JPH05255109A - Antibacterial agent and treating method for article using the same - Google Patents

Abstract

PURPOSE:To provide an antibacterial agent further enhanced in the antibacterial activity of a degradation product of lactoferrin and/or lactoferrin-related antibacterial peptide. CONSTITUTION:The objective antibacterial agent containing, as active ingredients, (A) a compound selected from degradation products of lactoferrins, lactoferrin-related antibacterial peptides, and any mixtures thereof and (B) a degradation product of casein.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to an antibacterial agent and a method for treating articles with the antibacterial agent. More specifically, the present invention relates to an antibacterial agent in which the antibacterial property of a decomposed product of lactoferrin and / or lactoferrin-related antibacterial peptide is further enhanced, and a method of treating an article using the antibacterial agent.

[0002]

BACKGROUND OF THE INVENTION Lactoferrin is a natural iron-binding protein contained in tears, saliva, peripheral blood, milk, etc. in vivo, and has an antibacterial action against harmful microorganisms such as Escherichia coli, Candida and Clostridia. Is known to indicate [Journal of Pediatrics (Journal of P
ediatrics), Vol. 94, p. 1: 1979]. Also,
0.5-30 mg / m for Staphylococcus and Enterococcus
It is known to have an anti-effect at a concentration of 1 [Nonnecke, Smith; KL; Journal of Dairy Scie
nce) 67, 606: 1984].

On the other hand, there are many inventions of peptides having antibacterial activity against various microorganisms, such as phosphonotripeptides which are effective against Gram-positive and Gram-negative bacteria (JP-A-57-106689) and phosphos. Nodipeptide derivative (Japanese Patent Laid-Open No. 58-13594),
Cyclic peptide derivatives (JP-A-58-213744), peptides exhibiting antibacterial and anti-viral effects (JP-A-59-51247), yeast-effective polypeptides (JP-A-60-130599), Glycopeptide derivatives effective against Gram-positive bacteria (JP-A-60-172998, JP-A-61-251699, JP-A-63-63)
44598), oligopeptides effective against Gram-positive bacteria (JP-A-62-22798), peptide antibiotics (JP-A-62-51697, JP-A-63-63).
17897) and other antibacterial peptides extracted from blood cells of horseshoe crabs produced in North America (JP-A-2-53799), antibacterial peptides isolated from hemolymph of bees (JP-A-2-500084), etc. Has been.

The inventors of the present invention intend to inexpensively isolate from nature the substance that has no adverse side effects (eg, antigenicity), is heat resistant, and has a strong antibacterial action, It was found that a decomposed product obtained by hydrolyzing lactoferrin with an acid or an enzyme has stronger heat resistance and antibacterial property than undecomposed lactoferrin, and a patent application has already been filed (Japanese Patent Application No. 3-171736).

Furthermore, the inventors of the present invention have no adverse side effects (eg, antigenicity) and have heat resistance,
As a lactoferrin-related antibacterial peptide having a strong antibacterial action, an antibacterial peptide having 20 amino acid residues (Japanese Patent Application No. 3-186260) and an antibacterial peptide having 11 amino acid residues (Japanese Patent Application No. Flat 3-48196
No.), an antimicrobial peptide having 6 amino acid residues (Japanese Patent Application No. 3-94492), an antimicrobial peptide having 5 amino acid residues (Japanese Patent Application No. 3-94493), amino acid residues Is an antimicrobial peptide having 3 to 6 residues (Japanese Patent Application No. 3-9
4494) and filed a patent application.

Various studies have been conducted so far to enhance the antibacterial activity of lactoferrin, and lysozyme, IgA and glycopeptides are known as such physiological activity enhancing auxiliary agents. For example, a method of synergistically enhancing antibacterial action by coexistence of lactoferrin and lysozyme [JP-A-62-249931], a method of synergistically enhancing antibacterial action by coexistence of lactoferrin and secretory IgA [ Immunology (Stephens,
S., et al; Immunology); 41, 597: 1980.
Year] etc. have been reported. In addition, spik etc.
F. Williams and J. D. Baum (A.
F.Williams & JDBaum, edited by "Human Milk Banking", Nestle Nutrition Workshop Series, Volume 5, (Nest
le Nutrition Workshop Series Volume 5), 133
Page, Raven Press Books,
Ltd.), 1984], lactoferrin has an action of preventing bacteria from adhering to the mucosal surface, and this action is enhanced by the presence of lysozyme or glycopeptides.

[0007]

However, a method for enhancing the antibacterial activity of the above-mentioned decomposed products of lactoferrins or lactoferrin-related antibacterial peptides, especially multi-component products and articles (for example, milk etc.), has the above-mentioned effects. There is no known method for enhancing the antibacterial activity of a lactoferrin degradation product or a lactoferrin-related antibacterial peptide.

The present invention uses the above-mentioned decomposed product of lactoferrin and / or lactoferrin-related antibacterial peptide, which the inventors of the present invention have already invented, particularly in multi-component products and articles (eg milk). To provide an antibacterial agent in which a decomposed product of lactoferrin and / or an antibacterial peptide related to lactoferrin is further enhanced, and a method for treating an article using the antibacterial agent I am trying.

[0009]

Means for Solving the Problems The inventors of the present invention have conducted extensive studies on the above problems, and as a result, by combining a degradation product of lactoferrin and / or an antibacterial peptide related to lactoferrin with a specific compound. , Compared to the case where the degradation product of lactoferrin and / or the lactoferrin-related antibacterial peptide is used alone,
They have found that they can increase antibacterial properties and completed the invention.

That is, the present invention comprises (A) a compound selected from the group consisting of lactoferrin degradation products, lactoferrin-related antibacterial peptides, or any mixture thereof, and (B) casein degradation product as active ingredients. An antibacterial agent is provided. Further, the present invention provides an antibacterial agent comprising (A) a lactoferrin degradation product, a lactoferrin-related antimicrobial peptide, or a compound selected from the group consisting of any mixture thereof, and (B) a casein degradation product as active ingredients. There is provided a method of treating an article with.

Next, the present invention will be described in detail. In the present invention, lactoferrins are commercially available lactoferrin, colostrum of mammals (eg, humans, cows, sheep, goats, horses, etc.), transitional milk, normal milk, end-stage milk, etc., or defatted products of these milks. Lactoferrin separated from milk, whey, etc. by a conventional method (for example, ion exchange chromatography), apolactoferrin deferred with hydrochloric acid, citric acid, etc., or apolactoferrin with metal such as iron, copper, zinc, manganese, etc. -Metal saturated or partially saturated lactoferrin, or a mixture thereof, which can be used as a commercially available product or prepared by a known method.

The decomposed product of lactoferrin used in the present invention can be obtained, for example, by the method described in the invention of Japanese Patent Application No. 3-171736, in which the above-mentioned lactoferrin is hydrolyzed with an acid or an enzyme. That is, in the case of performing hydrolysis with an acid, lactoferrin is made into an aqueous solution, an inorganic acid or an organic acid is added to this, and the mixture is heated to a predetermined temperature for a predetermined time for hydrolysis. When the enzyme is used for hydrolysis, lactoferrins are made into an aqueous solution, the pH of the enzyme used is adjusted to an optimum pH, enzymes such as pepsin and trypsin are added, and the mixture is kept at a predetermined temperature for a predetermined time for hydrolysis. After hydrolysis, the enzyme is inactivated by a conventional method. The degradation product obtained by hydrolysis with an acid or an enzyme is a mixture of degradation products containing peptides having antibacterial properties and having various molecular weights. The degree of decomposition by the above hydrolysis is 6 to
The range of 20% is desirable. The above-mentioned degree of decomposition was calculated by the following formula from these values by measuring the total nitrogen of the sample by the Kjeldahl method and the formal nitrogen of the sample by the formol titration method.

Decomposition degree = (formal nitrogen content / total nitrogen content) × 100 The above reaction solution obtained by hydrolysis with an acid or an enzyme (a solution of lactoferrin-decomposed product) is cooled by a conventional method. If necessary, it is neutralized, desalted, decolorized by a conventional method, and if necessary, fractionated by a conventional method, and the obtained solution is used as it is, or in a concentrated liquid form, or in a powder form after concentration and drying. , Mixed with casein hydrolyzate.

The lactoferrin-related antibacterial peptide used in the present invention is the same as the antibacterial peptide obtained from the decomposed product of lactoferrin by the separation means and the antibacterial peptide obtained from the decomposed product of the lactoferrin by the separation means. Peptide having the same chemical structure (amino acid sequence), a derivative of the peptide having the same chemical structure (amino acid sequence) as the peptide having antibacterial properties, which is obtained from the degradation product of lactoferrin by a separation means, and any mixture thereof. It is a compound selected from the group. These lactoferrin-related antibacterial peptides are disclosed, for example, in Japanese Patent Application Nos. 3-186260, 3-48196, 3-94492, and 3-94492.
-94493, Japanese Patent Application No. 3-94494, the method described in each invention, that is, lactoferrins are hydrolyzed with an acid or an enzyme, and have antibacterial properties by a separation means such as liquid chromatography from a decomposition product mixture. A method for obtaining a fraction containing a peptide, or the amino acid sequence of the peptide having antibacterial properties obtained as described above is determined by a known method (for example, a method using a gas phase sequencer, etc.) The peptide containing the peptide of the sequence can be prepared by a known method (for example, a method using an automatic peptide synthesizer) to obtain the desired peptide. These lactoferrin-related antibacterial peptides are, for example, the following peptides having the following amino acid sequences: Antibacterial peptides having the amino acid sequences of SEQ ID NOs: 1, 2 and 27 (Japanese Patent Application No. 3-311).
-48196 invention); antibacterial peptides having the amino acid sequences of SEQ ID NOs: 3, 4, 5 and 6 (Japanese Patent Application No. 3-94).
No. 492); an antibacterial peptide having the amino acid sequences of SEQ ID NOs: 7, 8, 9 and 31 (Japanese Patent Application No. 3-9449).
No. 3 invention); SEQ ID NOS: 10, 11, 12, 13, 14,
Antibacterial peptide having amino acid sequence of 15, 16, 17, 18, 19, 20 and 21 (Japanese Patent Application No. 3-9449)
No. 4 invention); SEQ ID NOs: 22, 23, 24, 25, 26,
An antibacterial peptide having an amino acid sequence of 28, 29 and 30 (Japanese Patent Application No. 3-186260).

These antibacterial peptides are mixed with the casein decomposition product in the form of a solution as it is, or in a concentrated liquid form, or in a powder form after concentration and drying. Whether to combine the casein degradation product with any of the degradation product of the lactoferrins, the lactoferrin-related antibacterial peptide, or any mixture thereof can be appropriately selected depending on the purpose of use. Further, the mixing ratio thereof can be appropriately selected depending on the kind of each component or the purpose of use. The above-mentioned mixing can be performed in a liquid state or in a powder state as described above, and in this case, a known diluent or excipient may be appropriately added.

Commercially available products of casein degradation products may be used, or they may be prepared and used by known methods.
For example, it is a mixture of degradation products obtained by hydrolyzing casein such as milk with proteolytic enzymes, acids, or alkalis by a conventional method, or a product obtained by fractionating specific components from the degradation products. Especially, the number average molecular weight is about 380 and the molecular weight distribution is 7
The use of a mixture of 5 or more and about 10,000 or less peptides (and amino acids) is preferred.

The antibacterial agent of the present invention comprises various bacteria, yeasts,
It has strong antibacterial properties against fungi and is not only used as a drug but also as a food, feed, cosmetic, etc. that is taken into the human or animal body or applied to the surface of the body, and generally it does not grow bacteria. The antibacterial agent of the present invention can be used by blending it with any product for which prevention or inhibition is desired, and can treat those products or raw materials. That is, the antibacterial agent of the present invention can be directly administered to humans or animals, or food (eg, chewing gum, etc.), drug (eg, eye drops, mastitis therapeutic agent, antidiarrheal drug, athlete's foot drug, etc.), pharmacy External products (for example, mouthwash, antiperspirant, hair nourishing agent, etc.), various cosmetics (for example, hair dressing, cream, emulsion, etc.), various toothpaste products (for example, toothpaste, toothbrush, etc.), various sanitary products, various Baby
Goods (for example, diapers), various elderly goods (for example,
Denture fixative, diapers, etc.), various detergents (eg soap,
Medicated soap, shampoo, rinse, laundry detergent, kitchen detergent, household detergent, etc.), various sterilization products (for example, kitchen sterilization paper, toilet sanitization paper, etc.),
Feed (e.g., pet hood, etc.), raw materials thereof, and other generally, the growth of microorganisms can be added, compounded, sprayed, attached, coated, impregnated, etc. to any article for which inhibition or suppression of microorganisms is desired, In addition, it can be generally used for treating any article for which the prevention or inhibition of the growth of microorganisms is desired.

Next, the present invention will be described in more detail with reference to test examples. Test example 1. Preparation of sample (1) Antibacterial peptide (powder) The sample was prepared by the method described in Example 3 and used.

(2) Casein degradation product: Prepared by the method described in Reference Example 1 and used. 2. Test method (1) Preparation of staphylococcal preculture liquid One platinum loop was taken from a stock slant of Staphylococcus aureus JCM-2151, and spread on standard agar medium (Nippon Pharmaceutical Co., Ltd.) at 37 ° C. After culturing for 16 hours, the colony grown on the surface of the standard agar medium is scraped off with a platinum loop.
% Peptone (manufactured by Difco) was cultivated at 37 ° C. for several hours, and the bacterial solution in the logarithmic phase grown to a bacterial concentration of 3 × 10 ⁸ / ml was used as a preculture liquid.

(2) Preparation of basal medium (milk medium) The above sample was added to commercially available milk, the concentration of milk was diluted to 1/2 with purified water, and sterilized at 115 ° C for 15 minutes to prepare a basal medium. .. (3) Preparation of test medium and control medium (3-1) Preparation of test medium The aqueous solution of the antimicrobial peptide prepared in (1) of the sample preparation described in 1 above and the aqueous solution of the casein degradation product in (2) were prepared. Sterilized with a sterilizing filter (manufactured by Advantech), added to the basic medium and adjusted to the final concentrations shown in Table 1,
The test medium was prepared.

(3-2) Preparation of Control Medium 1 Commercially available milk was diluted 2-fold with purified water and sterilized at 115 ° C. for 15 minutes to prepare Control Medium 1. (3-3) Preparation of Control Medium 2 The aqueous solution of the casein degradation product of (3) in the preparation of the sample described in 1 above was sterilized with a sterilizing filter (manufactured by Advantech),
Control medium 2 was then prepared by adding it to the basic medium and adjusting it to the same concentration as the test medium.

(4) Antibacterial effect test (4-1) Survival rate test 20 μl of the preculture liquid of Staphylococcus prepared in (1) above was added to
Add to 2 ml of test medium prepared in (3-1) above, and keep at 37 ℃
Incubate for 1 hour at room temperature, collect 200 μl from the culture and
After diluting 10 ⁿ times with a peptone aqueous solution, 110 μl of the solution was spread on a regular agar medium plate and cultured at 37 ° C. for 24 hours to measure the number of bacteria (test bacteria).

Separately, as Control 1, 20 μl of the preculture liquid of Staphylococcus prepared in (1) above was added to 2 ml of the control medium 1 prepared in (3-2) above to measure the number of test bacteria. The bacterial count (control bacterial count 1) was measured in the same manner as in (1). Further, as a control 2, 20 μl of the preculture liquid of Staphylococcus prepared in (1) above was added to 2 ml of the control medium 2 prepared in (3-3) above.
Then, the bacterial count (control bacterial count 2) was measured in the same manner as in the case of measuring the test bacterial count.

The survival rate was calculated by the following formula. Survival rate 1 = (Number of test bacteria / Number of control bacteria 1) × 100 Survival rate 2 = (Number of control bacteria 2 / Number of control bacteria 1) × 100 (Note: In each table below, the survival rate 2 is the antibacterial peptide. It is displayed on the line where the density is 0). Test results The test results are shown in Table 1. As is clear from Table 1, it was confirmed that the casein degradation product enhances the bactericidal effect of the antibacterial peptide. When the casein degradation product was added and the antibacterial peptide was not added, no bactericidal effect was observed. Therefore, it is clear that the above-mentioned enhancement of the bactericidal effect is a synergistic effect of the antibacterial peptide and the casein degradation product. Similar tests were conducted on antibacterial peptides other than the above antibacterial peptides and lactoferrin degradation products, and it was confirmed that the casein degradation products enhanced the bactericidal effect also in this case.

[0025]

[Table 1]

Reference Example 1 (Preparation of Casein Decomposition Product) 20 g of acid casein (manufactured by Merck & Co., Inc.) was swollen in water at 50 ° C. at a final concentration of 5%, and 1N aqueous sodium hydroxide solution was added to adjust the pH to 8.5. The acid casein was dissolved. It was sterilized at 80 ° C. for 5 to 10 minutes, cooled to 50 ° C., and an aqueous suspension of pancreatin F (manufactured by Amano Pharmaceutical Co., Ltd.) was added at a ratio of an enzyme amount of 5% to the substrate. It was hydrolyzed at 50 ° C. for 10 hours, kept at 85 ° C. for 10 minutes to inactivate the enzyme, cooled, filtered to remove insoluble matter, and the filtrate was freeze-dried to obtain about 16 g of casein degradation product. ..

Next, the present invention will be described more specifically and in detail by showing examples, but the present invention is not limited to the following examples.

[0028]

【Example】

Example 1 50 g of commercially available lactoferrin (manufactured by Oleofina Co., Belgium) as separated from milk was dissolved in 950 g of purified water, and 1N hydrochloric acid was added to the resulting solution to adjust the pH to 2, 120 Heat at 15 ° C. for 15 minutes, cool, and then lactoferrin degradation product solution (lactoferrin degradation product concentration: 5
%) About 1000 g was obtained. The degradation degree of this lactoferrin degradation product was 9%.

The above-mentioned lactoferrin degradation product solution was concentrated under reduced pressure and freeze-dried to give about 49 powders of the lactoferrin degradation product.
g was obtained. 10 g of this lactoferrin decomposition product powder was decomposed with casein (prepared by the method described in Reference Example 1).
An antibacterial agent was prepared by uniformly mixing 2.5 g. Example 2 1 kg of commercially available lactoferrin (manufactured by Oleofina Co., Belgium) as separated from milk was dissolved in 9 kg of purified water, and 2 molar citric acid was added to adjust the pH to 2.5. 30 g of pork pepsin (1: 10,000, manufactured by Wako Pure Chemical Industries, Ltd.) was added and mixed uniformly, and the mixture was heated to 37 ° C. for 1 hour.
The mixture was kept for 80 minutes, heated at 85 ° C. for 10 minutes to inactivate the enzyme, and then cooled to obtain about 10 kg of a lactoferrin degradation product solution (lactoferrin degradation product concentration: 10%). The decomposition degree of this lactoferrin decomposition product was 11.3%.

The above-mentioned lactoferrin decomposition product solution was concentrated under reduced pressure and freeze-dried to give about 9 powders of the lactoferrin decomposition product.
60 g was obtained. 100g of this lactoferrin decomposition product powder
An antibacterial agent was prepared by uniformly mixing with 25 g of a casein degradation product (prepared by the method described in Reference Example 1). Example 3 50 mg of commercially available bovine lactoferrin (manufactured by Sigma) was dissolved in 0.9 ml of purified water, and the pH was adjusted to 0.1 with hydrochloric acid of 0.1 standard.
Adjusted to 5, then commercial pig pepsin (manufactured by Sigma) 1 m
g, and hydrolyzed at 37 ° C. for 6 hours. Then 0.
Adjust the pH to 7.0 with 1 standard sodium hydroxide, and
The enzyme was inactivated by heating at 0 ° C for 10 minutes, cooled to room temperature, and centrifuged at 15,000 rpm for 30 minutes to obtain a transparent supernatant. 100 μl of this supernatant was added to TSK gel ODS-
After high-performance liquid chromatography using 120T (manufactured by Toso Co., Ltd.), 10 samples were injected at a flow rate of 0.8 ml / min.
20 minutes with 0.05% TFA (trifluoroacetic acid)
% Acetonitrile, then 0.05% T for 30 minutes
Elute with a gradient of 20-60% acetonitrile containing FA, collect fractions eluting between 24-25 minutes,
Vacuum dried. This dried product was dissolved in purified water at a concentration of 2% (W / V), and again subjected to high performance liquid chromatography using TSK gel ODS-120T (manufactured by Toso Corporation),
0.05% for 10 minutes after sample injection at a flow rate of 0.8 ml / min
Elute with 24% acetonitrile containing TFA, then 30
Fractions eluting with a gradient of 24-32% acetonitrile containing 0.05% TFA per minute and eluting between 33.5 and 35.5 minutes were collected. The above operation was repeated 25 times and dried in vacuum to obtain about 1.5 mg of antibacterial peptide.

The above-mentioned antibacterial peptide was hydrolyzed with 6N hydrochloric acid, and the amino acid composition was analyzed by an ordinary method using an amino acid analyzer. The same sample was subjected to Edman degradation 25 times using a gas phase sequencer (manufactured by Applied Biosystems) to determine the sequence of 25 amino acid residues. In addition, a disulfide bond analysis method using DTNB (5,5-dithio-bis (2-nitrobenzoic acid)) [Analytic Biochemistry (Analytic
al Biochemistry), 67, 493, 1975.
Year] confirmed the existence of disulfide bonds.

As a result, this peptide consisted of 25 amino acid residues, the 3rd and 20th cysteine residues were disulfide-bonded, and the 3rd cysteine residue resulted in N
It was confirmed to have the amino acid sequence of SEQ ID NO: 26 in which two amino acid residues were bound to the -terminal side and 5 amino acids were bound to the C-terminal side from the 20th cysteine residue, respectively. .

An antibacterial agent was prepared by uniformly mixing 300 mg of the casein degradation product (prepared by the method described in Reference Example 1) with 100 mg of the above-mentioned antibacterial peptide powder. Example 4 Using a peptide automatic synthesizer (Pharmacia LKB Biotechnology, LKBBiolynx4170), Shepard et al. [Journal of Chemical Society Parkin I (Journal of Chemical Societ)
y Perkin I), p. 538, 1981], and an antibacterial peptide was synthesized as follows.

N, N-dicyclohexylcarbodiimide was added to an amino acid whose amine functional group was protected by a 9-fluorenylmethoxycarbonyl group to produce the desired anhydride of the amino acid, and this Fmoc-amino acid anhydride was synthesized. Using. In order to produce a peptide chain, Fmoc-lysine anhydride corresponding to the C-terminal lysine residue was used as a catalyst for ultrocin A resin (Pharmacia LKB Biotechnology-
Fixed). The resin is then washed with dimethylformamide containing piperidine to remove the amine functional group protecting group of the C-terminal amino acid. Later C of amino acid sequence
The penultimate-corresponding Fmoc-lysine anhydride was coupled via the C-terminal amino acid residue to the deprotected amine functional group of lysine immobilized on the resin. In the same manner, methionine, arginine, tryptophan,
Glutamine, tryptophan, arginine, arginine, threonine, and lysine were fixed. After completion of coupling of all amino acids and formation of a peptide chain having a desired amino acid sequence, 94% TFA, 5% pheno-
Of the protective groups other than acetamidomethyl and elimination of the peptide with a solvent consisting of 1% ethanol and 1% ethanediol, the peptide was purified by high performance liquid chromatography, and the solution was concentrated and dried, Peptide powder was obtained.

The peptide thus obtained was analyzed for amino acid composition by an ordinary method using an amino acid analyzer, and it was confirmed to have the amino acid sequence of SEQ ID NO: 27. 100 mg of antibacterial peptide powder prepared by the above synthesis
Then, 200 mg of a casein degradation product (prepared by the method described in Reference Example 1) was uniformly mixed to prepare an antibacterial agent. Example 5 An eye drop (aqueous solution) having the following composition was produced by a conventional method.

Boric acid 1.60 (%) Antibacterial agent of Example 4 0.03 Methylcellulose 0.50 Example 6 A chewing gum having the following composition was produced by a conventional method.

Gum base 25.00 (%) Calcium carbonate 2.00 Fragrance 1.00 Antibacterial agent of Example 2 0.10 Sorbitol powder 71.90 Example 7 A dentifrice having the following composition was produced by a conventional method. did.

Dicalcium phosphate dihydrate 36.80 (%) sorbitol 45.00 glycerin 15.00 carbooxymethylcellulose sodium 1.50 sorbitan fatty acid ester 0.50 saccharin sodium 1.00 Example Antibacterial agent 1 of 0.20 Example 8 A skin cleansing solution having the following composition was produced by a conventional method. This skin cleansing solution is diluted 50 times with water before use.

Sodium chloride 8.00 (%) Antibacterial agent of Example 3 0.70 Purified water 91.30

[0040]

The effects of the present invention are as follows. (1) An antibacterial agent having an excellent antibacterial action against a wide range of microorganisms is provided. (2) An extremely safe antibacterial agent is provided for use in foods and pharmaceuticals. (3) Since the antibacterial agent of the present invention exhibits an antibacterial effect even in a small amount, it has almost no effect on the flavor when used for treating articles such as foods.

[0041]

[Sequence list]

SEQ ID NO: 1 Sequence length: 11 Sequence type: Amino acid Topology-: Linear Sequence type: Peptide Sequence characteristics: This peptide, and a peptide containing this peptide as a fragment (In the above sequence, Xaa represents any amino acid residue except Cys) SEQ ID NO: 2 Sequence length: 11 Sequence type: Amino acid Topology-: Linear sequence type: Peptide Sequence characteristics: This peptide , And a peptide containing this peptide as a fragment (In the above sequence, Xaa represents any amino acid residue except Cys) SEQ ID NO: 3 Sequence length: 6 Sequence type: Amino acid Topology-: Linear sequence type: Peptide Sequence characteristics: This peptide , And a peptide containing this peptide as a fragment (In the above sequence, Xaa represents any amino acid residue except Cys) SEQ ID NO: 4 Sequence length: 6 Sequence type: Amino acid Topology-: Linear sequence type: Peptide Sequence characteristics: This peptide , And a peptide containing this peptide as a fragment (In the above sequence, Xaa represents any amino acid residue except Cys) SEQ ID NO: 5 Sequence length: 6 Sequence type: Amino acid Topology-: Linear sequence type: Peptide Sequence characteristics: This peptide , And a peptide containing this peptide as a fragment (In the above sequence, Xaa represents any amino acid residue except Cys) SEQ ID NO: 6 Sequence length: 6 Sequence type: Amino acid Topology-: Linear sequence type: Peptide Sequence feature: This peptide , And a peptide containing this peptide as a fragment (In the above sequence, Xaa represents any amino acid residue except Cys) SEQ ID NO: 7 Sequence length: 5 Sequence type: Amino acid Topology-: Linear sequence type: Peptide Sequence characteristics: This peptide , And a peptide containing this peptide as a fragment (In the above sequence, Xaa represents any amino acid residue except Cys) SEQ ID NO: 8 Sequence length: 5 Sequence type: Amino acid Topology-: Linear sequence type: Peptide Sequence characteristics: This peptide , And a peptide containing this peptide as a fragment (In the above sequence, Xaa represents any amino acid residue except Cys) SEQ ID NO: 9 Sequence length: 5 Sequence type: Amino acid Topology-: Linear sequence type: Peptide Sequence characteristics: This peptide , And a peptide containing this peptide as a fragment (In the above sequence, Xaa represents any amino acid residue except Cys) SEQ ID NO: 10 Sequence length: 6 Sequence type: Amino acid Topology-: Linear sequence type: Peptide Sequence characteristics: This peptide , And a peptide containing this peptide as a fragment SEQ ID NO: 11 Sequence length: 5 Sequence type: Amino acid Topology-: Linear sequence type: Peptide Sequence characteristics: This peptide and a peptide containing this peptide as a fragment SEQ ID NO: 12 Sequence length: 4 Sequence type: Amino acid Topology-: Linear Sequence type: Peptide Sequence characteristics: This peptide and a peptide containing this peptide as a fragment SEQ ID NO: 13 Sequence length: 3 Sequence type: Amino acid Topology-: Linear Sequence type: Peptide Sequence characteristics: This peptide and a peptide containing this peptide as a fragment SEQ ID NO: 14 Sequence length: 5 Sequence type: Amino acid Topology-: Linear Sequence type: Peptide Sequence characteristics: This peptide and a peptide containing this peptide as a fragment SEQ ID NO: 15 Sequence length: 4 Sequence type: Amino acid Topology-: Linear Sequence type: Peptide Sequence characteristics: This peptide and a peptide containing this peptide as a fragment SEQ ID NO: 16 Sequence length: 4 Sequence type: Amino acid Topology-: Linear Sequence type: Peptide Sequence characteristics: This peptide and a peptide containing this peptide as a fragment SEQ ID NO: 17 Sequence length: 3 Sequence type: Amino acid Topology-: Linear Sequence type: Peptide Sequence characteristics: This peptide, and a peptide containing this peptide as a fragment SEQ ID NO: 18 Sequence length: 6 Sequence type: Amino acid Topology-: Linear Sequence type: Peptide Sequence characteristics: This peptide and a peptide containing this peptide as a fragment SEQ ID NO: 19 Sequence length: 5 Sequence type: Amino acid Topology-: Linear Sequence type: Peptide Sequence characteristics: This peptide and a peptide containing this peptide as a fragment SEQ ID NO: 20 Sequence length: 4 Sequence type: Amino acid Topology-: Linear sequence type: Peptide Sequence characteristics: This peptide and a peptide containing this peptide as a fragment SEQ ID NO: 21 Sequence length: 3 Sequence type: Amino acid Topology-: Linear Sequence type: Peptide Sequence characteristics: This peptide and a peptide containing this peptide as a fragment SEQ ID NO: 22 Sequence length: 20 Sequence type: Amino acid Topology-: Linear Sequence type: Peptide Sequence characteristics: Peptide containing this peptide and this peptide as a fragment In the following sequence, Cys No. 2 Cys 19 is disulfide-bonded. Sequence: Lys Cys Arg Arg Trp Gln Trp Arg Met Lys Lys Leu Gly Ala Pro 1 5 10 15 Ser Ile Thr Cys Val 20 SEQ ID NO: 23 Sequence length: 20 Sequence type: Amino acid Topology: Linear sequence Type: Peptide Sequence characteristics: Peptide containing this peptide, and a peptide containing this peptide as a fragment In the following sequence, Cys * represents cysteine in which a thiol group is chemically modified in order to prevent the formation of a disulfide bond.

Sequence: Lys Cys * Arg Arg Trp Gln Trp Arg Met Lys Lys Leu Gly Ala Pro 1 5 10 15 Ser Ile Thr Cys * Val 20 SEQ ID NO: 24 Sequence Length: 20 Sequence Type: Amino Acid Topology: Type of linear sequence: Peptide Sequence characteristics: Peptide containing this peptide and this peptide as a fragment In the following sequence, Cys 2 and 19 are disulfide-bonded.

Sequence: Lys Cys Phe Gln Trp Gln Arg Asn Met Arg Lys Val Arg Gly Pro 1 5 10 15 Pro Val Ser Cys Ile 20 SEQ ID NO: 25 Sequence Length: 20 Sequence Type: Amino Acid Topology: Linear Type of sequence: Peptide Sequence characteristics: Peptide containing this peptide and this peptide as a fragment In the following sequence, Cys * represents cysteine in which a thiol group is chemically modified to prevent the formation of a disulfide bond. ..

Sequence: Lys Cys * Phe Gln Trp Gln Arg Asn Met Arg Lys Val Arg Gly Pro 1 5 10 15 Pro Val Ser Cys * Ile 20 SEQ ID NO: 26 Sequence Length: 25 Sequence Type: Amino Acid Topology: Type of linear sequence: Peptide Sequence characteristics: This peptide and peptides containing this peptide as a fragment In the following sequence, Cys 3 and 20 are disulfide-bonded.

Sequence: Phe Lys Cys Arg Arg Trp Gln Trp Arg Met Lys Lys Leu Gly Ala 1 5 10 15 Pro Ser Ile Thr Cys Val Arg Arg Ala Phe 20 25 SEQ ID NO: 27 Sequence Length: 11 Sequence Type: Amino acid Topology-: Linear Sequence type: Peptide Sequence characteristics: This peptide and a peptide containing this peptide as a fragment SEQ ID NO: 28 Sequence length: 38 Sequence type: Amino acid Topology-: Linear Sequence type: Peptide Sequence feature: Peptide containing this peptide and this peptide as a fragment Cys of 16 in the following sequence No. 33 Cys is disulfide-bonded.

Sequence: Lys Asn Val Arg Trp Cys Thr Ile Ser Gln Pro Glu Trp Phe Lys 1 5 10 15 Cys Arg Arg Trp Gln Trp Arg Met Lys Lys Leu Gly Ala Pro Ser 20 25 30 Ile Thr Cys Val Arg Arg Ala Phe 35 SEQ ID NO: 29 Sequence length: 32 Sequence type: Amino acid Topology-: Linear Sequence type: Peptide Sequence characteristics: Peptide containing this peptide and this peptide as a fragment Cys number 10 in the following sequence And Cys 27 are disulfide-bonded.

Sequence: Thr Ile Ser Gln Pro Glu Trp Phe Lys Cys Arg Arg Trp Gln Trp 1 5 10 15 Arg Met Lys Lys Leu Gly Ala Pro Ser Ile Thr Cys Val Arg Arg 20 25 30 Ala Phe SEQ ID NO: 30 of the sequence Length: 47 Sequence type: Amino acid Topology-: Linear Sequence type: Peptide Sequence characteristics: Peptide containing this peptide and this peptide as a fragment In the following sequence, the length of sequence 36 is number 9, 26
Nos. 9 and Cys of peptides having Cys at Nos. 35 and 35
And Cys at No. 26 are disulfide-bonded, and the Cys at No. 35 of the peptide having a sequence length of 36 and the Cys at No. 10 of the peptide having a sequence length of 11 and having Cys at 10 are disulfide-bonded. is doing.

Sequence: Val Ser Gln Pro Glu Ala Thr Lys Cys Phe Gln Trp Gln Arg Asn 1 5 10 15 Met Arg Lys Val Arg Gly Pro Pro Val Ser Cys Ile Lys Arg Asp 20 25 30 Ser Pro Ile Gln Cys Ile 35 Gly Arg Arg Arg Arg Ser Val Gln Trp Cys Ala 1 5 10 SEQ ID NO: 31 Sequence length: 5 Sequence type: Amino acid Topology-: Linear Sequence type: Peptide Sequence characteristics: This peptide and this peptide Peptides included as fragments (In the above sequence, Xaa represents any amino acid residue except Cys)

 ─────────────────────────────────────────────────── ─── Continuation of the front page (72) Inventor Mitsunori Takase 138-10 Minami-Nakamaru, Omiya City, Saitama Prefecture (72) Inventor Wayne Berami-3-22-6 Higashihara, Zama City, Kanagawa Prefecture Villa Sagamino West C-5 (72) Inventor Tsuneharu Yamauchi 4-2-2 Tamawano, Kamakura City, Kanagawa Prefecture 405 Garden Heights Kamakura Tamawata (72) Inventor Hiroyuki Wakabayashi 118 Minami Kibogaoka, Asahi-ku, Yokohama-shi, Kanagawa Morinaga Kibogaoka Dormitory (72) Inventor Yukiko Tokita 3-6 Asahi-cho, Sagamihara-shi, Kanagawa Flat Stone No. 201 Sagami

Claims (2)

[Claims] 1. An antibacterial agent comprising (A) a lactoferrin degradation product, a lactoferrin-related antimicrobial peptide, or a compound selected from the group consisting of any mixture thereof and (B) a casein degradation product as active ingredients. .. 2. An antibacterial agent comprising (A) a lactoferrin degradation product, a lactoferrin-related antimicrobial peptide, or a compound selected from the group consisting of any mixture thereof and (B) a casein degradation product as active ingredients. A method of treating an article using.