ES2545494T3 - Proteasas - Google Patents

Proteasas Download PDF

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Publication number
ES2545494T3
ES2545494T3 ES10180267.6T ES10180267T ES2545494T3 ES 2545494 T3 ES2545494 T3 ES 2545494T3 ES 10180267 T ES10180267 T ES 10180267T ES 2545494 T3 ES2545494 T3 ES 2545494T3
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ES
Spain
Prior art keywords
dsm
nocardiopsis
proteases
protease
stable
Prior art date
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Active
Application number
ES10180267.6T
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English (en)
Inventor
Soeren Flensted Lassen
Carsten Sjoeholm
Peter Rahbek Oestergaard
Morten Fischer
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Novozymes AS
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Novozymes AS
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Publication of ES2545494T3 publication Critical patent/ES2545494T3/es
Active legal-status Critical Current
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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)
    • C12N9/48—Hydrolases (3) acting on peptide bonds (3.4)
    • C12N9/50—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25)
    • C12N9/52—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from bacteria or Archaea
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23J—PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
    • A23J3/00—Working-up of proteins for foodstuffs
    • A23J3/30—Working-up of proteins for foodstuffs by hydrolysis
    • A23J3/32—Working-up of proteins for foodstuffs by hydrolysis using chemical agents
    • A23J3/34—Working-up of proteins for foodstuffs by hydrolysis using chemical agents using enzymes
    • A23J3/346—Working-up of proteins for foodstuffs by hydrolysis using chemical agents using enzymes of vegetable proteins
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K20/00—Accessory food factors for animal feeding-stuffs
    • A23K20/10—Organic substances
    • A23K20/189—Enzymes
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K50/00—Feeding-stuffs specially adapted for particular animals
    • A23K50/30—Feeding-stuffs specially adapted for particular animals for swines
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K50/00—Feeding-stuffs specially adapted for particular animals
    • A23K50/60—Feeding-stuffs specially adapted for particular animals for weanlings
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K50/00—Feeding-stuffs specially adapted for particular animals
    • A23K50/70—Feeding-stuffs specially adapted for particular animals for birds
    • A23K50/75—Feeding-stuffs specially adapted for particular animals for birds for poultry
    • 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
    • C11D3/38609—Protease or amylase in solid compositions only
    • 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
    • C11D3/38618—Protease or amylase in liquid compositions only

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  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Polymers & Plastics (AREA)
  • Zoology (AREA)
  • Food Science & Technology (AREA)
  • Health & Medical Sciences (AREA)
  • Wood Science & Technology (AREA)
  • Organic Chemistry (AREA)
  • Birds (AREA)
  • Animal Husbandry (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Genetics & Genomics (AREA)
  • Biochemistry (AREA)
  • Oil, Petroleum & Natural Gas (AREA)
  • Molecular Biology (AREA)
  • Microbiology (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Biomedical Technology (AREA)
  • Medicinal Chemistry (AREA)
  • Biotechnology (AREA)
  • General Chemical & Material Sciences (AREA)
  • Nutrition Science (AREA)
  • Enzymes And Modification Thereof (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Detergent Compositions (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
  • Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
  • Fodder In General (AREA)

Abstract

Polipéptido aislado con actividad de proteasa, que muestra i) una actividad relativa a 15ºC y pH 9 de al menos 0,8, ii) una actividad relativa a 25ºC y pH 9 de al menos 0,10; y/o una actividad relativa a 37ºC y pH de al menos 0,20 donde la actividad se compara con la actividad a óptima temperatura de la misma proteasa; seleccionado del grupo que consiste en: (a) un polipéptido con una secuencia de aminoácidos que tiene un grado de identidad para aminoácidos 1- 192 de la SEC ID nº: 4 o aminoácidos 1-192 de la SEC ID nº:6 de al menos un 80%; y (b) un fragmento de (a) que tiene actividad de proteasa.

Description

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E10180267
17-08-2015
Tabla 2: Perfil de estabilidad de pH
pH
Proteasa L1a Proteasa L2
2,0
0,50 1,00
2,5
0,81 0,95
3,0
0,93 0,97
3,5
0,94 1,01
4,0
0,97 0,98
5,0
0,96 0,97
6,0
0,95 0,98
7,0
0,99 0,96
8,0
0,97 0,99
9,0
0,93 0,99
10,0
0,94 0,96
11,0
0,94 0,94
12,0
0,92 0,84
9,0 y tras 2 horas a 5 °C
1,00 1,00
Tabla 3: Perfil de actividad de temperatura
Temperatura (°C)
Proteasa L1a Proteasa L2
15
0,11 0,01
25
0,17 0,01
37
0,30 0,03
50
0,58 0,09
60
0,90 0,19
70
1,00 0,63
80
0,34 1,00
90
- 0,35
5 Otras características
[0247] La proteasa L1a es una proteasa alfa-lítica como enzima (familia de peptidasa S1E, notación antigua S2A) que se descubre que se inhibe por fenil metil sulfonil fluoruro (PMSF) y por el inhibidor de subtilisina de
10 Streptomyces (SSI). Su peso molecular relativo como determinado por SDS-PAGE es Mr = 22kDa, y la secuencia Nterminal: ADIVGGEAY (SEC ID nº: 26).
Ejemplo 5: Actividad específica de la proteasa L1a
15 [0248] La preparación de proteasa purificada descrita en el ejemplo 4 se usó para determinación de la actividad específica. La pureza de la preparación fue de sobre 95% cuando se analizó por SDS-PAGE (determinado como se describe en el ejemplo 2A en la WO 01/58275). La muestra de proteasa se dividió en dos. Una parte se analizó para el contenido de proteína (mg/ml) por análisis de aminoácido, la otra parte se analizó para actividad de proteasa.
20 Análisis de aminoácido (AAA)/(mg/ml)
[0249] Los enlaces peptídicos de la muestra de proteasa se sometieron a hidrólisis ácida, seguido de separación y
28
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Claims (1)

  1. imagen1
    imagen2
ES10180267.6T 2004-06-21 2005-06-17 Proteasas Active ES2545494T3 (es)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
DKPA200400969 2004-06-21
DK200400969 2004-06-21

Publications (1)

Publication Number Publication Date
ES2545494T3 true ES2545494T3 (es) 2015-09-11

Family

ID=34969746

Family Applications (2)

Application Number Title Priority Date Filing Date
ES05748647T Active ES2380105T3 (es) 2004-06-21 2005-06-17 Proteasas de Nocardiopsis
ES10180267.6T Active ES2545494T3 (es) 2004-06-21 2005-06-17 Proteasas

Family Applications Before (1)

Application Number Title Priority Date Filing Date
ES05748647T Active ES2380105T3 (es) 2004-06-21 2005-06-17 Proteasas de Nocardiopsis

Country Status (11)

Country Link
US (4) US8357408B2 (es)
EP (3) EP1766001B1 (es)
JP (3) JP5623691B2 (es)
CN (1) CN101014704A (es)
AT (1) ATE541034T1 (es)
AU (2) AU2005254611B2 (es)
BR (1) BRPI0512341A (es)
CA (1) CA2591858C (es)
ES (2) ES2380105T3 (es)
MX (1) MXPA06014649A (es)
WO (1) WO2005123911A2 (es)

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