JPH072956B2 - Immobilized enzyme-containing detergent - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION (Field of Industrial Application) The present invention relates to an immobilized enzyme-containing detergent containing at least one immobilized enzyme among immobilized amylase, immobilized protease, and immobilized lipase. Regarding

(Prior Art) Conventionally, surfactants such as fatty acid salts, alkyl sulfates, alkylbenzene sulfonates, polyoxyethylene alkyl ethers, polyoxyethylene alkyl phenyl ethers, etc., and builders such as sodium carbonate, sodium silicate and zeolite. , Carboxymethyl cellulose, antioxidants, bleaching agents, optical brighteners, bactericides and other substances are mixed to make a cleaning agent, depending on its use, ingredients, etc.
It is classified into soaps for makeup, laundry, etc., detergents for clothes, houses, kitchens, etc. These detergents have a wide pH range from acidic to alkaline depending on their components. For example, soaps, clothes and household detergents are alkaline at pH 9-11. In order to accelerate the removal of sugars, lipids, proteins, etc. that cause stains, these detergents have recently been blended with enzymes to decompose and remove the causes of stains and improve the cleaning power. It was However, these detergents contain various surfactants, various ions, bleaching agents and other drugs, and have a wide pH range from acidic to alkaline ones. Does not exist stably even if it is added as it is, and there is a problem in that it is inactivated during long-term storage or cannot fully exhibit its activity during short-term washing. Further, since the enzyme is sensitive to temperature, there is a problem that it cannot be washed at a temperature higher than the enzyme action temperature. For this reason, in particular, the development of an alkali-resistant enzyme suitable for cleaning has been advanced, and an alkali-resistant protease has been used for cleaning. However, the chemical resistance and temperature resistance are still not sufficient, and the present situation is that enzymes suitable for detergents are still being investigated.

(Problems to be Solved by the Invention) In order to solve such a weak point of the enzyme for cleaning addition, as a result of various studies, a generally commercially available enzyme was immobilized and added to a detergent to obtain a conventional enzyme-added detergent. It has been confirmed that the disadvantages of the above are eliminated and a stable detergent containing an enzyme having good chemical resistance and temperature resistance can be prepared. That is, the present invention provides a composition of a detergent having a strong detergency, which is generally immobilized on commercially available enzymes and is stable even in the presence of alkalinity, warming, surfactants and other chemicals mixed in a detergent. To do.

(Means for Solving Problems) In the present invention, an enzyme that decomposes saccharides, proteins, lipids, etc., which are sources of stains, that is, amylase, protease, and lipase is immobilized by using a chemical binder and blended in a detergent. It is a thing.

Amylase is an enzyme that hydrolyzes starch and glycogen, and includes α-amylase, β-amylase, and saccharified amylases, pig pancreas, Bacillus, Aspergillus,
Barley buds, saliva, sweet potatoes, amylase from barley, etc. are commercially available, and proteases that decompose proteins are pepsin, trypsin, chymotrypsin and other animal proteases, plant proteases such as papain, promeline, and phydin, and fungi. , Bacterial proteases are commercially available, and lipases that decompose lipids are animal lipases such as triacetin, tristearin, and triolein, plant lipases such as wheat germ, castor seeds, extracts from rape seeds, candida, Molds such as extracts from Kumonosukubi, and bacterial and yeast lipases are commercially available. In the present invention, any of these commercially available enzymes may be selected and used.

As a method of immobilizing these enzymes, glutaraldehyde, maleic anhydride / ethylene copolymer, N, N′-polymethylenebisiodoacetamide, N, N′-ethylenebismaleimide, hexamethylenediisocyanate, hexamethylenediamine, Bis-azobenzidine, bisazobenzidine-2,2'-disulfonic acid, 3-
A chemical binder such as (dimethylamino) propyl isocyanite and N-ethyl-5-phenylisoxazolium-3'-sulfonate is used after being adsorbed and bound to hydroxyapatite which is an insoluble carrier.

The enzyme and the immobilization reagent can be appropriately selected depending on the pH of the detergent to which the enzyme is added. For example, in order to immobilize the enzyme added to an alkaline detergent, it is preferable to treat an enzyme that acts on the alkaline side such as bioburase or trypsin with an acidic binder such as bisdiazobenzidine-2,2'-disulfonic acid. When it is added to a detergent, it is preferable to treat an enzyme such as pepsin or papain that also acts on the acidic side with a basic binder such as hexamethylenediamine. Immobilization of these enzymes can be performed by a known method. That is, the binder and the insoluble carrier enzyme are added to a buffer solution in the vicinity of neutrality, and the mixture is reacted at low temperature (about 5 ° C.) for several hours while stirring. The obtained insoluble immobilized enzyme is thoroughly washed with the buffer solution used and then with cold water, and dried by freeze-drying.

The immobilized enzyme thus obtained is mixed with a surfactant, a builder, a bleaching agent, and other chemicals used in detergents by a conventional method to obtain a desired detergent composition.

(Action) When an immobilized enzyme is mixed with a detergent, the enzyme is immobilized, so it exhibits a wide range of activities with respect to temperature and pH, and is less susceptible to interference by ions, surfactants, bleaching agents, etc. Compared with, the Michaelis constant is increased, so the enzyme reaction does not take much time and the dirt is decomposed in a short time.

Hereinafter, the present invention will be specifically described with reference to Examples. The amylase of the immobilized enzyme used was barley amylase, the protease was bioprase, and the lipase was wheat germ lipase, all of which were crosslinked with glutaraldehyde. In each case, the solid in the additive component was finely pulverized,
The mixture was gradually added to the surfactant heated to ℃ with stirring, and after the addition was completed, stirring was continued for 30 minutes at that temperature and then left to cool to room temperature.

The apatite-immobilized enzyme used in the examples was produced by the following method.

At least one enzyme of amylase, protease and lipase is added to the apatite suspension.

The cross-linking agent is added while stirring the suspension.

The apatite-immobilized enzyme obtained by the reaction as described above is used in the state of suspension or after freeze-drying.

The reaction amount is shown in the reaction example.

Immobilized amylase (reaction example 1) Amylase 100mg Apatite 2g Glutaraldehyde (crosslinking agent concentration: 28mg / 100ml) 2ml Pure water 50ml Immobilized protease (reaction example 2) Protease 150mg Apatite 10g glutaraldehyde (crosslinking agent concentration: 28mg / 100ml) 8ml Phosphate buffer (0.05mM, pH6.8) 100ml Immobilized lipase (Reaction example 3) Lipase 100mg Apatite 2g Glutaraldehyde (Crosslinker concentration: 28mg / 100ml) 4ml Pure water 50ml Example 1. Cloth cleaning agent wt% excess Sodium carbonate 20 Sodium carbonate 30 Immobilized protease 20 Propylene glycol monostearate 30 Total 100 Example 2. Clothing detergent wt% Immobilized protease 6 Immobilized amylase 6 Immobilized lipase 6 Sodium lauryl benzene sulfonate 6 Cetyl trimethyl ammonium chloride 5 C ₁₄ Sodium olefin sulfonate 3 Sodium sulfate 12 pcs Sodium formate 12 Sodium aluminosilicate 12 Propylene glycol monostearate 27 Fluorescent agent 5 Total 100 Example 3. Clothes detergent detergent wt% immobilized protease 20 Sodium hypochlorite 30 Propylene glycol monostearate 50 Total 100 Example 4. Clothes detergent wt% Immobilized protease 20 Sodium oleyl ether sulfate 15 Polyoxyethylene lauryl ether 24 Propylene glycol monostearate 41 Total 100 Example 5. Facial cleanser wt% Immobilized protease 5 Immobilized lipase 5 Triclosan 1 Paraben 0.25 Dibutylhydroxytoluene 0.75 Propylene glycol 88 Yellow No. 4 Suitable amount Fragrance Suitable amount Example 6. Whole body cleaning agent wt% Immobilized protease 5 Immobilized lipase 5 Immobilized amylase 5 Triclosan 1 Dibutylhydroxytoluene 1 Sodium benzoate 3 Tri Tanolamine 10 Propylene glycol 70 Green No. 3 Proper amount Yellow No. 4 Proper amount Perfume Proper amount Reference example Using the detergent of Example 1 composition containing unfixed enzyme as a control, the enzyme activity of the composition of Example 1 was measured at 40 ° C and measured. The initial activity was set to 100, and the specific activity with time was shown. 1 g of each detergent was dissolved in 100 ml, and 1 ml thereof was collected after each lapse of time and used as a test solution. After using milk casein as a substrate and adding a test solution to act on it, trichloroacetic acid was added and the precipitate was filtered off, a fixed amount of the filtrate was added, and a certain amount of sodium carbonate solution and Holin reagent were added to develop the color. The enzyme activity was measured by the Hollin analysis method, and the following results were obtained.

This result indicates that the immobilized enzyme is relatively stable in the presence of the surfactant, salts and bleach.

Comparative Example Comparative test with other water-soluble immobilized enzyme-containing facial cleanser Using results using hydroxyapatite-immobilized enzyme-containing facial cleanser and protease cross-linked to carboxymethylcellulose as a group to be compared, and examining changes over time It was

The blending composition is the facial cleansing composition of Example 5.

20 inspectors used it twice a day for 7 consecutive days and obtained the following results.

Neither change in color nor odor with time was observed.

(Effects of the Invention) According to the present invention, by incorporating an enzyme in which an enzyme is cross-linked and immobilized to hydroxyapatite into a detergent, the enzyme which has not been used in the conventional detergent can be used in the detergent, and the chemical resistance can be improved. When a temperature-resistant detergent is produced and a facial cleanser containing the immobilized enzyme is used, the effect is superior to the conventional facial cleanser containing the immobilized enzyme.

 ─────────────────────────────────────────────────── ─── Continuation of front page (72) Inventor Kiminori Atsumi 2-11-10 Tsukiji, Chuo-ku, Tokyo (Tsukiji Central Building) Dental Chemical Co., Ltd. (72) Inventor Souki Saito 2 Tsukiji, Chuo-ku, Tokyo Inc. 11-10 (Tsukiji Central Building) Dental Kagaku Co., Ltd. (72) Inventor Keijiro Fujita 2-11-10 Tsukiji, Chuo-ku, Tokyo (Tsukiji Central Building) Dental Kagaku Co., Ltd. (72) Inventor Tsutomu Ishizaki 2-11-10 Tsukiji, Chuo-ku, Tokyo (Tsukiji Central Building) Dental Chemical Co., Ltd. (56) References JP 61-207499 (JP, A) JP 60-190497 (JP, A) ) JP-A-57-70199 (JP, A) JP-A-57-73098 (JP, A)

Claims (1)

[Claims] 1. An immobilized enzyme-containing detergent containing an enzyme immobilized by cross-linking at least one of amylase, protease and lipase to an inactive carrier of hydroxyapatite.