KR20040089693A - Sterilization methods, sterilizing/washing agent and washing method - Google Patents

Abstract

A solution containing chelating agent resistant to chelating agent at a concentration of at least 2.5 U / mL of lytic activity, having a calcium ion concentration of 10 ppm or less in terms of CaO, a pH of more than 8 and an aqueous solution in the range of 12 or less Disinfection treatment method characterized in that the treatment of the substance adhered to the cells. A bactericidal treatment method comprising lysing a lysing enzyme having a chelating agent at a concentration such that the lytic activity is 15 U / mL or more, and treating the substance to which the cells are adhered using an aqueous solution having a pH greater than 8 and 12 or less.

Description

Disinfection method, disinfectant detergent composition and washing method {STERILIZATION METHODS, STERILIZING / WASHING AGENT AND WASHING METHOD}

As a method of treating and disinfecting a substance with bacteria, there are a method using a bactericide or a bleach (Japanese Patent Laid-Open No. 63-131124, Japanese Patent Laid-Open 2000-26894, Japanese Patent Laid-Open 63-286158, Japanese Laid-Open Patent Publication). 11-29797, etc.).

As an enzyme reaction, a composition for disinfecting a contact lens containing an enzyme acting on a cell wall of a microorganism, an enzyme acting on a cell membrane, an antibacterial protein and an antifungal agent has been proposed (Japanese Patent Laid-Open No. 9-10288).

However, the above technique requires a strong irritant fungicide or a special chemical component to the human body, and a method and composition for efficiently disinfecting using a low irritant component have been desired. Furthermore, the above technique is intended for living cells, but since most of the cells causing odors, stickiness, fibers and other odors in the environment around water, such as a dishwasher and a bath, are killed, There has been a demand for a method and composition capable of efficiently decomposing and removing dead cells.

The present invention relates to a bactericidal treatment method, a bactericidal detergent composition, and a laundry method for efficiently sterilizing by treating a substance with a bacterial cell by using a lytic enzyme.

It is an object of the present invention to provide a bactericidal treatment method, a bactericidal detergent composition and a laundry method capable of efficiently decomposing and removing cells using a low irritant component.

The inventors of the present invention contain a lysing enzyme having a chelating agent resistance at a concentration at which a specific lytic activity is achieved, by setting the calcium ion concentration to a specific amount, and treating the substance with the bacteria in an aqueous solution having a pH of more than 8 and 12 or less. In addition, the present inventors have found that the killed cells can be efficiently decomposed and removed.

That is, this invention provides the following disinfection method, the disinfecting detergent composition, and the washing method of fibers.

(1) A solution containing a chelating agent having a chelating agent resistance at a concentration of at least 2.5 U / mL of lytic activity, having a calcium ion concentration of 10 ppm or less in terms of CaO, a pH of more than 8 and 12 or less Disinfection treatment method characterized in that the treatment of the substance adhered to the cells.

(2) A bactericidal treatment comprising lysing enzymes resistant to chelating agents at a concentration of 15 U / mL or higher, and treating the cells to which the cells are attached by using an aqueous solution having a pH greater than 8 and 12 or less. Way.

(3) A bactericidal detergent composition containing a bactericidal enzyme having a chelating agent resistance and containing no phosphate, wherein the bactericidal detergent composition or the diluent thereof is used at a concentration such that the bactericidal activity is 2.5 U / mL or higher. And a calcium ion concentration of 10 ppm or less in terms of CaO and a pH of more than 8 and 12 or less.

(4) A bactericidal detergent composition containing a bactericidal enzyme having a chelating agent resistance and containing no phosphate, wherein the bactericidal detergent composition or the diluent thereof contains the bactericidal enzyme at a concentration such that the bactericidal activity is 15 U / mL or more. And a pH of 8 or more and 12 or less solution.

(5) When washing fibers using a bactericidal detergent composition containing a lytic agent and a surfactant having a chelating agent resistance and containing no phosphate,

(a) A range of lysing enzymes at a concentration of lysine activity of 2.5 U / mL or more, wherein the calcium ion concentration is 10 ppm or less in terms of CaO and pH is more than 8 and 12 or less

(b) the lysing enzyme is contained at a concentration such that the lytic activity is 15 U / mL or more, and the pH is more than 8 and 12 or less

A method of washing fibers, wherein the fibers are sterilized and washed using an aqueous solution of a bactericidal detergent adjusted to any one of them, to suppress the occurrence of odors at the time of drying the fibers indoors or storing the fibers after drying.

In the present invention, a lytic enzyme having chelating agent resistance refers to a lytic enzyme whose lytic activity does not decrease in the presence of a chelating agent. Specifically, in "Measurement method of unit (U)" mentioned later, it measured by "measurement method of unit (U)" with respect to unit (U ') in the case of measuring without adding a chelating agent (EDTA, tetrasodium). The ratio (%) of the unit (U), that is, "(U / U ') x 100" refers to an enzyme having lytic activity (U) of 50% or more, preferably 80% or more. As the lytic enzyme, a protease having lytic activity is preferable. The unit U of lytic activity is defined by the following method.

<Measurement method of unit U>

25 ml of a 25 degreeC water bath was prepared by dissolving EDTA.sodium tetrahydrate, 111 ppm of sodium bicarbonate, 278 ppm of sodium bicarbonate and an enzyme in sterile water (distilled water was sterilized by heating at 121 ° C. for 20 minutes). Keep warm for 5 minutes. To this solution, 1 mL of heated cells of Escherichia coli (Eschericha coli ATCC8739: Escherichia coli), which became about 3.3 at a turbidity of OD 660 nm (turbidity at wavelength 660 nm in a spectrophotometer), were added and stirred, It is confirmed that turbidity of OD 660nm becomes 0.3 ± 0.03 within 30 seconds, and a U test solution is obtained. The U test solution was kept in a water bath at 25 ° C. for 10 minutes, stirred 10 minutes after the addition of the heated cells, and the turbidity of OD 660 nm was measured. The amount of protease required to reduce the turbidity of U test solution OD 660 nm to 0.05 in 10 minutes is 10 units (unit, U).

In addition, a lysis rate is measured by the following method.

<Method of lysis rate>

(Preparation of Test Bacteria Liquid)

Brain Hert Infusion Medium (manufactured by Diffco Corporation) is dissolved in prescribed amount (37 g / L) purified water and sterilized by placing 50 mL in a 500 mL Sakaguchi flask. Thereafter, Escherichia coli ATCC8739 (E. coli) is seeded in a platinum loop and shaken at 37 ° C for 18 hours. The cells were collected by centrifugation (3,000 rpm (about 1800 G) for 5 minutes), suspended in sterile water, centrifuged again, the cells were collected and suspended in sterile water, and the cells suspended in sterile water at 20 ° C. at 20 ° C. The mixture is heated for minutes, centrifuged, and the heated cells are collected and suspended in sterile water to obtain a cell suspension having a turbidity of OD 660 nm of 3.3.

(Lysis rate measurement)

9 mL of the sample for measuring the lysis rate is added to the test tube, and kept in a 25 ° C water bath for 5 minutes. 1 mL of the suspension obtained above was added and stirred to obtain a total solution of 10 mL of lysis rate, and the turbidity of OD 660 nm was measured at the start of the reaction (within 30 seconds). Then, the mixture is kept in a water bath at 25 ° C. for 10 minutes, and after stirring, a turbidity of OD 660 nm is measured. As a blank, 1 mL of sterile water is used instead of the cell suspension. In addition, it adjusts so that "the total 10 mL lysis rate test liquid" may be the density | concentration of the aqueous solution and the bactericidal detergent composition solution to measure.

The lysis rate is calculated by the formula shown below.

In the aqueous solution of the present invention (1), the bactericidal detergent composition (or its water-dilution liquid) solution (a) in (a) in (3) and (5) (hereinafter abbreviated as a solution of 2.5 U / mL or more), measured by the above measuring method. The aqueous solution of (2) of the present invention, (4), and the lysine activity are contained at a concentration of 2.5 U / mL or more, preferably 3 to 400,000 U / mL, more preferably 15 to 400,0 U / mL. In the solution of the bactericidal detergent composition (or its water-dilution solution) in (5) (hereinafter abbreviated as 15 U / mL or more solution), the lytic enzyme has a lytic activity of 15 U / mL or more, preferably 16 U / mL or more And especially at a concentration of 20 to 400,000 U / mL. The concentration of the solution is adjusted to make this lytic activity. In this case, when diluting for washing or the like, the upper limit is preferably set to 3,000 U / mL in a solution of 2.5 U / mL or more and 1,000 U / mL in a solution of 15 U / mL or more. In addition, the lysis activity of the solution of this invention is calculated | required by converting the density | concentration in the solution of each enzyme in U / mL based on the lysis activity value (U / unit weight) of an enzyme. In addition, a 2.5U / mL or more solution and a 15U / mL or more solution are collectively called only an aqueous solution or an antibacterial detergent composition solution below.

Specific examples of the lytic enzymes include savinase 12T, savinase 16L, esperase 4T, everase 8T, cannase 24TK, alcalase 2T (manufactured by Novozymes), Bacillus sp. Bacillus sp, a P enzyme produced from P (Microbiological Research Institute Accession No. 8090). K enzyme, Bacil1ussp, produced from K (Microbiological Research Institute Accession No. 8801). X enzyme, Bacillus sp. Produced from X (Microbiological Research Institute Accession No. 8082). Y enzyme produced from Y (Microbiological Research Institute Accession No. 8088), etc. are mentioned. Preferred enzymes among these are savinase 12T, savinase 16L, esperase 4T, everase 8T, cannase 24TK, alcalase 2T, deozyme, Y enzyme and the like.

In addition, when the lytic enzyme is used in combination with an enzyme having other effects, an increase in the lytic effect can be expected. For example, the lipase excellent in the washing effect is mentioned.

The measurement of the lipase activity is usually performed by measuring the amount of free fatty acids when the substrate is decomposed using the lipase. As a lipase to be incorporated into the cleaning composition of the present invention, "the amount of fatty acids produced from triolein after the substrate decomposition test relative to the amount of fatty acids produced from n-hexadecyl palmitic acid after the substrate decomposition test is twice or more (molar ratio). Lipase having an enzymatic activity to be obtained ”, and the enzymatic activity is measured by the following substrate decomposition test method.

Put a magnet sterling bar (diameter 8 mm, length 30 mm, columnar shape) in a beaker of 200 mL (63 mm in diameter and 80 mm in height), and 100 ± 0.2 mL of the base solution at 22.5 ° C. ± 0.5 ° C. [α-SF-Na (C14 / 16 = 200 ppm of sodium salt of α-sulfofatty acid methyl ester having 2/8 (mass ratio) alkyl group, sodium carbonate, German hardness 3 ° DH (adjusted to 30 ppm in terms of CaO by adding calcium chloride to ion-exchanged water)], and substrate 0.2 g of triolein (Fulka's reagent) or palmitic acid n-hexadecyl (Tokyo Kaase's reagent) was added and stirred for 15 to 20 minutes, and the pH became constant at the initial pH (about 10 to 10.5). Shall be. The apparatus used is set to 200 rpm using a titrator TS-980 (manufactured by Hiranuma Sangyo Co., Ltd.). In addition, since a pH electrode is also put in the titration apparatus, this electrode is used.

Subsequently, KOH (N / 10) solution is added dropwise while maintaining the initial pH value (about 10 to 10.5) while stirring at room temperature. One minute after the start of titration, various lipases are added so as to be in the range of 0.1 to 0.01 ppm of protein of the lipase in the base solution, and the dropwise amount of KOH (N / 10) after 61 minutes is measured. The protein concentration of the lipase is the same for each lipase with triolein and palmitic acid n-hexadecyl.

After adding the lipase thus obtained, the amount of fatty acid produced from each substrate (triolein or palmitic acid n-hexadecyl) was obtained with a KOH dropwise amount of 60 minutes. From the average value of 10 iterations of fatty acid production | generation amount, the fatty acid production | generation amount (molar ratio) from triolein with respect to the palmitic-acid n-hexadecyl fatty acid production | generation amount, ie, enzyme activity, is computed.

As a lipase which has the enzyme activity as mentioned above, the lipase derived from Humicola lanuginosa of Unexamined-Japanese-Patent No. 11-510699 is mentioned, for example. Such lipases are commercially available from Novozymes under the trade name of "Lipex."

As for content of a lipase, 0.01-5 mass%, especially 0.1-2 mass% are preferable as an enzyme preparation with respect to the aqueous solution or the whole amount of the disinfecting detergent composition solution.

It is also preferable to use starch degrading enzyme. For example, the optimum pH as shown in Japanese Patent Laid-Open No. 2-49584 is less than 60 ° C. or as shown in Japanese Patent Application Laid-Open No. 11-5030O3. As shown, calcium resistant starch degrading enzyme and the like are also preferable. In addition, starch degrading enzymes that decompose α-1,6 bonds of starch as shown in Japanese Patent Laid-Open No. 3-87176, Japanese Patent Laid-Open No. 3-87177, Japanese Patent Laid-Open No. Hei 3-108482 and Japanese Patent Laid-Open No. Hei 6-14775. Also preferred.

Also preferred is an amylase in which a wild type amylase derived from the genus Bacillus is modified by genetic mutation.

Examples of the amylase include amylase: Duramil 60T (manufactured by Novozymes), pullulanase (manufactured by Sigma), and the like, and these may be used alone or in combination of two or more thereof.

As for content of amylase, 0.01-5 mass%, especially 0.1-2 mass% are preferable as an enzyme preparation with respect to the aqueous solution or the whole amount of the disinfecting detergent composition solution.

The bactericidal detergent composition of the present invention does not contain phosphate. However, in some cases, 0.5 mass% or less of phosphate may be contained in a composition as an impurity from an enzyme preparation etc.

In a 2.5 U / mL or more solution, calcium ion concentration is 10 ppm or less in CaO conversion, Preferably it is 0.01-9 ppm, Especially preferably, it is 0.01-5 ppm. Although it may exceed 10 ppm in 15U / mL or more solution, Preferably it is 0.01-20 ppm, More preferably, it is 0.01-15 ppm, Especially it is suitable when it is set as 0.01-10 ppm. By carrying out calcium ion concentration in the said range, the lysis effect of a lytic enzyme will improve.

In addition, the calcium ion concentration of CaO conversion shall be based on the calcium ion concentration measuring method (absorbance method using pontachrome violet) described in the Example of Unexamined-Japanese-Patent No. 9-318615.

Calcium ion concentration of the said range can be obtained by mix | blending the chelating agent and general purpose chelating agent which have high chelating ability with respect to calcium ion in aqueous solution or detergent composition individually by 1 type or in combination of 2 or more types as appropriate. .

As a chelating agent which has a high chelating ability, an aminocarboxylic acid derivative type chelating agent, polycarboxylic acid, or its salt etc. are mentioned.

As aminocarboxylic acid derivative type chelating agent, aminomethyl glycidine acetate, (beta) -alanine diacetate, imino disuccinate, hydroxyimino disuccinate (HIDS), serine diacetate, isoserine diacet Acid salts, aspartic acid diacetate (ASDA), ethylenediamine tetraacetate (EDTA), nitro triacetate (NTA), hydroxyethylenediamine pentaacetate (HEDTA), diethylenetriamine pentaacetate (DTPA), etc. Can be mentioned. In addition, EGTA (ethylene glycol bis [2-aminoethyl ether] tetraacetate) and the like can also be used. Sodium salt and potassium salt are preferable for these chelating agents.

In the case of using an aqueous solution or a disinfectant detergent composition solution for the fibers, among the chelating agents described above, aminomethylglycine diacetate, β-alanine diacetate, imino disuccinate, hydroxyimino disuccinate (HIDS), Serine diacetate, isocerine diacetate, aspartic acid diacetate (ASDA) and the like are preferred.

Examples of the polycarboxylic acid or salts thereof include polymer electrolytes such as polyacrylic acid, copolymers of maleic acid and acrylic acid, and polysaccharide polycarboxylates. The average molecular weight of the copolymer of polyacrylic acid, maleic acid and acrylic acid becomes like this. Preferably it is 1000-1,000,000, More preferably, it is 1000-100,000. As polycarboxylic acid or its salt, sodium salt and potassium salt are preferable.

Among polycarboxylic acids or salts thereof, polysaccharide-based polycarboxylates are obtained by oxidatively cleaving a polysaccharide so that a carboxylic acid group is formed by an oxidizing agent (hypochlorite, periodate, chlorite, etc.), and polysaccharides include starch and dextrin. , Cellulose, hemicellulose, amylose, pectin and the like. The content of oxidized anhydroglucose units is preferably from 50 to 100 mol%, particularly preferably from about 70 to 95 mol%. Of the polysaccharide polycarboxylates, oxidized starch obtained by oxidative cleavage of starch is particularly preferred.

Examples of general chelating agents include crystalline alkali metal silicates such as crystalline aluminosilicates (A, P and X zeolites) and amorphous aluminosilicates, crystalline alkali metal silicates such as layered silicates, citric acid or salts thereof, and gluconic acid. Or low molecular weight oxycarboxylic acid chelating agents such as salts, glycolic acids or salts thereof, malic acid or salts thereof, 1-hydroxyethane-1, 1-diphosphonic acid (HEDP), aminotrimethylenephosphonic acid (ATMP Inorganic phosphates such as phosphonic acid type chelating agent, tripolyphosphate, and pyrophosphate.

Among the above chelating agents, particularly preferably, they are unmanned chelating agents.

The pH of the aqueous solution and the bactericidal detergent composition solution is more than 8 and 12 or less, preferably 8.1 to 11 in a 2.5 U / mL or more solution, and more than 8 and 12 or less, preferably 8.1 to 11 or more in a 15 U / mL or more solution. 11, especially 9-11. In this invention, by making pH into the said range, the activity of a lytic enzyme becomes high, and as pH increases, a lysis effect improves.

The antibacterial detergent composition of the present invention can be used as a medical detergent, a hand washing dish washing detergent, a detergent for a bath, a detergent for a washing machine, and a detergent for a toilet and a drain pipe.

In the case of a medical detergent, the content of the lytic enzyme in the disinfectant detergent composition is preferably 1,000 to 400,000 U / g, more preferably 4,500 to 60,000 U / g, 30,000 to 60,000 U / g, and an aqueous solution or a disinfectant detergent composition. The pH of a solution is suitable for it to be 8.1-11, especially 8.5-11.

In the case of dishwashing detergent, the content of the lytic enzyme in the disinfectant detergent composition is preferably 100 to 400,000 U / g, and particularly preferably 300 to 20,000 U / g. The pH of the aqueous solution or the bactericidal detergent composition solution is suitably set to 8.1 to 10, particularly to 8.1 to 9.

In the case of a bath detergent, the content of the lytic enzyme in the disinfectant detergent composition is preferably from 1,000 to 400,000 U / g, particularly preferably from 2,000 to 40,000 U / g. The pH of the aqueous solution or the bactericidal detergent composition solution is suitably set to 8.1 to 10, particularly to 8.1 to 9.

In the case of a washing machine detergent, 1,000-100,000 U / g is preferable and, as for content of the lytic enzyme in a disinfecting detergent composition, 2,000-20,000 U / g is especially preferable. The pH of the aqueous solution or the bactericidal detergent composition solution is suitably set to 8.1 to 12, particularly to 8.5 to 11.

In the case of the washing | cleaning agent for a toilet and a drainage pipe, content of the lytic enzyme in a disinfecting detergent composition is preferable 1,000-100,000U / g, and especially 2,000-20,000U / g is preferable. The pH of the aqueous solution or the bactericidal detergent composition solution is preferably 9 to 12, particularly 9 to 11.

In the disinfectant cleaning composition of the present invention, the chelating agent is effectively penetrated into the contaminants, and it is preferable to contain a surfactant in order to efficiently lyse the cells present therein.

As surfactant, various surfactant, such as an anionic surfactant, a nonionic surfactant, a cationic surfactant, and an amphoteric surfactant, etc. are mentioned, These can be used individually by 1 type or in combination of 2 or more types as appropriate. .

As for content of these surfactant, 3-50 mass% is preferable with respect to the whole amount of disinfecting detergent composition, and 5-40 mass% is preferable. Moreover, it is preferable that an anionic surfactant or a nonionic surfactant is a main surfactant (surfactant with the highest content among surfactant of an antibacterial detergent composition).

As an anionic surfactant, C10-C14 linear alkylbenzenesulfonate, C13-19 alpha- sulfofatty acid lower alkyl (C1-3) ester salt, C10-20 alkyl sulfate ester salt, olefin sulfonate, Alkanesulfonates, polyoxyethylene alkyl or alkenyl sulfate ester salts, saturated or unsaturated fatty acids having 12 to 22 carbon atoms, and the like can be suitably used. As these salts, alkali metal salts such as sodium salts and potassium salts, amine salts, ammonium salts and the like can be used.

Content of an anionic surfactant becomes like this. Preferably it is 5-45 mass%, More preferably, it is 10-35 mass% with respect to the disinfecting detergent composition whole quantity.

As a preferable nonionic surfactant, the following can be illustrated.

(1) EO addition type nonionic surfactant (alkyl ether ethoxylate) which added 5-30 mol of ethylene oxide (EO) to the average C10-C20 alcohol.

(2) An EO-PO addition type nonionic surfactant in which an average of 5 to 30 moles of ethylene oxide (EO) and propylene oxide (PO) are added to an alcohol having an average carbon number of 10 to 20.

(3) Fatty acid ester type nonionic surfactant represented by following formula (1).

R ¹ -CO (OR ² ) _n OR ³ . (One)

(In formula, R <^1> is a C5-C21, Preferably 9-17 alkyl group or alkenyl group, OR <^2> represents the addition unit of a C2-C4 alkylene oxide, EO is singly or EO and PO are mixed. N represents an average added mole number of OR ² , and represents 5 to 30, preferably 5 to 20. R ³ represents an alkyl group having 1 to 4 carbon atoms, preferably 1 to 2 carbon atoms. .)

As for content of a nonionic surfactant, 1-40 mass% is preferable with respect to the disinfecting detergent composition whole quantity, and 1-30 mass% is especially preferable.

Preferred amphoteric surfactants include betaine such as amido propyl betaine and amido ethyl betaine, and N- palm oil fatty acid amyl-N-carboxyethyl-N-hydroxyethyl ethylenediamine sodium. Dazoline derivatives.

In the case where the antibacterial detergent composition is a medical detergent, a hand washing dish washing detergent, a bath detergent, or a washing machine detergent, it is particularly preferable to contain an anionic surfactant.

In the case of a medical detergent, as anionic surfactant, a linear alkylbenzene sulfonate, the (alpha)-sulfo fatty acid alkyl ester salt, an olefin sulfonate, an alkane sulfonate, an alkyl sulfate, a fatty acid salt, etc. are preferable, Among these, especially alpha-sulfo fatty acid. Alkyl ester salts are preferable because of their excellent cleaning power in a small amount.

In the case of hand-washing dish detergent, as anionic surfactant, polyoxyethylene alkyl or alkenyl sulfate ester salt, linear alkylbenzene sulfonate salt, (alpha)-sulfo fatty acid alkyl ester salt, olefin sulfonate, alkane sulfonate, alkyl sulfate, Fatty acid salts are preferable.

In addition, the surfactant concentration of the aqueous solution or the disinfectant detergent composition solution is preferably 1 to 20% by mass in the case of a dishwasher / washing table detergent, and 0.5 to 1.5 in the case of a toilet, a drain pipe and a detergent for a bath (bath). Mass% is preferable, In the case of the detergent for washing machines, 0.05-0.5 mass% is preferable, and in the case of a medical detergent, 100-1200 ppm is preferable.

In the case of the medical detergent, the disinfectant detergent composition of the present invention is used as it is or as a diluent thereof. However, dilution is preferably a concentration of the disinfectant detergent composition, preferably 0.04 to 10% by mass, more preferably in the diluent detergent composition. It is preferable to dilute so that it may become 0.04-8 mass%, Especially preferably, it is 0.05-1 mass%. A suitable use concentration can be set according to the use of the antibacterial detergent composition.

The use concentration in the case of the dishwashing detergent is preferably in the range of 1 to 50% by mass for washing in a sink or dishwashing container and 5 to 50% by mass for washing in a sink as the disinfectant cleaning composition concentration (hereinafter the same).

The use density | concentration in the case of the detergent for bathtubs has the preferable range of 1,000-100,000 ppm.

In the case of a washing machine detergent, the range of 1,000-10,000 ppm is preferable.

The use density | concentration in the case of the detergent for toilets and a drainage pipe is 3-100 mass%, Especially the range of 5-50 mass% is preferable.

In the range which does not impair the objective of this invention, the disinfecting detergent composition of this invention can mix | blend the various components mix | blended with the dishwashing detergent, the bathroom bath detergent, the dishwasher and the drainage pipe detergent, the toilet cleaner, the medical detergent, etc. Can be.

The bactericidal cleaner composition of the present invention can be prepared in various formulations such as liquid, gel, spray, powder, tablet, granule detergent, sheet, and bar.

The preparation method of the bactericidal detergent composition of the present invention is not particularly limited, and can be produced in accordance with the conventional method of each of the above formulations.

The bactericidal detergent composition of the present invention is particularly suitably used as a detergent for clothing, and it is used for disinfecting and cleaning fibers, and for suppressing the occurrence of odor during the drying of the fibers or preserving the fibers after drying, that is, the drying of the disinfectant for indoor drying. It can be used especially as a composition. It is also suitable for the decomposition and washing of dead cells.

In addition, the function, effect, and the like of the composition may be described, promoted, or advertised as a letter or a picture in a package or pamphlet of the composition product.

In addition, the bactericidal treatment method of the present invention is capable of efficiently decomposing and removing dead cells, and the washing method of the present invention can suppress generation of odors at the time of indoor drying of fibers or storage of fibers after drying. have.

The detail about each component and aqueous solution in the disinfection method and washing method of this invention is based on the said disinfecting detergent composition.

In the present invention, the substance adhered to cells is a hard surface such as a dishwasher, a triangular corner (kitchen trash container), a washing container, a drainage pipe, a bath, a bath, a toilet, or other hard surfaces such as utensils, clothing, bedding, and curtains. And textile products such as carpets, floor mats, and cleaning tools.

Although it does not specifically limit as a bactericidal treatment method, The suitable treatment method by the substance to which a microbe adhered is shown below.

In the case of a hard surface or an apparatus, it is preferable to immerse a substance with bacteria in an aqueous solution or to apply, spray or pour an aqueous solution on the substance with bacteria.

In the case where the substance adhered with the cells is a textile product such as clothing, washing with a washing machine or washing after immersion treatment is preferable.

The lysis effect can be further enhanced by adjusting the temperature of the aqueous solution, the treatment time, the amount of the aqueous solution, and the like in the following preferred ranges.

In the case of a solution of 2.5 U / mL or more, the temperature is preferably 10 to 90 ° C, particularly preferably 10 to 60 ° C. 5 second-10 hours are preferable, and, as for processing time, 10 minutes-60 minutes are especially preferable.

On the other hand, in the case of a solution of 15 U / mL or more, the treatment time is preferably 5 seconds to 60 minutes in an aqueous solution of 10 to 90 ° C, and particularly preferably 5 seconds to 30 minutes at 10 to 60 ° C.

When the substance is a hard surface, the amount of the aqueous solution is preferably 0.01 to 0.5 mL per 1 cm ² surface, and particularly preferably 0.1 to 0.3 mL. In the case of fibers, 1-30 mL is preferable per 1 g of fibers.

According to the present invention, it is possible to provide a bactericidal treatment method, a bactericidal detergent composition and a washing method capable of efficiently decomposing and removing the cells by using a low irritant component.

EXAMPLES Next, the present invention will be described in more detail with reference to Examples, but the present invention is not limited to these Examples. In addition, the following% represents the mass%.

[Examples 1-17, Comparative Examples 1-5]

The aqueous solution of the composition shown to Table 1, 2 was prepared using the tap water adjusted to 3.3 degrees DH (33 mg / L as CaO) using calcium chloride dihydrate. The lysis rate of the obtained aqueous solution was measured based on the following measuring method. The results are written together in Tables 1 and 2. In addition, the measuring method of the lysis activity of the used enzyme is shown below. In addition, the enzyme activity in the table | surface converted the density | concentration in the solution of each enzyme to U / mL based on the lysis activity value (U / unit weight) of an enzyme sample.

Moreover, about thermolysine which is an enzyme which does not have a chelating agent tolerance, it melt | dissolved and used so that it might become 10 mg / L (* of Table 2).

<Measurement method of unit U>

25 ml of a 25 degreeC water bath was prepared by dissolving EDTA and sodium tetrahydrate 111 ppm, sodium bicarbonate 278 ppm and an enzyme in sterile water (distilled water was sterilized by heating at 121 ° C. for 20 minutes) at pH 8.5 with sodium hydroxide. I kept warm for five minutes. To this solution was added 1 mL of heated cells of Escherichia coli (Eschericha coli ATCC8739: Escherichia coli), which had a turbidity of OD 660 nm (turbidity at wavelength 660 nm in a spectrophotometer), which was about 3.3, and stirred, And the turbidity of OD 660 nm became 0.3 +/- 0.03 within 30 second, and the U test liquid was obtained. The U test solution was kept in a water bath at 25 ° C. for 10 minutes, and stirred 10 minutes after the addition of the heated cells, and the turbidity of OD 660 nm was measured. The amount of protease required to reduce the turbidity of the U test solution OD 660 nm to 0.05 in 10 minutes was 10 units (unit, U).

In addition, the lysis rate was measured by the following method.

[Measibility Rate Measurement]

(Preparation of Test Bacteria Liquid)

Brain Heart Infusion Medium (manufactured by Diffco Co., Ltd.) was dissolved in a prescribed amount (37 g / L) purified water, and 50 mL of a 500 mL Sakaguchi flask was sterilized. Thereafter, Escherichia coli ATCC8739 (E. coli) was plated in a platinum loop and shaken at 37 ° C for 18 hours. The cells were collected by centrifugation (3,000 rpm (about 1800 G) for 5 minutes) and suspended in 30 mL of sterile water, and again subjected to centrifugation (3,000 rpm (about 1800 G)) to collect the cells and suspended in sterile water. The suspended cells were heated at 90 ° C. for 20 minutes, centrifuged (3,000 rpm (about 1800 G)), and the heated cells were collected and suspended in sterile water to obtain a cell suspension having a turbidity of OD 660 nm of 3.3.

(Lysis rate measurement)

9 mL of the sample which measures a lysis rate was added to the test tube, and it kept warm for 5 minutes in 25 degreeC water bath. 1 mL of the cell suspension obtained above was added and stirred to obtain a total solution of 10 mL of lysis rate, and the turbidity of OD 660 nm was measured at the start of the reaction (within 30 seconds). Thereafter, the mixture was kept warm for 10 minutes in a 25 ° C water bath, and the turbidity of OD 660 nm was measured after stirring. As a blank, 1 mL of sterile water was used instead of the cell suspension. In addition, when turbidity of OD 660 nm exceeds 0.7 before adding a cell suspension to the sample which measures a lysis rate, it filtered beforehand by Millipore filter (0.45 micron), and added 1 mL of cell suspensions to 9 mL of the solution. In addition, it adjusted so that said "10 mL of lysis rate test liquids" may be set to the density | concentration of the aqueous solution and the disinfecting detergent composition solution to measure.

The lysis rate was calculated | required by the following formula.

<Calcium concentration measurement>

In the above "(lysis rate)", when preparing a "lysis rate test solution", Japanese Unexamined Patent Application Publication No. 9-318615 for a test solution in which 1 mL of sterile water was added to 10 mL in total instead of the "cell suspension". Calcium ion concentration was measured by the method according to the calcium ion concentration measuring method (absorbance method using a ponta chromium violet) described in the Example of the. When precipitates, such as a zeolite, exist in a solution, it performed after filtering by a Millipore filter (0.45 micron).

[Examples 18 to 35]

About an aqueous solution which prepared the antibacterial detergent composition of the composition shown to Tables 3-7, and adjusted it to the use concentration in the table with tap water adjusted to 3 * 3 degrees DH (33 mg / L as CaO) using calcium chloride dihydrate, The lysis rate was measured by the same method as described above. The results are shown in Tables 3-7. The compositions of Tables 3-7 are the following cleaning agents, respectively. In addition, the lysis activity (U / mL) in a table | surface is the conversion of the concentration in solution to U / mL from the use concentration in the table | surface based on the lysis activity value (U / unit weight) of an enzyme, and calcium ion concentration in a table | surface And about the lysis rate, it adjusted and measured so that the "test liquid and lysis rate test liquid" described in the said "calcium ion concentration measuring method and lysis rate measuring method" may be used concentrations in a table | surface. The same applies to the following tables.

Table 3: Dishwashing Detergents

Table 4: Detergents for Washing Machines

Table 5: Bath Detergents

Table 6: Drainage Detergent

Table 7: Toilet cleaners

Examples 36 to 38 and Comparative Example 6

After preparing the medical detergent of the composition shown in Table 8, washing and drying by the following evaluation method-1, odor evaluation was performed. About the aqueous solution diluted to the use concentration in table with tap water of 3 degrees DH, evaluation result is written together with the result of a lysis rate in Table 8.

Evaluation method-1

5L of tap water (3 ° DH) at 25 ° C is placed in a two-chamber washing machine (CW-C30A1-H manufactured by Mitsubishi Denki Co., Ltd.), and the detergent composition is dissolved to the use concentration shown in Table 8. 5 polo shirts and 5 kg of new 100% cotton towels used daily in the kitchen are added in total for 1 minute, soaked for 10 minutes, 25L of tap water is added and washed for 10 minutes, and dehydrated for 1 minute, followed by 30L of tap water. Two minutes of rinse off and two times of dehydration for one minute (during rinse and last). Subsequently, it dried with the standard course by the commercial dryer (DE-N5S3 by Hitachi Sekushoku Co., Ltd.).

After drying the polo shirt and towel in a constant temperature room at a temperature of 30 ° C. and a humidity of 80% RH for 5 days, the odor strength of the polo shirt and towel was evaluated by an experienced monitor based on the following criteria (average value of five people). ).

Evaluation standard

5: remarkably bad smell

4: quite stinky

3: slightly stinky

2: very little stinky

1: Almost no smell or odor

[Examples 40-47, Comparative Example 7]

After preparing the medical detergent of the composition shown to Table 9, 10, and washing and drying by the following evaluation method-2, odor evaluation was performed. About the aqueous solution diluted to the use concentration in table | surface with tap water of 3 degrees DH, evaluation result is written together in Table 9, 10 with the result of a lysis rate.

Evaluation method-2

1 kg of 5 polo shirts worn daily for direct contact with the skin and 5 sheets of 100% cotton towels used in the kitchen for 1 day are placed in a fully automatic washing machine (NA-F70AP, manufactured by Matsushita Denki Co., Ltd.), and the temperature is approximately 25 ° C. The tap water of about 3 degrees DH was poured, and it wash | cleaned by the standard course. In addition, detergent input amount followed the use concentration shown to Table 9, 10. After dehydration, it was dried in a room temperature of 30 ° C. and a humidity of about 80% RH, and dried for 12 hours. For the odor intensity of the towel after 12 hours, the skilled monitor performed odor evaluation based on the following criteria (average value of five people).

Evaluation standard

5: remarkably bad smell

4: quite stinky

3: slightly stinky

2: very little stinky

1: Almost no smell or odor

[Examples 48-51, Comparative Example 8]

After preparing the medical detergent of the composition shown in Table 11, washing and drying by the following evaluation method-3, odor evaluation was performed. In addition, the result is shown together in Table 11 with the lysis rate measured similarly to the above.

Evaluation method-3

After using 10 sheets of new 100% cotton towels for about 1 day each in normal life, put it in a drum type washing machine (ES-E61 manufactured by Sharp), adjust the temperature to about 20 ° DH by adding calcium chloride and dihydrate at a temperature of about 40 ° C. One tap was poured and the washing was done with a standard course. In addition, the detergent input amount was made into 5,000 ppm of detergent concentration, after dehydration, it was dried in room temperature of 30 degreeC, and about 80% RH of humidity, and it dried for 12 hours. The odor intensity of the towel after 12 hours was evaluated by a skilled monitor based on the following criteria (average value of five people).

Evaluation standard

5: remarkably bad smell

4: quite stinky

3: slightly stinky

2: very little stinky

1: Almost no smell or odor

In addition, the component used in Tables 1-11 is as follows. The compounding quantity in a table | surface is the value which carried out net conversion (except Na percarbonate and a bleach activator).

Sodium lauryl sulfate: 95% pure, Lion Co., Ltd., Sunol LM-1100NT

α-SF-Na: sodium salt of α-sulfofatty acid methyl ester having an alkyl group having 14 to 16 carbon atoms, a Lion Co., Ltd. product, 70% pure, 72 to 73% solid

LAS-K: Linear alkyl (10-14 carbon atoms) benzenesulfonic acid potassium (at the time of preparation of a detergent composition, neutralizing Lipon LH-200 (96% of LAS-H pure content) by 48% potassium hydroxide aqueous solution) Prepared)

LAS-Na: Straight-chain alkyl (C10-C14) benzenesulfonate sodium (formulated by neutralizing Lipon LH-200 (96% of LAS-H pure components) by 48% sodium hydroxide aqueous solution)

AOS-Na: α-olefin sulfonate sodium having an alkyl group having 14 to 18 carbon atoms (product manufactured by Lion Co., Ltd., 55% pure slurry)

AES-Na: polyoxyethylene (3 mol of EO) lauryl ether sulfate, pure 70%, Lion Co., Ltd. product

PTS: Paratoluene Sulphonic Acid

Cumenesulfonic acid: manufactured by Albright & Wilson

Soap: Fatty acid sodium having a C12-18 alkyl group (Lion Co., Ltd. product, C12: 0.9%, C18: 80.2%, unsaturated fatty acid 80.2%, molecular weight 289, pure content 67-68%, titer 47.0 ° C)

N-lauryldimethyl acetate betaine: Sanyo casein

Alkyl amidopropyl acetate betaine: Ipposha Yugyo Co., Ltd.

BRE15: straight chain primary alcohol ethoxylate of carbon chain 12 (average added mole number of ethylene oxide: 15) Lion Corporation

Aet1: ethylene oxide average 15 mole adduct of diamond 12 (made by mitsubishi kagaku) (product made by lion kagaku)

AEt2: Ethylene oxide average 8 mole adduct of linear alcohol (C12-C14) (made by Lion Kagaku)

AEs: Ethylene oxide average 15 mole adduct of C12-C18 fatty acid methyl ester (made by Lion Kagaku)

AEP: Average 15 mol of ethylene oxide and 3 mol of propylene oxide adducts (made by Lion Kagaku) of diamond 12 (made by Mitsubishi Kagaku)

APG: Alkylpolyglucoside (Cognis Trial)

LDE: lauryl diethanol amide (made by Kawaken Fine Chemicals)

Dicyadiamide: Nippon carbide

CDE: Palm fatty acid diethanolamide (made by Kawaken Fine Chemicals)

AX: lauryl dimethylamine oxide (made by Lion Co., Ltd.)

Enzyme (protease)

Savinase 12T, Esperase 4T, Everase 8T, Cannase 24TK, Alcalase 2T, Savinase 16L, and Deozyme are detergent preparations for Novozymes.

Penzyme: BaciI1us sp. It was produced from P (Microbiological Research Institute Accession No. 8090), and prepared by the culture purification described in JP-A-8-165493.

K enzyme: Bacil1us sp. It was produced from K (Microbiological Research Institute Accession No. 8801), and prepared by the culture purification described in Japanese Patent Application Laid-Open No. 8-165493.

X enzyme: Bacillus sp. It was produced from X (Microbiological Research Institute Accession No. 8802) and prepared by the culture purification described in Japanese Patent Application Laid-Open No. 8-165493.

Y enzyme: Bacillus sp. It was produced from Y (Microbiological Research Institute Accession No. 8088), and prepared by the culture purification described in Japanese Patent Application Laid-Open No. 8-165494.

Thermolysin: Sigma Priest

<Enzyme (others)>

Amylase: 10: 1 (weight ratio) mixture of Duramil 60T (made by Novozymes) and pullulanase (made by Sigma)

Cellulase: Cellulzyme 0.7T (manufactured by Novozymes)

Lipase: Lipex 50T (manufactured by Novozymes)

<Chelating agent>

ASDA: Aspartic acid sodium acetate (product of Nippon Shokubai Co., Ltd.)

HIDS: hydroxyiminodisuccinate sodium (product made by Nippon Shokubai Co., Ltd.)

EDTA.4Na: ethylenediamine sodium tetraacetate

Phenanthroline: 1, 10-phenanthroline (made by Nihon Kayaku Co., Ltd.)

Sodium tripolyphosphate: manufactured by Nippon Kagaku Co., Ltd.

Citric acid and its sodium salt: manufactured by Fuso Kagaku Kogyo Co., Ltd.

Zeolite: Zeolite A (Mizusawa Kagaku, Silton B)

AA / MA copolymer salt: Sodium salt of acrylic acid / maleic acid copolymer, trade name Socalan CP7 (manufactured by BASF)

Polyacrylic acid Na: Sodium polyacrylate (made by Nippon Kayaku, Aaron Bis S)

Malic acid; Fuso Kagaku High School Co., Ltd.

<Other Ingredients>

Na carbonate: sodium carbonate (made by Asahi Glass, granule)

K carbonate: potassium carbonate (made by Asahi Glass)

Silicate Na: JIS No. 1 sodium silicate (made by Nippon Kagaku Co., Ltd.)

Sulfuric acid Na: neutral anhydrous forage (Nippon Kagaku Co., Ltd. product)

Na chloride: Sodium chloride (reagent)

Sulfurous acid Na: Anhydrous sodium sulfite (made by Mitsui Chemical Co., Ltd.)

Ethanol: 99.5% pure (product made in Japan ethanol)

Diethylene glycol monobutyl ether: manufactured by Nippon Nuka Co., Ltd.

Fluorescent agent CBS: Cinopearl CBS-X (made by Ciba specialty chemicals company)

Fluorescent agent AMS: Cinopearl AMS-GX (made by Ciba specialty chemicals company)

Na percarbonate: Coated sodium percarbonate (made by Mitsubishi Chemical Corporation, SPC-D)

Bleach activator OBC: 4-decanoyloxy benzoic acid and polyethylene glycol (PEG) # 6000 (product made by Lion Co., Ltd.) and C14 alpha olefin sulfonic acid powder which are bleach activators for Extrude Ohmic EM 16 made by Hosokawa Micron Co., Ltd. The product (Lipola PB-800, Lion Co., Ltd.) was mixed so that it might become 70/25/5 by mass ratio, and it injected | threw-in and knead | extruded and obtained the noodle-shaped extruded product of 0.8 mm diameter in diameter. The extruded product (60 ° C.) was kneaded and extruded using a Pitsmill DKA-3 type manufactured by Hosokawa Micron Co., Ltd., and introduced in the same direction as the granulated product was introduced. And a bleaching activator granulated product having an average particle diameter of 700 µm obtained by pulverization.

Fragrance composition

Fragrance of Table 3, 4, 5, 7, 8, 9, 10, 11: Japanese Patent Laid-Open No. 2002-146399

The composition of description in Tables 11-18.

Fatty acid amidopropyl betaine: (manufactured by Lion Co., Ltd.)

Acylamidopropyl Acetate Betaine: (manufactured by Lion Co., Ltd.)

Alkylphenoxy phenyl disulfonic acid Na: (manufactured by Nikko Chemical Co., Ltd.)

Stearyl trimethyl ammonium chloride: (manufactured by Lion Arc Co., Ltd.)

Didecylmethyl ammonium chloride (made by Lion Arc Co., Ltd.)

Synthetic polymer: Methacrylic acid copolymer (manufactured by Ipposha New High School)

Paratoluene sulfonic acid: (manufactured by Kyowa Hakko Co., Ltd.)

Triethanolamine: (manufactured by Nihon Shokubai Co., Ltd.)

Silica: Silicon graft polymer (manufactured by Ipposha New High School)

0.1N sodium hydroxide: (reagent)

NRE2: (product made by Lion Co., Ltd.)

NRE5: (product made by Lion Corporation)

NRE7: (product made by Lion Corporation)

BRE15: (product made by Lion Co., Ltd.)

Polyethylene glycol (product of Nippon Nushi)

Diethanolamine (made by Nihon Shokubai Co., Ltd.)

Triethanolamine (made by Nippon Shokubai Co., Ltd.)

From the results in Tables 1 and 2, the lysing enzyme having chelating agent resistance was contained at a concentration such that the lytic activity was 2.5 U / mL or more, the calcium ion concentration was 10 ppm or less in terms of CaO, and the pH was greater than 8 It turns out that the aqueous solution of the following ranges shows the outstanding lysis rate. In addition, an aqueous solution containing a chelating agent resistant to chelating agent at a concentration such that the lytic activity is 15 U / mL or more, the pH of which is greater than 8 and 12 or less, exhibits excellent lysis rate even when the calcium ion concentration exceeds 10 ppm. It can be seen that.

From the results of Tables 3-7, the Example showed the excellent lysis effect. In addition, using the detergent of Table 3, the dishwashing dish was cleaned for one week using a dishwashing sponge (with stickiness and odor after 6 months of use). Significant improvement in stickiness and odor was observed. The aqueous solution of Example 2 was sprayed onto a kitchen sink, a triangular corner, and a drain hole (wire mesh part) and washed with water 10 minutes later, whereby a significant decrease in stickiness was observed.

From the result of Tables 8-11, the Example showed the outstanding malodor suppression effect.

Claims (10)

The chelating enzyme having a chelating agent resistance is contained at a concentration such that the lytic activity is 2.5 U / mL or more, the calcium ion concentration is 10 ppm or less in terms of CaO, the pH is greater than 8, Disinfection treatment method characterized in that to treat the substance attached. A bactericidal treatment method comprising lysing a lysing enzyme having a chelating agent at a concentration such that the lytic activity is 15 U / mL or more, and treating the substance to which the cells are adhered using an aqueous solution having a pH greater than 8 and 12 or less. A bactericidal detergent composition comprising a bactericidal enzyme having chelating agent resistance and not containing phosphate, wherein the bactericidal detergent composition or its dilution liquid contains the bactericidal enzyme at a concentration such that the bactericidal activity is 2.5 U / mL or more, A bactericidal detergent composition, wherein the calcium ion concentration is a solution of 10 ppm or less in terms of CaO and a pH of more than 8 and 12 or less. A bactericidal detergent composition containing a lysing enzyme having chelating agent resistance and containing no phosphate, wherein the bactericidal detergent composition or its dilution liquid contains the bactericidal enzyme at a concentration such that the bactericidal activity is 15 U / mL or more, Is a solution of greater than 8 and less than or equal to 12. The bactericidal cleaner composition according to claim 3 or 4, further comprising a surfactant. The fatty acid production amount from triolein after a substrate degradation test is twice or more (molar ratio) with respect to the fatty acid production amount from n-hexadecyl palmitate after a substrate decomposition test according to any one of claims 3 to 5. A sterile detergent composition comprising a lipase having enzymatic activity. The wild-type amylase derived from the genus Bacil1us further comprises a starch degrading enzyme that degrades the α-1, 6 bond of amylase or starch modified by genetic variation. Antibacterial detergent composition. 8. The bactericidal cleaner composition according to any one of claims 3 to 7, which is used for decomposition and removal of killed cells. 8. The bactericidal according to any one of claims 3 to 7, wherein the fibers are sterilized and cleaned by using the bactericidal detergent composition or the diluent thereof, and used for suppressing the occurrence of odor at the time of drying the fibers indoors or storing the fibers after drying. Detergent composition. When washing fibers using a bactericidal detergent composition containing a chelating agent resistant chelating agent and a surfactant, and containing no phosphate, (a) A lytic enzyme is contained at a concentration such that the lytic activity is 2.5 U / mL or more, and the calcium ion concentration is 10 ppm or less in terms of CaO and the pH is more than 8 and 12 or less. (b) The microbial cells are sterilized and cleaned using an aqueous solution of a microbial detergent containing a lysing enzyme at a concentration of at least 15 U / mL of lytic activity and adjusted to a pH of more than 8 and any one of 12 or less. A method for washing fibers, wherein the occurrence of odors during drying or indoor storage of fibers after drying is suppressed.