[go: up one dir, main page]

WO1999049010A2 - Enzymes d'oxydation de phenol et leurs utilisations - Google Patents

Enzymes d'oxydation de phenol et leurs utilisations Download PDF

Info

Publication number
WO1999049010A2
WO1999049010A2 PCT/EP1999/002042 EP9902042W WO9949010A2 WO 1999049010 A2 WO1999049010 A2 WO 1999049010A2 EP 9902042 W EP9902042 W EP 9902042W WO 9949010 A2 WO9949010 A2 WO 9949010A2
Authority
WO
WIPO (PCT)
Prior art keywords
phenol oxidizing
oxidizing enzyme
stachybotrys
enzyme
detergent composition
Prior art date
Application number
PCT/EP1999/002042
Other languages
English (en)
Other versions
WO1999049010A3 (fr
Inventor
Daniel Convents
Antoine Amory
Huaming Wang
Patrick Dhaese
Annick Lambrechts-Rongvaux
Cynthia Wang
Original Assignee
Unilever N.V.
Unilever Plc
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Priority claimed from US09/218,702 external-priority patent/US6426410B1/en
Application filed by Unilever N.V., Unilever Plc filed Critical Unilever N.V.
Priority to EP99917861A priority Critical patent/EP1066364A2/fr
Priority to CA002323092A priority patent/CA2323092A1/fr
Priority to BR9909043-0A priority patent/BR9909043A/pt
Priority to AU35995/99A priority patent/AU3599599A/en
Publication of WO1999049010A2 publication Critical patent/WO1999049010A2/fr
Publication of WO1999049010A3 publication Critical patent/WO1999049010A3/fr

Links

Classifications

    • 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
    • C12N9/0004—Oxidoreductases (1.)
    • C12N9/0055—Oxidoreductases (1.) acting on diphenols and related substances as donors (1.10)
    • 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
    • C11D3/38636—Preparations containing enzymes, e.g. protease or amylase containing enzymes other than protease, amylase, lipase, cellulase, oxidase or reductase
    • 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
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/14—Fungi; Culture media therefor
    • C12N1/145—Fungal isolates
    • 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
    • C12N9/0004—Oxidoreductases (1.)
    • C12N9/0055—Oxidoreductases (1.) acting on diphenols and related substances as donors (1.10)
    • C12N9/0057—Oxidoreductases (1.) acting on diphenols and related substances as donors (1.10) with oxygen as acceptor (1.10.3)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/645—Fungi ; Processes using fungi

Definitions

  • phenol oxidizing enzymes exhibit pH optima in the acidic pH range while being inactive in neutral or alkaline pHs.
  • the present invention also encompasses Stachybotrys phenol oxidizing enzyme mutants as long as the mutant is able to modify the color associated with dyes or colored compounds.
  • the present invention provides an isolated polynucleotide encoding a phenol oxidizing enzyme obtainable from Stachybotrys wherein said polynucleotide comprises a nucleic acid sequence having at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90% and at least 95% identity to SEQ ID NO:1 or SEQ ID NO:3 as long as the polynucleotide encodes a phenol oxidizing enzyme capable of modifying the color associated with dyes or colored compounds.
  • detergent compositions comprising a Stachybotrys phenol oxidizing enzyme of the present invention alone or in combination with an enhancer and other detergent ingredients, including proteases, amylases and/or cellulases.
  • Maximum stringency typically occurs at about Tm-5°C (5°C below the Tm of the probe); “high stringency” at about 5°C to 10°C below Tm; “intermediate stringency” at about 10°C to 20°C below Tm; and “low stringency” at about 20°C to 25°C below Tm.
  • a maximum stringency hybridization can be used to identify or detect identical polynucleotide sequences while an intermediate or low stringency hybridization can be used to identify or detect polynucleotide sequence homologs.
  • the vector or cassette contains sequences directing transcription and translation of the nucleic acid, a selectable marker, and sequences allowing autonomous replication or chromosomal integration.
  • Suitable vectors comprise a region 5' of the gene which harbors transcriptional initiation controls and a region 3' of the DNA fragment which controls transcriptional termination. These control regions may be derived from genes homologous or heterologous to the host as long as the control region selected is able to function in the host cell.
  • Initiation control regions or promoters, which are useful to drive expression of the phenol oxidizing enzymes in a host cell are known to those skilled in the art. Virtually any promoter capable of driving these phenol oxidizing enzyme is suitable for the present invention.
  • Anthocyanins have a high diversity in glycosidation patterns.
  • the phenol oxidizing enzymes of the present invention may be produced by cultivation of phenol oxidizing enzyme-producing Stachybotrys strains (such as S. parvispora MUCL 38996, S. chartarum MUCL 38898) under aerobic conditions in nutrient medium containing assimiable carbon and nitrogen together with other essential nutrient(s).
  • the medium can be composed in accordance with principles well-
  • the phenol oxidizing enzymes of the present invention may be formulated and utilized according to their intended application.
  • the phenol oxidizing enzyme may be formulated, directly from the fermentation broth, as a coated solid using the procedure described in United States Letters Patent No. 4,689,297.
  • the phenol oxidizing enzyme may be formulated in a liquid form with a suitable carrier.
  • the phenol oxidizing enzyme may also be immobilized, if desired.
  • the present invention also encompasses expression vectors and recombinant host cells comprising a Stachybotrys phenol oxidizing enzyme of the present invention and the subsequent purification of the phenol oxidizing enzyme from the recombinant host cell.
  • the rate of oxygen consumption was measured with each of the dyes, in a magnetically stirred chamber equipped with a Clark electrode (oxygraph K-IC from Gilson).
  • the oxygraph chamber contained, in a final volume of 2 ml, 200 mM Tris/HCI (pH 7.0), 5 mM of each of the dyes, and 100 ml (39 EU) of phenol oxidizing enzyme from S. parvispora MUCL 38996, obtained as described above in Example 5.
  • the reactions were started by the addition of the enzyme, and the dissolved oxygen concentration was recorded during 5 minutes. The slope of the curves were determined from their linear parts.
  • Table 3 The results of this experiment are also summarized below in Table 3.
  • PCR reaction was performed at 95°C for 1 minute, the primers were annealed to the template at 45°C for 1 minute and extension was done at 68°C for 1 minute. This cycle was repeated 30 times to achieve a gel-visible PCR fragment.
  • the PCR fragment detected by agarose gel contained a fragment of about 1 kilobase which was then cloned into the plasmid vector pCR-ll (Invitrogen). The 1 kb insert was then subjected to nucleic acid sequencing.
  • the expression plasmid for use in transforming Trichoderma reesei was constructed as follows. The ends of the Bglll to Xbal fragment shown in Figure 9 containing the gene encoding the Stachybotrys phenol oxidizing enzyme were blunted by T4 DNA polymerase and inserted into Pmel restriction site of the Trichoderma expression vector, pTrex, which is a modified version of pTEX, see PCT Publication No. WO 96/23928 for a complete description of the preparation of the pTEX vector, which discussion is herein incorporated by reference, which contains a CBHI promoter and terminator for gene expression and a Trichoderma pyr4 gene as a selection marker for transformants.

Landscapes

  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Organic Chemistry (AREA)
  • Wood Science & Technology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Zoology (AREA)
  • Genetics & Genomics (AREA)
  • Biotechnology (AREA)
  • General Health & Medical Sciences (AREA)
  • General Engineering & Computer Science (AREA)
  • Microbiology (AREA)
  • Medicinal Chemistry (AREA)
  • Biomedical Technology (AREA)
  • Biochemistry (AREA)
  • Molecular Biology (AREA)
  • Botany (AREA)
  • Mycology (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Virology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Oil, Petroleum & Natural Gas (AREA)
  • Enzymes And Modification Thereof (AREA)
  • Detergent Compositions (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)

Abstract

L'invention concerne des enzymes d'oxydation de phénol pouvant être obtenues à partir d'espèces de Stachybotrys et s'utilisant pour modifier la couleur associée aux colorants et composés colorés, ainsi que dans des applications anti-transfert de colorant. L'invention concerne également des cultures biologiquement pures de souches du genre Stachybotrys, désignées ci-après sous le nom de Stachybotrys parvispora MUCL 38996 et Stachybotrys chartarum MUCL 38898, capables de produire naturellement des enzymes d'oxydation de phénol. L'invention concerne aussi la séquence d'acides aminés et la séquence nucléotidique d'enzymes d'oxydation de phénol à partir de Stachybotrys, ainsi que des vecteurs d'expression et des cellules hôtes comprenant l'acide nucléique. L'invention concerne en outre des méthodes de production de l'enzyme d'oxydation de phénol ainsi que des méthodes d'élaboration d'hôtes d'expression. L'invention concerne enfin des compositions détergentes comprenant des enzymes d'oxydation de phénol pouvant être obtenues à partir d'espèces de Stachybotrys.
PCT/EP1999/002042 1998-03-24 1999-03-23 Enzymes d'oxydation de phenol et leurs utilisations WO1999049010A2 (fr)

Priority Applications (4)

Application Number Priority Date Filing Date Title
EP99917861A EP1066364A2 (fr) 1998-03-24 1999-03-23 Enzymes d'oxydation de phenol et leurs utilisations
CA002323092A CA2323092A1 (fr) 1998-03-24 1999-03-23 Enzymes d'oxydation de phenol et leurs utilisations
BR9909043-0A BR9909043A (pt) 1998-03-24 1999-03-23 Composição detergente
AU35995/99A AU3599599A (en) 1998-03-24 1999-03-23 Phenol oxidizing enzymes and their use

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
US4696998A 1998-03-24 1998-03-24
US09/046,969 1998-03-24
US09/218,702 1998-12-22
US09/218,702 US6426410B1 (en) 1998-12-22 1998-12-22 Phenol oxidizing enzymes

Publications (2)

Publication Number Publication Date
WO1999049010A2 true WO1999049010A2 (fr) 1999-09-30
WO1999049010A3 WO1999049010A3 (fr) 1999-12-29

Family

ID=26724487

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/EP1999/002042 WO1999049010A2 (fr) 1998-03-24 1999-03-23 Enzymes d'oxydation de phenol et leurs utilisations

Country Status (6)

Country Link
EP (1) EP1066364A2 (fr)
AU (1) AU3599599A (fr)
BR (1) BR9909043A (fr)
CA (1) CA2323092A1 (fr)
TR (1) TR200002713T2 (fr)
WO (1) WO1999049010A2 (fr)

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2000039306A3 (fr) * 1998-12-23 2000-10-26 Unilever Nv Compositions detergentes contenant des enzymes oxydant le phenol
WO2001047478A3 (fr) * 1999-12-24 2002-01-10 Henkel Kgaa Colorant enzymatique
US7160709B2 (en) * 1998-03-24 2007-01-09 Genencor International, Inc. Phenol oxidizing enzymes
US8067198B2 (en) 2003-06-25 2011-11-29 Prolume Ltd. Protein expression system

Family Cites Families (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH02160684A (ja) * 1988-12-14 1990-06-20 Yuukishitsu Hiryo Seibutsu Katsusei Riyou Gijutsu Kenkyu Kumiai コンポストの製造方法
JP3181660B2 (ja) * 1992-01-24 2001-07-03 天野エンザイム株式会社 ビリルビンオキシダーゼの製造法
DK77393D0 (da) * 1993-06-29 1993-06-29 Novo Nordisk As Aktivering af enzymer
ATE214750T1 (de) * 1994-10-20 2002-04-15 Novozymes As Bleichverfahren unter verwendung eines phenoloxidierenden enzyms, einer wasserstoffperoxidquelle und eines verstärkungsmittels

Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7160709B2 (en) * 1998-03-24 2007-01-09 Genencor International, Inc. Phenol oxidizing enzymes
WO2000039306A3 (fr) * 1998-12-23 2000-10-26 Unilever Nv Compositions detergentes contenant des enzymes oxydant le phenol
US6509307B1 (en) 1998-12-23 2003-01-21 Unilever Home & Personal Care Usa Division Of Conopco, Inc. Detergent compositions comprising phenol oxidizing enzymes from fungi
WO2001047478A3 (fr) * 1999-12-24 2002-01-10 Henkel Kgaa Colorant enzymatique
US8067198B2 (en) 2003-06-25 2011-11-29 Prolume Ltd. Protein expression system
US9115364B2 (en) 2003-06-25 2015-08-25 Prolume Ltd. Protein expression system

Also Published As

Publication number Publication date
AU3599599A (en) 1999-10-18
CA2323092A1 (fr) 1999-09-30
EP1066364A2 (fr) 2001-01-10
TR200002713T2 (tr) 2000-12-21
WO1999049010A3 (fr) 1999-12-29
BR9909043A (pt) 2000-12-05

Similar Documents

Publication Publication Date Title
EP1141321B1 (fr) Enzymes d'oxydation du phenol
US7144717B1 (en) Oxidizing enzymes
US6426410B1 (en) Phenol oxidizing enzymes
CA2384954C (fr) Nouveaux enzymes oxydant des phenols
WO1999049010A2 (fr) Enzymes d'oxydation de phenol et leurs utilisations
US6329332B1 (en) Pleurotus phenol oxidizing enzymes
US20020165113A1 (en) Detergent compositions comprising novel phenol oxidizing enzymes
US20020019038A1 (en) Phenol oxidizing enxymes
US20060024784A1 (en) Phenol oxidizing enzymes
WO2001021748A1 (fr) Compositions detergentes contenant des enzymes d'oxydation de phenol

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A2

Designated state(s): AL AM AT AU AZ BA BB BG BR BY CA CH CN CU CZ DE DK EE ES FI GB GD GE GH GM HR HU ID IL IN IS JP KE KG KP KR KZ LC LK LR LS LT LU LV MD MG MK MN MW MX NO NZ PL PT RO RU SD SE SG SI SK SL TJ TM TR TT UA UG US UZ VN YU ZW

AL Designated countries for regional patents

Kind code of ref document: A2

Designated state(s): GH GM KE LS MW SD SL SZ UG ZW AM AZ BY KG KZ MD RU TJ TM AT BE CH CY DE DK ES FI FR GB GR IE IT LU MC NL PT SE BF BJ CF CG CI CM GA GN GW ML MR NE SN TD TG

121 Ep: the epo has been informed by wipo that ep was designated in this application
DFPE Request for preliminary examination filed prior to expiration of 19th month from priority date (pct application filed before 20040101)
AK Designated states

Kind code of ref document: A3

Designated state(s): AL AM AT AU AZ BA BB BG BR BY CA CH CN CU CZ DE DK EE ES FI GB GD GE GH GM HR HU ID IL IN IS JP KE KG KP KR KZ LC LK LR LS LT LU LV MD MG MK MN MW MX NO NZ PL PT RO RU SD SE SG SI SK SL TJ TM TR TT UA UG US UZ VN YU ZW

AL Designated countries for regional patents

Kind code of ref document: A3

Designated state(s): GH GM KE LS MW SD SL SZ UG ZW AM AZ BY KG KZ MD RU TJ TM AT BE CH CY DE DK ES FI FR GB GR IE IT LU MC NL PT SE BF BJ CF CG CI CM GA GN GW ML MR NE SN TD TG

WWE Wipo information: entry into national phase

Ref document number: 1999917861

Country of ref document: EP

ENP Entry into the national phase

Ref document number: 2323092

Country of ref document: CA

Ref document number: 2323092

Country of ref document: CA

Kind code of ref document: A

WWE Wipo information: entry into national phase

Ref document number: 2000/02713

Country of ref document: TR

NENP Non-entry into the national phase

Ref country code: KR

WWP Wipo information: published in national office

Ref document number: 1999917861

Country of ref document: EP

REG Reference to national code

Ref country code: DE

Ref legal event code: 8642

NENP Non-entry into the national phase

Ref country code: CA

WWW Wipo information: withdrawn in national office

Ref document number: 1999917861

Country of ref document: EP