JP2010126710A - Biofilm removing agent composition - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a biofilm removing agent composition effectively removing biofilm. <P>SOLUTION: The biofilm removing agent composition contains a basic amino acid derivative expressed by general formula (1) or its salt, and an enzyme. In general formula (1), R<SP>1</SP>represents a linear or branched alkyl or alkenyl group having 4 to 18 carbon atoms; X and Y each represents a group selected from groups of following formulae; and m represents an integer of 1 to 5. <P>COPYRIGHT: (C)2010,JPO&INPIT

Description

The present invention relates to a biofilm removing agent, and more particularly, biofilm removal for effectively removing biofilm and preventing harm caused by biofilm in various fields involving microorganisms. The agent composition.

A biofilm is also called a biofilm or slime, and generally refers to an aqueous system in which microorganisms adhere to and grow on the surface of substances to produce macromolecular substances such as polysaccharides and proteins from within microbial cells to form structures. . When a biofilm is formed, harm caused by microorganisms occurs, causing problems in various industrial fields. For example, when a biofilm is formed in the piping of a food plant, the biofilm peels off and leads to contamination by foreign substances in the product, and causes food poisoning due to microorganism-derived toxins. Furthermore, biofilm formation on the metal surface causes metal corrosion and promotes aging of the equipment.
Furthermore, compared to microorganisms that are dispersed and suspended in an aqueous system, microbial control agents such as bactericides and bacteriostatic agents are often unable to exert sufficient effects on microbial aggregates that have formed biofilms. . For example, in the medical field, many cases of nosocomial infections have been reported in recent years due to microorganisms remaining in narrow gaps and holes in medical devices to form biofilms. In the human oral cavity, it is well known that biofilms formed on teeth, so-called dental plaque (plaque), cause caries and periodontal disease, and these problems have been studied for a long time.

Until now, in order to prevent the harm of biofilm, the idea of not allowing bacteria to grow by giving bactericidal or bacteriostatic action to microorganisms, particularly bacteria, has been generally studied. Patent Document 1 describes an antibacterial preparation containing arginine hydrochloride, arginine ethyl ester, arginine glutamic acid or other arginine or a derivative thereof and a compound exhibiting antibacterial activity, but the effect is not yet satisfactory. In addition, this document shows an antibacterial effect on microbial aggregates and is not intended to remove biofilms.
Furthermore, as disclosed in Patent Document 2, a method for reducing harm by removing a biofilm using an enzyme has been studied, but it has not been completely removed.
JP-A-8-151324 JP-T-2001-508677

  Accordingly, an object of the present invention is to provide a biofilm remover composition that effectively removes biofilms composed of microorganisms and microorganism-producing substances in various regions.

  The present inventor conducted intensive research to obtain a biofilm remover that effectively removes biofilm and microorganisms therein, and a biofilm remover composition that combines a specific amino acid derivative and an enzyme is The inventors have found that the film and the microorganisms therein can be effectively removed, and have completed the present invention.

  That is, the present invention provides the following general formula (1)

(In the formula, R ¹ represents a linear or branched alkyl or alkenyl group having 4 to 18 carbon atoms, and X and Y represent the following formulae:

And m represents an integer of 1 to 5. )
A biofilm remover composition comprising a basic amino acid derivative represented by the formula (1) or a salt thereof and an enzyme is provided.

  The present invention also provides a biofilm remover composition containing an arginine derivative or a salt thereof obtained by reacting arginine or a salt thereof with glycidyl ether and an enzyme.

  According to the present invention, biofilms composed of microorganisms and microorganism-producing substances can be effectively removed in various regions.

  The biofilm remover composition of the present invention has the following general formula (1):

(In the formula, R ¹ represents a linear or branched alkyl or alkenyl group having 4 to 18 carbon atoms, and X and Y represent the following formulae:

And m represents an integer of 1 to 5. )
The basic amino acid derivative represented by these, its salt, and an enzyme are contained.

In general formula (1), the alkyl group or alkenyl group represented by R ¹ may be linear or branched, but has 4 to 18 carbon atoms and 6 to 14 carbon atoms in terms of biofilm removal effect. Are preferable, those having 8 to 14 carbon atoms are more preferable, and those having 10 to 14 carbon atoms are more preferable. Specific examples include an n-hexyl group, a 2-ethylhexyl group, an n-octyl group, a decyl group, a dodecyl group, and a myristyl group. The alkyl group or alkenyl group represented by R ¹ may be single or mixed. Moreover, the mixed alkyl composition derived from natural origin, for example, palm oil or palm kernel oil, may be sufficient.
In general formula (1), m represents an integer of 1 to 5, preferably 2 to 4, and more preferably 3. X is preferably —OCH ₂ —CH (—OH) CH ₂ —, and Y is preferably —NH—C (—NH ₂ ) ═NH.

  Examples of the salt of the basic amino acid derivative (1) include salts of inorganic acids such as hydrochloride, sulfate and phosphate, and salts of organic acids such as acetate, lactate, citrate and acidic amino acid salt. Preferable examples include hydrochloride, acetate, lactate and citrate.

  The basic amino acid derivative (1) can be obtained by reacting a basic amino acid such as arginine, lysine, ortinine, histidine, hydroxyhistidine and the like with a compound such as glycidyl ether, fatty acid chloride, fatty acid anhydride, or epoxyalkane. Preferred is a compound obtained by reacting arginine with a compound such as glycidyl ether, acid chloride, or epoxyalkane, more preferably an arginine derivative obtained by reacting arginine with glycidyl ether, that is, the following general formula ( 2).

(R ² represents a linear or branched alkyl or alkenyl group having 4 to 18 carbon atoms.)
These reaction products may contain unreacted substances and by-products as long as they do not impair the cleaning effect and the storage stability of the preparation.

  The concentration of the basic amino acid derivative (1) in the biofilm remover composition of the present invention can be appropriately determined depending on the use and dosage form. However, in the situation where it acts on the biofilm, it is usually in the state of an aqueous solution. The concentration is preferably 0.001 to 10% by weight, more preferably 0.002 to 7% by weight, and still more preferably 0.005 to 5% by weight from the viewpoint of cost and handleability.

The biofilm remover composition of the present invention further contains an enzyme.
Examples of the enzyme used in the present invention include one or more enzymes selected from oxidoreductase (oxidoreductase), transferase (transferase), hydrase (hydrolase), lyase and isomerase (isomerase). .

Here, examples of the oxidoreductase (oxidoreductase) include peroxidase, superoxide dismutase, and lactate dehydrogenase. Examples of the transferase (transferase) include serine hydroxymethyltransferase, aspartate aminotransferase, and hexokinase. Examples of hydrase (hydrolase) include amylase, cellulase, dextranase, glucosidase, galactosidase, mannosidase, agarase, lactose, carbohydrases such as mutanase, lysozyme, chitinase, chitosanase, pepsin, trypsin, chymotrypsin, collagenase, gelatinase. And proteolytic enzymes such as keratinase, elastase, subtilisin, papain, promeline, carboxypeptidase, aminopeptidase, thermolysin and achromopeptidase, and lipases such as lipases, cutinase and phospholipase. Examples of the lyase include lyases such as alginate lyase, aldolase, and serine dehydratase. Examples of the isomerase include nuclease, UDP-glucose-4-epimerase, and glucose phosphate isomerase.
Of these, preferred are hydrase, lyase, and isomerase, more preferred are proteolytic enzyme, alginate lyase, and nucleolytic enzyme, and still more preferred is proteolytic enzyme.

These enzymes can be used alone or in combination of two or more, and the concentration thereof is not particularly limited and can be appropriately selected depending on the type and enzyme activity, but from the viewpoint of economy and effect The biofilm remover action system is preferably 0.01 to 20,000 ppm, more preferably 0.1 to 2,000 ppm, and particularly preferably 1 to 200 ppm.
The content ratio of the basic amino acid derivative (1) and the enzyme is 50/1 to 1/5, more preferably 30/1 to 1/3, particularly 10/1 to 1/2, from the viewpoint of achieving the effect of the present invention. preferable.

  The biofilm remover composition of the present invention is other than the basic amino acid derivative (1) for the purpose of increasing the solubility of the basic amino acid derivative (1), improving the biofilm removal performance, and further enhancing the cleaning effect. These surfactants can be used in combination. As the said surfactant, 1 or more types chosen from anionic surfactant, a nonionic surfactant, an amphoteric surfactant, and a cationic surfactant can be used.

  Examples of the anionic surfactant include lignin sulfonate, alkylbenzene sulfonate, alkyl sulfonate, polyoxyethylene (hereinafter referred to as POE) alkyl sulfonate, POE alkylphenyl ether sulfonate, and POE alkylphenyl. Ether phosphate ester salt, POE aryl phenyl ether sulfonate, alkyl sulfate ester salt, POE alkyl ether sulfate ester salt, POE aryl phenyl ether phosphate ester salt, naphthalene sulfonate, naphthalene sulfonate formalin condensate, POE tribenzyl Phenyl ether sulfonate, alkyl phosphate, POE alkyl phosphate, POE tribenzyl phenyl ether phosphate ester salt, dialkyl sulfosuccinate, fatty acid salt (soap), P E alkyl ether acetate and the like, it is more preferable to use inter alia alkyl sulfate and POE alkyl ether sulfate or POE alkyl ether acetates.

  Nonionic surfactants include monohydric alcohol derivative type nonionic surfactants such as POE alkyl ether, POE alkyl phenyl ether, POE aryl phenyl ether, POE styrenated phenyl ether, POE tribenzyl phenyl ether, (poly ) Glycerin fatty acid ester, sucrose fatty acid ester, sorbitan fatty acid ester, POE sorbitan fatty acid ester, alkylpolyglycoside, polyhydric alcohol derivative type nonionic surfactants such as fatty acid alkanolamide, etc., among them POE alkyl ether, ( It is more preferable to use poly) glycerin fatty acid ester, alkyl polyglycoside, sorbitan fatty acid ester or POE sorbitan fatty acid ester.

  Examples of amphoteric surfactants include alkylcarboxybetaines, alkylsulfobetaines, alkylhydroxysulfobetaines, fatty acid amide betaines, and alkyldimethylamine oxides. Among them, alkyldimethylamine oxide is preferably used.

  Examples of the cationic surfactant include alkyltrimethylammonium salts and dialkyldimethylammonium salts. Among them, alkyltrimethylammonium salts are preferable. As said salt, a halide is preferable and a chloride and a bromide are more preferable.

  These surfactants can be used in combination with the basic amino acid derivative (1) at any ratio depending on the purpose. A preferred ratio is component (A): component (C) = 90: 10 to 1:99 in terms of weight ratio in terms of solubility of the basic amino acid derivative (1) and cleaning effect, and more preferably 50 : 50 to 10:90.

  As a dosage form of the biofilm remover composition of the present invention, a solution dissolved in a solvent such as water, ethanol, isopropanol, or a solid, gel, emulsified / dispersed, powder, aerosol, depending on the application and purpose These can be selected appropriately from these, and in addition to the product form that matches the working concentration, leave it in a high-concentration product form and dilute it in the scene of use, or incorporate a surfactant in the scene of use It can also be used.

  The biofilm remover composition of the present invention is a thickener, a viscosity modifier, a pH adjuster, a solvent, a fragrance, a colorant, an antioxidant, an antiseptic, a fluorescent agent, as long as the object of the present invention is not impaired. An excipient, a soil release agent, a bleaching agent, a bleaching activator, a powdering agent, a granulating agent, a coating agent and the like can be blended.

  The biofilm remover composition of the present invention can be used in a wide range of fields where there is a risk of biofilm harm. For example, the present invention can be applied to detergents for food production or beverage production plants with a high risk of bacterial contamination, kitchens, kitchens, bathrooms, toilets, drains for kitchens, kitchens, etc., drain pipes. It can also be applied to cooling water systems such as industrial cooling towers, desalination equipment, pulp and paper manufacturing systems, and circulating water systems such as bathtubs, pools, and artificial ponds. The present invention can also be applied to medical devices in which biofilms are easily formed, such as cleaning agents such as endoscopes, catheters, and artificial dialysis machines. Furthermore, it can also be used for denture care agents, contact lens cleaners, and the like.

  The biofilm remover composition of the present invention exerts its effect by bringing an aqueous solution into contact with the surface on which the biofilm is formed. Examples of the method include dipping, coating or spraying. Further, physical force such as sponge, towel, brush, water flow, etc. may be applied. In addition, the time for which the biofilm remover composition is allowed to act varies depending on the amount of attached biofilm, the concentration at which the biofilm remover composition acts, the working temperature, and the presence or absence of physical force, but usually several seconds. To several hours. In addition, it is desirable that the removed biofilm is immediately rinsed away with running water after the action.

Production Example 1: Production method of basic amino acid derivative (1)
The synthesis and synthesis of N- [2-hydroxy-3- (2-ethylhexyl) oxypropyl] -L-arginine hydrochloride was carried out with reference to Production Example 1 described in JP-A-9-271655.
In a 200 mL four-necked flask equipped with a reflux condenser, a dropping funnel, a thermometer, and a stirring blade, 9.4 g (53.7 mmol) of L (+)-arginine, 50.0 g of water, and 50.0 g of ethanol were placed. The temperature was raised to 78 ° C. while stirring. Next, 10.0 g (53.7 mmol) of 2-ethylhexyl glycidyl ether was added dropwise while maintaining the reaction system at 78 to 80 ° C. Then, after aging at 78-80 ° C. for 4 hours, the temperature was returned to room temperature. Next, neutralization was performed by adding 9.8 g (53.7 mmol) of 6 mol / L hydrochloric acid aqueous solution. Water and ethanol were completely distilled off from the reaction solution under reduced pressure to obtain 21.6 g of N- [2-hydroxy-3- (2-ethylhexyl) oxypropyl] -L-arginine hydrochloride as a white solid.
Similarly, those having R ¹ of C10 (isodecyl), C12 (straight chain), and C16 (straight chain) were synthesized.

The enzymes used in the examples are as follows.
(Enzyme A) Alcalase 2.5L, TypeDX [Proteolytic enzyme, manufactured by Novozymes, Inc.]
(Enzyme B) Sabinase 16L, Type EX [Proteolytic enzyme, manufactured by Novozymes)
(Enzyme C) Dextranase L “Amano” [Amano Enzyme Co., Ltd.]
(Enzyme D) Duramil [α-Amylase, manufactured by Novozymes, Inc.]

Example 1
<Biofilm removal ability test>
Pseudomonas aeruginosa NBRC13275, Serratia marcescens NBRC12648, Staphylococcus epidermidis NBRC12773, soybean-Casein Digest Agar (SCD agar medium: manufactured by Nippon Pharmaceutical Co., Ltd.) ], A very small amount of bacterial mass from a colony formed by pre-culturing at 37 ° C. for 24 hours, and using a sterilized bamboo skewer, 1.5 mL each of Mueller Hinton medium was added to each well. Inoculated into plates. After culturing at 37 ° C. for 24 hours, the culture solution was discarded, and each well was rinsed 5 times with 2 mL of purified water to form and attach a biofilm to the microplate wall. Immediately, 2 mL of the prepared biofilm remover composition shown in Table 2 was added and allowed to act at room temperature for 10 minutes, and then the biofilm remover composition in each well was discarded. Each well was rinsed twice with 2 mL of purified water, and then 2 mL of 0.1% crystal violet was added to stain the biofilm remaining on the microplate wall. After rinsing the excess staining solution with water, 2 mL of 80% ethanol was added, and the crystal violet stained biofilm was uniformly dissolved, and then the absorbance was measured at 570 nm to obtain a measured value. Similarly, after the well not treated with the biofilm remover composition was treated with 0.1% crystal violet, the absorbance was measured to obtain an initial value. In addition, 1.5 mL each of Mueller Hinton medium in 24 wells but not inoculated with the bacterial mass was performed in the same manner, and the absorbance was measured to obtain a blank value. Each test was performed five times and the average value was used. The removal rate was calculated by the following formula. Concentrations in the table are shown as effective concentrations (% by weight) based on the total amount, and pH was adjusted using potassium hydroxide or hydrochloric acid as necessary.

Removal rate (%) = 100 × [{(initial value−blank value) − (measured value−blank value)} / (initial value−blank value)]

The obtained results are shown in Table 2.
The compounds A to D in the table are those shown in the following Table 1.

  From the above results, it can be seen that the biofilm can be effectively removed by using the biofilm remover composition of the present invention.

Claims (4)

The following general formula (1)

(In the formula, R ¹ represents a linear or branched alkyl or alkenyl group having 4 to 18 carbon atoms, and X and Y represent the following formulae:

And m represents an integer of 1 to 5. )
A biofilm remover composition comprising a basic amino acid derivative represented by the formula: In Formula (1), m is 3, and Y is -NH-C (-NH ₂₎ = biofilm remover composition of claim 1 wherein is NH. The biofilm remover composition according to claim 1, wherein X in the general formula (1) is —OCH ₂ —CH (—OH) CH ₂ —.   A biofilm remover composition comprising an arginine derivative or a salt thereof obtained by reacting arginine or a salt thereof with glycidyl ether and an enzyme.