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WO1996006930A1 - Laccases obtenues a partir de coprinaceae - Google Patents

Laccases obtenues a partir de coprinaceae Download PDF

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Publication number
WO1996006930A1
WO1996006930A1 PCT/DK1995/000344 DK9500344W WO9606930A1 WO 1996006930 A1 WO1996006930 A1 WO 1996006930A1 DK 9500344 W DK9500344 W DK 9500344W WO 9606930 A1 WO9606930 A1 WO 9606930A1
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WO
WIPO (PCT)
Prior art keywords
laccase
enhancing agent
obtainable
coprinus
detergent
Prior art date
Application number
PCT/DK1995/000344
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English (en)
Inventor
Ture Damhus
Palle Schneider
Lisbeth Bech
Marion Heinzkill
Original Assignee
Novo Nordisk A/S
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
Application filed by Novo Nordisk A/S filed Critical Novo Nordisk A/S
Priority to AU32536/95A priority Critical patent/AU3253695A/en
Publication of WO1996006930A1 publication Critical patent/WO1996006930A1/fr

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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
    • C12N9/0004—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)
    • C12N9/0061—Laccase (1.10.3.2)
    • 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
    • C11D3/38654—Preparations containing enzymes, e.g. protease or amylase containing oxidase or reductase

Definitions

  • the present invention relates to a method of oxidiz ⁇ ing a substrate in an alkaline solution with a laccase.
  • the invention also relates to a detergent additive and to a detergent composition.
  • Laccases are enzymes that catalyze the oxidation of a substrate with dioxygen, reducing dioxygen to water. Such enzymes are known from microbial, plant and animal origins.
  • oxidizable substrates e.g., metal ions and phenolic compounds such as 7-hydroxycouma in, vanillin, and p- hydroxybenzenesulfonate
  • accelerators or enhancing agents able to enhance bleaching reactions cf. e.g. WO 92/18683, WO 92/18687, and Kato M and Shimizu S. Plant Cell Physiol. 1985 26 (7), pp. 1291-1301 (cf. Table 1 in particu ⁇ lar) ) .
  • enhancing agents e.g., phenothiazines and phenoxazines.
  • the invention provides a method of oxidizing a substrate in a solution with a pH at or above 7, comprising contacting the substrate with an effective amount of a laccase obtainable from Coprinaceae.
  • the invention al provides use of these laccases in bleaching compositions a detergent compositions.
  • Fig. 1 shows the bleaching of gradual added Acid Blue 45 in phosphate/borate buffer pH 10 at 35°
  • I Only dye addition
  • II Dye addition in the presence Laccase
  • III Dye addition in the presence of Laccase + 1 ethylphenothiazine-4-carboxylic acid
  • IV Dye addition in t presence of Laccase + phenoxazine-10-propionic acid; t experiment conducted as described in Example .
  • the present invention provides a method of oxidizi a substrate in a solution with a pH at or above 7, preferab in a solution with a pH at or above 8, even more preferably a solution with a pH at or above 9, in particular in a soluti with a pH around 10, comprising contacting said substrate wi an effective amount of a laccase obtainable from Coprinacea optionally in the presence of an enhancing agent.
  • Laccases (EC 1.10.3.2) are oxidoreductases th function with molecular oxygen as electron acceptor. Molecul oxygen from the atmosphere will usually be present in su ficient quantity, so normally it is not necessary to add ext oxygen to the process medium.
  • laccases th function at a high pH are obtainable from laccase-produci strains of the family Coprinaceae.
  • the family Coprinaceae comprises the followi genera: Coprinus. Podaxis, Montaqnea. Macrometrul Psathyrella, Panaeolina, Panaeolus, Copelandia, Anellaria, Limnoperdon, Panaeolopsis and Polvplocium.
  • the laccase employed in the method of the present invention is obtainable from Coprinus. Panaeolus or 5 Psathyrella, in particular Coprinus cinereus. Coprinus comatus. Coprinus friesii. Coprinus plicatilis. or Psathyrella condolleana. or Panaeolus papilionaceus; most preferred strain is Coprinus cinereus (IFO30116) . The most preferred strain is freely available to the public from Institute of Fermentation,
  • the laccase according to the invention may furthe more be one which is producible by a method comprising cu tivating a host cell transformed with a recombinant DNA vect which carries a DNA sequence encoding said laccase as well DNA sequences encoding functions permitting the expression the DNA sequence encoding the laccase, in a culture medi under conditions permitting the expression of the lacca enzyme, and recovering the laccase from the culture.
  • LACU Laccase Activity
  • Laccase activity is determined by oxidizing syringa dazin under aerobic conditions. The violet colour produced photo etered at 530 nm.
  • the analytical conditions are 19 syringaldazin, 23.2 mM acetate buffer, pH 5.5, 30°C, 1 m reaction time.
  • 1 laccase unit (LACU) is the amount of enzyme th catalyses the conversion of 1.0 ⁇ mole syringaldazin per minu under these conditions.
  • the immunochemical properties can be determin immunologically by cross-reaction identity tests.
  • the identi tests can be performed by the well-known Ouchterlony doub immunodiffusion procedure or by tandem crossed im unoelectr phoresis according to I. M. Roitt; Immunology, Gower Medic Publishing (1985) and N. H. Axelsen; Handbook of Immunopr cipitation-in-Gel Techniques; Blackwell Scientific Publicatio (1983) , Chapters 5 and 14.
  • laccases displayi immunochemical cross-reactivity with an antibody raised again a laccase obtainable from Coprinus cinereus IFO30116 are also included.
  • the laccases of the invention show excellent results. If additionally an enhancing agent is added, the result may be improved.
  • an enhancing agent is any compound that enhances the oxidation.
  • the enhancing agent will typically be an oxidizable compound, e.g., a metal ion or a phenolic compound such as 7-hydroxycoumarin, vanillin, or p- hydroxybenzenesulfonate, (for reference see WO 92/18683, WO 92/18687, and Kato M and Shimizu S. Plant Cell Physiol. 198526 (7), pp. 1291-1301 (cf. Table 1 in particular)).
  • R 1 is H, OH, C n H 2n+1 or OC n H 2n+1 , in which n is an integer of from 1 to 10; and R 2 and R 3 are the same or different and selected from C m H 2m+1 , in which m is an integer of from 1 to 10.
  • R 1 , R 2 and R 3 may also contain double bonds or cyclic groups.
  • the enhancing agent is acetosyringone, syringaldehyde, methylsyringate or syringic acid.
  • formula X represents (-0-) or (-S-)
  • substituent groups R 1 -R 9 which may be identical or differen independently represent any of the following radical hydrogen, halogen, hydroxy, formyl, carboxy, and esters a salts hereof, carbamoyl, sulfo, and esters and salts hereo sulfamoyl, nitro, amino, phenyl, C 1 -C u -alkyl, C.-C 5 -alkox carbonyl- ⁇ -C j -alkyl, aryl- ⁇ -C j -alkyl; which carbamoyl, sulfam yl, and amino groups may furthermore be unsubstituted substituted once or twice with a substituent group R 10 ; a which phenyl may furthermore be unsubstituted or substitut with one or more substituent groups R 10 ; and which C.-C u -alkyl C ⁇
  • the enhancing agent is 10- methylphenothiazine, phenothiazine-10-propionic acid, N- hydroxysuccinimide phenothiazine-10-propionate, 10-ethylpheno- thiazine-4-carboxylic acid, 10-ethylphenothiazine, 10-propyl- phenothiazine, 10-isopropylphenothiazine, methyl phenothiazine- 10-propionate, 10-phenylphenothiazine, 10-allylphenothiazine, 10-(3-(4-methylpiperazin-l-yl)propyl)phenothiazine, 10-(2- pyrrolidin-1-yl-ethyl) phenothiazine, 2-methoxy-10-methyl- phenothiazine, l-methoxy-10- ⁇ rtethylphenothiazine, 3-methoxy-10- methylphenothia
  • the enhancing agents may be obtained from Sigma- Aldrich, Janssen Chimica, Kodak, Tokyo Kasai Organic Chemicals, Daiichi Pure Chemicals Co. or Boehringer Mannheim; N-methylated derivatives of phenothiazine and phenoxazine may be prepared by methylation with methyliodide as described by Cornel Bodea and loan Silberg in "Recent Advances in the Chemistry of Pheno- thiazines" (Advances in heterocyclic chemistry, 1968, Vol. 9, pp. 321-460) ; B. Cardillo & G. Casnati in Tetrahedron, 1967, Vol. 23, p. 3771.
  • Phenothiazine and phenoxazine propionic aci may be prepared as described in J. Or . Che . 15, 1950, p 1125-1130. Hydroxyethyl and hydroxypropyl derivatives phenothiazine and phenoxazine may be prepared as described G. Cauquil in Bulletin de la Society Chemique de France. 196 p.1049.
  • the enhancing agent may be present in concentratio of from 0.01 to 500 ⁇ M, preferably in concentrations of fr 0.1 to 250 ⁇ M.
  • the method of the inve tion finds application for bleaching of a dye or dyes solutions.
  • the dye may be a synthetic dye such as an azo or anthraquinone dye, or a natural or nature-identical dy
  • dyes are Acid Red 151, Acid blue 45, Direct Blue and indigo carmine, where Acid Red 151, Acid blue 45 and Dire
  • the method of the invention finds application for dye transfer inhibition, e.g., during processing of dyed textiles (cf. e.g. WO 92/18687) or during laundering (cf. e.g. WO 91/05839) .
  • the invention provides a method•for inhibiting the transfer of a textile dye from a dyed fabric to another fabric when said fabrics are rinsed together or washed together in a wash liquor, the method comprising treatment of the wash liquor with a laccase obtainable from Coprinaceae in the presence or absence of an enhancing agent.
  • the method of the invention finds application in treatment of waste water, e.g., waste water from the chemical or pharmaceutical industry, from dye manufacturing, from dye-works or from the textile industry.
  • the laccase may be present in concentrations of from 0.001-500 LACU/liter, preferably in concentrations of from 0.5-100 LACU/liter.
  • a laccase of t invention may be added as a component of a detergent compos tion. As such, it may be included in the detergent compositi in the form of a detergent additive.
  • the detergent compositi as well as the detergent additive may additionally comprise o or more other enzymes, such as proteases, Upases, a ylase cutinases, cellulases and other oxidoreductases than laccase e.g., peroxidases and hydrogen peroxide generating oxidases
  • the invention provides detergent additive.
  • the enzymes may be included in a deterge composition by adding separate additives containing one or mo enzymes, or by adding a combined additive comprising all these enzymes.
  • a detergent additive of the invention i.e. separated additive or a combined additive, can be formulat e.g. as granulates, liquids, slurries, etc.
  • Preferred deterge additive formulations are granulates, in particular non-dusti granulates, liquids, in particular stabilized liquids, slu ries, or protected enzymes.
  • Non-dusting granulates may be produced, e.g., disclosed in US 4,106,991 and 4,661,452 (both to Novo Indust A/S) and may optionally be coated by methods known in the ar Examples of waxy coating materials are poly(ethylene oxid products (polyethyleneglycol, PEG) with mean molecular weigh of 1000 to 20000; ethoxylated nonylphenols having from 16 to ethylene oxide units; ethoxylated fatty alcohols in which t alcohol contains from 12 to 20 carbon atoms and in which the are 15 to 80 ethylene oxide units; fatty alcohols; fatty acid and mono- and di- and triglycerides of fatty acids.
  • PEG poly(ethylene oxid products
  • PEG polyethyleneglycol, PEG
  • ethoxylated nonylphenols having from 16 to ethylene oxide units
  • Liqu enzyme preparations may, for instance, be stabilized by addi a polyol such as propylene glycol, a sugar or sugar alcoho lactic acid or boric acid according to established method Other enzyme stabilizers are well known in the art.
  • Protect enzymes may be prepared according to the method disclosed in 238,216.
  • the detergent composition of the invention may be in any convenient form, e.g., as powder, granules, paste or liquid.
  • a liquid detergent may be aqueous, typically containing up to 70% water and 0-30% organic solvent, or nonaqueous.
  • the detergent composition comprises one or more surf ⁇ actants, each of which may be anionic, nonionic, cationic, or zwitterionic.
  • the detergent will usually contain 0-50% of anionic surfactant such as linear alkylbenzenesulfonate (LAS) , alpha-olefinsulfonate (AOS) , alkyl sulfate (fatty alcohol sulfate) (AS) , alcohol ethoxysulfate (AEOS or AES) , secondary alkanesulfonates (SAS) , alpha-sulfo fatty acid methyl esters, alkyl- or alkenylsuccinic acid, or soap.
  • anionic surfactant such as linear alkylbenzenesulfonate (LAS) , alpha-olefinsulfonate (AOS) , alkyl sulfate (fatty alcohol sulfate) (AS) , alcohol ethoxysulfate (AEOS or AES
  • Nonionic surfactant such as alcohol ethoxylate (AEO or AE) , carboxylated alcohol ethoxylates, nonylphenol ethoxylate, alkylpolyglycoside, alkyldi ethylamine oxide, ethoxylated fatty acid monoethanolamide, fatty acid monoethanolamide, or polyhydroxy alkyl fatty acid amide (e.g. as described in WO 92/06154) .
  • the detergent composition may additionally comprise one or more other enzymes, such as amylases, lipases, cuti- nases, proteases, cellulases, and other oxidoreductases than laccases, e.g., peroxidases and hydrogen peroxide generating oxidases.
  • enzymes such as amylases, lipases, cuti- nases, proteases, cellulases, and other oxidoreductases than laccases, e.g., peroxidases and hydrogen peroxide generating oxidases.
  • the detergent may contain 1-65% of a detergent builder or complexing agent such as zeolite, diphosphate, triphosphate, phosphonate, citrate, nitrilotriacetic acid (NTA) , ethylenediaminetetraacetic acid (EDTA) , diethylenetri- aminepentaacetic acid (DTMPA) , alkyl- or alkenylsuccinic acid, soluble silicates or layered silicates (e.g. SKS-6 from Hoechst).
  • a detergent builder or complexing agent such as zeolite, diphosphate, triphosphate, phosphonate, citrate, nitrilotriacetic acid (NTA) , ethylenediaminetetraacetic acid (EDTA) , diethylenetri- aminepentaacetic acid (DTMPA) , alkyl- or alkenylsuccinic acid, soluble silicates or layered silicates (e.g. SKS-6 from Hoech
  • the detergent may comprise one or more polymers. Examples are carboxymethylcellulose (CMC) , poly(vinyl- pyrrolidone) (PVP) , polyethyleneglycol (PEG), poly(vinyl alcohol) (PVA) , polycarboxylates such as polyacrylates, maleic/acrylic acid copolymers and lauryl methacrylate/acrylic acid copolymers.
  • the detergent may contain a bleaching system whi may comprise a H 2 0 2 source such as perborate or percarbona which may be combined with a peracid-forming bleach activat such as tetraacetylethylenedia ine (TAED) or nonanoyloxybe 5 zenesulfonate (NOBS) .
  • the bleaching system m comprise peroxyacids of, e.g., the amide, imide, or sulfo type.
  • the enzymes of the detergent composition of t invention may be stabilized using conventional stabilizi
  • composition may be formulated as described in, e.g., 92/19709 and WO 92/19708.
  • the detergent may also contain other convention detergent ingredients such as, e.g., fabric conditioners i cluding clays, foam boosters, suds suppressors, anti-corrosi agents, soil-suspending agents, anti-soil-redeposition agent dyes, bactericides, optical brighteners, or perfume.
  • fabric conditioners i cluding clays foam boosters
  • suds suppressors anti-corrosi agents
  • soil-suspending agents anti-soil-redeposition agent dyes
  • bactericides bactericides
  • optical brighteners or perfume.
  • the pH (measured in aqueous solution at use co centration) will usually be neutral or alkaline, e.g. in t range of 7-11.
  • compositions within t scope of the invention include:
  • a detergent composition formulated as a granulate having bulk density of at least 600 g/1 comprising
  • Polymers e.g. maleic/acrylic acid copolymer, PVP, PEG 0 - 3%
  • Enzymes (calculated as pure enzyme 0.0001 - 0.1% protein)
  • Minor ingredients e.g. suds suppressors, perfume, optical 0 - 5% brightener, photobleach
  • a detergent composition formulated as a granulate having a bulk density of at least 600 g/1 comprising
  • a detergent composition formulated as a granulate having bulk density of at least 600 g/1 comprising
  • Alcohol ethoxylate e.g. C 12 15 alco ⁇ hol, 7 - 14% 7 EO
  • Soap as fatty acid e.g. C 16.22 fatty 1 - 3% acid
  • Zeolite (as NaAlSiO 23 - 33%
  • Phosphonate e.g. EDTMPA 0 - 1%
  • Polymers e.g. maleic/acrylic acid copolymer, PVP, PEG 0 - 3%
  • Minor ingredients e.g. suds suppressors, perfume, optical 0 - 5% brightener
  • a detergent composition formulated as a granulate having bulk density of at least 600 g/1 comprising
  • Alcohol ethoxylate e.g. C 12 15 alco ⁇
  • Soluble silicate (as Na ? 0,2Si0 7 ) 1 - 5%
  • Zeolite (as NaAlSiO 25 - 35%
  • Carboxymethylcellulose 0 - 2%
  • Polymers e.g. maleic/acrylic acid copolymer, PVP, PEG 1 - 3%
  • Enzymes (calculated as pure enzyme 0. 0001 - 0.1% protein)
  • Minor ingredients e.g. suds 0 - 5% suppressors, perfume
  • An aqueous liquid detergent composition comprising
  • An aqueous structured liquid detergent composition compriing
  • a detergent composition formulated as a granulate having bulk density of at least 600 g/1 comprising
  • Soluble silicate (as Na.0,2Si0-.) 1 - 4%
  • Zeolite (as NaAlSiO 20 - 40%
  • Enzymes (calculated as pure enzyme 0.0001 - 0.1% protein)
  • Minor ingredients e.g. optical brightener, suds suppressors, per ⁇ 0 - 5% fume
  • a detergent composition formulated as a granulate comprising
  • a detergent composition formulated as a granulate comprising
  • Polymers e.g. polycarboxylate or 1 - 5% PEG
  • Enzymes (calculated as pure enzyme 0.0001 - 0.1% protein)
  • An aqueous liquid detergent composition comprising
  • Alcohol ethoxysulfate e.g. C 12 . l5 alcohol, 2-3 EO 8 - 15%
  • Alcohol ethoxylate e.g. C 12 . 15 al ⁇ cohol, 7 EO, or C 12 . l5 alcohol, 5 3 - 9% EO
  • Soap as fatty acid e.g. lauric 0 - 3% acid
  • Hydrotrope e.g. sodium 2 - 6% toluensulfonate
  • Enzymes (calculated as pure enzyme 0.0001 - 0.1% protein)
  • Minor ingredients e.g. polymers, dispersants, perfume, optical 0 - 5% brighteners
  • An aqueous liquid detergent composition comprising
  • Alcohol ethoxylate e.g. C 12 15 alco ⁇ hol, 6 - 12% 7 EO, or C 17 . 1c; alcohol, 5 EO
  • Polymer e.g. maleic/acrylic acid copolymer, anchoring polymer such as, e.g. , lauryl 0 - 3% ethacrylate/acrylic acid copolymer
  • Enzymes (calculated as pure enzyme 0.0001 - 0.1% protein)
  • a detergent composition formulated as a granulate having a bulk density of at least 600 g/1 comprising
  • Anionic surfactant linear alkylbenzenesulfonate, alkyl sulfa ⁇ te, alpha-olefinsulfonate, alpha- 25 - 40% sulfo fatty acid methyl esters, alkanesulfonates, soap
  • Nonionic surfactant e.g. alcohol 1 - 10% ethoxylate
  • Soluble silicates (as Na.,0, 2Si0 7 ) 5 - 15%
  • Zeolite (as NaAlSiO 15 - 28%
  • Enzymes (calculated as pure enzyme 0.0001 - 0.1% protein)
  • a detergent composition formulated as a granulate having a bulk density of at least 600 g/1 comprising
  • a detergent composition formulated as a granulate havi a bulk density of at least 600 g/1 comprising
  • Soluble silicate (as Na ? 0,2Si0 7 ) 0 - 4%
  • Polymers e.g. polycarboxylates and 0 - 3% PVP
  • Enzymes (calculated as pure enzyme 0.0001 - 0.1% protein)
  • the manganese catalyst may, e.g., be one of the compounds o described in "Efficient manganese catalysts for low-temperature bleaching", Nature 369, 1994, pp. 637-639.
  • Detergent composition formulated as a nonaqueous detergent liquid comprising a liquid nonionic surfactant such as, e.g., linear alkoxylated primary alcohol, a builder system (e.g. phosphate), enzyme and alkali.
  • a liquid nonionic surfactant such as, e.g., linear alkoxylated primary alcohol, a builder system (e.g. phosphate), enzyme and alkali.
  • the detergent may also comprise anionic surfactant and/or a bleach system.
  • the laccases of the invention may be incorporated at an enzyme protein level conventionally employed for other enzymes in detergents. It is at present contemplated that, in detergent compositions of the invention, the laccases may be added at a level corresponding to 0.00001-5 mg, preferably at a level corresponding to 0.0001-1 mg (calculated as pure enzyme protein) per liter of wash liquor.
  • Coprinus friesii Coprinus plicatilis, Panaeolus papilionace or Psathyrella condolleana and one of the following enhanci agents:
  • PPT phenothiazine-10-propionic acid
  • PPO phenoxazine-10-propionic acid
  • the strains were inoculated on PDA agar plates (PDA: g/1 potato dextrose agar) and grown at 26°C for 3 days. Sha flasks were then inoculated with 6-8 small squares ( " 0.5 cm
  • A soja meal 30 g/1 maltodextrin 15 g/1 bacto peptone 5 g/1 pluronic 0.2 g/1
  • B potato meal 50 g/1 barley meal 25 g/1 BAN 800MG* 0.025 g/1
  • the bleaching rate of DBl was determined using the following conditions:
  • Reagents were mixed in a 1 ml thermostated cuvette at 30°C and the bleaching was started by addition of the laccase.
  • the bleaching was followed spectrophoto etrically at 610 nm, which is the wavelength of the absorption peak of DBl, with readings every 5 sec. for a period of 5 minutes.
  • the initial bleaching rate was determined from the first linear part of the absorbance curve. The following results were obtained with PPT:
  • PPT phenothiazine-10-propionic acid
  • the laccase was obtained in the following way: Coprinus cinereus (IFO 30116) was inoculated from a PDA agar slant (PDA: 39 g/1 potato dextrose agar) into a 100 ml shake flask containing medium A (Medium A is described in Example 1) . The culture was cultivated for 6 days at 26°C and 100 rp . A 10-liter fermentor containing medium A was inoculated with the 100 ml culture broth. The fermentation ran for 6 days at 26°C and 100 rpm. The culture broth was filtrated and concentrated by ultrafiltration. Further purification was carried out using hydrophobic interaction chromatography followed by anionic exchange chromatography. This process resultated in a preparation with a laccase activity of 3.6 LACU/ml. The estimated purity was >80% on a protein basis.
  • the bleaching rate of DBl was determined using the following conditions: Final concentration 400 ⁇ l 50 mM Britton-Robinson buffer*,
  • the bleaching was followed spectrophotometrically at 61 nm, which is the wavelength of the absorption peak of DBl, wit readings every 5 sec. for a period of 5 minutes.
  • the initia bleaching rate was determined from the first linear part of th
  • Bleaching of the dye Direct Blue 1 at various pH values 5 was conducted using Coprinus cinereus laccase and the enhancing agent acetosyringone.
  • the laccase was obtained as described in Example 2.
  • Acetosyringone was obtained from Aldrich.
  • the bleaching rate of DBl was determined using the ⁇ o following conditions:
  • Reagents were mixed in a 1 cm thermostated cuvette at 20 30"C and the bleaching was started by addition of the laccase.
  • the bleaching was detected spectrophotometrically at 610 nm, which is the wavelength of the absorption peak of DBl. After 5 sec. bleaching was followed for 4 minutes.
  • dye transfer inhibition systems for laundry ap plications should be tested in a real wash where dyed fabric give off dyes to the wash solution as a result of the combine action of the detergent, temperature and mechanical agitatio taking place.
  • a magneticall stirred beaker was used as the reaction vessel and dye wa added gradually from a stock solution (using a Metrohm 72 dosimat) .
  • the solution was monitored spectrophotometricall using a a Zeis ⁇ multichannel spectrometer (MCS) equipped wit a fibre-optics immersion probe.
  • MCS Zeis ⁇ multichannel spectrometer
  • Stock solutions of the enhancing agents were prepare either in a suitable water/ethanol mixture or by first dissol ving the enhancing agent in a small amount of dimethylformamid and then diluting with a pH 10.0 phosphate/borate buffer. Stoc solutions of the dye Acid Blue 45 were made with water.
  • the laccase was obtained as described in Example 2.
  • Enhancing agent when applicable: 10 ⁇ M Laccase: 0.04 LACU/ml
  • Dye addition program linear addition at rates of ca 0.3 abs/40 in, referring to the absorbance of the dye at it maximum absorbance wavelength (590 nm for Acid Blue 45) .
  • enhancing agents were tested: phenoxazine 10-propionic acid and 10-ethylphenothiazine-4-carboxylic acid.
  • Fig. 1 shows the results of the bleaching tests. The following symbols are used: (I) : Only dye addition; (II) : Dye addition in the presence of Laccase; (III): Dye addition in the presence of Laccase + 10-ethylphenothiazine-4-carboxylic acid; (IV) : Dye addition in the presence of Laccase + phenoxazine-10- propionic acid.
  • Liquid detergent and powder detergent as typically met in the North American market place; both detergents contained no bleaching system.
  • Coprinus cinereus laccase obtained as described in Example 2. Washing procedure:
  • the washing processes were carried out in beakers wit magnetical stirring at 35°C for 15 min. , after which the tes fabrics were rinsed thoroughly in tap water and air-drie overnight in the dark before the Hunter readings were taken b using a Datacolor Elrephometer 2000 reflectance spectrometer
  • Treatment 1 was in each case a wash with laccase at level of 40 LACU/1 with the enhancing agent 10 ethylphenothiazine-4-carboxylic acid at a level of 10 ⁇ M.
  • Treatment 2 was in each case a wash with laccase at level of 40 LACU/1 with the enhancing agent acetosyringone a a level of 10 ⁇ M.

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Abstract

Procédé d'oxydation d'un substrat dans une solution dont le pH est de 7 ou plus, qui consiste à mettre en contact ledit substrat avec une quantité efficace d'une laccase obtenue à partir de Coprinaceae.
PCT/DK1995/000344 1994-08-26 1995-08-25 Laccases obtenues a partir de coprinaceae WO1996006930A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
AU32536/95A AU3253695A (en) 1994-08-26 1995-08-25 Coprinaceae laccases

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
DK0982/94 1994-08-26
DK98294 1994-08-26

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WO1996006930A1 true WO1996006930A1 (fr) 1996-03-07

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WO1997008325A3 (fr) * 1995-08-25 1997-04-17 Novo Nordisk Biotech Inc Laccases de coprin purifiees et acides nucleiques les codant
WO1997043384A1 (fr) * 1996-05-13 1997-11-20 The Procter & Gamble Company Detergents comportant des enzymes, protease et laccase
WO1997043382A1 (fr) * 1996-05-13 1997-11-20 The Procter & Gamble Company Composition detergente comportant une enzyme, une laccase, et un polymere s'opposant au transfert pigmentaire
WO1998027198A1 (fr) * 1996-12-19 1998-06-25 Novo Nordisk A/S Laccases mutantes
WO1998038287A1 (fr) * 1997-02-28 1998-09-03 Novo Nordisk A/S Mutants de laccases
WO1998023716A3 (fr) * 1996-11-25 1998-10-08 Unilever Nv Procede d'oxydation enzymatique
WO1999023887A1 (fr) * 1997-11-10 1999-05-20 Novo Nordisk A/S Activite antimicrobienne des lacasses
US5912405A (en) * 1994-09-27 1999-06-15 Novo Nordisk A/S Enhancers such as acetosyringone
US5998353A (en) * 1996-12-19 1999-12-07 Novo Nordisk A/S Laccase mutants
WO2000039263A1 (fr) * 1998-12-23 2000-07-06 Genencor International, Inc. Enzymes de pleurote oxydant le phenol
WO2001021748A1 (fr) * 1999-09-22 2001-03-29 Unilever N.V. Compositions detergentes contenant des enzymes d'oxydation de phenol
US6228128B1 (en) 1997-11-10 2001-05-08 Charlotte Johansen Antimicrobial activity of laccases
WO2001084937A1 (fr) * 2000-05-08 2001-11-15 Novozymes A/S Activite antimicrobienne induite par oxydoreductase
US6905853B1 (en) 2000-09-07 2005-06-14 Genencor International, Inc. Phenol oxidizing enzyme variants
US7144717B1 (en) 1998-03-24 2006-12-05 Genecor International, Inc. Oxidizing enzymes
US7183090B2 (en) 2000-05-23 2007-02-27 Valtion Teknillinen Tutkimuskeskus Laccase enzyme and the gene encoding the enzyme
EP2258837A1 (fr) 2004-09-10 2010-12-08 Novozymes North America, Inc. Procédés permettant de détruire, de réduire, d'éliminer ou d'empêcher la formation d'un film biologique
EP2431048A2 (fr) 2002-10-08 2012-03-21 Genencor International, Inc. Peptides de liaison phénolique

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WO1992018687A1 (fr) * 1991-04-12 1992-10-29 Novo Nordisk A/S Extraction des colorants excedentaires dans des matieres textiles neuves
WO1992018683A1 (fr) * 1991-04-12 1992-10-29 Novo Nordisk A/S Procede de blanchiment de textiles colores
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Cited By (32)

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US5912405A (en) * 1994-09-27 1999-06-15 Novo Nordisk A/S Enhancers such as acetosyringone
US6008029A (en) * 1995-08-25 1999-12-28 Novo Nordisk Biotech Inc. Purified coprinus laccases and nucleic acids encoding the same
US6242232B1 (en) 1995-08-25 2001-06-05 Novozymes Biotech, Inc. Purified Coprinus laccases and nucleic acids encoding same
US6207430B1 (en) 1995-08-25 2001-03-27 Novo Nordisk Of Biotech, Inc. Nucleic acids encoding polypeptides having laccase activity
WO1997008325A3 (fr) * 1995-08-25 1997-04-17 Novo Nordisk Biotech Inc Laccases de coprin purifiees et acides nucleiques les codant
WO1997043381A1 (fr) * 1996-05-13 1997-11-20 The Procter & Gamble Company Composition detergente comportant des enzymes, cellulase et laccase
WO1997043383A1 (fr) * 1996-05-13 1997-11-20 The Procter & Gamble Company Compositions detergentes comportant une enzyme, la laccase
WO1997043384A1 (fr) * 1996-05-13 1997-11-20 The Procter & Gamble Company Detergents comportant des enzymes, protease et laccase
WO1997043382A1 (fr) * 1996-05-13 1997-11-20 The Procter & Gamble Company Composition detergente comportant une enzyme, une laccase, et un polymere s'opposant au transfert pigmentaire
WO1998023716A3 (fr) * 1996-11-25 1998-10-08 Unilever Nv Procede d'oxydation enzymatique
US6080573A (en) * 1996-11-25 2000-06-27 Lever Brothers Company, Division Of Conopco, Inc. Enzymatic oxidation process
WO1998027198A1 (fr) * 1996-12-19 1998-06-25 Novo Nordisk A/S Laccases mutantes
US6277611B1 (en) 1996-12-19 2001-08-21 Novozymeo A/S Laccase mutants
US5998353A (en) * 1996-12-19 1999-12-07 Novo Nordisk A/S Laccase mutants
US6140092A (en) * 1996-12-19 2000-10-31 Novo Nordisk A/S Laccase mutants
WO1998038287A1 (fr) * 1997-02-28 1998-09-03 Novo Nordisk A/S Mutants de laccases
US6184015B1 (en) 1997-02-28 2001-02-06 Novo Nordisk A/S Laccase mutants
US7622287B2 (en) 1997-02-28 2009-11-24 Novozymes A/S Myceliophthora thermophila laccase variants
US5985818A (en) * 1997-02-28 1999-11-16 Novo Nordisk A/S Laccase mutants
WO1999023887A1 (fr) * 1997-11-10 1999-05-20 Novo Nordisk A/S Activite antimicrobienne des lacasses
US6228128B1 (en) 1997-11-10 2001-05-08 Charlotte Johansen Antimicrobial activity of laccases
US7144717B1 (en) 1998-03-24 2006-12-05 Genecor International, Inc. Oxidizing enzymes
US6329332B1 (en) 1998-12-23 2001-12-11 Genencor International, Inc. Pleurotus phenol oxidizing enzymes
WO2000039263A1 (fr) * 1998-12-23 2000-07-06 Genencor International, Inc. Enzymes de pleurote oxydant le phenol
WO2001021748A1 (fr) * 1999-09-22 2001-03-29 Unilever N.V. Compositions detergentes contenant des enzymes d'oxydation de phenol
WO2001084937A1 (fr) * 2000-05-08 2001-11-15 Novozymes A/S Activite antimicrobienne induite par oxydoreductase
US7183090B2 (en) 2000-05-23 2007-02-27 Valtion Teknillinen Tutkimuskeskus Laccase enzyme and the gene encoding the enzyme
US6905853B1 (en) 2000-09-07 2005-06-14 Genencor International, Inc. Phenol oxidizing enzyme variants
EP2431048A2 (fr) 2002-10-08 2012-03-21 Genencor International, Inc. Peptides de liaison phénolique
US8293702B2 (en) 2002-10-08 2012-10-23 Danisco Us Inc. Phenolic binding peptides
EP2258837A1 (fr) 2004-09-10 2010-12-08 Novozymes North America, Inc. Procédés permettant de détruire, de réduire, d'éliminer ou d'empêcher la formation d'un film biologique
EP2258836A1 (fr) 2004-09-10 2010-12-08 Novozymes North America, Inc. Procédés permettant de détruire, de réduire, d'éliminer ou d'empêcher la formation d'un film biologique

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