JPS59206497A - Stabilized aqueous enzyme composition - Google Patents

Abstract

alpha -Amylase derived from B. licheniformis is stabilized in aqueous liquid detergent compositions containing surfactants and sequestering agents by the inclusion of a calcium-containing component that provides at least about 1 x 10<-4.5> millimole of enzyme-accessible calcium per liter of composition.

Description

TECHNICAL FIELD 1, FIELD OF THE INVENTION The present invention relates to stabilized aqueous enzyme compositions containing detergent ingredients.

2. Description of the Technology The formulation of enzyme-containing aqueous liquid detergent compositions is difficult due to the tendency of enzyme activity to rapidly decrease within the aqueous medium upon storage. U.S. Patent No. 4,111,855, U.S. Patent No. 4,287,082, U.S. Patent No. 4,3
No. 05,837 and U.S. Patent No. 4.318,8
No. 18 discloses an approach to the stabilization of enzymes, all in aqueous media.

DISCLOSURE OF THE INVENTION The stabilized aqueous enzyme compositions of the present invention contain (a) from about 1% to about 75% of a non-soap detergent surfactant.1. (blPanrus licheniformis (Bacillus lichenifor)
α-amylolytic enzyme derived from approximately 0. '0
1% to about 5%, (C) Kinzan Io/Sequestering agent about 0.25% to
about 40%, (at least about 1 x 1 per d1 composition 1)
0 mmol of enzyme-available calcium salt (
enzyme-acce-ssible calciu
m 1 on)) a Ll sium-containing component, and (e) about 10% to about 80% water.

Stabilization of enzymes in aqueous media has been the subject of substantial research, with particular effort directed toward compositions such as liquid detergent compositions that face relatively long storage periods and sometimes extreme temperatures before use. is the subject of.

Proteolytic enzymes (proteases) and α-amylases are very useful cleaning agents due to their ability to recycle or degrade soils containing tough bacterial and carbohydrate components, respectively. It has been proven to be a force aid.

Although the use of calcium to stabilize enzymes in aqueous media is well known, this stabilization method is incompatible with the incorporation of detergency builder ingredients into liquid detergent compositions. Effective Detergency The primary function of Burmé ingredients is to remove metal ions other than alkali metal ions from the cleaning solution by metal ion/sequestration or precipitation. These undesirables 1
Metal ions, especially calclum ions and magnesium ions, are commonly referred to as water hardness. The incorporation of such detergency builder ingredients into liquid detergent compositions clearly complicates enzyme stability issues.

By the practice of the present invention, the α-amylase derived from B. licheniformis and the below-mentioned very low amount of carunium ionic power; It is surprising that α-amylase can be stabilized in liquid detergent compositions containing carunium sequestering agents when given the formulation of carunum-containing acids according to a mathematical formula that includes an equilibrium constant of complexation. Especially now.

ing.

Non-soap detergent surfactants include non-ionic surfactants, anionic surfactants, cationic/surfactants, zwitterionic surfactants, amphoteric surfactants, semipolar medio/surfactants and the like. may be selected from a mixture of The surfactant preferably comprises from about 10 to about 65%, more preferably from about 10% to about 65% by weight of the formulation.

Nonionic Surfactants One useful nonionic surfactant is to combine ethylene oxide with a reactive hydrogen atom, such as a hydroxyl group, a carboxyl group,
It is produced by condensation with a hydrocarbon having an amino group or an amide group in the presence of an acid or base catalyst. Such nonionic surfactants have the general formula RA(CH
2CH20)nH, where R represents a hydrophobic moiety, A represents a group having a reactive hydrogen atom, and n represents the average number of ethylene oxide moieties. R typically has about 8 to 22 carbon atoms, but can also be produced by condensation of propylene oquinide with a low molecular weight compound. n can vary from about 2 to about s depending on the desired physical and detergency properties. The hydrophobic part of a nonionic compound is
Preferred are primary or secondary straight or slightly branched aliphatic alcohols having a carbon number of from about 8 to about 1, preferably from about 12 to about 1,000.

Alkylamides/oquinodos and other semipolar nonionic surfactants are discussed below. Another class of useful nonionic surfactants include hydrophobic groups having from about 8 to about 1 carbon atoms and about 1
．． It is an alkyl polysaccharide having a polysaccharide hydrophilic group containing from 5 to about 10 sugar units.

A more complete disclosure of suitable nonionic surfactants is found in the aforementioned US Pat. No. 4,111,855.

Anionic surfactant synthesis The anionic surfactant has the general formula R803M (wherein,
R1 represents a hydrocarbon group selected from the group consisting of a straight or branched alkyl group having from about 8 to about 15 carbon atoms and an alkylphenyl group having from about 9 to about 15 carbon atoms in the alkyl group. can be expressed. M is a salt-forming cation typically selected from the group consisting of sodium, potassium, ammonium, monoalkanol ammonium, dialkanol ammonium, trialkanol/monium and magnesium cations and mixtures thereof.

Preferred synthetic anionic surfactants are, for example, water-soluble salts of alkylbenzene sulfonic acids having from about 9 to about 15 carbon atoms in the alkyl group and water-soluble alkyl sulfates having from about 10 to about 18 carbon atoms.

Another preferred synthetic anionic surfactant has an alkyl group having from about 8 to about 18 carbon atoms, preferably from about 10 to about 18 carbon atoms, and from about 1 to about 20, preferably from about 1 to about 12, Etkyne groups. It is a water-soluble salt of alkyl polyethoxycrate ether sulfuric acid. Other suitable anionic surfactants are:
Disclosed in US Pat. No. 4,170,565.

Cationic Surfactants Suitable cationic surfactants have the general formula %5, where each R is optionally substituted with up to 3 phenyl groups or hydroxy7 groups and optionally has the following formula: an organic group containing a straight or branched alkyl group f, or alkenyl group interrupted by up to four selected structures, each R having from about 8 to about 22 carbon atoms.
and can additionally contain up to about 12 ethylene oquinodo groups, m being a number from 1 to 3, and each R3 being an alkyl group having 1 to 4 carbon atoms or a hydroquinoalkyl group or 1 This is a benzyl group, 831 or fewer in the molecule are perzyl, X is a number from O to 11, the rest of the carbon atom positions are filled by hydrogen, and Y is fil-N+-.

e f3+-P+-.

■ (41S+- ■ selected from the group consisting of (C2H40), H (C2H40)', H (9) and mixtures thereof, and 2 is an anion, such as halide, methyl sulfate or hydroxide).

One R3 can also be a proton. The resulting tertiary amines can have properties similar to cationic surfactants at wash liquor pH values below about 8.5.

A more complete disclosure of cationic surfactants can be found in U.S. Pat.
, 228,044, "Laundry Detergent Compositions with Enhanced Particulate Soil Removal and Anti-Redeposition Performance."

Compatibility must be considered when cationic surfactants are used in combination with anionic surfactants. Cationic surfactants of the type that are generally compatible with anionic surfactants include C8-□8 alkyl tri-C1-3
Alkylammonium chloride or 08-□8 alkyl triC□-3 alkylammonium methyl sulfate.

Zwitterionic Surfactants Zwitterionic surfactants are those in which the aliphatic portion can be straight or branched and one of the aliphatic substituents has about 8 carbon atoms.
-24 and one containing an anionic water-solubilizing group, derivatives of aliphatic quaternary ammonium compounds, phosphonium compounds and sulfonium compounds. Particularly preferred zwitterionic surfactants are the ethoquinated ammonium sulfonates and ethoquinated ammonium sulfates disclosed in U.S. Pat. Nos. 3,925,262 and 3,929,678.

Amphoteric Surfactants Amphoteric surfactants are those in which the aliphatic portion can be straight or branched and one of the aliphatic substituents has about 8 carbon atoms.
- an aliphatic heterocyclic secondary having an aqueous properties and in which at least one aliphatic substituent contains an anionic hydration group;
and tertiary amine derivatives.

Semi-polar non-ionic surfactants Semi-polar non-ionic surfactants are made from alkyl or hydroquinoalkyl moieties with a carbon number of about 8 to about 1 and an alkyl group having from 1 to about 3 carbon atoms and a hydroquinoalkyl group. a water-soluble amine oxide containing two moieties selected from the group consisting of (optionally combined to form a ring structure); one alkyl or hydroquinoalkyl moiety with a carbon number of about 8 to about a water-soluble phosphine oxyto containing two moieties selected from the group consisting of an alkyl group having about 1 to about 3 carbon atoms and a hydroquinoalkyl group; and an alkyl moiety having a carbon number of about 8 to about a or 1 hydroquine alkyl moiety and 1 or more carbon atoms
water-soluble sulfoquinodo containing one moiety selected from the group consisting of alkyl moieties of 3 and hydroquinoalkyl moieties.

A more complete disclosure of compounds suitable for inclusion in detergent compositions is found in U.S. Pat. No. 4,056,481, no.
Specification No. 049,586, Specification No. 4.040,988, Specification No. 4,035,257 1゛, No. 4,033,
Specification No. 718, Specification No. 4,019,999, No. 4
,019,998 specification and No. 3,985,669
found in the specification.

Enzymes Compositions of the present invention contain from about 0.01 to about 5% by weight, preferably from about 0.05 to about 5%, based on standard enzymes, of α-de/pun-degrading enzyme (α-amylase) derived from B. licheniformis. 2 Mm%, most preferably about 0.1 to about 1.0% by weight
Contains. Commercial enzyme preparations, especially those prepared for use in detergent compositions, are rarely pure enzymes, and in each case are generally mixed with various inert substances. Commercial formulations will typically have a small amount of calcium metal uncomplexed with the enzyme as part of the inactive portion of the formulation. As discussed below, this calcium content must be recognized for stability considerations. For purposes of describing the present invention and preferred enzyme amounts, the standard enzyme reference is K11o Novo.
) Measured in α-amylase units (KNU) 120
Defined as an active preparation of KNUL&. One kilo novo alpha-amylase unit (IKNU) is calculated by the following conditions:
Rum content 0.0043M Reaction time 7
Starch 5 + 269 hours (Merck, Amylum S
olubileErg, B, 6, Batch
This is the amount of enzyme that destroys 9947275). As described in Patent No. 1,296,839, the Novo method (Novo I/$Stree A of Pagspard, Denmark)
/S) is a modification of the SKB method achieved at 37°C without the addition of β-amylase.

The SKB method is based on Cereal Chemistry 16
, 712 (1939).

α-Amylase derived from B. licheniformis is
It has been characterized as a carbohydrolase, more specifically as a 1,4-α-D-gluca/glucanohydrolase. α-Amylases used in detergent compositions, particularly granular detergent compositions, are generally derived from Bacillus subtilis.
), but suitable B. licheniformis-derived alpha-amylases for use in the compositions of the present invention are available from a number of sources. Termamyl 120 L, registered trademark, manufactured by Novo Industries A/S, Nonogesvaard, Denmark, and Taka-Therm L-3, manufactured by Miles Riboradores, Elkhard, Indiana.
40. Rovarase (registered trademark) manufactured by Rohm & De Haas, Estophila Terfia, Pe/Nornonia.
halase) AT, a registered trademark maki-length mill (Maxam
yl) HT is an α-amylase derived from B. licheniformis and is suitable for use in the compositions of the invention.

The report deposit numbers of Noce/Las licheniformis that can produce α-amylase are NCI B 8061, NCIB
8059, ATCC6634, ATCC6598, AT
CC11945, ATCC8480 and ATCC99
It is 458. GB 1,296,839 discloses a process for the preparation of α-amylase derived from B. licheniformis.

Sequestering agents The compositions of the present invention contain approximately 0.25 Cal/Ram sequestrants.
% to about 40%, preferably about 3% to about 30%, most preferably about 5% to about 25%. These metal ion sequestrants act as detergents to improve cleaning, especially when the cleaning solution contains metal ions other than alkali metal ions. Sequestration involves the formation of coordination complexes between the sequestering agent and the total condensed ions in solution to reduce the interaction of Cal/Ram with other substances in the wash solution. As used herein, the term "sequestering agent" includes polydentate ligands that can act as chelating agents, and can include some ion exchange materials, but only by precipitation reactions. Does not include cleaning power buildup substances that are removed from the solution.

Sequestering agents used in the compositions of the invention are, for example, polyphosphates, polyphosphonates and polycarpokylates in the form of soluble salts or acids.

Polyphosphates capable of sequestering cal/lam ions are characterized by the general formula (-valent cations) n+2Pn03n+□ and include the acid forms of birophosphate and triboriphosphate as well as alkali metal salts, ammonium salts and substituted ammonium salts and water-soluble Consists of polymetaphosphate. For use in aqueous liquid detergent compositions, birophosphoric acid and its salts are preferred because of their stability. Aqueous solutions of triphosphate and polymetaphosphate tend to be mixtures of pyrophosphate and orthophosphate. The latter has the ability to precipitate calcium/ium, but does not sequester metal ions.

Polyphosphonates contain two or more of the following groups -C-PO3M
It consists of a wide range of organic compounds having a basic structure (where M is a frozen or salt-forming group). Suitable phosphonates are, for example, ethane-1-hydroquine-1,1-diphosphonate,
ethanehydroquine-1°1.2-) diphosphonates, and their oligomeric ester chain condensates. Particularly suitable polyphosphonates for use in the compositions of the invention are nitrogen-containing polyphosphonates such as ethylenediaminetetramethylenephosphonic acid and diethylenetriaminepentamethylenephosphonic acid and their alkali metal, ammonium and substituted ammonium salts. .

Suitable polycarboxylates are, for example, citric acid, ascorbic acid, phytic acid, mellitic acid, pe/zenpentacarboxylic acid, oxynediacetic acid, carboxy/methyloxine succinic acid, carboxylmethyloxy7malo7, cis-7chloro The acid forms and alkali metal, ammonium and substituted ammonium salts of hexanehexacarpo7 acid, cis-7 clopentanetetracarboxylic acid and oxydisuccinic acid. Polycarboxylate materials described in U.S. Pat. No. 3,364,103 and U.S. Pat.
．． Also suitable are the polycarphokyflate polymers and copolymers described in US Pat. No. 308,067.

Due to its stability in aqueous media, U.S. Pat. No. 4,144,226 and U.S. Pat. No. 4.14
The polyacecool carboxylates disclosed in US Pat. No. 6,495 may be incorporated into the compositions of the present invention.

Particularly suitable polycarboxylates are those containing nitrogen, such as ethylenediaminetetraacetic acid, hydroxyethylenediaminetriacetic acid, diethylenetriami/pe/triacetic acid and nitrilotriacetic acid and their alkali metal, ammonium and substituted ammonium salts. .

As detailed by the examples below, combinations of metal ion M-chain agents with varying degrees of calcium ion sequestering power are particularly useful in the practice of this invention. Nitrogen-containing compound selected from the group consisting of ethylenediaminetetramethylenephosphonic acid, diethylenetriaminepe/tamethylenenophosphonic acid, nitrilotriacetic acid, ethylenediaminetetraacetic acid, hydroquineethylenediaminetriacetic acid, diethylenetriaminepe/lyacetic acid or mixtures and suitable salts thereof. Particularly preferred is the combination of sequestering agents and citric acid. Citric acid has a relatively lower equilibrium constant for complexing with calcium than the nitrogen-containing sequestering agents. A preferred weight ratio of citric acid to nitrogen-containing sequestering agent is about 50=1 to about 1:2, more preferably about 30=1 to about 2:1.

Fermentation-available Cal/Rum Ion The stabilized aqueous enzyme compositions of the present invention contain a calfuram-containing acid content that provides at least about 1X10-'5 mmol of enzyme-available calcium (described below) per liter of said composition. Preferably, sufficient calcium-containing ingredients are included to provide at least about 1X10-' mmol of enzyme-available calcium, more preferably at least about 1X10-' mmol. Most preferably, the composition contains at least about IX],0; mol of enzyme-available calcium per composition.

The calcium-containing component can be or be part of other essential or optional components, such as anionic surfactants in the form of calcium salts or calcium-sequestering agent complexes. can. More typically, water-soluble or solubilizing calcium salts, such as calcium chloride, are used. In the aqueous liquid detergent compositions of the present invention, the different forms of insoluble, sequestrant and enzyme-available calcium will reach equilibrium as a function of composition ingredients and pH, regardless of the initial state of the calcium-containing ingredients. Dew.

The upper limit of enzyme-available calcium within the compositions of the invention may be set by practical considerations unrelated to enzyme stability. The stability of α-amylase derived from B. licheniformiscala in the compositions of the present invention is hardly improved at enzyme-available calcium levels of I×10 ζ mol/or more. In preferred compositions of the invention, this amount of enzyme-available calcium is provided by a total lucium content of about 1 to about 3% by weight of the composition. Composition i in a washing machine with a washing liquid capacity of 18 gal.
In the recommended usage amount of the cup, total carunium 0.18
% (from 120-5% CaC) increases the water hardness of the cleaning fluid by about 1 grain/gallon (as CaCO3).

Calcium is generally detrimental to detergency, but the grains per gallon are almost negligible. The calfuram-containing acid content partially reduces the sequestering capacity of the sequestering agent of the composition as far as calcium is concerned. but,
In the cleaning solution, the calcium-sequestering agent complex is nevertheless capable of sequestering heavy metal ions/ions associated with several types of staining problems, such as iron ions, manganese ions, and copper ions. This is force/L/V
This is the result of a higher complexation and formation constant of the sequestering agent with heavy metal ions compared to the constant with Lamb.

Enzyme-available calcium resembles free or ionic calcium in a less complex aqueous solution than the compositions of the present invention.

For the purposes of the present invention, the amount of enzyme-available calcium is determined from the following publications: 51 Morgan, F. Morel, Env.
, Sci, Tech, 6, 58-67 (1972
) It is measured by the method described in ``Method of subtracting equilibrium in an aqueous chemical system''.

For the purposes of the present invention, the method is based on the following assumptions: 1) 0
．． 5) a fixed ionic strength, 2) a pH fixed to the pH of the composition, and 3) divalent (e.g., calcium) metals and polydentate ligands (sequestering agents of the invention) only considered. and 4) used to calculate the enzyme-available calcium in the compositions of the invention by using the free or uncomplexed calcium concentration to be the enzyme-available calcium concentration.

Other methods of calculating the equilibrium concentration of all species in an aqueous solution give essentially equivalent results and are also known to those skilled in the art.

It should be noted that alkaline earth metals other than calcium are substantially less effective in stabilizing the compositions of the present invention.

The compositions of the present invention contain from about 10% to about 60% by weight water, preferably from about 15% to about 60% by weight. A preferred composition is
It has a pH of about 6.5 to about 9.0.

Optionally Incorporated Components The composition of the present invention may contain components other than those disclosed as essential.

The compositions of the present invention may contain enzymes other than α-amylase derived from B. licheniformis, particularly enzymes degrading tanx. Examples of suitable tan-A degrading enzymes are the many enzyme preparations that are suitable for use in detergent compositions and are actually used in detergent compositions. Enzyme sources are available, for example, from commercial enzyme preparations such as the registered open-collared alcalase (A1calase) sold by Novo Industries A/S, Copenhage/Denmark, and by Gist Brokers, Delft, The Netherlands. Registered trademark Maxatase
). Another preferred enzyme composition is manufactured and sold by Novo Industrie, eA/S, Copenhageno, Denmark under the trade name Esperase.
ase) and “Az-Protease (
AZ-Protease), which is commercially available.

Enzyme-available calcium in amounts greater than or equal to about 1×10−2 mmol provides excellent stability to proteases incorporated into the compositions of the present invention;
It should be noted that the combination of tannoki degrading enzyme is a preferred embodiment.

However, over a narrow range of enzyme-available Cal/Ram amounts from IX10'' to lXl0-4, the α-amylase (derived from B. licheniformis) A more complete disclosure of suitable enzymes can be found in U.S. Patent No. 4.101.
, No. 457.

Compositions of the invention can contain solvents other than water.

Low molecular weight imaginary primary or secondary alcohols are preferred, exemplified by methanol, ethanol, sol/ni-nor and isopro, 6-nor. Monohydric alcohols are preferred for solubilizing surfactants, but contain 2 to about 6 carbon atoms.
and polyols containing from 2 to about 6 hydroquino groups can be used and provide improved enzyme stability. Examples of polyols are propylene glycol, ethylene glycol, glycerin and 1,2-propanediol. Ethanol is a particularly preferred alcohol. The composition contains 0% to about 30% alcohol, preferably about 1% to about 15%.

Short-chain carboxylates can be used to stabilize enzymes, particularly lysiolytic enzymes, as disclosed in US Pat. No. 4,318,818. Short chain canopoquinates are preferably formates, such as formic acid and its salts. Formate is surprisingly much more effective than other short chain calpholates such as acetate and propionate. When the short-chain carpoquinoletate has a pH of about 8.5 or lower, it is about 0.1% to about 10%, preferably about 0.3% to about 3%, more preferably about 0. 5% to about 2.0%, and when the product pH is about 8.5 to about 10, about 3% to about 10%, preferably about 4% to about 8%. used.

In preferred embodiments, the fatty acid component is about 3 to about 2
It is blended in an amount of 5M weight %, fE- or about 5 to about eggplant weight %. Fatty acids have 10 to 22 carbon atoms in the alkyl chain,
Preferably it has 12-18. The fatty acid containing composition will have a pH of about 6.5 to about 9.0.

The compositions of the present invention include fabric brighteners, antifoam agents, hydrotropes such as sodium toluene/l/sulfonate or sodium xylene sulfonate, fragrances, colorants, opacifiers,
Materials such as anti-redeposition agents and alkalinity control agents or buffers such as monoethanolamine and triethanolamine can be included.

The use of these substances is well known in the detergent art.

Substances or anions that tend to precipitate calcium should preferably be limited to minimum amounts. An example is
carbonates, sulfates and orthophosphates. The Ifi fatty acids of the preferred compositions act to precipitate Cal/Rum in the wash solution, but the undiluted compositions of the present invention have no such effect if the Cal content is carefully controlled. Not yet. At a pH of at least 8.5 or less, the fatty acid content of the compositions of the present invention does not substantially affect the enzyme-available calcium content of the composition and the enzyme-available cal/lam content of the compositions. can be ignored when calculating as above.

The following examples illustrate and facilitate the understanding of the invention.

All parts, percentages and ratios in the specification are by weight unless otherwise specified.

Example ■ A liquid detergent composition was prepared by mixing the ingredients listed below in the following proportions.

Fatty acid lauric acid 10.1 7.5 Myristic acid L, l 2-5 Oleic acid -5,0 Sequestering agent Citric acid 4.0 6.9 Noethylenetriaminepentafil 0.6 Sodium acetate (DTPA) Enzyme Maxatase ■ 0.75 0.7
5Termmamy4'12OL O,370
,37 solvent water 30.7 1
9.4 Ethanol 7.0 1.01.
2-Propanediol 5.0 5.0 Other components Monoethanolamine y 2.0 12.0 Triethanolamine/4.0 6.7 NaOH and KOH 4,9- Toluenesulfo/? -) -) Liu-5,0 sodium malformate 1.0 1. OCa
, Cl 2 (Ij
(itl) H8,48,4 (() Listed in the table below (the amount added is deduced from the total water) Composition A has the amount of CaCl2 and sodium diethylenetriaminepentaacetate (DTPA) described below. Thirteen variants were prepared. Variants were stored for one week at 100°F (approximately 37.8°C) in an airtight container. α-Amylase activity was then determined on September 12, 1977.
Technique Industrial Systems, Inc., Leak County, New York, 10591), as measured by the Technique Industrial Method +500-77P, dated September 1979, revised September 1979, is the preferred method, e.g., U.S. Pat. What is described in the specification will give equivalent results. α− at the end of the storage period
The amylase activity was divided by the activity immediately after composition preparation to calculate the retained activity percentage.

(2) In addition to a small amount of calcium in the enzyme preparation (3
) Composition B, expressed as the negative log of the enzyme available calfuram concentration in millimoles, was prepared in seven variants with the amounts of CaC1□ described below. Variants are below 90 degrees (approximately 32.2 degrees Celsius) and 70 degrees Fahrenheit
(approximately 21.1°C) for one month and evaluated for α-amylase activity in the same manner as the Composition A variant.

(2) In addition to the small amount of calcium in the enzyme preparation (the amount added is worked out from the total water) (3) expressed as the negative logarithm of the enzyme available calcium concentration in mmol' (4) Precipitated calcium citrate 7 um. These results demonstrate that the amount of calcium in an aqueous liquid detergent composition when combined with a calfurum-containing acid that provides at least about I x 10'-'5 mmol of enzyme-available calcium alpha-amylase (derived from B. licheniformis) shows a dramatic improvement in stability. The results also point out the advantage of mixtures of sequestering agents. Sequestering agents with high equilibrium constants of complexation, e.g. The metal ion sequestering agent contained is
Although there will be a tendency to sequester all available calcium up to the point of theoretical capacity, such calcium will be dissociated in the wash solution to form heavy metal ions such as copper, iron and manganese. can be blocked. Sequestering agents with low equilibrium constants of complexation, such as alkali metal citrates or citric acid,
The compositions of the present invention may be formulated without adding calcium-containing ingredients in amounts that satisfy the theoretical capacity. Thus, such sequestering agents can be utilized to act as effective detergent builders in the presence of alkaline earth metal ions in cleaning solutions.

Apparently, the α-amylase derived from B. licheniformis in fact has an equilibrium constant of complexation with calcium that is slightly larger than that of citrate, and more precisely at least about lXl0-'°5 mmol of enzyme. It has an equilibrium constant that allows it to retain enough calcium for stabilization in aqueous solutions containing available calcium.

Example ■ Commercial enzyme, registered trademark TaKa-Therm L-34
0, registered trademark Rohalase AT and registered trademark M
axamyl HT is a registered trademark Termamyl 1
2OL is incorporated into the composition of Example I. Comparable alpha-amylase stability results are obtained with equivalent amounts of enzyme-available calcium.

Example■ α-amylase derived from Bacillus subtilis, namely the registered trademark Ban from Novo Industrie and the registered trademark Rapidase from Gist Procard, is
mamyl 120 L is added to the composition of Example (2). Without precipitation of calcium citrate within the aqueous liquid detergent composition, it is not possible to add a sufficient amount of calcium-containing ingredients to stabilize the amylase derived from Bacillus subtilis.

EXAMPLE ■ The C□2 alkyltrimethylammonium chloride in the composition of Example I is replaced by dode/rudimethylamine oquinide. Equivalent results are obtained.

Example V The following composition is prepared by mixing the ingredients listed below.

2← Niji -9- 013 Linear alkyl pen 12.1% 10.5
6% -Zensulfonic acid monotrolide CI4-15A'"-""-7,812,020-0
Toquinlate (7) U'7 acid 9.08 7-5 myristic acid 3-03 2,5-oleic acid
2-4 5-0 -citric acid
5.3 0.2 0.05 Maxas6
■ 0.97 0.97 1.0Term
amyl■120L 0.3 0-3 0-
3 Monoethanolamine 1.9 Triethanolamine 7 3-6 4.5NaO
H and KOH4,191,65 ethanol
4.8 8.63 7.01.2-Propa/Geo 7.2 3.0-Sodium Ruformate 0.97 1.0. 2.
0 Calunium chloride 0.02 0.02 0
-02 Wednesday 29.2 3
5.5 58.0 The incorporation of 02% calcium chloride into each composition provides an enzyme-available calcium content of at least 1 x 10-4°5 mmol/l.

EXAMPLES ■ Potassium hyrophosphate, carpoquine methyl ochinemalon potassium and potassium carboquinomethyl ochinsuccinate are used in place of citric acid in each of the four examples I, 1 and 2. Composition pH is adjusted to 8.0. 0
．． Cal/rum amount of 3% by weight is I x 10-'
It is sufficient to provide an enzyme-available calcium content of greater than 5 mmol and to stabilize the α-amylase derived from B. licheniformis incorporated into the composition.

Applicant's agent Inomata Kiyoshi procedure amendment writing method) June F, 1980 Commissioner of the Patent Office Kazuo Wakasugi 1 Indication of the case 1981 Patent application No. 24920 2 Name of the invention Stabilized aqueous enzyme composition 3 Amendment Relationship with the patent applicant The Brocter, End, 'Gamble, Company 4 agent May 9, 1980 (Shipping date - May 29, 1980)

Claims (1)

Claims: 1. (a) from about 1% to about 75% by weight of a non-soap detergent surfactant or mixture thereof; (b) on a standard enzyme basis, an alpha-denzonolytic enzyme derived from Nonanrus licheniformis; Approximately 0, O1 to approximately 5
% by weight, (C) Sequestering agent or mixture thereof about 0
．． 25-about 40% by weight, (di at least about lXl0-' per composition 1)
- A stabilized aqueous enzyme composition comprising: a calcium-containing component that provides 5 mmol of enzyme-available calcium ions; and (e) from about 10 to about 80% by weight water. 2. The composition according to claim 1, wherein the composition is an aqueous liquid detergent composition. 3. The composition of claim 2, wherein said calcium-containing component provides at least about 1X108 lmol of enzyme-available calcium ions per composition. 4. The composition of claim 2, wherein the calcium-containing component provides at least about IX] ON remol of enzyme-available calcium ions per 1 part of the composition. 5. The non-soap detergent surfactant comprises about 1% of the composition.
The composition of claim 2 comprising 0% to about 65%. 6. The composition of claim 5, wherein the sequestering agent comprises from about 3 to about 3% by weight of the composition. 7. The composition according to claim 6, wherein the sequestering agent comprises citric acid, a water-soluble salt of citric acid, or a mixture thereof. 8. The composition according to claim 7, wherein the metal ion sequestering agent comprises a nitrogen-containing metal ion sequestering agent. 9. The composition according to claim 2, wherein the composition contains a kuku degrading enzyme. 10. The composition according to claim 9, wherein the composition contains formic acid or a water-soluble salt of formic acid or a mixture thereof. 11. The composition of claim 6, wherein said composition has a pH of about 6.5 to about 9.0. 12. The composition of claim 11, wherein the composition contains from about 3 to about 5% by weight fatty acids having from about 10 to about n carbon atoms. 13. The composition of claim 12, wherein water comprises from about 15 to about (a) percent by weight of the composition. 14. The composition according to claim 2, wherein the calcium-containing component comprises a calcium salt or a mixture thereof. 15. The composition according to claim 2, wherein the calcium-containing component comprises a carnoum sequestering agent complex or a mixture thereof. 16. The composition according to claim 15, wherein the calcium-sequestrant complex comprises a nitrogen-containing sequestrant.