EP0702712B1 - Synergistically stabilized liquid enzymatic compositions - Google Patents

Synergistically stabilized liquid enzymatic compositions Download PDF

Info

Publication number
EP0702712B1
EP0702712B1 EP94919433A EP94919433A EP0702712B1 EP 0702712 B1 EP0702712 B1 EP 0702712B1 EP 94919433 A EP94919433 A EP 94919433A EP 94919433 A EP94919433 A EP 94919433A EP 0702712 B1 EP0702712 B1 EP 0702712B1
Authority
EP
European Patent Office
Prior art keywords
enzyme
liquid enzymatic
poly
formulation
ether
Prior art date
Legal status (The legal status 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 status listed.)
Revoked
Application number
EP94919433A
Other languages
German (de)
English (en)
French (fr)
Other versions
EP0702712A1 (en
Inventor
Percy A. Jaquess
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Buckman Laboratories International Inc
Original Assignee
Buckman Laboratories International Inc
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
Family has litigation
First worldwide family litigation filed litigation Critical https://patents.darts-ip.com/?family=22108388&utm_source=google_patent&utm_medium=platform_link&utm_campaign=public_patent_search&patent=EP0702712(B1) "Global patent litigation dataset” by Darts-ip is licensed under a Creative Commons Attribution 4.0 International License.
Application filed by Buckman Laboratories International Inc filed Critical Buckman Laboratories International Inc
Publication of EP0702712A1 publication Critical patent/EP0702712A1/en
Application granted granted Critical
Publication of EP0702712B1 publication Critical patent/EP0702712B1/en
Anticipated expiration legal-status Critical
Revoked legal-status Critical Current

Links

Classifications

    • 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
    • C11D3/38663—Stabilised liquid enzyme compositions

Definitions

  • the present invention relates to novel formulations for stabilizing at least one enzyme contained in a liquid enzymatic composition.
  • the unique rheological properties of the components of these stabilizing formulations preferably afford synergistic stabilizing capacity over other water-based mixtures of enzymatic dispersions, even under conditions of moderate to high heat and wide pH ranges.
  • the invention therefore, also relates to stabilized liquid enzymatic compositions. Additionally, the invention relates to novel methods for the preparation of stabilized liquid enzymatic compositions, and methods using the stabilizing formulations with liquid enzymatic compositions.
  • enzymes and liquid enzymatic compositions in industry and in the commercial marketplace has grown rapidly over the last several years.
  • enzymes can be acid, alkaline or neutral, depending upon the pH range in which they are active. All of these types of enzymes are contemplated to be useful in connection with the invention disclosed herein.
  • enzymes and liquid enzymatic compositions have been associated with liquid detergents and have shown utility as solubilizing and cleaning formulations. In addition to their association with liquid detergents, enzymes and liquid enzymatic compositions have also shown utility in a number of different commercial and industrial areas in which a wide variety of enzyme classes are now used.
  • Proteases are a well-known class of enzymes frequently utilized in a wide variety of industrial applications where they act to hydrolyze peptide bonds in proteins and proteinaceous substrates.
  • Commercially, the greatest uses of proteases are made in the laundry detergent industry, where they help to remove protein-based stains such as blood or egg stains, and in the cheese-making industry, where they aid in curdling milk.
  • Proteases are also used as meat tenderizers, for softening leather, for modifying food ingredients, and for flavor development.
  • Liquid enzymatic compositions containing alkaline proteases have also shown to be useful as dispersants of bacterial films and algal and fungal mats in cooling tower waters and metalworking fluid containment bays.
  • Proteases can be characterized as acid, neutral, or alkaline proteases depending upon the pH range in which they are active.
  • the acid proteases include the microbial rennets, rennin (chymosin), pepsin, and fungal acid proteases.
  • the neutral proteases include trypsin, papain, bromelain/ ficin, and bacterial neutral protease.
  • the alkaline proteases include subtilisin and related proteases.
  • Amylases another class of enzymes, have also been utilized in many industrial and commercial processes in which they act to catalyze or accelerate the hydrolysis of starch.
  • Amylases are used largely in the corn syrup industry for the production of glucose syrups, maltose syrups, and a variety of other more refined end products of starch hydrolysis such as high fructose syrups.
  • they include ⁇ -amylase, ⁇ -amylase, amyloglucosidase (glucoamylase), fungal amylase, and pullulanase.
  • liquid enzymatic compositions containing amylases are available under the names BAN, Termamyl®, AMG, Fungamyl®, and PromozymeTM, which are supplied by Novo Nordisk, and Diazyme L-200, a product of Solvay Enzyme Products.
  • cellulases are enzymes that degrade cellulose, a linear glucose polymer occurring in the cell walls of plants.
  • Hemicellulases are involved in the hydrolysis of hemicellulose which, like cellulose, is a polysaccharide found in plants.
  • the pectinases are enzymes involved in the degradation of pectin, a carbohydrate whose main component is a sugar acid.
  • ⁇ -glucanases are enzymes involved in the hydrolysis of ⁇ -glucans which are also similar to cellulose in that they are linear polymers of glucose. In a commercial context, these enzymes have utility to a greater or lesser degree in manufacturing processes dependent on fiber degradation.
  • Cellulases have reported utility in the de-inking process of old newsprint (ONP) wastepaper, eliminating the need for any surfactants and alkaline chemicals.
  • the enzymes dislodge inks from fiber surfaces and disperse ink particles to a finite size. See S. Say-Kyoun Ow, Biological De-Inking Methods of Newsprint Wastepaper, World Pulp and Paper Technology, pp. 63, 64 (1992).
  • cellulases include endocellulase, exocellulase, exocello-biohydrolase, mannase, and cellobiase.
  • Commercial liquid enzymatic compositions containing cellulases are available under the names Celluclast® and Novozym®188 which are both supplied by Novo Nordisk.
  • Hemicellulases are also used in the de-inking process to dislodge ink particles from the fiber surface of ONP. See D. Y. Prasad et al., Enzyme Deinking of Black and White Letterpress Printed Newsprint Waste, Progress in Paper Recycling, May 1992, pp. 21, 22. Additionally, hemicellulases, such as the xylanases, are employed in the pulp bleaching process. Xylanase pretreatment of kraft pulps has resulted in major reductions in bleaching chemical requirements, such as molecular chlorine, and has also improved pulp quality as reflected by higher brightness ceilings. See D. J.
  • hemicellulases include hemicellulase mixture and galactomannanase.
  • Commercial liquid enzymatic compositions containing hemicellulases are available as PULPZYM® from Novo, ECOPULP® from Alko Biotechnology and Novozym®280 and GamanaseTM, which are both products of Novo Nordisk.
  • pectinases are used commercially to weaken cell walls and enhance extraction of fruit juice, as well as to aid in decreasing viscosity and preventing gelation in these extracts.
  • Pectinases consist of endopolygalacturonase, exopoly-galacturonase, endopectate lyase (transeliminase), exopectate lyase (transeliminase), and endopectin lyase (transeliminase).
  • Commercial liquid enzymatic compositions containing pectinases are available under the names PectinexTM Ultra SP and PectinexTM*, both supplied by Novo Nordisk.
  • ⁇ -glucanases play an important role in the malting and brewing industries where modification of barley cell walls containing ⁇ -glucans is necessary.
  • ⁇ -glucanases are comprised of lichenase, laminarinase, and exoglucanase.
  • Commercial liquid enzymatic compositions containing ⁇ -glucanases are available under the names Novozym®234, Cereflo®, BAN, Finizym®, and Ceremix®, all of which are supplied by Novo Nordisk.
  • Lipases and phospholipases are esterase enzymes which hydrolyze fats and oils by attacking the ester bonds in these compounds. Lipases act on triglycerides, while phospholipases act on phospholipids. In the industrial sector, lipases and phospholipases represent the commercially available esterases, and both currently have a number of industrial and commercial applications.
  • liquid enzyme preparations containing lipases have proven to be particularly useful in reducing pitch deposits on rolls and other equipment during the production process.
  • the treatment of unbleached sulfite pulp with lipases prior to bleaching with chlorine to reduce the content of chlorinated triglycerides, which are reportedly the cause of pitch deposition during the paper manufacturing process has been reported.
  • Novo Nordisk markets two liquid lipase preparations under the names ResinaseTM A and ResinaseTM A 2X, both of which, under certain conditions, reportedly reduce pitch deposits significantly by breaking down wood resins in pulp.
  • lipases Another important use of lipases is to degrease hides and pelts in the leather-making process.
  • Alkaline lipases are used in conjunction with special proteases and emulsifying systems to aid degreasing, as well as to improve the soaking and liming effect in leather-making. See J. Christner, The Use of Lipases in the Beamhouse Processes, 87 J.A.L.C.A. 128 (1992).
  • Lipases have also been used for the development of flavors in cheese and to improve the palatability of beef tallow to dogs. In nonaqueous systems, lipases have been employed to synthesize esters from carboxylic acids and alcohols.
  • liquid enzymatic compositions containing lipases are available.
  • lipases are available under the trade names Lipolase 100, Greasex 50L, PalataseTMA, PalataseTMM, and LipozymeTM which are all supplied by Novo Nordisk.
  • pancreatic phospholipase A 2 has been used to convert lecithin into lysolecithin.
  • Lysolecithin reportedly is an excellent emulsifier in the production of mayonnaise and the baking of bread.
  • phospholipase A 2 is available in a liquid enzymatic composition sold as LECITASETM by Novo Nordisk.
  • the isomerases are particularly important in the high fructose corn syrup industry.
  • the aldose-ketose isomerase reaction catalyzed by glucose isomerase, involves the conversion of glucose to fructose and is just one of three key enzyme reactions in the industry.
  • Sweetzyme® product is a liquid enzymatic composition containing glucose isomerase which is supplied by Novo Nordisk.
  • Redox enzymes are enzymes that act as catalysts in chemical oxidation/reduction reactions and, consequently, are involved in the breakdown and synthesis of many biochemicals.
  • redox enzymes have not gained a prominent place in industry since most redox enzymes require the presence of a cofactor.
  • cofactors are an integral part of an enzyme or do not have to be supplied, redox enzymes are commercially useful, particularly in the food processing industry.
  • the redox enzyme glucose oxidase
  • Glucose oxidase is also used as an "oxygen scavenger" to prevent the development of off-flavors in juices and to preserve color and stability in certain sensitive food ingredients.
  • the redox enzyme, catalase has been utilized to decompose residual hydrogen peroxide used as a sterilizing agent.
  • redox enzymes have possible applications ranging from the enzymatic synthesis of steroid derivatives to use in diagnostic tests. These redox enzymes include peroxidase, superoxide dismutase, alcohol oxidase, polyphenol oxidase, xanthine oxidase, sulfhydryl oxidase, hydroxylases, cholesterol oxidase, laccase, alcohol dehydrogenase, and steroid dehydrogenases.
  • enzymes When enzymes, such as those described above, are prepared or sold for use in industrial processes, they generally are formulated as water-based or aqueous liquid enzymatic compositions designed for a particular process. Water-based liquid enzymatic compositions may contain additional solvents depending upon the particular enzyme or use of the composition. These liquid enzymatic compositions, however, have historically been plagued with problems such as chemical instability which can result in the loss of enzymatic activity, particularly upon storage. This critical problem of loss of enzymatic activity upon storage has particularly affected the liquid detergent industry.
  • the stabilization of an aqueous enzyme preparation using certain esters has been described in U.S. Patent No. 4,548,727.
  • the ester used as a stabilizer has the formula, RCOOR', where R is an alkyl of from one to three carbons or hydrogen, and R' is an alkyl of from one to six carbons.
  • the ester is present in the aqueous enzyme preparation in an amount from 0.1 to about 2.5% by weight.
  • U.S. Patent No. 4,318,818 describes a stabilizing system for aqueous enzyme compositions where the stabilizing system comprises calcium ions and a low molecular weight carboxylic acid or its salt.
  • the pH of the stabilizing system is from about 6.5 to about 10.
  • U.S. Patent No. 4,243,543 teaches the stabilization of liquid proteolytic enzyme-containing detergent compositions.
  • the detergent compositions are stabilized by adding an antioxidant and a hydrophilic polyol to the composition while stabilizing the pH of the composition.
  • U.S. Patent No. 4,169,817 teaches a liquid cleaning composition containing stabilized enzymes.
  • the composition is an aqueous solution containing from 10% to 50% by weight of solids and including detergent builders, surface active agents, an enzyme system derived from Bacillus subtilis and an enzyme stabilizing agent.
  • the stabilizing agents comprise highly water soluble sodium or potassium salts and/or water soluble hydroxy alcohols and enable the solution to be stored for extended periods without deactivation of the enzymes.
  • European Patent No. 0 352 244 A2 describes stabilized liquid detergent compositions using an amphoteric surfactant.
  • the present invention provides a formulation capable of synergistically stabilizing one or more enzymes contained in a liquid enzymatic composition.
  • the invention thus, also provides stabilized liquid enzymatic compositions.
  • the invention provides methods for the preparation of stabilized liquid enzymatic compositions.
  • a formulation for stabilizing a liquid enzymatic composition comprising:
  • the inventive stabilizing formulation can be used with a wide variety of enzymes utilized in liquid enzymatic compositions performing a wide variety of functions.
  • the enzymes and classes of enzymes with which this stabilizing formulation can be used include, but are not limited to, those discussed above.
  • the invention also relates to a stabilized liquid enzymatic composition
  • a stabilized liquid enzymatic composition comprising:
  • the invention further relates to a method for the preparation of a stabilized liquid enzymatic composition by combining an enzyme with the stabilizing formulation above. Additionally, the invention relates to a method of using the stabilizing formulation to stabilize a liquid enzymatic formulation comprising the step of combining the stabilizing formulation with a liquid enzymatic composition.
  • the invention provides a formulation for stabilizing a liquid enzymatic composition comprising:
  • a water-soluble, or at least partially water-soluble, polymer is used in a formulation for stabilizing a liquid enzymatic composition or a stabilized liquid enzymatic compositions of the invention. That is, the polymer must have sufficient solubility to be miscible with water and form a single phase. Having this solubility, the polymer should not separate out when combined with the C 2 -C 6 polyhydric alcohol and water of a stabilizing formulation or with a liquid enzymatic composition.
  • the formulation is not required to be a clear solution.
  • the formulation is an emulsion having an apparent homogeneous texture.
  • the amount of polymer present also depends on the molecular weight of the particular polymer used. The higher the molecular weight of the polymer used, the lower the amount of polymer generally required to stabilize an enzyme.
  • the polymer may preferably be used in amounts up to about 50% by weight of the stabilizing formulation. More preferably, the polymer is present from 0.05 to 30% by weight, and most preferably from 1% to 10% by weight. In a preferred embodiment, the polymer, of course, is present in an amount that gives the desired synergistic stabilization of a liquid enzymatic composition when combined with the polyhydric alcohol in a water-based or aqueous formulation.
  • the polymer employed in the present invention is poly(cellulosic)ethers.
  • the polymer may be substituted or unsubstituted.
  • the polymer may have a slight ionic charge, but is preferably non-ionic in nature.
  • the polymer is preferably selected from poly(carboxymethylcellulose)ether, poly(hydroxypropylmethylcellulose)ether, poly(hydroxyethylmethylcellulose)ether, poly(hydroxybutylmethylcellulose)ether, poly(hydroxypropylcellulose)ether, and poly(ethylhydroxyethylcellulose)ether. More preferably, the polymer is poly(carboxymethylcellulose)ether.
  • Certain poly(cellulosic)ethers used in the present invention, such as poly(carboxymethylcellulose)ether, are sold as salts, i.e. sodium salts, and possess a slight anionic charge.
  • Preferable poly(cellulosic)ethers are those with molecular weights ranging from 15,000 to 100,000, but more preferred are those with molecular weights ranging from 20,000 to 75,000.
  • the stabilizing formulation also contains a C 2 -C 6 polyhydric alcohol as a second component.
  • the C 2 -C 6 polyhydric alcohol acts synergistically with the polymer, described above, to stabilize an enzyme in a liquid enzyme composition.
  • the C 2 -C 6 polyhydric alcohol is preferably selected from a glycol and a trihydric alcohol. More preferably, the C 2 -C 6 polyhydric alcohol is glycerol, sorbitol, propylene glycol, butylene glycol, hexylene glycol, or ethylene glycol. Most preferably, the C 2 -C 6 polyhydric alcohol is glycerol.
  • the stabilizing formulation contains the C 2 -C 6 polyhydric alcohol in an amount sufficient, with the polymer, to stabilize at least one enzyme in a liquid enzymatic composition.
  • the C 2 -C 6 polyhydric alcohol present is 0.50 to 60% by weight of the stabilizing formulation, more preferably, 5 to 50% by weight and, even more preferably, between 10 and 30%.
  • a stabilizing formulation or enzymatic composition contains the C 2 -C 6 polyhydric alcohol in a combined amount with the polymer to achieve synergistic stabilization.
  • the formulations of the invention are water-based or aqueous formulations containing sufficient water to allow the polymer to be miscible with the formulation and not separate out.
  • the C 2 -C 6 polyhydric alcohol is soluble in water, but sufficient water should be present to allow the formulation to form a single phase.
  • the formulation is not required to be a clear solution and preferably is an emulsion having an apparent homogeneous texture.
  • Water-based formulations may contain additional solvents other than water.
  • the stabilizing formulations of the invention can be prepared by mixing the components in any order, the formulations are preferably prepared by adding the desired amount of polymer to a C 2 -C 6 polyhydric alcohol/water mixture.
  • the C 2 -C 6 polyhydric alcohol/water mixture can be prepared by means known in the art.
  • the mixture can be prepared by simply mixing the desired polyhydric alcohol with an appropriate amount of water, or diluting a previously prepared mixture.
  • the preferred mixture of the C 2 -C 6 polyhydric alcohol and water may contain any percentage of C 2 -C 6 alcohol sufficient with the polymer discussed above to stabilize, preferably synergistically, at least one enzyme in a liquid enzymatic composition.
  • the mixture is 1-95% by weight of the polyhydric alcohol, or water-soluble polymer thereof; more preferably 10-50% by weight; and most preferably, 30-50% by weight.
  • the polyhydric alcohol is glycerol
  • the mixture is preferably a 50% by weight glycerol/ water mixture.
  • applicant believes that the mixture acts to wet the polymer used in the present invention and the C 2 -C 6 polyhydric alcohol in the mixture, to synergistically stabilize an enzyme.
  • the enzyme should possess at least 90% activity after 30 days at 25°C.
  • the examples below demonstrate the preferred synergistic stabilization of various enzymes at 50°C after 30 days.
  • the stabilizing formulation described here can be employed with a wide variety of enzymes and industrial processes or commercial products.
  • the enzymes, industrial processes and commercial products with which this stabilizing formulation can be used include, but are not limited to, those previously discussed.
  • the invention also relates to a stabilized liquid enzymatic composition
  • a stabilized liquid enzymatic composition comprising at least one poly(cellulosic)ether; a C 2 -C 6 polyhydric alcohol; water; and at least one enzyme.
  • the polymer and C 2 -C 6 polyhydric alcohol are present in a combined amount effective to stabilize, preferably to synergistically stabilize, at least one enzyme contained in the liquid enzymatic composition.
  • Preferred compositions according to the invention are capable of developing greater viscosities than quantitatively proportional aqueous mixtures with the same polymers.
  • the liquid enzymatic composition of this invention can be practiced with a wide variety of enzymes.
  • enzymes include, but are not limited to, the enzyme classes and specific enzymes heretofore discussed.
  • Enzymes that may be used are derived from animal, plant, fungal, bacterial, and synthetic sources.
  • Preferable water dispersible enzymes for this system are proteases, including acid, alkaline, and neutral proteases, which are widely used in the laundry detergent and cheese making industries; amylases, including acid, alkaline, and neutral amylases, used, for example, in the corn syrup industry; lipases, used in developing flavors in cheese, and in the pulp and paper and leather making industries; cellulases, and xylases.
  • enzymes are often packaged and sold in concentrated liquid enzymatic compositions to be diluted prior to use. Enzymes can also be supplied in powdered or desiccated form. The amount of enzyme present after dilution of the concentrated enzyme depends on the form in which the enzyme is supplied. In general, the amount of enzyme preferably may range from 0.05 to 40% by weight of a concentrated liquid enzymatic composition, more preferably 0.5 to 25%, and most preferably 10 to 20%.
  • the stabilizing formulation of this invention is specifically contemplated for use in concentrated liquid enzymatic compositions as well as with compositions already diluted for use.
  • the amount of stabilizing formulation needed to stabilize, or to synergistically stabilize, a concentrated solution can, and most likely will, differ from that for a diluted composition. Determining the appropriate quantity of stabilizing formulation or its components can be readily ascertained by one of ordinary skill in the art using the method set out in the examples below. As known in the art, the amount of enzyme present, however, is dependent upon the activity of the particular enzyme and the desired end use.
  • the pH of the final stabilized liquid ⁇ enzymatic composition is preferably from 5.0 to 10.0, but more preferably around 7.0. Most preferably, the system should be allowed to seek its own pH, generally around neutral. But, as understood in the art, adjustment of pH may be necessary with a small amount of acidic or alkaline material.
  • the stabilized liquid enzymatic composition may be water-based or aqueous and contain other solvents or additives directed toward the use of the composition in a particular industrial process.
  • the stabilized liquid enzymatic composition can contain additives such as surfactants, defoamers, and the like, as are known in the art.
  • additives such as surfactants, defoamers, and the like, as are known in the art.
  • synergistic stabilization such additives may be added in amounts not interfering with the synergistic stabilization of the liquid enzymatic composition.
  • the stabilizing formulation may also act as a dispersant aid for the enzyme in industrial process waters.
  • the invention also relates to a method for the preparation of a stabilized liquid enzymatic composition comprising the step of combining at least one enzyme with the inventive stabilizing formulation.
  • the invention further relates to a method of using the stabilizing formulation to stabilize a liquid enzymatic composition comprising the step of combining a liquid enzymatic composition with the stabilizing formulation.
  • Illustrative and preferred components, as well as the amounts of the components used in the method, are the same as discussed above.
  • An alternative method for the preparation of a stabilized liquid enzymatic composition comprises the steps of:
  • the polymer added in step (b) may be added alone, as an aqueous dispersion, or as a solution where the polymer is dissolved in water or a suitable organic solvent.
  • additives may be added at any time, but preferably prior to step (c), or in a separate step after the enzyme is added.
  • Synergism was demonstrated by testing glycerol, designated as Compound A, and poly(carboxymethylcellulose)ether, (CMC), having a 250,000 molecular weight, designated as Compound B.
  • CMC poly(carboxymethylcellulose)ether
  • Tables 1 and 2 experiments were set up by varying ratios of Compound A to Compound B over a range of concentrations and assaying various types of enzymes for enhanced stabilization of enzymatic activity at 50°C for 30 days. The concentration of each compound required for an assay of 90% of the original enzymatic activity was taken as an end point. The concentrations are expressed as percent by weight of the compound in the final composition including the added enzyme with the balance being water. Water was added to compound A, the glycerol to form a glycerol-water mixture. End points for the compositions containing Compound A and Compound B were then compared with the end points for Compound A alone and Compound B alone.
  • Synergism was determined by the method described by Kull, F.C., Eisman, P.C., Sylwestrowicz, H.D., and Mayer, R.L., Applied Microbiology 9: 538-541 (1961) employing the ratio determined by QA Qa + QB Qb where
  • Stabilizing formulations of the invention as in Example 1 were tested with another enzyme, Diazyme L-200, a glucoamylase sold by Solvay Enzymes, Inc.
  • Diazyme L-200 a glucoamylase sold by Solvay Enzymes, Inc.
  • the results, set forth in Tables 3 and 4 also demonstrate enhanced stabilization with this enzyme.
  • the sums of QA/Qa + QB/Qb for all compositions containing glycerol (Compound A) and CMC (Compound B) were calculated. Sample calculations are shown for some end points where synergism was evident in this example. As set forth in Table 2 and the sample calculations, these end point values were 0.59, 0.67, 0.67, and 0.84, respectively, indicating the existence of synergism.
  • Enzymes with esterase activity were tested with stabilizing formulations according to the invention as in Example 1.
  • the results obtained with the enzyme, Lipolase, a lipase sold by Novo Nordisk Bioindustries, Inc., are set forth in Table 3 and indicate the enhanced stabilization of this enzyme.
  • the sums of QA/Qa + QB/Qb for all compositions containing aqueous glycerol (Compound A) and CMC (Compound B) were calculated. Sample calculations are shown for some end points where synergism was evident in this example. As set forth in Table 3 and the sample calculations, these end point values were 0.43, 0.37, 0.57, and 0.77, respectively, indicating the existence of synergism.
  • the hemicellulase, Pulpzyme HB, a xylanase sold by Novo Nordisk Bioindustrials, Inc. was selected for enhanced stability formulation testing as in Example 1.
  • the following results obtained for this example are set forth in Table 4.
  • the sums of QA/Qa + QB/Qb for all compositions containing glycerol (Compound A) and CMC (Compound B) were calculated. Sample calculations are shown for those end points where synergism was evident in this example. As set forth in Table 7 and the sample calculations, these end point values were 0.43, 0.53, 0.57, and 0.77, respectively, indicating the existence of synergism.

Landscapes

  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Oil, Petroleum & Natural Gas (AREA)
  • Wood Science & Technology (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Enzymes And Modification Thereof (AREA)
  • Medicines Containing Plant Substances (AREA)
  • Medicines Containing Material From Animals Or Micro-Organisms (AREA)
  • Cosmetics (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Detergent Compositions (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
EP94919433A 1993-06-07 1994-06-06 Synergistically stabilized liquid enzymatic compositions Revoked EP0702712B1 (en)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
US7256093A 1993-06-07 1993-06-07
US72560 1993-06-07
PCT/US1994/006522 WO1994029424A1 (en) 1993-06-07 1994-06-06 Synergistically stabilized liquid enzymatic compositions

Publications (2)

Publication Number Publication Date
EP0702712A1 EP0702712A1 (en) 1996-03-27
EP0702712B1 true EP0702712B1 (en) 1998-12-23

Family

ID=22108388

Family Applications (1)

Application Number Title Priority Date Filing Date
EP94919433A Revoked EP0702712B1 (en) 1993-06-07 1994-06-06 Synergistically stabilized liquid enzymatic compositions

Country Status (18)

Country Link
EP (1) EP0702712B1 (sv)
JP (1) JPH08510786A (sv)
CN (1) CN1128543A (sv)
AT (1) ATE174956T1 (sv)
AU (1) AU7058694A (sv)
BR (1) BR9407029A (sv)
CA (1) CA2164615A1 (sv)
CZ (1) CZ323095A3 (sv)
DE (1) DE69415524T2 (sv)
DK (1) DK0702712T3 (sv)
ES (1) ES2126764T3 (sv)
FI (1) FI955851A0 (sv)
NO (1) NO954957L (sv)
NZ (1) NZ267909A (sv)
SG (1) SG104908A1 (sv)
SK (1) SK153695A3 (sv)
WO (1) WO1994029424A1 (sv)
ZA (1) ZA943640B (sv)

Cited By (16)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7569226B2 (en) 2001-12-22 2009-08-04 Henkel Kommanditgesellschaft Auf Aktien (Henkel Kgaa) Alkaline protease from Bacillus sp. (DSM 14392) and washing and cleaning products comprising said alkaline protease
US7666618B2 (en) 2004-07-16 2010-02-23 Danisco A/S Lipolytic enzyme: uses thereof in the food industry
US7718204B2 (en) 1998-07-21 2010-05-18 Danisco A/S Foodstuff
US7718408B2 (en) 2003-12-24 2010-05-18 Danisco A/S Method
US7803604B2 (en) 2000-07-28 2010-09-28 Henkel Ag & Co. Kgaa Amylolytic enzyme extracted from Bacillus sp. A 7-7 (DSM 12368) and washing and cleaning agents containing this novel amylolytic enzyme
US7807398B2 (en) 2003-01-17 2010-10-05 Danisco A/S Method of using lipid acyltransferase
US7888104B2 (en) 2000-11-28 2011-02-15 Henkel Ag & Co. Kgaa Cyclodextrin glucanotransferase (CGTase), obtained from<I>Bacillus agaradherens<λ>(DSM 9948) and detergents and cleaning agents containing said novel cyclodextrin glucanotransferase
US7906307B2 (en) 2003-12-24 2011-03-15 Danisco A/S Variant lipid acyltransferases and methods of making
US7955814B2 (en) 2003-01-17 2011-06-07 Danisco A/S Method
US7960150B2 (en) 2007-01-25 2011-06-14 Danisco A/S Production of a lipid acyltransferase from transformed Bacillus licheniformis cells
US8012732B2 (en) 2004-03-12 2011-09-06 Danisco A/S Fungal lypolytic and amylase enzyme composition and methods using the same
US8030044B2 (en) 2003-12-24 2011-10-04 Danisco A/S Lipid acyltransferases
USRE43135E1 (en) 2001-05-18 2012-01-24 Danisco A/S Method of improving dough and bread quality
USRE43341E1 (en) 1995-06-07 2012-05-01 Danisco A/S Method of improving the properties of a flour dough, a flour dough improving composition and improved food products
US8652809B2 (en) 2007-08-17 2014-02-18 Dupont Nutrition Biosciences Aps Method for producing ultra-heat treatment milk
US8753858B2 (en) 2009-12-31 2014-06-17 Shenzhen Mindray Bio-Medical Electronics Co., Ltd. Reagents and processes for stabilizing alkaline phosphatase or conjugates thereof

Families Citing this family (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6342381B1 (en) 1998-02-27 2002-01-29 Buckman Laboratories Internationals, Inc. Enzyme stabilization with pre-superpolyamide or pre-fiber-forming polyamide oligomers
WO1999057986A1 (en) * 1998-05-13 1999-11-18 Novo Nordisk Biotech, Inc. Methods for using dehydrogenases in baking
ATE487796T1 (de) 2003-01-17 2010-11-15 Danisco Verfahren zur in-situ-herstellung eines emulgators in einem nahrungsmittel
EP2505642B1 (en) * 2009-11-24 2016-09-21 YFY Biopulp Technology Limited Xylanase composition and method for production thereof
CN102269761A (zh) 2010-06-04 2011-12-07 深圳迈瑞生物医疗电子股份有限公司 一种碱性磷酸酶结合物的合成工艺
CN102269762B (zh) 2010-06-04 2014-12-10 深圳迈瑞生物医疗电子股份有限公司 结合物的制备方法及相关试剂盒
MX2016005852A (es) 2013-11-11 2016-07-13 Ecolab Usa Inc Detergente de lavado de utensilios altamente alcalino con control de incrustaciones y dispersion de suciedad mejorados.
CN105705625A (zh) 2013-11-11 2016-06-22 艺康美国股份有限公司 多用途的酶洗涤剂和稳定使用溶液的方法
EA036683B1 (ru) 2015-04-10 2020-12-08 Комет Биорефайнинг Инк. Способы и составы для обработки целлюлозной биомассы и продукты на ее основе
AU2019265921A1 (en) 2018-05-10 2020-11-26 Comet Biorefining Inc. Compositions comprising glucose and hemicellulose and their use

Family Cites Families (12)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
GB758049A (en) * 1953-04-06 1956-09-26 Armour & Co Improvements in or relating to prolonged-action local anaesthetics
GB942305A (en) * 1958-12-23 1963-11-20 Parachem Corp Improvements in or relating to antiseptic film and method and apparatus for producing the same
DE2038103A1 (de) * 1970-07-31 1972-02-10 Henkel & Cie Gmbh Waessrige Reinigungsmittelkonzentrate mit einem Gehalt an stabilisierten Enzymen
JPS5536317B2 (sv) * 1972-04-22 1980-09-19
SU889689A1 (ru) * 1979-09-05 1981-12-15 Всесоюзный научно-исследовательский и проектный институт химической промышленности Моющее средство дл стирки
IT1129814B (it) * 1980-07-02 1986-06-11 Unilever Nv Composizione detergente enzimatica liquida
FR2488797A1 (fr) * 1980-08-19 1982-02-26 Lhd Lab Hygiene Dietetique Composition dermatologique, son procede de preparation et son application dans le domaine des pansements
FR2519552B1 (fr) * 1982-01-12 1987-02-27 Mechin Jean Claude Procede pour stabiliser le lysozyme dans une composition liquide ou semi-liquide, et composition stabilisee a base de lysozyme
FR2531337A1 (fr) * 1982-08-04 1984-02-10 Hoechst Lab Nouvelles formes galeniques administrables par voie orale, des derives de la dimethylxanthine solubles
SE468518B (sv) * 1985-07-26 1993-02-01 Colgate Palmolive Co Stabiliserad textilmjukgoerande enzyminnehaallande foerstaerkt flytande detergentkomposition och dess anvaendning vid tvaett av textilier
JPH03127988A (ja) * 1989-10-13 1991-05-31 Central Glass Co Ltd 酵素の安定化方法
DE4211883C1 (en) * 1992-04-09 1993-03-04 Desitin Arzneimittel Gmbh, 2000 Hamburg, De Stable, heat-sterilisable carbamazepine soln. for parenteral admin. - comprises carbamazepine dissolved in tetra:hydro:furfuryl alcohol, polyethylene glycol ether, water and polyvinyl pyrrolidone

Cited By (26)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
USRE43341E1 (en) 1995-06-07 2012-05-01 Danisco A/S Method of improving the properties of a flour dough, a flour dough improving composition and improved food products
US7972638B2 (en) 1998-07-21 2011-07-05 Danisco A/S Foodstuff
US7718204B2 (en) 1998-07-21 2010-05-18 Danisco A/S Foodstuff
US7781001B2 (en) 1998-07-21 2010-08-24 Danisco A/S Foodstuff
US8163315B2 (en) 1998-07-21 2012-04-24 Danisco A/S Foodstuff
US7803604B2 (en) 2000-07-28 2010-09-28 Henkel Ag & Co. Kgaa Amylolytic enzyme extracted from Bacillus sp. A 7-7 (DSM 12368) and washing and cleaning agents containing this novel amylolytic enzyme
US7888104B2 (en) 2000-11-28 2011-02-15 Henkel Ag & Co. Kgaa Cyclodextrin glucanotransferase (CGTase), obtained from<I>Bacillus agaradherens<λ>(DSM 9948) and detergents and cleaning agents containing said novel cyclodextrin glucanotransferase
USRE43135E1 (en) 2001-05-18 2012-01-24 Danisco A/S Method of improving dough and bread quality
US7569226B2 (en) 2001-12-22 2009-08-04 Henkel Kommanditgesellschaft Auf Aktien (Henkel Kgaa) Alkaline protease from Bacillus sp. (DSM 14392) and washing and cleaning products comprising said alkaline protease
US8278062B2 (en) 2003-01-14 2012-10-02 Dupont Nutrition Biosciences Aps Method of using lipid acyltransferase
US8003095B2 (en) 2003-01-17 2011-08-23 Danisco A/S Method of using lipid acyltransferase
US7955814B2 (en) 2003-01-17 2011-06-07 Danisco A/S Method
US7807398B2 (en) 2003-01-17 2010-10-05 Danisco A/S Method of using lipid acyltransferase
US7955813B2 (en) 2003-01-17 2011-06-07 Danisco, A/S Method of using lipid acyltransferase
US8440435B2 (en) 2003-12-24 2013-05-14 Dupont Nutrition Biosciences Aps Method for reducing 1,2-diglyceride content of an edible oil
US7718408B2 (en) 2003-12-24 2010-05-18 Danisco A/S Method
US8030044B2 (en) 2003-12-24 2011-10-04 Danisco A/S Lipid acyltransferases
US7906307B2 (en) 2003-12-24 2011-03-15 Danisco A/S Variant lipid acyltransferases and methods of making
US8012732B2 (en) 2004-03-12 2011-09-06 Danisco A/S Fungal lypolytic and amylase enzyme composition and methods using the same
US7666618B2 (en) 2004-07-16 2010-02-23 Danisco A/S Lipolytic enzyme: uses thereof in the food industry
US8192782B2 (en) 2004-07-16 2012-06-05 Danisco A/S Enzymatic oil-degumming method
US8535900B2 (en) 2004-07-16 2013-09-17 Dupont Nutrition Biosciences Aps Lipolytic enzyme uses thereof in the food industry
US8889371B2 (en) 2004-07-16 2014-11-18 Dupont Nutrition Biosciences Aps Lipolytic enzyme: uses thereof in the food industry
US7960150B2 (en) 2007-01-25 2011-06-14 Danisco A/S Production of a lipid acyltransferase from transformed Bacillus licheniformis cells
US8652809B2 (en) 2007-08-17 2014-02-18 Dupont Nutrition Biosciences Aps Method for producing ultra-heat treatment milk
US8753858B2 (en) 2009-12-31 2014-06-17 Shenzhen Mindray Bio-Medical Electronics Co., Ltd. Reagents and processes for stabilizing alkaline phosphatase or conjugates thereof

Also Published As

Publication number Publication date
NZ267909A (en) 1996-09-25
DE69415524D1 (de) 1999-02-04
ATE174956T1 (de) 1999-01-15
CN1128543A (zh) 1996-08-07
BR9407029A (pt) 1996-03-19
FI955851A (sv) 1995-12-05
ES2126764T3 (es) 1999-04-01
NO954957L (no) 1995-12-07
AU7058694A (en) 1995-01-03
CZ323095A3 (en) 1996-05-15
SG104908A1 (en) 2004-07-30
CA2164615A1 (en) 1994-12-22
DE69415524T2 (de) 1999-05-20
JPH08510786A (ja) 1996-11-12
NO954957D0 (no) 1995-12-06
DK0702712T3 (da) 1999-08-23
SK153695A3 (en) 1996-04-03
FI955851A0 (sv) 1995-12-05
EP0702712A1 (en) 1996-03-27
WO1994029424A1 (en) 1994-12-22
ZA943640B (en) 1995-01-26

Similar Documents

Publication Publication Date Title
EP0702712B1 (en) Synergistically stabilized liquid enzymatic compositions
EP0670894B1 (en) Stabilized liquid enzymatic compositions
US5780283A (en) Enzyme stabilization by oxygen-containing block copolymers
CA2180366C (en) Enzymes for recreational water
US6342381B1 (en) Enzyme stabilization with pre-superpolyamide or pre-fiber-forming polyamide oligomers
NZ314287A (en) Stabilized liquid enzyme compositions

Legal Events

Date Code Title Description
PUAI Public reference made under article 153(3) epc to a published international application that has entered the european phase

Free format text: ORIGINAL CODE: 0009012

17P Request for examination filed

Effective date: 19960105

AK Designated contracting states

Kind code of ref document: A1

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

RAX Requested extension states of the european patent have changed

Free format text: SI PAYMENT 960105

17Q First examination report despatched

Effective date: 19960610

GRAG Despatch of communication of intention to grant

Free format text: ORIGINAL CODE: EPIDOS AGRA

GRAG Despatch of communication of intention to grant

Free format text: ORIGINAL CODE: EPIDOS AGRA

GRAH Despatch of communication of intention to grant a patent

Free format text: ORIGINAL CODE: EPIDOS IGRA

GRAH Despatch of communication of intention to grant a patent

Free format text: ORIGINAL CODE: EPIDOS IGRA

GRAA (expected) grant

Free format text: ORIGINAL CODE: 0009210

AK Designated contracting states

Kind code of ref document: B1

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

AX Request for extension of the european patent

Free format text: SI PAYMENT 960105

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: LI

Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES

Effective date: 19981223

Ref country code: CH

Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES

Effective date: 19981223

REF Corresponds to:

Ref document number: 174956

Country of ref document: AT

Date of ref document: 19990115

Kind code of ref document: T

REG Reference to a national code

Ref country code: CH

Ref legal event code: NV

Representative=s name: A. BRAUN, BRAUN, HERITIER, ESCHMANN AG PATENTANWAE

Ref country code: CH

Ref legal event code: EP

REF Corresponds to:

Ref document number: 69415524

Country of ref document: DE

Date of ref document: 19990204

REG Reference to a national code

Ref country code: IE

Ref legal event code: FG4D

ITF It: translation for a ep patent filed
REG Reference to a national code

Ref country code: ES

Ref legal event code: FG2A

Ref document number: 2126764

Country of ref document: ES

Kind code of ref document: T3

ET Fr: translation filed
REG Reference to a national code

Ref country code: PT

Ref legal event code: SC4A

Free format text: AVAILABILITY OF NATIONAL TRANSLATION

Effective date: 19981231

PGFP Annual fee paid to national office [announced via postgrant information from national office to epo]

Ref country code: NL

Payment date: 19990630

Year of fee payment: 6

Ref country code: DK

Payment date: 19990630

Year of fee payment: 6

PGFP Annual fee paid to national office [announced via postgrant information from national office to epo]

Ref country code: PT

Payment date: 19990702

Year of fee payment: 6

Ref country code: IE

Payment date: 19990702

Year of fee payment: 6

PGFP Annual fee paid to national office [announced via postgrant information from national office to epo]

Ref country code: GR

Payment date: 19990705

Year of fee payment: 6

PGFP Annual fee paid to national office [announced via postgrant information from national office to epo]

Ref country code: LU

Payment date: 19990706

Year of fee payment: 6

PGFP Annual fee paid to national office [announced via postgrant information from national office to epo]

Ref country code: MC

Payment date: 19990707

Year of fee payment: 6

PGFP Annual fee paid to national office [announced via postgrant information from national office to epo]

Ref country code: CH

Payment date: 19990709

Year of fee payment: 6

REG Reference to a national code

Ref country code: DK

Ref legal event code: T3

PLBQ Unpublished change to opponent data

Free format text: ORIGINAL CODE: EPIDOS OPPO

PLBI Opposition filed

Free format text: ORIGINAL CODE: 0009260

PLBQ Unpublished change to opponent data

Free format text: ORIGINAL CODE: EPIDOS OPPO

PLBI Opposition filed

Free format text: ORIGINAL CODE: 0009260

PLBF Reply of patent proprietor to notice(s) of opposition

Free format text: ORIGINAL CODE: EPIDOS OBSO

26 Opposition filed

Opponent name: HENKEL KGAA

Effective date: 19990923

26 Opposition filed

Opponent name: UNILEVER N.V.

Effective date: 19990922

Opponent name: HENKEL KGAA

Effective date: 19990923

NLR1 Nl: opposition has been filed with the epo

Opponent name: HENKEL KGAA

NLR1 Nl: opposition has been filed with the epo

Opponent name: UNILEVER N.V.

Opponent name: HENKEL KGAA

PLBF Reply of patent proprietor to notice(s) of opposition

Free format text: ORIGINAL CODE: EPIDOS OBSO

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: LU

Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES

Effective date: 20000606

Ref country code: IE

Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES

Effective date: 20000606

Ref country code: DK

Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES

Effective date: 20000606

PGFP Annual fee paid to national office [announced via postgrant information from national office to epo]

Ref country code: GB

Payment date: 20000629

Year of fee payment: 7

PGFP Annual fee paid to national office [announced via postgrant information from national office to epo]

Ref country code: SE

Payment date: 20000630

Year of fee payment: 7

Ref country code: FR

Payment date: 20000630

Year of fee payment: 7

Ref country code: AT

Payment date: 20000630

Year of fee payment: 7

PGFP Annual fee paid to national office [announced via postgrant information from national office to epo]

Ref country code: ES

Payment date: 20000705

Year of fee payment: 7

PGFP Annual fee paid to national office [announced via postgrant information from national office to epo]

Ref country code: BE

Payment date: 20000714

Year of fee payment: 7

PGFP Annual fee paid to national office [announced via postgrant information from national office to epo]

Ref country code: DE

Payment date: 20000727

Year of fee payment: 7

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: PT

Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES

Effective date: 20001231

Ref country code: MC

Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES

Effective date: 20001231

Ref country code: GR

Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES

Effective date: 20001231

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: NL

Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES

Effective date: 20010101

REG Reference to a national code

Ref country code: DK

Ref legal event code: EBP

REG Reference to a national code

Ref country code: CH

Ref legal event code: PL

NLV4 Nl: lapsed or anulled due to non-payment of the annual fee

Effective date: 20010101

RDAH Patent revoked

Free format text: ORIGINAL CODE: EPIDOS REVO

REG Reference to a national code

Ref country code: IE

Ref legal event code: MM4A

REG Reference to a national code

Ref country code: PT

Ref legal event code: MM4A

Free format text: LAPSE DUE TO NON-PAYMENT OF FEES

Effective date: 20001231

RDAG Patent revoked

Free format text: ORIGINAL CODE: 0009271

STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: PATENT REVOKED

27W Patent revoked

Effective date: 20010419

GBPR Gb: patent revoked under art. 102 of the ep convention designating the uk as contracting state

Free format text: 20010419

REG Reference to a national code

Ref country code: PT

Ref legal event code: MF4A

Effective date: 20010903

PLAB Opposition data, opponent's data or that of the opponent's representative modified

Free format text: ORIGINAL CODE: 0009299OPPO