JPH0530880B2 - - Google Patents

Description

[Detailed description of the invention]

[Industrial Field of Application] The present invention relates to a cleaning composition. More specifically, the present invention relates to a cleaning composition characterized by containing a special cellulase produced from a specific cellulase-producing bacterium and having high enzymatic activity even in alkaline conditions. [Prior Art] In recent years, there has been significant progress in washing clothes. That is, washing clothes has become significantly easier due to the development of raw materials suitable for detergents, improvements in water quality, improvements and spread of washing machines, improvements in fibers, etc. In particular, improvements in raw materials for detergents have been remarkable, and it appears that the composition of laundry detergents has almost reached the point of perfection through modifications of surfactants, builders, dispersants, fluorescent dyes, bleaching agents, etc. be. However, the idea behind the development of laundry detergents is that (1) surfactants and builders adsorb to the dirt and/or fiber surface, thereby reducing the bond between the dirt and/or fiber and water; (2) Disperse and solubilize dirt with surfactants and inorganic builders; (3) Chemically decompose dirt with enzymes such as protease; (4) Bleaching colored stains with bleach, etc., (5) Adsorbing fluorescent dye, etc. on the fiber surface,
(6) Using a chelating agent to prevent precipitation of divalent metal ions in ingredients effective for cleaning. That is, the basis of conventional clothing cleaning has been how to effectively incorporate components that directly attack dirt or components that assist in the attacking power of such components as one component of the detergent composition. [Problems to be Solved by the Invention] However, at present, the cleaning performance of cleaning compositions based on this basic principle has almost reached a saturation point in a sense, and there is a need for cleaning compositions with even higher cleaning power. development was desired. [Means for Solving the Problems] The present inventors have discovered that the activity of cellulase produced by a specific cellulase-producing bacterium decreases slowly even in an alkaline region, and has superior cleaning power compared to conventional cellulase-containing products. The present invention was completed based on the discovery that this is significantly superior. The special cellulase used in the present invention is a cellulase produced by an alkalophilic actinomycete belonging to the genus Streptomyces, and is a special cellulase that maintains high activity even in an alkaline region and has alkaline tolerance. The microorganism used to produce the enzyme used in the present invention is an alkalophilic actinomycete belonging to the genus Streptomyces, and any microorganism or mutant strain thereof that has the ability to produce the cellulase of the present invention may be used. Examples of strains of alkalophilic actinomycetes belonging to the genus Streptomyces include Streptomyces sp. KSM-2.
(Feikoken Bibori No. 7621) and Streptomyces sp. These two strains were isolated by the present inventor from soil in Haga District, Tochigi Prefecture, and have the mycological properties shown in Tables 1 and 2 below.

【table】

【table】

【table】

[Table] Strains KSM-2 and KSM-9 have morphological characteristics of actinomycetes. Also, it is classified into the genus Streptomyces because its cell wall contains L-L-diaminopimelic acid. The KSM-2 strain belongs to the yellow series of strains based on the color of aerial mycelia on various agar media, the surface of the spores is smooth, the spore chains are slightly curved or generally straight, and they resemble melanin. Based on the fact that no pigment is produced and the results of a carbon source assimilation test, we searched for similar strains to this strain among known strains according to the 8th edition of Virgies Manual, and found that Streptomyces canescens
Canescens) is mentioned as a similar strain. However, the two differ in that the KSM-2 strain produces alkaline-resistant cellulase, whereas Streptomyces canescens does not produce alkaline-resistant cellulase. Furthermore, when searching for KSM-9 according to the 8th edition of Virgie's Manual, Streptomyces Puniceus is listed as a similar strain. However, whereas Streptomyces puniceus is characterized by producing purple to red pigments, KSM-9 does not have this property, and only KSM-9 produces alkaline-resistant cellulase. The two are different. Therefore, the present inventor developed the KSM-9 strain and KSM-2 strain.
The strain was identified as a new bacterial species and deposited at the Institute of Microbiology, Agency of Industrial Science and Technology as described above. Isolation of this bacterial strain from soil as an isolation source can be carried out by a conventional method using an agar medium, for example, as shown in Reference Example 1 below. The culture medium may be any medium as long as it is utilized by the strain used and contains carbon sources, nitrogen sources, organic nutrient sources, inorganic salts, etc. necessary for the successful production of alkaline cellulase. Examples of carbon sources include soluble cellulose derivatives such as carboxymethylcellulose (CMC), cellulose such as pulp powder, paper powder, and solid fibers such as Avicel; carbohydrates such as glucose, fructose, sucrose, or sorbitol; citric acid. , organic acids such as succinic acid; n-dodecane, n
- Any hydrocarbon such as hexadecane can be used as long as it can be assimilated. Among these carbon sources, a medium using cellulose, especially a soluble fiber derivative, is suitable because it produces a large amount of alkaline cellulase. Examples of the nitrogen source or organic nutrient source include nitrates such as sodium nitrate, potassium nitrate, and ammonium nitrate; yeast extract, meat extract, and peptone. Examples of inorganic salts include carbonates such as sodium carbonate, potassium carbonate, sodium hydrogen carbonate, and potassium hydrogen carbonate, sodium phosphate, sodium hydrogen phosphate, sodium dihydrogen phosphate, potassium phosphate, potassium pyrophosphate, and pyrophosphate. Phosphates such as sodium, sodium tripolyphosphate and sodium hexametaphosphate, and inorganic salts such as magnesium sulfate can be used. Among them, carbonate is 0.1
A medium containing ~1.5% by weight is suitable because it produces a large amount of alkaline cellulase. In addition, trace amounts of heavy metal salts are used, but their addition is not necessarily required in media containing natural products. Furthermore, when using a mutant strain that requires nutritional sources other than those mentioned above, a substance that satisfies the nutritional requirements must be added to the medium. For culturing, the culture medium may be sterilized by heating or the like, inoculated with bacteria, and shaken or stirred with aeration at 25 to 35°C for 2 to 4 days. Good results can be obtained by adjusting the pH to around 8 to 11. When using a carbon source that is poorly soluble in water, it is also possible to add various surfactants such as polyoxyethylene sorbitan to the medium. As the cellulase used in the present invention, the above-mentioned culture solution itself may be used, or a crude enzyme solution obtained by removing bacterial cells by centrifuging the culture solution or the like may be used. Alternatively, an enzyme powder obtained by purifying the enzyme by ammonium sulfate fractionation or precipitation with an organic solvent such as acetone or ethanol may be used. As described above, Streptomyces sp.
The alkaline cellulase KSM-2 obtained from KSM-2 and the alkaline cellulase KSM-9 obtained from Streptomyces sp. KSM-9 have the following physicochemical properties.

[Effect]

As mentioned above, the present invention is characterized in that a special cellulase is used as a component of a cleaning composition, but surprisingly, it contains inorganic stains that are completely unrelated to cellulase activity, especially inorganic stains and skin. It exhibits remarkable cleaning properties for collar stains that change over time due to the mixture of oil secreted on the surface. As mentioned above, the use of enzymes in the technical field of detergents is known, but only enzymes that are particularly effective against dirt are known. That is, protease is known to fight protein stains, amylase is known to fight starch stains, and lipase is known to fight oil stains, all of which are enzymes that directly attack stains. The cleaning mechanism of cellulase in the present invention has not yet been completely elucidated, but it is not based on a mere swelling effect on the fibers, the essence of which can be seen in the surfactant. [Effects of the Invention] Thus, according to the present invention, the cleaning agent of the present invention contains a special cellulase having high activity in an alkaline region and having alkali resistance produced by live alkalophilic actinomycetes belonging to the genus Streptomyces. By using the agent composition, the cleaning bath
Excellent cleaning effects can be obtained over a wide range of pH. Furthermore, this effect more than compensates for the decrease in detergency caused by the decrease in alkaline ability among the builder effects accompanying the decrease in the pH of the cleaning bath during cleaning. [Reference example] Reference example 1 One cup of soil spatula from Haga District, Tochigi Prefecture (approximately 0.5
g) was suspended in 10 ml of sterile water, thoroughly stirred, and left to stand. 0.1 ml of the soil suspension supernatant obtained in this way was
It was applied to a separation agar medium having the following composition. Composition: CMC 20g Peptone 10g Meat extract 10g KH ₂ PO ₄ 1g NaCl 10g Na ₂ CO ₃ 10g Agar 7.5g Water 1000ml PH10.5 Next, this was cultured at 30℃ for 3 days, and the area around the village was grown to dissolve CMC. After confirming the appearance of bacteria with a clear zone, the colonies forming a clear zone were collected and inoculated onto a slanted agar medium with the same composition as the above isolation agar medium, and cultured at 30°C for 3 days. It was confirmed macroscopically and microscopically that the bacterial strain on the slant medium was the same strain. It was also confirmed that the properties and physiological properties of these 10 strains on each medium were the same. The properties and physiological properties of the above strain on each medium are shown in Tables 1 and 2 above, and the present inventor named it Streptomyces sp. KSM-2. . Streptomyces sp. KSM-9 was similarly isolated from soil in Haga District, Tochigi Prefecture. As a result of the above test, it was found that each of the 10 cultured bacteria was a single strain isolated from nature. Next, a loopful of the pure cultured bacterial strain on the slant medium was suspended in a cryopreservation vial containing a sterilized 10% glycerin aqueous solution (2 ml), and -
Store frozen at 80℃. After 3 months of frozen storage, a loopful of the suspension obtained by rapid thawing was placed on an ordinary agar medium, and the properties and physiological properties were examined on each medium under the same conditions as above. No change was observed. Furthermore, when the properties and physiological properties of the strains on each culture medium were similarly examined after the above-mentioned freezing and thawing was repeated five times every month, no changes were observed. Reference example 2 Avicel 1.0% in a 500ml Sakaguchi flask,
Add 1.5% meat extract, 0.5% yeast extract, 0.1% potassium dihydrogen phosphate, and 0.5% sodium carbonate (PH9.5), and add Streptomyces sp.
KSM-2 strain (Feikoken Deposit No. 7621) was inoculated through a slant and cultured with shaking at 30°C. After culturing for 2 days, the cells were removed by centrifugation, the resulting culture solution was fractionated with ammonium sulfate, and the resulting solid fraction was freeze-dried to obtain enzyme powder. 0.6 g of enzyme powder was obtained per culture solution. The CMC degrading activity of the obtained enzyme was 1.84 mg and 1.02 mg glucose equivalent/mg min at pH 7.0 and 9.0, respectively, and the Avicel degrading activity was similarly
They were 0.083 mg and 0.056 mg glucose equivalent/mg min. Reference example 3 CMC1.0 instead of Avicel 1.0% in reference example 2
Enzyme powder was obtained from the culture solution obtained by culturing Streptomyces sp. of the obtained enzyme
CMC degrading activity is 0.90 mg and 0.44 at PH7.0 and 9.0, respectively.
mg glucose equivalent/mg·min. Reference example 4 Streptomyces sp.KSM as in reference example 2
Enzyme powder was obtained from the culture solution obtained by culturing the -9 strain (Feikoken Bacteria Deposit No. 7620). of the obtained enzyme
CMC degrading activity is 3.85 mg and 2.50 at PH7.0 and 9.0, respectively.
mg glucose equivalent/mg·min. Reference example 5 Streptomyces sp.KSM as in reference example 3
Enzyme powder was obtained from the culture solution obtained by culturing -9 strain. The CMC decomposition activity of the obtained enzyme was 2.01 mg and 1.49 mg glucose equivalent/mg·min at pH 7.0 and 9.0, respectively. [Example] In the following examples, studies were conducted under the following experimental conditions. 1) Contaminated natural collar cloth: Cotton gold cloth #2023 was sewn onto the collar of a shirt and an adult male was allowed to wear it for 3 days. 25 days after wearing
After leaving it for one month at ℃ and 65% RH, the degree of staining is divided into three levels, and from among the most heavily soiled fabrics, the fabric with the stains symmetrical to the center point is selected, and the fabric is cut in half at the symmetrical point of the stains. It was used for experiments. 2) Cleaning conditions and method When cleaning naturally contaminated fabrics, cut a 9cm x 30cm naturally contaminated fabric in half at symmetrical positions, and wash one of the 9cm x 15cm pairs of contaminated fabrics with a standard detergent that does not contain enzymes. Then, one side was washed with the detergent of the present invention, which is a comparison detergent. First, a piece of naturally contaminated cloth.
Sewn 15 sheets onto a 50cm x 50cm cotton cloth, add 1kg of the contaminated cloth and cotton underwear to a 0.665% detergent solution (6) in the case of powdered detergent, soak it at 30℃ for 2 hours, and then use Toshiba's Transfer to washing machine "Ginga"
After adjusting the total volume to 30%, wash with strong inversion for 10 minutes,
In the case of the liquid detergent used for the evaluation after drying, 20 c.c. of the detergent solution was evenly applied to the contaminated cloth, and after 10 minutes, the cloth was weighed to a total weight of 1 kg with cotton underwear.
The total volume was adjusted to 30,000 ml, and washed with strong inversion for 10 minutes. After drying, it was used for evaluation. A half-cut fabric washed with the standard detergent and a half-cut fabric washed with the detergent of the present invention were evaluated by pairwise comparison by visual judgment. Cleaning cloths were ranked based on standard soiling, which is ranked in 10 levels to indicate the degree of soiling. The detergency was expressed as a score for the detergency of the detergent of the present invention, with the detergency of the reference detergent set at 100. A difference in detergency index of 0.5 or more can be considered a significant difference. 3) Enzyme used Cellulase of the present invention (obtained in Reference Example 2, diluted 20 times with Glauber's salt and granulated) Cellulase of the present invention (obtained in Reference Example 3, diluted 20 times with Glauber's salt) Cellulase of the present invention (obtained in Reference Example 4, diluted 20 times with Glauber's salt and granulated) Cellulase of the present invention (obtained in Reference Example 5, granulated 20 times with Glauber's salt) cellulase (SIGMA Type I, origin:
Aspergillus niger) lipase (Gist Procades NV,
Origin: R.Oryzae) Amylase (Novo Industries, Inc.)
Termamil 60G) Protease (Novo Industries, Alcalase 2.0M) Example 1 A highly alkaline powder laundry detergent was prepared using the following detergent formulation. In addition, the PH of a 0.665% aqueous solution of detergent is
It was 11.3. Composition: Linear sodium dodecylbenzene sulfonate 20% Soap (sodium tallow fatty acid) 2 Sodium orthophosphate 20 Sodium metasilicate 10 Sodium carbonate 15 Carboxymethyl cellulose 1 Polyethylene glycol 1 Fluorescent dye 0.4 Salmon salt Balance enzyme 0 or 2 Water 5 Table 1 shows the results of cleaning tests using various detergents.
Shown below. The detergent numbers in the table are expressed as Example number minus Enzyme number used. (However, those that do not use enzymes are indicated as Example number -.)

[Table] Example 2 A weakly alkaline powder laundry detergent was prepared using the following formulation. The pH of a 0.665% aqueous solution of detergent is
It was 10.5. Composition: Sodium alpha olefin sulfonate 10% Linear sodium dodecylbenzenesulfonate 10 Soap 1 Sodium tripolyphosphate 20 Sodium silicate (JIS No. 2 Keiso) 10 Sodium carbonate 5 Carboxymethylcellulose 1 Polyethylene glycol 1 Fluorescent dye 0.4 Salt Nitrogen Balanced fermentation Base: 0 or 2 Moisture: 10 As in Example 1, a cleaning test was conducted and the results are shown in Table 2.

[Table] Example 3 A neutral powder laundry detergent was prepared using the following formulation. The pH of a 0.665% aqueous detergent solution was 7.2. Composition: Straight chain alcohol (=14) Sodium sulfate 30% Polyethylene glycol 1 Sodium phosphate 1 Fluorescent dye 0.2 Glauber's salt Balance enzyme 0 or 2 Moisture 5 The results of the cleaning test for each detergent are shown in Table 3.

[Table] Example 4 A phosphorus-free, weakly alkaline detergent was prepared using the following formulation. Composition: Linear sodium dodecylbenzene sulfonate 15% sodium alkyl ethoxy sulfate ( _C14 - _C15 ,
EO = 3 moles) 5 Builders and enzymes (see Table 4) 20 Sodium silicate 15 Sodium carbonate 15 Sodium polyacrylate 1.5 Polyethylene glycol 1.5 Fluorescent dye 0.5 Salmonella balance moisture 5 Table 4 shows the results of the cleaning test.

[Table] Example 5 Detergents were prepared using various combinations of enzymes in the formulation of Example 2. Table 5 shows the results of the cleaning test for each detergent obtained.

【table】

[Table] Example 6 A weakly alkaline powder laundry detergent was prepared using the following formulation. Composition: Sodium alkyl sulfate (=14.5) 15% Sodium alkyl ethoxy sulfate (=
14.5, = 3) 5 Soap (tallow-based) 2 Sodium pyrophosphate 18 Sodium silicate 13 Sodium carbonate 5 Polyethylene glycol 2 Fluorescent dye 0.2 Salmon sulfate Balanced magnesium silicate 1 Moisture 5 Enzyme 2 Sodium percarbonate 15 Obtained Table 6 shows the results of the cleaning test for each detergent.

[Table] Example 7 A weakly alkaline liquid laundry detergent was prepared using the following formulation. The pH of the detergent stock solution was 9.5. Composition: Secondary alcohol ethoxylate (=13.5,
EO=7.0) 10% Linear sodium dodecylbenzenesulfonate 20 Coconut fatty acid diethanolamide 3 Carboxymethyl cellulose 1 Polyethylene glycol (6000) 2 Potassium pyrophosphate 14 Sodium formate 1 Calcium chloride 0.01 Sodium meta-xylene sulfonate 5 Enzyme 2 Water Balance Profit Table 7 shows the results of the cleaning tests for each detergent.

[Table] Example 8 A neutral liquid laundry detergent was prepared using the following formulation. The pH of the detergent stock solution was 7.0. Composition: Sodium alkyl ethoxy sulfate ( _C14 - _C15 ,
EO=3.0 mol) 20% Secondary alcohol (=13.5) Ethoxylate (=7) 25 Triethanolamine 1 Polyethylene glycol (6000) 2 Carboxymethyl cellulose 1 Citric acid 1 Fluorescent dye 0.3 Blue tinting agent 0.05 EtOH 8 Water Balance Enzyme 2 Table 8 shows the results of the cleaning test for each detergent obtained.

【table】 [Brief explanation of drawings]

Figures 1 and 2 are diagrams showing the PH dependence of the alkaline cellulase activity obtained in Reference Example 5, and Figures 3 and 4 are diagrams showing the PH dependence of the alkaline cellulase activity obtained in Reference Example 3. FIG.

Claims (1)

[Scope of Claims] 1. A cleaning composition containing alkaline cellulase produced by alkaline cellulase-producing bacteria belonging to the genus Streptomyces. 2 The alkaline cellulase producing bacterium is Streptomyces sp. KSM-
The cleaning composition according to claim 1, which is No. 2 (Keikoken Kyoiyori No. 7621). 3 The alkaline cellulase producing bacterium is Streptomyces sp. KSM-
The cleaning composition according to claim 1, which is No. 9 (Keikoken Kyoiyori No. 7620).