Stabilized liquid enzyme containing compositions

A liquid concentrate which has a content of proteases and/or amylases, non-ionic and optionally anionic surfactants, water and optionally solvents selected from mono- and poly-valent alcohols and ethers thereof, which concentrate contains an alkoxylated alkylamine of the formula ##STR1## wherein R is alkyl of 4 to 20 carbon atoms, R' is hydrogen or alkyl of 1 to 10 carbon atoms, provided that the sum of the carbon atoms in R and R' is from 9 to 19, R" is hydrogen, methyl or hydroxymethyl, x is an integer from 1 to 5, and y is 0 or an integer from 1 to 5, provided that the sum of x and y is from 1 to 10; as well as its use as a washing and cleaning agent and washing and cleaning formulations containing the same.

RELATED ART 
Liquid, enzyme-containing washing agent concentrates have a number of 
advantages as compared with pulverulent products. Substantial 
simplifications ensue particularly during manufacture, since the 
preparations, which are obtained during the production of the enzymes, and 
which are separated from the nutrient solutions by filtration or 
centrifuging, do not have to be subjected to a drying process and may be 
further processed in a moist state, thus avoiding any possible trouble 
caused by enzyme dust. Furthermore, liquid preparations can be metered in 
a particularly satisfactory manner during use and make it possible to use 
automatic metering devices. However, problems can arise owing to the fact 
that enzymes decompose substantially more rapidly in solution, 
particularly in the presence of water, than in a dried state. 
U.S. Pat. No. 3,627,688 describes aqueous enzyme preparations which contain 
dialkyl polyglycol ether in addition to non-ionic or zwitterionic 
surfactants. However, the stabilizing action of these compounds is 
inadequate, particularly when anionic surfactants are additionally added. 
Liquid, enzyme-containing concentrates having substantially improved 
stability are described in German Offenlegungsschrift No. 2,038,107. The 
concentrates contain, as stabilizers, a first component comprising 
polyvalent aliphatic alcohols, and alkyl- or hydroxyalkylamides or 
dialkylsulfoxides derived from carboxylic acids of phosphoric acid, and a 
second component in the form of hydroxyalkylamines which are derived from 
aliphatic or heterocyclic amines which optionally contain hydroxyl groups 
and which have a maximum of 8 carbon atoms. Low molecular weight 
carboxylic acids can also be present as a further stabilizing agent. 
German Offenlegungsschrift No. 2,058,826 describes agents of similar 
composition in which the aliphatic or heterocyclic amines are entirely or 
partially replaced by diamines or polyamines which contain aliphatic or 
alicyclic hydroxyalkyl groups and which have a maximum of 8 carbon atoms 
in the molecule. These stabilizers develop their optimum effect in a 
slightly acid to neutral environment or in the absence of anionic 
surfactants. However, since there is a special interest in concentrates 
for use in an alkaline environment, which also optionally contain anionic 
surfactants, the problem arose of further improving the stability of 
agents of this type. 
OBJECTS OF THE INVENTION 
An object of the invention is, therefore, to provide a stabilized liquid 
enzyme preparation which substantially retains its enzyme activity upon 
storage. 
Another object of the invention is to provide a liquid proteolytic and/or 
amylolytic enzyme-containing composition stabilized against loss of 
activity by the presence of an enzyme-stabilizing compound. 
A further object of the invention is to provide a stabilized liquid 
enzyme-containing concentrate which is usable as a washing and cleaning 
agent. 
These and other objects of the present invention will become more apparent 
as the description thereof proceeds.

DESCRIPTION OF THE INVENTION 
These and other objects of the present invention have been achieved by the 
discovery of a stabilized liquid enzyme-containing concentrate which can 
be used as a washing and cleaning agent and which comprises a content of 
proteases and/or amylases, non-ionic and, optionally, anionic surfactants, 
water and, optionally, solvents or additional substances promoting the 
stability of the enzymes selected from the class of mono- and polyvalent 
alcohols and ethers thereof, and an effective amount of an alkoxylated 
alkylamine of the formula 
##STR2## 
wherein R is an alkyl group having 4 to 20 carbon atoms, R' is hydrogen or 
an alkyl group having 1 to 10 carbon atoms, provided that the total carbon 
atoms of R and R' is from 9 to 19 inclusive, R" is hydrogen, methyl or 
hydroxymethyl, x is an integer of from 1 to 5 inclusive, and y is 0 or an 
integer from 1 to 5 inclusive, provided that the sum of x and y is from 1 
to 10 inclusive. 
More particularly, the invention provides liquid concentrates, usable as 
washing and cleaning agents, comprising an enzyme preparation containing 
an enzyme selected from the group consisting of proteases, amylases, and 
mixtures thereof, non-ionic and, optionally anionic, surfactants, water 
and, optionally solvents or additional substances promoting the stability 
of the enzymes selected from the group consisting of mono- or polyvalent 
alcohols and ethers thereof, and, relative to 1 part by weight of said 
enzyme preparation having an activity of 10 to 10,000 SKBE/gm amylase or 
of 1,000 to 1,500,000, perferably 10,000 to 800,000, PE/gm protease, 2 to 
500 parts by weight of an alkoxylated alkylamine of the formula 
##STR3## 
wherein R is an alkyl group having 4 to 20 carbon atoms, R' is a member 
selected from the group consisting of hydrogen and an alkyl group having 1 
to 10 carbon atoms, provided that the total of the carbon atoms of R and 
R' is from 9 to 19 inclusive, R" is a member selected from the group 
consisting of hydrogen, a methyl group, and a hydroxymethyl group, x is an 
integer from 1 to 5 inclusive, and y is 0 or an integer from 1 to 5 
inclusive, provided that the sum of x and y is from 1 to 10 inclusive. 
Preferably, the quantity of alkoxylated N-alkylamines, relative to 1 part 
by weight of the above enzyme preparation of the invention, is 5 to 200 
parts by weight. 
Especially suitable alkoxylated N-alkylamines of the invention contain 
straight-chain or branched alkyl groups having a total of 10 to 20 carbon 
atoms as their 
##STR4## 
residues. The alkylamines containing straight-chain alkyl groups having a 
total of 12 to 18 carbon atoms are preferred. In the last-mentioned 
preferred alkoxylated alkylamines, the group 
##STR5## 
is thus a straight-chain alkyl group. The amino group can be located at 
the terminal position as well as at an inner position. Suitable starting 
materials are, for example, the primary C.sub.10- C.sub.20 amines produced 
from fatty acid nitriles of natural origin by hydrogenation, such as 
decylamine, dodecylamine, tetradecylamine, hexadecylamine, octadecylamine, 
eicosylamine and mixtures thereof, such as coconut alkyl amine and tallow 
alkyl amine. The amines can also be of synthetic origin, as for example, 
derived from oxo compounds or obtained from mineral oil hydrocarbons. This 
group includes, in addition to the above-mentioned amines, undecylamine, 
tridecylamine, pentadecylamine and heptadecylamine, wherein the amino 
group may be optionally located on any carbon atom of the alkyl chain. 
The alkylene oxide groups, on which the polyether radicals of the 
alkoxylated alkylamines of the invention are based, can be derived from 
ethylene oxide (R" = H), propylene oxide (R" = CH.sub.3) or glycidol (R" = 
--CH.sub.2 OH) or mixtures thereof. Preferably, the derivatives of 
ethylene oxide are used in the invention. The alkoxylated alkylamines 
having a total of 1 to 6 ethylene glycol ether groups have proved to be 
particularly successful. 
Examples of suitable alkoxylated alkylamines (the abbreviation EO 
representing added ethylene oxide groups) are dodecylamine-1 EO, 
dodecylamine-2 EO, dodecylamine-3 EO, dodecylamine-4 EO, dodecylamine-5 
EO, tetradecylamine-1 EO, tetradecylamine-2 EO, tetradecylamine-3 EO, 
tetradecylamine-4 EO, tetradecylamine-5 EO, hexadecylamine-2 EO, 
hexadecylamine-3 EO, hexadecylamine-4 EO, hexadecylamine-5 EO, 
hexadecylamine-6 EO, octadecylamine-2 EO, octadecylamine-3 EO, 
octadecylamine-4 EO, octadecylamine-5 EO, octadecylamine-6 EO, coconut 
alkylamine-1 EO, coconut alkylamine-2 EO, coconut alkylamine-3 EO, coconut 
alkylamine-4 EO, coconut alkylamine-5 EO, tallow alkylamine-2 EO, tallow 
alkylamine-3 EO, tallow alkylamine-4 EO, tallow alkylamine-5 EO, tallow 
alkylamine-6 EO. 
The enzymes contained in the agents in accordance with the invention can be 
of vegetable, animal or bacterial origin. The activity of the proteases 
involved in the present invention is given in "PE" (Protease units) which 
can be determined by the method given by H. B. van Raay, H. Saran and H. 
Verbeek in the Journal "Surfactants", 7 (1970), pages 125-132. 
The following species of bacteria may act as the source of bacterial 
amylases and proteases: bacillus types such as B. subtilis, B. cereus, B. 
licheniformus, B. mesentericus, B. brevis, lactic acid bacteria, 
micrococci, Pseudomonas, Arthrobacter, Klebsiella, coli-, proteus- and 
serratia types. The following may act as the source of fungus amylases and 
proteases: actinomycetes streptomycetes, alternaries, sporangies, 
microspora-, pencillium-, cephalosporium, rhizopus-, keratinomyces, 
aspergillus-, mucro-, gliocladeum-, mortierella types, as well as yeasts 
(Candida, Saccharomyces). 
The amylases are preferably amylases which liquify starch and/or 
saccharogenic amylases, both of which can be produced from cereal grain. 
Commercially available amylases which are usable in accordance with the 
invention are, for example, the following commercial products (the names 
of suppliers are also included): 
______________________________________ 
amylase Midwest Biochemical Company, 
Milwaukee, Wisconsin/USA 
.alpha.-amylase Nagase & Co., Osaka/Japan 
.alpha.-amylase Wallerstein Company, Staten 
Island, New York/USA 
Amylase 1000 Societe Rapidase, France 
Amylase preparation 2205/2209 
Rohm & Haas GmbH., Darmstadt 
Anamyl Organon 
Bacterial .alpha.-amylase, BAN 
Novo Industri A/S, Bagsvaerd, 
Denmark 
Bacteria amylase A 
Rohm & Haas GmbH., Darmstadt 
Biokleistase M16 Daiwa Kasei KK, Osaka 
Clarase Miles Laboratories, Inc, Elk- 
hart, Indiana/USA 
Fungamyl Novo Industri A/S, Bagsvaerd. 
Denmark 
Maxazyme FA Gist Brocades NV., Delft, 
Holland 
Maxamyl Gist Brocades NV., Delft, 
Holland 
Fungus amylase Schweizerische Ferment AG., 
Basel 
Rapidase 75 Societe Rapidase, France 
Rhozyme H 39 Rohm & Haas, Philadelphia 
Speedase K & G Nagase & Co., Osaka/Japan 
Termamyl Nova Industri A/S, Bagsvaerd, 
Denmark 
______________________________________ 
Usable as proteases are peptidylpeptidhydrolases such as 
leucineaminopeptidase, aminopeptidases, carboxypeptidases, di- and 
tripeptidases, pepsin, rennin, trypsin, chymotrypsin, pancreaspeptidase, 
enteropeptidase, cathepsins, collagenase, elastase, papain, chymopapain, 
ficin, subtilopeptidases, aspergillopeptidases, streptococcuspeptidases 
chlostridiopeptidases, thermolysin, streptomycespeptidases, bromelain and 
keratinase. Other examples of proteases which are usable for the purposes 
of the invention are aspergillopeptidase A and aspergillopeptidase B, as 
well as those proteolytic enzymes which are isolated from streptomycose 
types. Subtilisin, BPN' and the proteases isolated from streptomyces types 
are preferably used as proteases. 
Commercially available protease products can also be used for the purposes 
of the invention. The commercially available enzyme products which contain 
proteases and frequently certain quantities of amylases, are generally 
marketed in a pulverulent form. They are composed of active enzymes 
combined with relatively inert constituents such as sodium-or calcium 
sulfate or sodium chloride. By way of example, the following proteases are 
commercially available (the names of suppliers are also included): 
______________________________________ 
Alcalase Novo Industri A/S, Bagsvaerd, 
Denmark 
Alkaline protease 
N.V. Organon-Oss-Niederlande 
Alkaline protease 200+/290 
Takeda Chemical Industries, 
Japan 
Bioprase Nagase & Co. Ltd., Osaka/Japan 
Bromelain Takamine, Clifton, New Jersey 
(Miles) 
CRD protease Monsanto Company, St. Louis, 
(contains some .alpha.-amylase) 
Missouri/USA 
Enzyme AG-22 Monsanto Company, Missour/USA 
Enzyme AP densified 
Monsanto Company, Missouri/USA 
Esperase Novo Industri A/S, Bagsvaerd, 
Denmark 
Ficin Takamine, Clifton, New Jersey 
(Miles) 
HT proteolytic enzyme 200 
Miles Laboratories, Inc. Elk- 
hart, Indiana/USA 
HAT proteolytic enzyme 7XB 
Miles Laboratories, Inc. Elk- 
hart, Indiana/USA 
Matsulase MGI-10+/20 
Matsutanie Chemical & Co. Ltd., 
Japan (Mitsubishi) 
Maxatase Gist Brocades NV., Delft 
Optimase Miles Kali-Chemie GmbH, 
Nienburg 
P-11 concentrate 
Rohn & Haas, Philadelphia, 
Pennsylvania/USA 
Pronase Kaken Chemical Company, Japan 
Protease 2200 A Rohm & Haas, GmbH, Darmstadt 
Protease AP 10 .times. 
Schweizerische Ferment AG, 
Basel 
Protease B-4000 Sandoz, Basel/Switzerland 
Protease Hoechst 1549-1 
Farbwerke Hoechst, Frankfurt 
Protin AS Daiwa Kasei KK, Osaka/Japan 
Rapidase 75 Societe Rapidase, France 
Rapidase 400 Rapidase, Seclin, France 
Rhozym J-25, PF Rohm & Haas, Philadelphia, 
Pennsylvania/USA 
Sandoz AP Sandoz, Basel/Switzerland 
SP 88 Novo Industri A/S, Bagsvaerd, 
Denmark 
Tasinase B-11-100 
Kyowa Hakko Co., Japan 
Thermoase Daiwa Kasei KK, Osaka/Japan 
Wallerstein 627-P 
Wallerstein Company, Staten 
Island, New York/USA 
______________________________________ 
All the above enzymes can be used as technically produced, raw aqueous 
solutions, or in a further processed form, to produce the liquid, 
stabilized enzyme preparations in accordance with the invention. Thus, for 
example, it is possible to directly use the aqueous solutions which are 
produced as secretions of organs or as obtained after separation of the 
enzyme producing microorganisms. 
Preferably, however, the enzymes will be purified and concentrated by, for 
example, ultrafiltration methods or ion exchange methods. However, it is 
also possible to directly process the still wet filter cakes, resulting 
during the filtering of precipitated enzymes, to form the solutions in 
accordance with the invention. If the processing method is based on solid 
enzyme preparations, they are dissolved in water or aqueous buffer 
solutions. 
Alternatively, however, the enzymatically ineffective accompanying 
substances, which are usually present in the commercially available enzyme 
preparations in addition to the actual enzymatically effective substance, 
and which may be albumen substances or other substances originating from 
production or preparation, as well as, for example, inorganic salts, can 
remain in the solution, since they are not detrimental to most of the 
purposes for which the enzymes are used. 
The above-mentioned amylase activities of 10-10,000 SKBE/gm or protease 
activities of 1,000-1,500,000 PE/gm represent the activities of those 
enzyme preparations which, at the filing date of this application, appear 
to be economical for use in washing and cleaning agents and in the 
cosmetic field. The liquid compositions of the invention containing these 
enzymes of high activity represent a preferred embodiment of the 
invention. From the chemical-technical standpoint, the enzyme activities 
of the preparations can be increased as required, so that the activities 
of proteases and amylases can be increased to, for example, fivefold the 
maximum values given above. Thus, if preparations having such high 
activities should become readily obtainable in future and appear to be 
suitable, from an economic point of view, for use in the above-mentioned 
fields of application, liquid enzyme-containing compositions having 
correspondingly higher enzyme activities will be similarly readily 
obtainable. 
Suitable solvents or additional substances promoting the stability of the 
enzymes are water-miscible mono- and polyvalent alcohols, water-miscible 
polyethers of polyvalent alcohols and those water-miscible partial ethers 
of polyvalent alcohols which contain at least one hydroxyl group. 
Suitable monovalent alcohols are water-soluble alkanols such as methanol, 
ethanol, propanol and isopropanol. Suitable as the polyvalent alcohol 
additives are the bi- to hexavalent aliphatic alcohols (especially alkane 
polyols containing 2 to 6 carbon atoms), which include, for example, 
alkane diols such as ethylene glycol, 1,2-propylene glycol, 1,3-propylene 
glycol, 1,2-butylene glycol, 1,3-butylene glycol and 1,4-butylene glycol, 
dihydroxypentanes such as neopentyl glycol; glycerin; sugar alcohols such 
as dulcitol, mannitol, xylitol, and sorbitol, etc. Ethers or polyethers of 
these polyvalent alcohols useful in the invention are, for example, ethers 
of ethylene glycol, diethylene glycol, triethylene glycol, or polyethylene 
glycols and the polyethers of glycerin. Those polyethers are especially 
useful which are still liquid at room temperature. The alkyl ethers of 
alkyl alcohols are particularly suitable. The partial ethers of the 
polyvalent alcohols containing at least one hydroxyl group in the 
molecule, or the ethers or polyethers thereof with monovalent alcohols 
containing 1 to 4 carbon atoms, include, for example, the methyl-, ethyl-, 
propyl-, or butylethers of ethyleneglycol, di- or triethyleneglycol, or 
glycerin; the corresponding ethers of glycerin, particularly 
glycerin-.alpha.-monomethylether or glycerin isopropylidene ether, have 
proved to be particularly effective. 
Additives which are also usable in the compositions of the invention are, 
for example, hydroxyalkylamines having 2 to 8, preferably 2 to 6, carbon 
atoms per hydrocarbon radical, such as mono-, di- and triethanolamine, 
mono-, di-, or triisopropanolamine, as well as derivatives of these 
hydroxyalkylamine compounds which have at least one hydroxyl group in the 
molecule and which have structures wherein the said alkylol radicals are 
replaced by C.sub.1-4 -alkyl radicals or by cyclohexyl or cyclopentyl 
groups, such as N-methyldiethanolamine, N,N-dibutyl-ethanolamine, 
N-2-hydroxypropylbutylamine-1, N,N-di-(2-hydroxypropyl)-butylamine, 
N,N-di-(2-hydroxyethyl)-cyclohexylamine, N-2-hydroxypropyl-butylamine, 
N-1-hydroxypropyl-dimethylamine, and N-2-hydroxyethylcyclohexylamine. 
Other suitable amine additives usable in the compositions of the invention 
are those substituted on nitrogen by groups of the formula 
##STR6## 
wherein R.sub.1 is alkylene, preferably alkylene of 2 or 3 carbon atoms, 
and R.sub.2 is alkyl, preferably alkyl of 1 or 2 carbon atoms, the 
remaining 2 substituents on nitrogen being selected from the group 
consisting of hydrogen and alkyl groups, preferably alkyl groups of 1 to 3 
carbon atoms. Examples of such suitable amines are methoxyethylamine, 
3-methoxy- or 3-ethoxy-propylamine, ethoxyethylamine, methoxyethyl- or 
ethoxyethylpropylamine and 1-diethylaminopentanone-4. 
The amount of the above-mentioned amines in the compositions of the 
invention can be chosen so that the pH value of the concentrates, 
including other optional acid or alkaline components contained therein, is 
7 to 11, preferably 8 to 10. 
Further suitable mixture components of the compositions of the invention 
are preferably non-ionic and, if required, anionic surfactants. 
Suitable non-ionic surfactants are the polyglycol ether derivatives of 
alcohols, diols, fatty acids, fatty acid amides and alkyl phenols which 
contain 3 to 30 glycol ether groups and 8 to 20 carbon atoms in the 
hydrocarbon radical. Those non-ionic surfactants containing ethylene 
glycol ether groups are particularly useful. Among the latter polyglycol 
ether derivatives, those are particularly suitable in which the number of 
ethylene glycol ether groups is 5 to 15 and in which the hydrocarbon 
radicals are derived from straight-chain, primary alcohols having 12 to 18 
carbon atoms or from alkyl phenols having a straight-chain alkyl chain 
having 6 to 14 carbon atoms. Included among the straight-chain primary 
alcohols are alkanols and alkenols of both synthetic and natural origin. 
Advantageously, mixtures of lower and higher ethoxylated compounds can 
also be added to the compositions of the invention in their use as washing 
agents. Furthermore, non-ionic compounds of the amine oxide and sulfoxide 
type, which may optionally be ethoxylated, are also usable in the 
compositions of the invention. 
Further non-ionic surfactants, particularly suitable for dish-washing 
agents for use in machines, are the water-soluble addition products of 
ethylene oxide onto polyoxypropylene glycol and alkylene diamine 
polyoxypropylene glycol. Particularly suitable are the water-soluble 
polyethylene oxide adducts to polypropylene glycol and ethylene diamine 
polypropylene glycol, which contain 20 to 250 ethylene glycol ether groups 
and 10 to 100 propylene glycol ether groups. Said compounds normally 
contain 1 to 5 ethylene glycol units per propylene glycol unit. 
Ethoxylated and subsequently propoxylated fatty alcohols, secondary 
alcohols and alkyl phenols having 5 to 35 ethylene glycol or propylene 
glycol ether groups in each case, are also suitable for the same purpose. 
Also suitable are ethoxylated primary and secondary alcohols and alkyl 
phenols having a terminal hydroxyl group which is alkylated, acylated or 
acetalized. Agents of the invention having an increased foaming action can 
contain, instead of the aforesaid substances, fatty acid monoalkanolamides 
and dialkanolamides, such as the mono- or diethanolamide or mono- or 
diisopropanolamide of lauric-, myristic-, palmitic- and oleic acid or of 
coconut fatty acids. 
Suitable anionic surfactants for use in the compositions of the invention 
are those of the sulfonate or sulfate type, such as alkylbenzene 
sulfonates, particularly n-dodecylbenzene sulfonate, as well as olefin 
sulfonates, alkyl sulfonates, .alpha.-sulfo fatty acid esters, primary and 
secondary alkyl sulfates, and the sulfates of ethoxylated or propoxylated 
higher molecular weight alcohols. 
Further compounds of this class, which may be optionally present in the 
agents of the invention, are the higher molecular weight sulfated partial 
ethers and partial esters of polyvalent alcohols, such as the alkali metal 
salts of monoalkylethers or the mono- fatty acid esters of glycerin 
monosulfuric acid ester or of 1,2-dihydroxypropane sulfonic acid. Also 
suitable are sulfates of ethoxylated or propoxylated fatty acid amides and 
alkyl phenols, as well as fatty acid taurides and fatty acid isethionates. 
Eminently suitable anionic surfactants have proved to be the alkali metal 
(e.g. sodium or potassium) or triethanolamine soaps of fatty acids of 
natural or synthetic origin, such as the soaps of coconut, palm kernel or 
tallow fatty acids. 
In the same way as the soaps, the other anionic surfactants can also be 
present in the form of sodium, potassium and ammonium salts as well as 
salts of organic bases such as mono-, di- or triethanolamine. When the 
above anionic compounds have an aliphatic hydrocarbon radical, the latter 
is preferably straight-chain and has 8 to 22, particularly 12 to 18, 
carbon atoms. When the above anionic compounds have an araliphatic 
hydrocarbon radical, the hydrocarbon chains in said compounds are 
preferably unbranched alkyl chains and have 6 to 16, preferably 9 to 14, 
carbon atoms. 
Zwitterionic washing substances which are also usable in the compositions 
of the invention are alkyl betaines and particularly alkyl sulfobetaines, 
such as 3-(N,N-dimethyl-N-alkylammonium)-propane-1-sulfonate and 
3-(N,N-dimethyl-N-alkylammonium)-2-hydroxypropane-1-sulfonate, having 8 to 
22, preferably 12 to 18, carbon atoms in the alkyl group. 
In order to protect the enzymes present in the products in accordance with 
the invention from the detrimental influence of microorganisms, it is 
advantageous to add anti-microbial substances to the liquid enzyme 
concentrates. By way of example, the following substances may be mentioned 
from among the large number of antimicrobial substances which may be used: 
2,4,6- or 2,4,5-trichlorophenol, 2-hydroxydiphenyl, p-benzylphenol, 
p-phenylphenol, p-chloro-m-cresol, 1-hydroxypyridinethion-2 (Zn- or 
Na-salt), 2,2'-dihydroxydichlorodiphenylmethane, 4-hydroxybenzoic acid, 
bis-(2-hydroxy-3,5,6-trichloro-phenyl)-methane, tribromosalicylanilide, 
sodium azide, S-ethylmercuric-4-carboxythiophenol, and 
1,6-bis-(4-chlorophenylbiguanido)-hexane. 
As one skilled in the art will see from the above enumeration, the 
efficacies of the antimicrobial substances usable for this purpose vary to 
a considerable extent. Therefore, the quantities used can vary within wide 
limits of, for example, 0.0005 to 1 percent by weight, preferably from 
0.05 to 0.5 percent by weight of the agent of the invention. 
Furthermore, solubilizing intermediaries (hydrotropic substances) can be 
present in the compositions of the invention, such as toluene- xylene- or 
cumene sulfonate or alkyl sulfates or alkane sulfonates having 6 to 8 
carbon atoms in the hydrocarbon radical, or urea. Said sulfonates and 
sulfates can be present in the form of their sodium-, potassium- or 
ammonium salts or as salts of organic ammonium bases. 
The concentrates of the invention can also contain complex-forming builder 
substances. Suitable inorganic builder substances are acid or neutral 
pyrophosphates, polyphosphates, particularly pentasodium-tripolyphosphate 
and metaphosphates. Usable organic builder substances are the alkali metal 
salts of nitrilotriacetic acid and ethylenediamine-tetraacetic acid. Also 
suitable are the salts of diethylenetriamine pentaacetic acid and the 
higher homologs of the above-mentioned aminopolycarboxylic acids, the 
alkali metal salts of aminopolyphosphonic acids, particularly 
aminotri-(methylenephosphonic acid), 1-hydroxyethane-1,1-diphosphonic 
acid, methylenediphosphonic acid, ethylenediphosphonic acid and salts of 
the higher homologs of the above-mentioned polyphosphonic acids. Mixtures 
of the aforesaid complexing agents are likewise suitable. 
The above-mentioned builder substances may be entirely or partially 
replaced by nitrogen- and phosphorus-free polycarboxylic acids which form 
complex salts with calcium ions, as well as by polymers containing 
carboxyl groups. Citric acid, tartaric acid, benzene hexacarboxylic acid 
and tetrahydrofurantetracarboxylic acid are suitable. Polycarboxylic acids 
containing carboxymethylether groups are also usable, such as 
2,2'-oxydisuccinic acid as well as polyvalent alcohols, partially or fully 
etherified with glycolic acid, or hydroxycarboxylic acids such as 
triscarboxymethyl glycerin, biscarboxymethyl glycerin acid and 
carboxymethylated or oxygenated polysaccharides. Also suitable are 
polymeric carboxylic acids having a molecular weight of at least 350 in 
the form of water-soluble sodium- or potassium salts, such as polyacrylic 
acid, polymethacrylic acid, poly-.alpha.-hydroxyacrylic acid, polymaleic 
acid, polyitaconic acid, polymesaconic acid, polybutenetricarboxylic acid, 
and the copolymers of such monomeric carboxylic acids with one another or 
with ethylenically unsaturated compounds such as ethylene, propylene, 
isobutylene, vinylmethylether or furan. 
Also, so-called wash alkalis are suitable as constituents of the mixture, 
such as bicarbonates, carbonates, borates and silicates of sodium and 
potassium, wherein the silicates can have a ratio of Na.sub.2 O:SiO.sub.2 
of 1:2 to 1:3.5. Neutral salts, such as sodium sulfate, sodium chloride, 
and sodium acetate can also be present as well as optical brighteners of 
conventional constitution, and dyes, opacifiers and perfumes. 
The quantitative composition of the agents of the invention can fluctuate 
within wide limits, preferably within the following limits (in percent by 
weight): 
0.01 to 5% of enzyme preparation, the total quantity of said enzyme 
preparation being dimensioned so that the activity is preferably 100 to 
10,000 PE/gm, relative to the agent of the invention, 
0.05 to 20% alkoxylated alkylamine, which corresponds in the ratios 
indicated above to the quantity of the enzyme preparation, 
0 to 50%, preferably 0.5 to 30%, of mono- or polyvalent alcohols and/or 
partial ethers thereof, 
0.5 to 40%, preferably 5 to 30%, of nonionic surfactant, 
0 to 30%, preferably 1 to 20%, of soap and/or sulfate or sulfonate 
surfactant, 
0 to 25%, preferably 0.5 to 10%, of hydroxyalkylamine, particularly 
triethanolamine, 
0 to 1%, preferably 0.05 to 0.5%, of antimicrobial substances, 
0 to 20% of complex-forming builder substances, 
0 to 10% of solubilizing intermediary, 
0 to 1% of optical brighteners, dyes, perfumes and opacifiers 
20 to 50%, preferably 25 to 40%, of water. 
EXAMPLES 
Typical formulations of washing agent concentrates containing enzymes and 
of their enzyme activity determined after 12 and 16 weeks are listed in 
the following Tables. 
In the following TABLE I are listed the enzymes and ethoxylated alkylamines 
used in ten typical liquid concentrates of the invention. In this TABLE I 
coconut alkylamine is an amine produced by hydrogenation of coconut fatty 
acid nitriles having chain lengths of C.sub.12 -C.sub.18 with an average 
chain length of 13.6. The tallow alkylamine, produced in a corresponding 
manner, had chain lengths of C.sub.16 -C.sub.18 with an average chain 
length of 17.2. The C.sub.11-14 -alkylamine, produced from olefins having 
an average chain length of C.sub.12.5, had amino groups statistically 
distributed over the hydrocarbon chain. The general formula of the 
C.sub.11-14 -alkylamine-2EO is as follows: 
##STR7## 
The alkylamine of TABLE I were ethoxylated, the abbreviation in the table 
consisting of a number followed by EO indicating the number of ethylene 
oxide groups added. 
In the ten comparison liquid concentrates of TABLE I, designated in each 
case by the letter "a", the ethoxylated amines of the invention are 
replaced by ethoxylated alcohols (non-ionic surfactants). 
Enzyme I in the following tables is the product "Maxatase" of the firm Gist 
Brocades, Delft, Holland. Enzyme II is the preparation "Esperase" of the 
firm Novo Industri, Bagsvaerd, Denmark. Enzyme I has an activity of 
440,000 PE/gm and Enzyme II has an activity of 35,000 PE/gm, which results 
in an activity of the liquid concentrate of 440 PE/gm (when using Enzyme 
I) and 350 PE/gm (when using Enzyme II). 
The other components of the liquid concentrates (used in both the liquid 
concentrates of the invention and the comparison liquid concentrates) are 
listed in Table II. The C.sub.16-18 fatty alcohol is derived from tallow 
fatty alcohols. The quantity of triethanolamine is chosen so that, after 
neutralization of the fatty acid or of the toluenesulfonic acid, there is 
still an adequate excess to ensure an alkaline pH value (pH 9.5 to 10). 
The concentrates were stored in sealed bottles at a temperature of 
23.degree. C. The enzyme activity was determined after 12 and 16 weeks, by 
the method given by H. B. van Raay, H. Saran and H. Verbeek in the Journal 
"Surfactants", 7 (1970) pages 125-132. The enzyme activity is given in % 
of the initial activity of Table III. 
TABLE I 
______________________________________ 
Example 
1 2 3 4 5 6 7 8 9 10 
______________________________________ 
Enzyme I 0.1 -- 0.1 0.1 0.1 -- -- -- -- 0.1 
Enzyme II 
-- 1.0 -- -- -- 1.0 1.0 1.0 1.0 -- 
Coconut 
alkylamine- 
2 EO 20 10 -- -- -- -- 10 -- -- -- 
Tallow alkyl- 
amine-5 EO 
-- -- 10 10 -- -- -- 10 -- 10 
C.sub.11-14 -al- 
kylamine 2 
EO -- -- -- -- 10 10 -- -- 10 -- 
______________________________________ 
Comparison 
1a 2a 3a 4a 5a 6a 7a 8a 9a 10a 
______________________________________ 
Enzyme I 0.1 -- 0.1 0.1 0.1 -- -- -- 1 0.1 
Enzyme II 
-- 1.0 -- -- -- 1.0 1.0 1.0 -- 5.0 
C.sub.16-18 - 
fatty al- 
cohol-5 EO 
-- -- -- 10 -- 10 5.0 10 10 5.0 
C.sub.16-18 - 
fatty al- 
cohol-10 EO 
20 10 10 -- 10 -- 5.0 -- -- -- 
______________________________________ 
TABLE II 
__________________________________________________________________________ 
Other Com- 
ponents of 
Example 
Concentrates 
1 2 3 4 5 6 7 8 9 10 
__________________________________________________________________________ 
C.sub.16-18 -fatty 
alcohol-5 EO 
20 20 20 10 20 10 5.0 
-- -- 5 
C.sub.16-18 -fatty 
alcohol-10 
-- 10 10 20 10 20 5.0 
10 10 5 
EO 
Coconut fatty 
acid di- 
ethanolamide 
-- -- -- -- -- -- 5.0 
5.0 
5.0 
5.0 
Na-n-do- 
decylben- 
zenesul- 
fonate -- -- -- -- -- -- 10 10 10 10 
Fatty acid 
(10% coco- 
nut, 90% 
tallow 
fatty acid) 
5.5 
5.5 
5.5 
5.5 
5.5 
5.5 
3.5 
3.5 
3.5 
3.5 
Triethanol- 
amine 5.0 
5.0 
5.0 
5.0 
5.0 
5.0 
18 18 18 18 
Ethanol 
3.0 
3.0 
3.0 
3.0 
3.0 
3.0 
5.0 
5.0 
5.0 
5.0 
1,2-propy- 
leneglycol 
10 10 10 10 10 10 -- -- -- -- 
Diethylene- 
glycolmono- 
butyl ether 
12 12 12 12 12 12 -- -- -- -- 
Toluene 
sulfonic 
acid -- -- -- -- -- -- 7.0 
7.0 
7.0 
7.0 
Water 24.4 
23.5 
24.4 
24.4 
24.4 
24.4 
30.5 
30.5 
30.5 
31.4 
__________________________________________________________________________ 
TABLE III 
______________________________________ 
Residual activity in % of the initial value 
Example 12 weeks 16 weeks 
______________________________________ 
1 95 65 
1a 20 0 
2 75 60 
2a 58 0 
3 90 63 
3a 20 0 
4 95 60 
4a 20 0 
5 100 66 
5a 20 0 
6 65 45 
6a 58 0 
7 80 50 
7a 65 20 
8 73 52 
8a 65 20 
9 75 50 
9a 65 20 
10 90 83 
10a 58 37 
______________________________________ 
These results show that the stability of the enzymes is substantially 
increased by the addition, in accordance with the invention, of 
alkoxylated alkylamines. 
The preceding examples are illustrative of the practice of the invention. 
It is to be understood, however, that other expedients known to those 
skilled in the art or disclosed herein, may be employed without departing 
from the spirit of the invention or the scope of the appended claims.