ES2533469T3 - Método de preparación de un producto tratado con calor - Google Patents

Método de preparación de un producto tratado con calor Download PDF

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Publication number
ES2533469T3
ES2533469T3 ES07115541.0T ES07115541T ES2533469T3 ES 2533469 T3 ES2533469 T3 ES 2533469T3 ES 07115541 T ES07115541 T ES 07115541T ES 2533469 T3 ES2533469 T3 ES 2533469T3
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Prior art keywords
preparing
treated product
heat treated
aspergillus
asparaginase
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Gitte Budolfsen
Morten Tovborg Jensen
Hans Peter Heldt-Hansen
Mary Ann Stringer
Lene Lange
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Novozymes AS
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Novozymes AS
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    • A—HUMAN NECESSITIES
    • A21—BAKING; EDIBLE DOUGHS
    • A21D—TREATMENT OF FLOUR OR DOUGH FOR BAKING, e.g. BY ADDITION OF MATERIALS; BAKING; BAKERY PRODUCTS
    • A21D8/00—Methods for preparing or baking dough
    • A21D8/02—Methods for preparing dough; Treating dough prior to baking
    • A21D8/04—Methods for preparing dough; Treating dough prior to baking treating dough with microorganisms or enzymes
    • A21D8/042—Methods for preparing dough; Treating dough prior to baking treating dough with microorganisms or enzymes with enzymes
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • A23L19/00—Products from fruits or vegetables; Preparation or treatment thereof
    • A23L19/10—Products from fruits or vegetables; Preparation or treatment thereof of tuberous or like starch containing root crops
    • A23L19/12—Products from fruits or vegetables; Preparation or treatment thereof of tuberous or like starch containing root crops of potatoes
    • A23L19/18—Roasted or fried products, e.g. snacks or chips
    • A23L19/19—Roasted or fried products, e.g. snacks or chips from powdered or mashed potato products
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • A23L5/00—Preparation or treatment of foods or foodstuffs, in general; Food or foodstuffs obtained thereby; Materials therefor
    • A23L5/20—Removal of unwanted matter, e.g. deodorisation or detoxification
    • A23L5/25—Removal of unwanted matter, e.g. deodorisation or detoxification using enzymes
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • A23L7/00—Cereal-derived products; Malt products; Preparation or treatment thereof
    • A23L7/10—Cereal-derived products
    • A23L7/117—Flakes or other shapes of ready-to-eat type; Semi-finished or partly-finished products therefor
    • C—CHEMISTRY; METALLURGY
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    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14—Hydrolases (3)
    • C12N9/78—Hydrolases (3) acting on carbon to nitrogen bonds other than peptide bonds (3.5)
    • C12N9/80—Hydrolases (3) acting on carbon to nitrogen bonds other than peptide bonds (3.5) acting on amide bonds in linear amides (3.5.1)
    • C12N9/82—Asparaginase (3.5.1.1)
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    • C12P1/00—Preparation of compounds or compositions, not provided for in groups C12P3/00 - C12P39/00, by using microorganisms or enzymes
    • C—CHEMISTRY; METALLURGY
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    • C12Y101/00—Oxidoreductases acting on the CH-OH group of donors (1.1)
    • C12Y101/01—Oxidoreductases acting on the CH-OH group of donors (1.1) with NAD+ or NADP+ as acceptor (1.1.1)
    • C12Y101/01118—Glucose 1-dehydrogenase (NAD+) (1.1.1.118)
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    • C12Y101/01—Oxidoreductases acting on the CH-OH group of donors (1.1) with NAD+ or NADP+ as acceptor (1.1.1)
    • C12Y101/01119—Glucose 1-dehydrogenase (NADP+) (1.1.1.119)
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    • C12Y101/01—Oxidoreductases acting on the CH-OH group of donors (1.1) with NAD+ or NADP+ as acceptor (1.1.1)
    • C12Y101/01124—Fructose 5-dehydrogenase (NADP+) (1.1.1.124)
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    • C12Y101/00—Oxidoreductases acting on the CH-OH group of donors (1.1)
    • C12Y101/01—Oxidoreductases acting on the CH-OH group of donors (1.1) with NAD+ or NADP+ as acceptor (1.1.1)
    • C12Y101/01147—16-Alpha-hydroxysteroid dehydrogenase (1.1.1.147)
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    • C12Y101/01—Oxidoreductases acting on the CH-OH group of donors (1.1) with NAD+ or NADP+ as acceptor (1.1.1)
    • C12Y101/01148—Estradiol 17alpha-dehydrogenase (1.1.1.148)
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    • C12Y101/00—Oxidoreductases acting on the CH-OH group of donors (1.1)
    • C12Y101/03—Oxidoreductases acting on the CH-OH group of donors (1.1) with a oxygen as acceptor (1.1.3)
    • C12Y101/03009—Galactose oxidase (1.1.3.9)
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    • C12Y101/99—Oxidoreductases acting on the CH-OH group of donors (1.1) with other acceptors (1.1.99)
    • C12Y101/9901—Glucose dehydrogenase (acceptor) (1.1.99.10)
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    • C12Y101/99011—Fructose 5-dehydrogenase (1.1.99.11)
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    • C12Y101/99—Oxidoreductases acting on the CH-OH group of donors (1.1) with other acceptors (1.1.99)
    • C12Y101/99018—Cellobiose oxidase (1.1.99.18)
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    • C12Y101/00—Oxidoreductases acting on the CH-OH group of donors (1.1)
    • C12Y101/99—Oxidoreductases acting on the CH-OH group of donors (1.1) with other acceptors (1.1.99)
    • C12Y101/99028—Glucose-fructose oxidoreductase (1.1.99.28)
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    • C12Y102/00—Oxidoreductases acting on the aldehyde or oxo group of donors (1.2)
    • C12Y102/01—Oxidoreductases acting on the aldehyde or oxo group of donors (1.2) with NAD+ or NADP+ as acceptor (1.2.1)
    • C12Y102/01003—Aldehyde dehydrogenase (NAD+) (1.2.1.3)
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    • C12Y102/01005—Aldehyde dehydrogenase [NAD(P)+] (1.2.1.5)
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    • C12Y104/03—Oxidoreductases acting on the CH-NH2 group of donors (1.4) with oxygen as acceptor (1.4.3)
    • C12Y104/03002—L-Amino-acid oxidase (1.4.3.2)
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    • C12Y110/00—Oxidoreductases acting on diphenols and related substances as donors (1.10)
    • C12Y110/03—Oxidoreductases acting on diphenols and related substances as donors (1.10) with an oxygen as acceptor (1.10.3)
    • C12Y110/03002—Laccase (1.10.3.2)
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    • C12Y111/00—Oxidoreductases acting on a peroxide as acceptor (1.11)
    • C12Y111/01—Peroxidases (1.11.1)
    • C12Y111/01007—Peroxidase (1.11.1.7), i.e. horseradish-peroxidase
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    • C12Y301/00—Hydrolases acting on ester bonds (3.1)
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    • C12Y304/13—Dipeptidases (3.4.13)
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    • C12Y305/01026—N4-(Beta-N-acetylglucosaminyl)-L-asparaginase (3.5.1.26)
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    • C12Y305/01052—Peptide-N4-(N-acetyl-beta-glucosaminyl)asparagine amidase (3.5.1.52), i.e. glycopeptidase

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Abstract

Método para preparar un producto tratado con calor, que comprende las fases secuenciales de: a) proveer una materia prima que comprende hidrato de carbono, proteína y agua b) tratar la materia prima con una asparaginasa y c) tratar con calor hasta alcanzar un contenido de agua final por debajo del 35% en peso, donde el tratamiento con calor implica fritura a temperaturas de 150-180ºC, horneado en aire caliente a 160-310ºC durante 2-10 minutos, calentamiento en placa caliente y/o secado de la malta verde.

Description

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E07115541
23-03-2015
Soluciones concentradas
[0032] 50 mM tampón Tris, pH 8.6
189mM Solución de L-Asparraguina
1,5 M Ácido tricloroacético (ATC)
Reactivo de Nessler, Aldrich Stock No. 34,514-8 (Sigma-Aldrich, St. Louis, Mo. USA)
Asparaginasa, Sigma Stock No. A4887 (Sigma-Aldrich, St. Louis, Mo. USA)
Ensayo
Reacción enzimática:
[0033]
500 micro-I tampón
100 micro-I Solución de L-asparraguina
350 micro-I agua
son mezclados y equilibrados a 37ºC.
se añaden 100 micro-I de solución enzimática y las reacciones son incubadas a 37ºC durante 30 minutos.
[0034] Las reacciones son detenidas colocándolas en hielo y añadiendo 50 micro-I de 1.5M ATC.
[0035] Las muestras son mezcladas y centrifugadas durante 2 minutos a 20.000 g
Medición de amonio libre:
[0036] Se mezclan 50 micro-I de la reacción enzimática con 100 micro-I de agua y 50 micro-I de reactivo de Nessler. La reacción es mezclada y la absorbencia es medida a 436nm después de 1 minuto.
Estándar:
[0037] El concentrado de asparaginasa (Sigma A4887) es diluido 0,2, 0,5, 1, 1,5, 2, y 2,5 U/ml.
Ejemplo 1: expresión de una asparaginasa de Aspergillus oryzae en Aspergillus oryzae
[0038] Se generaron bibliotecas de ADNc de ARNm de Aspergillus oryzae, se ordenaron y almacenaron en una base de datos de ordenador como se describe en WO 00/56762.
[0039] Se comparó la secuencia péptida de asparaginasa de Saccharomyces cerevisiae (Kim,K.-W.; Kamerud, J.Q.; Livingston,D.M.; Roon,R.J., (1988) Asparaginase II of Saccharomyces cerevisiae. Characterization of the ASP3 gene. J. Biol. Chem. 263:11948), con traducciones de las secuencias de ADNc parcial de Aspergillus oryzae usando el programa TFASTXY, versión 3.2t07 (Pearson et al. Genomics (1997) 46:24-36). Se identificó una secuencia de A. oryzae traducida como teniendo una identidad del 52% para asparaginasa II de levadura a través de un recubrimiento de 165 aminoácidos. Se determinó la secuencia completa del inserto de ADNc del clon correspondiente (depositado como DSM 15960) y se presentó como SEC ID NO: 1, y el péptido traducido de esta secuencia, AoASP, es presentado como SEC ID NO: 2. Esta secuencia fue usada para designar cebadores para la amplificación PCR del gen de codificación AoASP de DSM 15960, con sitios de restricción apropiados añadidos a las extremidades del cebador para facilitar la subclonación del producto PCR (cebadores AoASP7 y AoASP8, SEC ID Nos: 14 y 15). La amplificación PCR fue realizada usando Extensor Hi-Fidelity PCR Master Mix (ABgene, Surrey, U.K.) siguiendo las instrucciones del fabricante y usando una temperatura de hibridación de 55ºC para los primeros 5 ciclos y 65ºC para un ciclo adicional y un tiempo de extensión de 1,5 minutos.
[0040] El fragmento de PCR fue restringido con BamHl y Hindlll y clonado en el vector de expresión Aspergillus pMStr57 usando técnicas estándares. El vector de expresión pMStr57 contiene los mismos elementos que pCaHj483 (WO 98/00529), con modificaciones menores hechas en el promotor NA2 de Aspergillus como se describe para el vector pMT2188 en WO 01/12794, y tiene secuencias para la selección y propagación en E. coli, y selección y expresión en Aspergillus. Específicamente, la selección en Aspergillus es facilitada por el gen amdS de Aspergillus nidulans, que permite el uso de acetamida como única fuente de nitrógeno. La expresión en Aspergillus es mediada por un promotor (NA2) de amilasa II neutra modificada de Aspergillus niger que es fusionado en la secuencia líder 5' del gen de codificación de triosa fosfato isomerasa (tpi) de Aspergillus nidulans, y el terminador del gen de codificación de amiloglucosidasa de Aspergillus niger. El gen de codificación de asparaginasa del constructo de expresión de Aspergillus resultante, pMStr90, fue secuenciado y la secuencia coincidía completamente con aquella determinada previamente para el inserto de DSM 15960.
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  1. imagen1
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EP1553848B1 (en) 2007-09-05
DE60316168D1 (de) 2007-10-18
US20060275879A1 (en) 2006-12-07
EP1886582A2 (en) 2008-02-13
ES2293012T3 (es) 2008-03-16
WO2004032648A1 (en) 2004-04-22
EP2294928A1 (en) 2011-03-16
CA2501741C (en) 2012-04-17
US8507246B2 (en) 2013-08-13
DK1886582T3 (en) 2015-04-20
EP1886582A3 (en) 2010-07-14
DE60316168T2 (de) 2008-05-29
US20130236598A1 (en) 2013-09-12
US7396670B2 (en) 2008-07-08
EP1553848A1 (en) 2005-07-20
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US8859023B2 (en) 2014-10-14
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US20080096260A1 (en) 2008-04-24
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US20120114793A1 (en) 2012-05-10
US20080095883A1 (en) 2008-04-24
DK2294928T3 (da) 2014-11-10
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ATE372059T1 (de) 2007-09-15
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