pasterizada (2 en cada una de las estaciones del año) y 4 a partir de leche cruda en la Quesería Picos de Europa S.L. (Posada de Valdeón, León, España). El estudio de la primera parte del objetivo principal, es decir, de la influencia del tiempo de maduración se abordó en cuatro de los artículos de la presente Tesis Doctoral. En primer lugar, se observó que las bacterias acido lácticas, especialmente lactococci, fueron la microbiota predominante durante las primeras etapas de la maduración, siendo gradualmente reemplazadas por los mohos y levaduras. Los recuentos de enterococci y Enterobacteriaceae fueron muy bajos o, incluso, no determinados. Esta variedad se caracterizó por un contenido en sólidos totales de 61,80 g/100 g de queso, un ratio sal/humedad de 8,92 g/100 g de humedad, un pH de 6,4-7,6 y una actividad de agua de 0,917. Al final de la maduración, la proteólisis primaria y secundaria fueron muy elevadas lo que dio lugar a una degradación casi total tanto de la αs1- como de las β- caseínas (aproximadamente el 90%). Asimismo, como consecuencia
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