ES2334590T3 - Alfa-amilasa mutante, detergente y agente de lavado de vajilla. - Google Patents
Alfa-amilasa mutante, detergente y agente de lavado de vajilla. Download PDFInfo
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- ES2334590T3 ES2334590T3 ES93914652T ES93914652T ES2334590T3 ES 2334590 T3 ES2334590 T3 ES 2334590T3 ES 93914652 T ES93914652 T ES 93914652T ES 93914652 T ES93914652 T ES 93914652T ES 2334590 T3 ES2334590 T3 ES 2334590T3
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- amylase
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- 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)
- C12N9/24—Hydrolases (3) acting on glycosyl compounds (3.2)
- C12N9/2402—Hydrolases (3) acting on glycosyl compounds (3.2) hydrolysing O- and S- glycosyl compounds (3.2.1)
- C12N9/2405—Glucanases
- C12N9/2408—Glucanases acting on alpha -1,4-glucosidic bonds
- C12N9/2411—Amylases
- C12N9/2414—Alpha-amylase (3.2.1.1.)
- C12N9/2417—Alpha-amylase (3.2.1.1.) from microbiological source
-
- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11D—DETERGENT COMPOSITIONS; USE OF SINGLE SUBSTANCES AS DETERGENTS; SOAP OR SOAP-MAKING; RESIN SOAPS; RECOVERY OF GLYCEROL
- C11D3/00—Other compounding ingredients of detergent compositions covered in group C11D1/00
- C11D3/16—Organic compounds
- C11D3/38—Products with no well-defined composition, e.g. natural products
- C11D3/386—Preparations containing enzymes, e.g. protease or amylase
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- 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)
- C12N9/24—Hydrolases (3) acting on glycosyl compounds (3.2)
- C12N9/2402—Hydrolases (3) acting on glycosyl compounds (3.2) hydrolysing O- and S- glycosyl compounds (3.2.1)
- C12N9/2405—Glucanases
- C12N9/2408—Glucanases acting on alpha -1,4-glucosidic bonds
- C12N9/2411—Amylases
- C12N9/2414—Alpha-amylase (3.2.1.1.)
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- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
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- Zoology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Genetics & Genomics (AREA)
- Molecular Biology (AREA)
- Biotechnology (AREA)
- Biomedical Technology (AREA)
- Medicinal Chemistry (AREA)
- Microbiology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Oil, Petroleum & Natural Gas (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Detergent Compositions (AREA)
- Enzymes And Modification Thereof (AREA)
Abstract
EN LA (ALFA)-AMILASA MUTANTE UNO O MAS DE LOS RESIDUOS DE AMINO ACIDO DE METIONINA ES INTERCAMBIADO CON CUALQUIER RESIDUO AMINO ACIDO EXCEPTO POR CYS Y MET. LA (ALFA)-AMILASA MUTANTE ASI PRODUCIDA EXHIBE UNA ESTABILIDAD MEJORADA EN PRESENCIA DE AGENTES DE OXIDACION Y UNA TERMOACTIVIDAD MEJORADA CON VALORES MODERADAMENTE BAJOS DE PH. LA (ALFA)-AMILASA MUTANTE ESTA BIEN INDICADA COMO UN ADITIVO PARA DETERGENTES, AGENTES DE LAVADO DE VAJILLA Y AGENTES DE LICUEFACION.
Description
\alpha-amilasa mutante,
detergente y agente de lavado de vajilla.
La invención comprende una
\alpha-amilasa mutante, un detergente y un agente
de lavado de vajilla.
Las amilasas mutantes con estabilidad a la
oxidación mejorada, donde una o más metioninas han mutado en
cisteínas o en cisteínas químicamente modificadas están descritas
en WO 91/16423. La razón para la investigación en este campo es la
necesidad de \alpha-amilasas de oxidación estable
como aditivos para detergentes y agentes para el lavado de vajilla,
debido a la presencia de agentes oxidantes fuertes en los
detergentes y en los agentes para el lavado de vajilla, como se
explica con más detalle en WO 91/16423.
Se ha descubierto que el nivel de actividad y la
estabilidad en presencia de agentes oxidantes de las amilasas
mutantes de la técnica anterior se puede mejorar, y así, el
propósito de la invención es proporcionar una
\alpha-amilasa mutante con un nivel de actividad
mejorada y una estabilidad mejorada en presencia de agentes
oxidantes en comparación con las amilasas mutantes de la técnica
anterior. En este contexto el término "estabilidad en presencia
de agentes oxidantes" se refiere tanto a la estabilidad de
almacenamiento de la \alpha-amilasa durante el
almacenamiento del producto de \alpha-amilasa y
durante el almacenamiento del detergente conteniendo
\alpha-amilasa como al agente para el lavado de
vajilla conteniendo \alpha-amilasa, y a la
estabilidad en la solución de lavado o solución para el lavado de
vajilla durante el proceso de lavado o el proceso de lavado de
vajilla, y además a la estabilidad de la
\alpha-amilasa durante la hidrólisis de almidón en
presencia de peróxido de hidrógeno u otros blanqueadores, p. ej.
durante el desencolado en la industria textil.
Según la invención, se proporciona una
alfa-amilasa mutante tal y como se define en la
reivindicación 1, un detergente tal y como se define en la
reivindicación 3 y un agente para el lavado de vajilla tal y como
se define en la reivindicación 4.
Sorprendentemente se ha descubierto que la
\alpha-amilasa mutante según la invención muestra
un mejor nivel de actividad y una mejor estabilidad en presencia de
agentes oxidantes que las amilasas mutantes de la técnica
anterior.
La \alpha-amilasa mutante
según la invención es una \alpha-amilasa de B.
licheniformis, una \alpha-amilasa de B.
amyloliquefaciens o una \alpha-amilasa de
B. stearothermophilus. Todas estas
\alpha-amilasas están bien caracterizadas y se
describe su secuencia de aminoácidos completa. Debe entenderse que
una \alpha-amilasa, la cual es idéntica a la
\alpha-amilasa de B. licheniformis
identificada en FR 2665178, excepto por el hecho de que ésta muestra
un residuo de arginina en la posición 23 en vez del residuo de
lisina, también pertenece a las \alpha-amilasas,
las cuales son las bases para las \alpha-amilasas
mutantes según la invención.
Una forma de realización preferida de la
\alpha-amilasa mutante reivindicada según la
invención se caracteriza por el hecho de que el residuo de
aminoácidos de metionina se sustituye por una Leu. En esta forma de
realización se obtiene un nivel de actividad y estabilidad muy
satisfactorio en presencia de agentes oxidantes.
Además, la invención comprende un detergente, el
cual se caracteriza por el hecho de que éste comprende la
\alpha-amilasa mutante según la invención. Así,
según la invención, la \alpha-amilasa mutante
puede añadirse como un componente de una composición detergente.
Como tal, puede incluirse en la composición detergente en forma de
un aditivo detergente. La composición detergente al igual que el
aditivo detergente puede comprender adicionalmente otra u otras
enzimas usadas de forma convencional en detergentes, tales como
proteasas, lipasas, celulasas, oxidasas o peroxidasas.
En un aspecto específico, la invención
proporciona un aditivo detergente. Las enzimas pueden incluirse en
una composición detergente añadiendo aditivos separados que
contengan una o más enzimas, o añadiendo un aditivo combinado
comprendiendo todas estas enzimas.
Preferiblemente, el aditivo detergente, es decir
un aditivo separado o un aditivo combinado, está provisto en forma
de granulado, preferiblemente un granulado que no desprenda polvo,
un líquido, en particular un líquido estabilizado, una suspensión, o
en una forma protegida.
Se pueden producir granulados libres de polvo,
p. ej. según se describe en US 4.106.991 y US 4.661.452 (ambas para
Novo Industri A/S) y opcionalmente pueden revestirse por métodos
conocidos en la técnica. Las enzimas de detergente pueden mezclarse
antes o después de la granulación.
Las preparaciones enzimáticas líquidas pueden,
por ejemplo, estabilizarse añadiendo un poliol tal como p. ej.
propilenoglicol, azúcar o alcohol de azúcar, ácido láctico o ácido
bórico, según métodos establecidos. Otros estabilizadores
enzimáticos se conocen bien en la técnica.
Las enzimas protegidas pueden prepararse según
el método descrito en EP 238.216 A.
La composición detergente de la invención puede
estar en cualquier forma conveniente, p. ej. en polvo, gránulos o
líquido. Un detergente líquido puede ser acuoso, conteniendo
normalmente hasta un 90% de agua y un 0-20% de
solvente orgánico.
La composición detergente comprende un
tensioactivo que puede ser aniónico, no-iónico,
catiónico, anfotérico o una mezcla de estos tipos. El detergente
normalmente contendrá un 0-50% de tensioactivo
aniónico tal como sulfonato de alquilbenceno lineal (SAL),
sulfonato de alfa-olefinas (SAO), sulfato de alquilo
(SA), sulfato de etoxialcohol (SEA) o jabón. También puede contener
un 0-40% de tensioactivo no-iónico
tal como etoxilato de nonilfenol o etoxilato de alcohol. Además,
puede contener un tensioactivo de amida de ácido graso de
polihidroxi (p. ej. como se describe en WO 92/06154).
El pH (medido en una solución de detergente
acuosa) será normalmente neutral o alcalino, p. ej. de
7-11. El detergente puede contener un
1-40% de un constructor de detergente tal como
zeolita, fosfato, fosfonato, citrato, NTA, EDTA o DTPA, anhídrido
succínico de alquenilo, o silicato, o puede no estar construido (es
decir, esencialmente libre de un constructor detergente). También
puede contener otros ingredientes de detergente convencionales, p.
ej. acondicionadores de tela, reforzadores de espuma,
blanqueadores, p. ej. perborato, percarbonato,
tetra-acetil etilenodiamina (TAED), o sulfonato de
nonanoiloxibenceno (NOBS), agentes anticorrosivos, agentes de
supensión de suciedad, agentes secuestrantes, agentes de reposición
antisuciedad, agentes estabilizantes para la(s)
enzima(s), depresores de espuma, tintes, bactericidas,
blanqueadores ópticos o perfumes.
Formas particulares de composiciones detergentes
dentro del campo de la invención incluyen:
- a)
- Una composición detergente formulada como un detergente en polvo que contiene constructor de fosfato, tensioactivo aniónico, tensioactivo no iónico, silicato, álcali para ajustar a un pH en uso deseado, y sal inorgánica neutral.
- b)
- Una composición detergente formulada como un detergente en polvo que contiene constructor de zeolita, tensioactivo aniónico, tensioactivo no iónico, acrílico o polímero equivalente, silicato, álcali para ajustar a un pH en uso deseado, y sal inorgánica neutral.
- c)
- Una composición detergente formulada como un líquido de detergente acuoso comprendiendo tensioactivo aniónico, tensioactivo no iónico, humectante, ácido orgánico, álcali, con un pH en uso ajustado a un valor entre 7 y 10.5.
- d)
- Una composición detergente formulada como un líquido detergente no acuoso que comprende un tensioactivo no iónico líquido que consiste esencialmente en alcohol primario lineal alcoxilado, constructor de fosfato, álcali, con un pH en uso ajustado a un valor entre aproximadamente 7 y 10.5.
- e)
- Una composición detergente formulada como un detergente en polvo en forma de un granulado con una densidad en masa de al menos 600 g/I, que contiene tensioactivo aniónico y tensioactivo no iónico, sal inorgánica de neutralidad baja o sustancialmente cero, constructor de fosfato, y silicato sódico.
- f)
- Una composición detergente formulada como un detergente en polvo en forma de un granulado con una densidad en masa de al menos 600 g/l, que contiene tensioactivo aniónico y tensioactivo no iónico, sal inorgánica de neutralidad baja o sustancialmente cero, constructor de zeolita, y silicato sódico.
- g)
- Una composición detergente formulada como un detergente en polvo que contiene tensioactivo aniónico, tensioactivo no iónico, polímero acrílico, jabón de ácido graso, carbonato sódico, sulfato de sodio, partículas de arcilla, y silicato sódico.
- h)
- Un líquido detergente compacto comprendiendo un 5-65% en peso de tensioactivo, un 0-50% en peso de constructor y un 0-30% en peso de electrolito.
Se contempla actualmente que, en la composición
detergente de la invención, la \alpha-amilasa
mutante puede añadirse en una cantidad correspondiente a
0.001-100 mg de enzima por litro de solución de
lavado.
Además, la invención comprende un agente de
lavado de vajilla, que se caracteriza por el hecho de que éste
comprende la \alpha-amilasa mutante según la
invención. Todas las formulaciones de agente de lavado de vajilla
pueden usarse en combinación con la
\alpha-amilasa mutante según la invención.
El detergente según la invención y el agente de
lavado de vajilla según la invención puede comprender agentes
oxidantes, los cuales pueden ser un activador, un blanqueante o una
enzima oxidante. El blanqueante puede ser un agente conteniendo
clorina, preferiblemente un agente generador de hipoclorito, un
percarbonato, o un perborato.
En la revista de bioquímica "Journal of
Biological Chemistry"; vol. 260, nº. 11, páginas
6518-6521 se puede leer que la mutagénesis dirigida
puede emplearse para alterar residuos críticos en proteínas los
cuales son susceptibles de oxidación química, y que la metionina
222 es un sitio primario para la inactivación oxidante de la
subtilisina. Esta citación, no obstante, no describe mutantes de
\alpha-amilasa, mientras que la invención está
estrictamente limitada a mutantes de
\alpha-amilasa.
La secuencia de aminoácidos para la
\alpha-amilasa de B. licheniformis aparece
en J. Bacteriol. 166, 635-643, 1986, FR 2665178 ó
EP 410498. Así, los números de metionina son: 8, 15, 197, 256, 304,
366, y 438.
La secuencia de aminoácidos para la
\alpha-amilasa de B. amyloliquefaciens
aparece en J. Biol. Chem. 258, 1007-1013, 1983. Así,
los números de metionina son: 6, 197, 256, 304, 366, y 438.
La secuencia de aminoácidos para la
\alpha-amilasa de B. stearothermophilus
aparece en J. Bacteriol. 166, 635-643, 1986. Así,
los números de metionina son: 8, 9, 97, 200, 206, 284, 307, 311,
316, y 437.
Una vez se ha decidido un modelo de sustitución
sobre la proteína creada genéticamente puede sintetizarse sin
ningún esfuerzo ingenioso según métodos de la técnica anterior.
FR 2665178 describe una variante termoestable de
la \alpha-amilasa de B. licheniformis.
Esta variante, no obstante, no implica una sustitución de metionina,
sino una sustitución en los alrededores de la histidina 133, y
además, la estabilidad oxidante de esta
\alpha-amilasa de la técnica anterior es inferior
en comparación con la estabilidad oxidante del mutante de
\alpha-amilasa según la invención.
Documentación para las propiedades sorprendentes
de la \alpha-amilasa mutante según la invención,
p. ej. el nivel de actividad mejorada y la estabilidad mejorada en
presencia de agentes oxidantes, se presentará a continuación.
Caldos de cultivo filtrados crudos con
diferentes mutantes de Termamyl® indicados abajo se diluyen hasta
una actividad amilasa de 100 NU/ml (la unidad de actividad de
amilasa NU se define en AF 207/1, la cual está disponible bajo
pedido a Novo Nordisk A/S, Novo Allé, DK-2880
Bagsvaerd, Dinamarca) en 50 mM de un tampón
Britton-Robinson a pH 9.0 y se incuba a 40ºC.
Posteriormente se añade H_{2}O_{2} hasta una concentración de
200 mM, y el valor de pH se reajusta a 9.0. La actividad se mide
ahora pasados 15 segundos y pasados 5, 15, y 30 minutos. Los
resultados se muestran a partir de la siguiente tabla 1, donde el
mutante de amilasa se identifica mediante el sistema de aminoácido
escrito. Así, M197A significa el mutante de Termamyl®, donde la
metionina en la posición 197 se sustituye por la alanina. En la
Tabla 1 aparece claramente que el mutante de cisteina de la técnica
anterior (M197C) muestra una estabilidad muy baja, incluso inferior
a la estabilidad de Termamyl®.
Los valores en la siguiente Tabla 1 son los
valores de absorción de OD_{620}, que se toman como indicación de
la actividad.
Todos los mutantes se purifican hasta la
homogeneidad, y la absorción A_{280} se toma como indicación del
contenido de proteína. Las muestras purificadas se diluyeron hasta
un valor A_{280} de 0.0014, correspondiente a 4 NU/ml en relación
a Termamyl®. Bajo estas circunstancias la actividad específica de
cada mutante se midió en presencia de un agente oxidante fuerte
(ácido peracético) y como comparación en ausencia del agente
oxidante fuerte. Las actividades se determinaron como se describe
en AF 207/1 pero con una curva estándar construida a pH 9.0, a 60ºC
en vez de a pH 7.3, a 37ºC. Termamyl® se usó como referencia en la
curva estándar. Las unidades de actividad están referidas por tanto
como NU*. Se muestra claramente en la Tabla 2, donde los valores de
las actividades específicas muestran que el nivel de actividad en
el medio ambiente comprendiendo el agente oxidante fuerte en
comparación con el control es superior para todos los mutantes, en
comparación con Termamyl®.
\vskip1.000000\baselineskip
\vskip1.000000\baselineskip
Las variantes de Termamyl® M197A y M197T y
Termamyl®, respectivamente, se purificaron hasta la homogeneidad, y
la dilución se realizó según A_{280} en el tampón
Britton-Robinson de 50 mM. Las diluciones se
realizaron para alcanzar los valores de absorción A_{620} entre
0.5 y 2 se midieron en el ensayo Phadebas descrito en Af 207/1. Los
valores de pH respectivos se ajustaron a los valores de pH indicados
entre 4 y 10.5. Después de la medición de los valores A_{620}
todos resultados se ajustaron a las unidades A_{620}/A_{280}
(contenido enzimático), véase la Tabla 3.
Los valores en la siguiente Tabla 3 son
absorciones A_{620}, que se toman como indicaciones de las
actividades.
\vskip1.000000\baselineskip
Las muestras mutantes purificadas se
liofilizaron. Se añadieron detergente y Berol 08 caliente a 75ºC
bajo fuerte agitación. Después de endurecer la cera se transfiere
el producto a un congelador y después de un par de horas el producto
se muele y se mezcla con detergente.
El almacenamiento se realizó en frascos cerrados
a 37ºC.
El porcentaje de actividades residuales después
del almacenamiento bajo las condiciones descritas se da en la
siguiente Tabla 4.
- 9% de monohidrato de perborato sódico
- 6% de TAED
- 2% de AEO
- 25% de disilicato de sodio
- 2.0% de fosfonato
- 1.2% de policarboxilatos
- 30% de citrato de trisodio
- 14.8% de carbonato sódico
- 10% de sulfato de sodio
Las muestras de mutantes purificados se
incubaron con una concentración de proteína de 1 mg/ml en una
suspensión del 5% p/p de detergente DLVA (DLVA es una abreviatura
para Detergente de Lavado de Vajilla Automático) a 30ºC y el pH se
ajustó a 10.5. El DLVA contiene agente oxidante (perborato sódico)
y TAED.
La reducción en la actividad de los diferentes
mutantes continuó durante 270 minutos.
El porcentaje de actividades residuales durante
la incubación se da en la siguiente Tabla 5 ("0" minutos es
una medición inicial después de la adición de la suspensión
detergente).
\vskip1.000000\baselineskip
Esta lista de documentos citados por el
solicitante ha sido recopilada exclusivamente para la información
del lector y no forma parte del documento de patente europea. La
misma ha sido confeccionada con la mayor diligencia; la OEP sin
embargo no asume responsabilidad alguna por eventuales errores u
omisiones.
- - WO 9116423 A [0002] [0002]
- - EP 238216 A [0013]
- - FR 2665178 [0006] [0022] [0026]
- - WO 9206154 A [0015]
- - US 4106991 A [0011]
- - EP 410498 A [0022]
- US 4661452 A [0011]
- Journal of Biological Chemistry, vol.
260 (11), 6518-6521 [0021]
- J. Bacteriol., 1986, vol. 166,
635-643 [0022] [0024]
- J. Biol. Chem., 1983, vol. 258,
1007-1013 [0023]
Claims (4)
1. Mutante de Bacillus licheniformis,
Bacillus amyloliquefaciens o alfa-amilasa de
Bacillus stearothermophilus caracterizado por el hecho
de que el residuo de aminoácido de metionina en la posición 197 en
el Bacillus licheniformis o la alfa-amilasa
de Bacillus amyloliquefaciens o en la posilion 200 en la
alfa-amilasa de Bacillus stearothermophilus
se sustituye por un Leu, Ile, Asn, Ser, Gln, Asp o Glu.
2. Alfa-amilasa mutante según la
Reivindicación 1, caracterizada por el hecho de que el
residuo de aminoácido de metionina se sustituye por Leu.
3. Detergente caracterizado por el hecho
de que éste comprende la alfa-amilasa mutante según
la Reivindicación 1 o la Reivindicación 2.
4. Agente de lavado de vajilla,
caracterizado por el hecho de que éste comprende una
alfa-amilasa mutante según la Reivindicación 1 o la
Reivindicación 2.
Applications Claiming Priority (6)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| DK946/92 | 1992-07-23 | ||
| DK94692A DK94692D0 (da) | 1992-07-23 | 1992-07-23 | Alfa-amylase-mutant, detergent og opvaskemiddel |
| DK1503/92 | 1992-12-16 | ||
| DK150392A DK150392D0 (es) | 1992-12-16 | 1992-12-16 | |
| DK29293A DK29293D0 (es) | 1993-03-15 | 1993-03-15 | |
| DK292/93 | 1993-03-15 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| ES2334590T3 true ES2334590T3 (es) | 2010-03-12 |
Family
ID=27220568
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| ES93914652T Expired - Lifetime ES2334590T3 (es) | 1992-07-23 | 1993-07-06 | Alfa-amilasa mutante, detergente y agente de lavado de vajilla. |
Country Status (8)
| Country | Link |
|---|---|
| EP (1) | EP0651794B1 (es) |
| JP (1) | JP3678309B2 (es) |
| KR (1) | KR100294361B1 (es) |
| AT (1) | ATE444356T1 (es) |
| DE (1) | DE69334295D1 (es) |
| ES (1) | ES2334590T3 (es) |
| FI (1) | FI120098B (es) |
| WO (1) | WO1994002597A1 (es) |
Families Citing this family (749)
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| US6197070B1 (en) | 1996-05-15 | 2001-03-06 | The Procter & Gamble Company | Detergent compositions comprising alpha combination of α-amylases for malodor stripping |
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| DE19732749A1 (de) | 1997-07-30 | 1999-02-04 | Henkel Kgaa | Glucanasehaltiges Waschmittel |
| DE19732751A1 (de) | 1997-07-30 | 1999-02-04 | Henkel Kgaa | Neue Beta-Glucanase aus Bacillus |
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| DE19819187A1 (de) | 1998-04-30 | 1999-11-11 | Henkel Kgaa | Festes maschinelles Geschirrspülmittel mit Phosphat und kristallinen schichtförmigen Silikaten |
| MXPA00012241A (es) | 1998-06-10 | 2002-06-04 | Novozymes As | Mananasas novedosas. |
| DE19850100A1 (de) | 1998-10-29 | 2000-05-04 | Henkel Kgaa | Polymer-Granulate durch Wirbelschichtgranulation |
| ATE504651T1 (de) | 1998-12-18 | 2011-04-15 | Novozymes As | Subtilase enzyme der i-s1- und i-s2-untergruppen mit einem zusätzlichen aminosäurerest in einer aktiven schleifenregion |
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| ES2322426T3 (es) | 1999-03-31 | 2009-06-22 | Novozymes A/S | Polipeptidos con actividad alfa-amilasa y acidos nucleicos que codifican a los mismos. |
| DE60040282D1 (de) | 1999-05-20 | 2008-10-30 | Novozymes As | SUBTILASE ENZYME DER I-S1 UND I-S2 UNTERGRUPPEN MIT MINDESTENS EINER ZUSäTZLICHEN AMINOSäURE ZWISCHEN POSITION 132 UND 133 |
| WO2000071687A1 (en) | 1999-05-20 | 2000-11-30 | Novozymes A/S | Subtilase enzymes of the i-s1 and i-s2 sub-groups having at least one additional amino acid residue between positions 129 and 130 |
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Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| NZ208612A (en) * | 1983-06-24 | 1991-09-25 | Genentech Inc | Method of producing "procaryotic carbonyl hydrolases" containing predetermined, site specific mutations |
| PT89702B (pt) * | 1988-02-11 | 1994-04-29 | Gist Brocades Nv | Processo para a preparacao de novos enzimas proteoliticos e de detergentes que os contem |
| DK97190D0 (da) * | 1990-04-19 | 1990-04-19 | Novo Nordisk As | Oxidationsstabile detergentenzymer |
-
1993
- 1993-07-06 JP JP50389894A patent/JP3678309B2/ja not_active Expired - Lifetime
- 1993-07-06 KR KR1019950700148A patent/KR100294361B1/ko not_active Expired - Fee Related
- 1993-07-06 DE DE69334295T patent/DE69334295D1/de not_active Expired - Lifetime
- 1993-07-06 ES ES93914652T patent/ES2334590T3/es not_active Expired - Lifetime
- 1993-07-06 AT AT93914652T patent/ATE444356T1/de not_active IP Right Cessation
- 1993-07-06 WO PCT/DK1993/000230 patent/WO1994002597A1/en not_active Ceased
- 1993-07-06 EP EP93914652A patent/EP0651794B1/en not_active Expired - Lifetime
-
1995
- 1995-01-20 FI FI950263A patent/FI120098B/fi not_active IP Right Cessation
Also Published As
| Publication number | Publication date |
|---|---|
| FI950263L (fi) | 1995-01-20 |
| JPH08500243A (ja) | 1996-01-16 |
| WO1994002597A1 (en) | 1994-02-03 |
| DE69334295D1 (de) | 2009-11-12 |
| EP0651794B1 (en) | 2009-09-30 |
| ATE444356T1 (de) | 2009-10-15 |
| FI120098B (fi) | 2009-06-30 |
| JP3678309B2 (ja) | 2005-08-03 |
| FI950263A0 (fi) | 1995-01-20 |
| KR100294361B1 (ko) | 2001-09-17 |
| EP0651794A1 (en) | 1995-05-10 |
| KR950702624A (ko) | 1995-07-29 |
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