[go: up one dir, main page]

WO1994010298A1 - Stabilization of hemoproteins - Google Patents

Stabilization of hemoproteins Download PDF

Info

Publication number
WO1994010298A1
WO1994010298A1 PCT/DK1993/000345 DK9300345W WO9410298A1 WO 1994010298 A1 WO1994010298 A1 WO 1994010298A1 DK 9300345 W DK9300345 W DK 9300345W WO 9410298 A1 WO9410298 A1 WO 9410298A1
Authority
WO
WIPO (PCT)
Prior art keywords
peroxidase
stabilizer
alkyl
activity
hydrogen peroxide
Prior art date
Application number
PCT/DK1993/000345
Other languages
English (en)
French (fr)
Inventor
Kaj André HOLM
Niels Krebs Lange
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 EP93924531A priority Critical patent/EP0669975A1/en
Priority to JP6510574A priority patent/JPH08502411A/ja
Publication of WO1994010298A1 publication Critical patent/WO1994010298A1/en

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.)
    • C12N9/0065—Oxidoreductases (1.) acting on hydrogen peroxide as acceptor (1.11)
    • 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
    • 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
    • C12N9/96—Stabilising an enzyme by forming an adduct or a composition; Forming enzyme conjugates
    • D—TEXTILES; PAPER
    • D06—TREATMENT OF TEXTILES OR THE LIKE; LAUNDERING; FLEXIBLE MATERIALS NOT OTHERWISE PROVIDED FOR
    • D06L—DRY-CLEANING, WASHING OR BLEACHING FIBRES, FILAMENTS, THREADS, YARNS, FABRICS, FEATHERS OR MADE-UP FIBROUS GOODS; BLEACHING LEATHER OR FURS
    • D06L4/00—Bleaching fibres, filaments, threads, yarns, fabrics, feathers or made-up fibrous goods; Bleaching leather or furs
    • D06L4/10—Bleaching fibres, filaments, threads, yarns, fabrics, feathers or made-up fibrous goods; Bleaching leather or furs using agents which develop oxygen
    • D06L4/12—Bleaching fibres, filaments, threads, yarns, fabrics, feathers or made-up fibrous goods; Bleaching leather or furs using agents which develop oxygen combined with specific additives

Definitions

  • This invention relates to the stabilization of hemoproteins, particularly oxidoreductases such as peroxidase or catalase.
  • oxidoreductases such as peroxidase or catalase are known to be hemoproteins. They can be obtained from many sources, e.g. from horseradish and microorganisms such as Coprinus (EP 179,486), Humicola or Scytalidium (WO 92/17571). Peroxidase can be added to a detergent together with a hydrogen peroxide precursor to improve the bleaching of stains (WO 89/09813) or to inhibit the transfer of dye from one textile to another (WO 91/05839). Other known uses of peroxidase are in the pulp and paper industry (EP 418,201 , EP 429,422, EP 433,258) and in production of particle boards (US 4,432,921).
  • Catalase may be used for removing residual hydrogen peroxide after bleaching, e.g. of textiles. It is in some cases desirable to improve the stability of hemoproteins against denaturation and loss of enzyme activity, and it is the object of this invention to provide a method of stabilizing hemoproteins in solution.
  • alkoxylated non-ionic compounds such as polyethylene glycol and alkylphenol ethoxylate, have a strong stabilizing effect on hemoproteins such as peroxidase in solution.
  • US-A-4,414,326 and EP-A-0 272 578 describe the use of alkoxylated compounds of the above type in an enzymatic reagent composition for the determination of cholesterol.
  • the compositions contained cholesterol esterase, cholesterol oxidase and peroxidase. Said documents describe activation and stabilization of cholesterol esterase and activation of cholesterol oxidase (US-A- 4,414,326 at col. 9 I. 26-28; EP-A-0 272 578 at col. 2 I. 18-20). No effect of the alkoxylated compound on peroxidase was described. Use of cholesterol oxidase and cholesterol esterase is not contemplated in the present invention.
  • the invention provides a method of stabilizing a hemoprotein in an aqueous solution, characterized by incorporating a stabilizer of the formula:
  • R-0-(CHR'-CH 2 -0-) n H (I) wherein R H or C ⁇ C ⁇ alkyl, aryl or alkylaryl
  • the invention also provides a process comprising addition of peroxidase and hydrogen peroxide or a precursor thereof, characterized by incorporating said stabilizer.
  • the invention also provides a peroxidase preparation for use in said bleaching composition or said process, in the form of a slurry, a non-dusting granulate or a stabilized liquid, characterized by comprising said stabilizer.
  • the method of the invention may be applied to any hemoprotein, e.g. an oxidoreductase.
  • hemoprotein e.g. an oxidoreductase.
  • peroxidase EC 1.11.1.7
  • Coprinus particularly C. cinereus, vide EP 179,486
  • lignin peroxidase ligninase
  • Phanerochaete chrysosporium or catalase e.g. from Humicola or Scytalidium
  • haloperoxidases i.e. peroxidases that utilize halide ions and hydrogen peroxide to form carbon-halogen bonds in substrates, e.g.
  • the hemoprotein may be produced by recombinant DNA technology (WO 92/16634).
  • Other examples are cellobiose oxidase or cytochrome oxidase
  • PODU Peroxidase Unit
  • the invention provides increased activity, in addition to increased stability.
  • the chain length (n) is preferably 50-500, particularly 100-300.
  • the stabilizer may be alcohol ethoxylate or alkyl-phenol ethoxylate (propoxylate), where R is a C 10 -C 18 linear or branched alkyl or alkyl-phenyl group.
  • the chain length (n) is preferably 5-50, particularly 5-20.
  • the amount of said stabilizer should be such that the weight ratio of stabilizer to hemoprotein is 1-50.
  • Suitable amounts of stabilizer are 0.1-5 g/l of aqueous solution, 1-30% by weight of bleaching composition or 1 -10% by weight of peroxidase preparation.
  • a suitable ratio of stabilizer and peroxidase corresponds to 0.2-5 mg of stabilizer per PODU of peroxidase activity (unit defined below).
  • the stabilizer is typically used at pH in the range 4 - 10, more preferably 6 - 8. Hydrogen peroxide source
  • the source of hydrogen peroxide can be hydrogen peroxide itself, a precursor thereof or an enzymatic system capable of generating it.
  • the precursor of hydrogen peroxide may be an inorganic or organic peracid or salt thereof, e.g. sodium perborate or sodium percarbonate.
  • the enzymatic system may be a combination of a suitable oxidase and a substrate thereof, e.g. glucose and glucose oxidase.
  • peroxidase or haloperoxidase may be used together with hydrogen peroxide or a precursor thereof e.g. in a process for bleaching of pulp for paper production (SE 88/0673, EP 418,201), in treatment of waste water from pulp production (US 4,623,465, JP-A 2-31887), in production of mechanical pulp from a fibrous product (EP 429,422, EP 433,258) and for lignin modification, e.g. in particle board production (US 4,432,921).
  • Catalase may be used to remove hydrogen peroxide from fabric which is bleached by an alkaline hydrogen peroxide treatment before dyeing (GB 2,216,149; JP 2-104781).
  • the peroxidase preparation of the invention may be provided as a slurry, a granulate or stabilized liquid, in a form suited for storage and shipping.
  • Enzyme granulates may be made by known methods, e.g. by extrusion or drum granulation according to CA 974,907, US 4,106,991, US 4,435,307 or US 4,661 ,452.
  • the stabilizer may be mixed with other ingredients in the core of the granules, preferably at above 1% by weight, preferably above 2%, particularly above 5%, and generally below 40%.
  • the stabilizer may be incorporated by coating a preformed enzyme granulate. The amount of the stabilizer in the coating should make up at least 1% by weight of the coated particles.
  • the coating may be applied by known methods e.g. as described in British Patent No. 1 ,362,365, page 1 , line 82 to page 2, line 34, and British Patent application No. 34973/73 and 10842/74 corresponding to Belgium patent No. 146,802.
  • the coating material may comprise, in addition to the ethoxylated compound, other waxy coating materials and particulate material, e.g. magnesium silicate, alumino silicates, CaSO 4I CaCO 3 , bentonite, zeolite or TiO 2 .
  • a liquid enzyme preparation according to the invention may be stabilized against microbial infection by incorporation of a stabilizing agent known in the art.
  • a stabilizing agent known in the art.
  • examples are inorganic salts (such as NaCI), sugars (such as sucrose and glucose), polyols (such as glycerol, propylene glycol and sorbitol) and alcohols
  • organic acids such as benzoic, sorbic, propionic, lactic and formic
  • stabilizing agents are antioxidants (such as sulphur dioxide), 1 ,2-benz-iso-thiazolin-3-one (BIT) and parabens.
  • the amount of the dispersant in the liquid enzyme preparation is preferably 1-10% by weight.
  • An enzyme slurry may be produced according to WO 91/09941.
  • Peroxidase activity is determined by an assay based on the oxidation of 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonate) (ABTS ® ) by hydrogen peroxide (as described by Bergmeyer H.U., Methods of enzymatic analysis, 3rd edition, vol III, p. 286-293, 1983, modified). The greenish-blue colour produced is photometered at 418 nm.
  • the analytical conditions are 0.88 mM hydrogen peroxide, 1.67 mM ABTS, 0.1 M phosphate buffer, pH 7.0, 30°C incubated for 3 minutes.
  • 1 peroxidase unit is the amount of enzyme that catalyzes the conversion of 1 xmol hydrogen peroxide per minute at these conditions.
  • the concentration of Triton X 405 in the peroxidase samples was varied from 0 to 1.5 g/l before incubation under the conditions described in Example 2, except that the temperature was 0°C.
  • the enzyme activity was determined at 0, 1.5 and 3.5 hour.
  • Table 3 It appears from Table 3 that an improved peroxidase stability is obtained by addition of 1 or 1.5 g/l of Triton X 405 to the 0.2 ml peroxidase sample (corresponding to a concentration of Triton X 405 of about 0.1 g/l during incubation).
  • the concentration of Coprinus peroxidase in the samples was varied from 0.3 - 0.6 PODU/ml to 3 - 6 PODU/ml.
  • the concentration of Triton X 405 in the peroxidase sample was 1.5 g/l.
  • Incubation with ABTS ® was carried out as described in Example 2, except that the temperature was 22°C.
  • the enzyme activity was determined at 0, 1 , 2, and 3 hours. The results are shown in Table 4.
  • Triton X 405 stabilizes the peroxidase activity for up to 3 hours at both the lower and the higher peroxidase concentrations.
  • a catalase solution was obtained from a culture broth filtrate prepared according to WO 92/17571 by adding 15% propylene glycol and adjusting to pH 7.0. Stabilizer was added as shown in Table 5. Each solution was stored at 25 or 37°C for 2 or 4 weeks, the catalase activity was determined before and after storage, and results were expressed as residual activity in % of the initial activity. The same stabilizers were used as in Example 1.

Landscapes

  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Health & Medical Sciences (AREA)
  • Organic Chemistry (AREA)
  • Wood Science & Technology (AREA)
  • Genetics & Genomics (AREA)
  • Zoology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Microbiology (AREA)
  • Biotechnology (AREA)
  • Biomedical Technology (AREA)
  • Molecular Biology (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Oil, Petroleum & Natural Gas (AREA)
  • Textile Engineering (AREA)
  • Enzymes And Modification Thereof (AREA)
PCT/DK1993/000345 1992-10-28 1993-10-28 Stabilization of hemoproteins WO1994010298A1 (en)

Priority Applications (2)

Application Number Priority Date Filing Date Title
EP93924531A EP0669975A1 (en) 1992-10-28 1993-10-28 Stabilization of hemoproteins
JP6510574A JPH08502411A (ja) 1992-10-28 1993-10-28 ヘムタンパク質の安定化

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
DK1317/92 1992-10-28
DK921317A DK131792D0 (ja) 1992-10-28 1992-10-28

Publications (1)

Publication Number Publication Date
WO1994010298A1 true WO1994010298A1 (en) 1994-05-11

Family

ID=8103498

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/DK1993/000345 WO1994010298A1 (en) 1992-10-28 1993-10-28 Stabilization of hemoproteins

Country Status (4)

Country Link
EP (1) EP0669975A1 (ja)
JP (1) JPH08502411A (ja)
DK (1) DK131792D0 (ja)
WO (1) WO1994010298A1 (ja)

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN109294547A (zh) * 2018-10-23 2019-02-01 天津大港油田滨港集团博弘石油化工有限公司 一种驱油用阴非离子两性表面活性剂及其制备方法
CN113813372A (zh) * 2021-09-26 2021-12-21 云锦华彰(北京)生物科技有限公司 血红素组合物、药物组合物及其应用

Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6136578A (en) * 1997-03-12 2000-10-24 Novo Nordisk A/S Storage-stable liquid formulation comprising a laccase
WO2014034822A1 (ja) * 2012-08-31 2014-03-06 協和メデックス株式会社 カタラーゼの安定化方法

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4414326A (en) * 1978-04-24 1983-11-08 Goldberg Jack M Diagnostic agents
EP0152847A1 (en) * 1984-02-03 1985-08-28 Abbott Laboratories Stabilized enzyme conjugate composition

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4414326A (en) * 1978-04-24 1983-11-08 Goldberg Jack M Diagnostic agents
EP0152847A1 (en) * 1984-02-03 1985-08-28 Abbott Laboratories Stabilized enzyme conjugate composition

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
PATENT ABSTRACTS OF JAPAN, Vol. 9, No. 78, C-274; & JP,A,59 210 885 (Toyo Boseki K.K.), 29 November 1984 (29.11.84). *

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN109294547A (zh) * 2018-10-23 2019-02-01 天津大港油田滨港集团博弘石油化工有限公司 一种驱油用阴非离子两性表面活性剂及其制备方法
CN109294547B (zh) * 2018-10-23 2020-11-20 天津大港油田滨港集团博弘石油化工有限公司 一种驱油用阴非离子两性表面活性剂及其制备方法
CN113813372A (zh) * 2021-09-26 2021-12-21 云锦华彰(北京)生物科技有限公司 血红素组合物、药物组合物及其应用

Also Published As

Publication number Publication date
DK131792D0 (ja) 1992-10-28
EP0669975A1 (en) 1995-09-06
JPH08502411A (ja) 1996-03-19

Similar Documents

Publication Publication Date Title
US5700769A (en) Enhancement of enzyme reactions
US5885304A (en) Enhancement of laccase reactions
EP0424398B1 (en) A detergent additive for bleaching fabric
KR100303620B1 (ko) 효소반응의증강법
US5912405A (en) Enhancers such as acetosyringone
JPH08503371A (ja) 酵素反応の強化
FI118341B (fi) Entsyymin stabilointi segmenttikopolymeereilla
US5474701A (en) Enzymes for recreational water
US5451337A (en) Dye transfer inhibition system containing a peroxidase/accelerator system
PT95553A (pt) Processo para inibir a transferencia de corantes
US6409770B1 (en) Bleaching and washing agents with enzyme bleaching system
JPH07504694A (ja) 洗剤組成物
EP0669975A1 (en) Stabilization of hemoproteins
US5286404A (en) Liquid enzymatic detergent composition
DE3369792D1 (en) Improved time-stable liquid cholesterol assay compositions
US6380142B1 (en) Storage-stable formulations comprising mediators, peroxide compounds and pH stabilizers and their use in enzymatic bleaching systems
Sutherland et al. The effect of manganese on the oxidation of chemicals by lignin peroxidase
JP3589870B2 (ja) 酵素によるピッチトラブル防止法
KR930002967B1 (ko) 변기 세정제 조성물
CA2201334C (en) Enhancers such as acetosyringone
Hirai et al. In vitro reduction of manganese dioxide by a ferrireductase system from the white-rot fungus Phanerochaete sordida YK-624
CN1154139A (zh) 含有过氧化物酶/促进剂体系的染料转移抑制体系

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A1

Designated state(s): JP US

AL Designated countries for regional patents

Kind code of ref document: A1

Designated state(s): AT BE CH DE DK ES FR GB GR IE IT LU MC NL PT SE

DFPE Request for preliminary examination filed prior to expiration of 19th month from priority date (pct application filed before 20040101)
121 Ep: the epo has been informed by wipo that ep was designated in this application
WWE Wipo information: entry into national phase

Ref document number: 1993924531

Country of ref document: EP

ENP Entry into the national phase

Ref country code: US

Ref document number: 1995 411837

Date of ref document: 19950403

Kind code of ref document: A

Format of ref document f/p: F

WWP Wipo information: published in national office

Ref document number: 1993924531

Country of ref document: EP

WWW Wipo information: withdrawn in national office

Ref document number: 1993924531

Country of ref document: EP