JPH02237905A - Anti-fungus agent - Google Patents

Abstract

PURPOSE:To provide an anti-fungus agent of high safety useful as a growth inhibitor for Gram-positive bacteria and Gram-negative bacteria, containing, as active ingredient, alpha-1,4-polygalactosamine, its degradation product or fractionated product of said degradation product. CONSTITUTION:The objective anti-fungus agent having the above-mentioned effects, containing, as active ingredient, (A) alpha-1,4-polygalactosamine produced, as PF-101 or PF-102, by culture of imperfect fungi, Paecilomyces I-1 (FERM BP-1180), (B) a degradation product obtained by hydrolysis of the ingredient A with an acid or polygalactosamine degradation enzyme produced by Pseudomonas spH 881 (FERM P-8955), or (C) a fractionated product >=50000, 10000-<50000, or <10000 in molecular weight, obtained by fractionation of the ingredient B through a ultrafiltration membrane.

Description

DETAILED DESCRIPTION OF THE INVENTION (Field of Industrial Application) The present invention relates to an antibacterial agent against Gram-positive and Gram-negative bacteria. More specifically, the present invention provides α-1
, 4-bond polygalactosamine as an active ingredient.

The antibacterial agent of the present invention can inhibit the growth of Gram-positive bacteria and Durum-negative bacteria by adding or spraying it, such as Escher, which is a Durum-negative bacteria.
Ichia spp., Pseudomonas spp. and Durham-positive bacteria Staphyrococcus spp., Bac.
(Prior art and problems) Conventional antibacterial preparations include chemically synthesized preparations such as sulfa drugs, or antibiotics mainly produced by actinomycetes. substances are known. However, in recent years, there has been interest in the acute toxicity, chronic toxicity, and carcinogenicity of drugs, as well as their genotoxicity.
There is a demand for safer formulations.

Reflecting these facts, research has been conducted on natural substances such as pectin decomposition products, protamine, spices, and chitin and chitosan, which are components of shrimp and crab shells, and their mild decomposition products, and some of them have been put into practical use. are being considered.

α-1,4-borigalac{-samine is also a natural polysaccharide produced by a living organism, and is structurally similar to the sugar constituting glucosamine and chitosan, and is expected to have similar physiological activities.

However, polygalactosamine (α-1,4 galactosaminogalactan) is very rare in the natural world, and only PF-Lot and PF-102 derived from Deuteromyces bacterium are known (Japanese Patent Publication No. 12639/1983). Unexamined Japanese Patent Publication 1986
-294093).

In addition, it is generally known that when polysaccharides are reduced in molecular weight, their physicochemical properties change, making them easier to handle than in a polymer state, and making them more physiologically active. Regarding the present borigalac l-samine, methods to reduce the molecular weight not only by treatment with acid or alkali but also by enzymatic decomposition, that is, polygalactosamine-degrading enzyme produced by Pseudomonas bacteria, have already been revealed. (Japanese Patent Application Laid-Open No. 63-164884, JP-A No. 63-164885). (Means for Solving the Problems) An object of the present invention is to provide an antibacterial preparation with higher safety.

The present invention relates to an antibacterial agent containing α-1.4-linked galac-1/samine as an active ingredient. The α-1,4-linked galacyl-zamine, which is the active ingredient of the antibacterial agent in the present invention, is derived from the Deuteromyces bacterium Paecilomyces I-1 (Feikokenjoyori No. 1).
180 FERM BP-1180), PF-101 and PF402 can be obtained.

In addition, decomposition products obtained by hydrolyzing PF-101 and PF-102 with a polygalac-1-samine degrading enzyme produced by Pseudomonas sp 8881 (WL Koken Yuyori No. 8955) or by acid hydrolysis By fractionating with an ultrafiltration membrane, α-1,4-linked polygalactosamines with fractional molecules R of 50,000 or more, 10,000 or more to less than 50,000, and less than 10,000 can be obtained, respectively.

The antibacterial agents of the present invention include + PF-101, PF-10
2. All α-1,4-linked galac I/samines such as these decomposition products and fractions can be used alone or in combination.

Next, α-1.4 polygalactosamine. PF-101
The bacteria producing PF-102 and PF-102 will be explained below.

The Deuteromyces II bacterium isolated from a humus layer in Wakayama Prefecture was recognized as belonging to the Paecilomyces genus and was named Baecilomyces I-1, and the strain has been deposited at the National Institute of Fine Technology as FERM BP-1180. .

Next, Paecijolllye published by T.
es I-1).

(a) Observation under microscope F This fungus lacks conidiophores, and conidia are individual phialids (phial.id) that grow directly from vegetative hyphae or vegetative hyphal bundles.
It is derived from the tip of e) in a long chain. Phialide is translucent and has a length of 20-45μ, with a slightly thick base (1.0-1.5μ) and a slightly tapered tip (0.5μ).
~1.0μ), and some have a straight or slightly curved tip. The conidia are cigar-shaped (or rod-shaped) by electron microscopy, and their size is 4 to 6 x 1.
It is 0 to 1.4μ.

Conidia are in chains of 25 to 35 conidia, but in rare cases long chains are formed. This chain of conidia is extremely fragile and can be easily broken by the slightest shock.

(b) Growth status in each medium (25°C flat culture) (1
) The colony grows well on Zuavec agar medium and reaches a diameter of about 45 mm on the 14th day. The flora is white, velvety to woolly, with a tuft-like bulge in the center, and a circular periphery of the colony. No water drops or wrinkles. The underside of the colony is white in the early stage of culture, and the central part is pale yellow in the later stage of culture.

No pigment production was observed on the agar.

(2) The malt agar medium colony grew well, with a diameter of approximately 54 m + n on the 14th day.
The area around the colony is not circular but rather plum-shaped.

The central part of the bacterial flora is white, but the peripheral part is pale yellow. The thickness of the flora is medium, and the central part is slightly concave. Water drops/
No wrinkles were observed, and the entire underside of the colony was pale yellow. A pale yellow pigment was produced on the agar medium.

(3) Potato dextrose agar medium Colonies grew very well and had a diameter of about 60 mI on the 14th day.
Reach I1. Forms a fairly thick white velvet-like or wool-like bacterial flora, with a slightly swollen central area, a slightly thinner flora with a pale yellow color in the sub-central area, and a relatively thick white bacterial flora in the peripheral area. . There are no wrinkles on the surface, but a few light brown water droplets can be seen. There are several radial wrinkles on the back of the colony, and concentric yellow shading can be observed. There is diffusion of pale yellow pigment into the agar.

(4) YpSs agar medium (composition starch 1.5%, yeast extract 0.4%, K2HP040.1%, MgS
(0.05% O, 2% agar) Colonies grew well and reached a diameter of about 50 mm on the 14th day. It is a fluffy, woolly, thick bacterial flora all over its white color. No water drops or wrinkles. The underside of the colony has no noteworthy features. No pigment production.

(5) MY2. Agar medium (composition: glucose 20%, polypeptone 0.5%, yeast extract 0.3%, malt extract 0.3%, agar 2%) Colony growth was not very good, with a diameter of about 30 mm on the 14th day.
It is mm. Aerial mycelium does not form much and there are many fine wrinkles.
The periphery is pale yellow and the center is pale brown. The underside of the colony is pale yellow with fine wrinkles. No pigment production.

Paecilomyces I-1 can be cultured by a conventional liquid culture method for filamentous fungi.

Spores or hyphae of Paecilomyces I-1 are inoculated into a liquid medium and cultured aerobically. Glucose as a carbon source,
Maltose, sucrose, starch, blackstrap molasses, etc. can be used, but glucose is preferably used. As a nitrogen source, inorganic nitrogen such as ammonium sulfate and sodium nitrate, and organic nitrogen such as peptone and yeast extract can be used.

Although the culture temperature can be changed as appropriate within the range in which the flocculating active substance-producing bacteria produce the flocculating active substance, it is usually preferable to culture at 20 to 25°C. The culture time varies depending on the culture conditions, but is usually about 4 to 5 days, and the culture may be terminated at an appropriate time by estimating the time when the agglutinating active substance reaches its maximum level.

Here, if the culture filtrate is concentrated by a method such as vacuum concentration or ultrafiltration, and an organic solvent such as ethanol is added to precipitate the concentrated liquid, PF-1 described in Japanese Patent Publication No. 56-12639 can be obtained.
01 is obtained.

In addition, PF-101-producing bacterium Paecilomycesto 1
is cultured and filtered, various salts are added to the obtained culture filtrate or filtrate concentrate, and if precipitation does not occur, an alkali is added as necessary to adjust the pH to 7 to 9 to precipitate.
Separate the precipitate, wash it with water, dissolve it in a dilute acid aqueous solution,
Add salt again or adjust the pH to 7 to 9 by adding alkali, etc., and precipitate.
PF-102 described in the publication can be obtained.

The physical and chemical properties of PF-101 are as follows.

(1) Agglomeration activity: Collects suspended fine particles in extremely small amounts.

(2) Aggregation activity pH range: Stably exhibits aggregation activity at p82-9.

(3) Aggregation activity temperature range; aggregation activity is observed at 0 to 100°C.

(4) Coagulation active ionic strength; carbonate ions and Fe,
(so4)3 inhibits aggregation activity, but other ions and ionic strength have no effect on aggregation activity, and NaCl. K2So4 has no effect on IM at all.

(5) Elemental analysis: Nitrogen 5.44%, carbon 37.45%,
Hydrogen 6.3%, phosphorus 0.27%.

(6) Ultraviolet absorption spectrum; as shown in Figure 1.

(7) Infrared absorption spectrum; as shown in Figure 2.

(8) Color reaction; Ninhydrin reaction, xanthoprotein reaction, Ehrlich reaction, Molisch reaction, ten-phenol sulfuric acid reaction, ten-rerosen test (9) Hydrolysis with acid: 6N hydrochloric acid, 110", 20
Galactosamine and ammonia are obtained by time resolution,
The sample 7 was decomposed with 4N hydrochloric acid at 100℃ for 16 hours.
4% galactosamine and a small amount of ammonia are obtained. (10) Electrophoresis: Confirmed as a single substance by density gradient isoelectric focusing, and the isoelectric point (ρ) is 8.5. (11) Color of substance: pale yellowish white (12) Distinction between basic, acidic, and neutral: Slightly basic with an isoelectric point of 8.5. (pH of 0.1% aqueous solution is 6.2, pH of deionized water is 5.8) (13) Solubility in solvents: Soluble in hot water, does not precipitate even when cooled after dissolution. Slightly soluble in cold water.

Dissolves lotus in dilute acid and dilute alkali.

The physical and chemical properties of 7PF-102, which is insoluble in alcohols, acetone, chloroform, benzene, ethyl acetate, and lobentane, are as follows.

(1) Agglomeration activity: Agglomerates turbid fine matter in extremely small amounts. (2) Aggregation activity pH range: Stably exhibits aggregation activity at p}12-9.

(3) Aggregation activity temperature range; aggregation activity is observed at 0 to 100°C.

(4) Coagulation active ionic strength: carbonic acid and Fe, (So.
)] inhibits aggregation activity, but other ions and ionic strength do not affect aggregation activity.
NaCl. K, So4 has no effect on IM at all.

(5) Elemental analysis; Nitrogen 8.64% & Carbon 4: 180%
, hydrogen 6.87% General formula: (CG +{1 x NO4 ・XH2 0
)n(6)2 external absorption spectrum 1-; as shown in FIG. S3.

(7) Infrared absorption spectrum: as shown in FIG.

(8) Color reaction; Protein reaction to ninhydrin reaction xan 1 - Lich reaction Molisch reaction phenol sulfuric acid method + ± Rerosentes j - (9) Electrophoresis; Confirmed as a single substance by density gradient isoelectric focusing, The isoelectric point (pI) is 8.5.

(10) Color of substance; light yellow (11) Distinction between basic, acidic, and neutral pH when suspended in water at 0.5% ν/V is 7.5 (pH 5.8 of deionized water) (12) Soluble in solvents Slightly soluble in hot water Slightly soluble in cold water Slightly soluble in dilute acids Slightly soluble in dilute alkalis Alcohols, acetone, gloloborm, benzene, n
-Insoluble in pentane.

(l3) average molecular weight of 160,000 or more.'' Both PF-101 and PF-102 have an average molecular weight of α-1.
4 polygalactosamine, and all of them were found to have antibacterial properties in the present invention, and their decomposition products and fractions of the decomposition products were also imbued with antibacterial properties, and the present invention was completed. be.

When α-1,4 polygalac-1-samine is decomposed, ie, reduced in molecular weight, it is hydrolyzed by an acid such as hydrochloric acid or sulfuric acid, or by a polygalactosamine-degrading enzyme.

Next, one example of polygalactosamine degrading enzyme will be explained. This polygalactosamine degrading enzyme is Pseudomonas s.
pH881 FERN P-8955.

(1) Action and substrate specificity This enzyme acts on oligo and polygalactosamines (α-1,4 borigalac 1 hesamine) with a degree of polymerization n=4 (tetragalactosamine) or higher to produce oligogalata 1 and samine.

Other polysaccharides, starch (α-1.4 glucan), glycogen (α-1,4 glucan), punorelan (α-1,
4 glucan), dextran (α-1.6 glucan)
, laminaran (β-1.3-gnolecan), carboxylcellulose (β-1.4-gnolecan), chitosan (β-
1,4 glucosaminoglucan), ethylene glycol chitin (β-1,4N-acetylglucosaminoglucan)
, Pseudomonas solanacearum
Poly N-acetylgalactosaminogalactan (poly β
-1,3N-acetylgalactosaminogalactan) (Y
．． Akiyama. , etc. al. , Agric
．． Biol. Chem. , 50(3)747.
1936) etc. have no effect at all.

It also does not act on α-1,4 galactosaminooligosaccharides with a degree of polymerization of n=3 (trigalactosamine) or less.

(2) Optimal pH and stable PH range When using citrate phosphate buffer, the optimal ρH is 4.5
~7.0. In addition, the stable ρH range is pH 4.5 to 8.
．． It is 0. This measurement showed the residual activity of the enzyme after being left at 37°C for 1 hour as a relative value.

(3) Enzyme activity measurement method Enzyme activity was determined using PF-101 or PF-102 produced by Paecilomyces I-1 bacteria (the main constituent sugar being α-1,4 galactosaminogalactan). The enzyme solution was added to 0.5ml of this 0.5% 70.1M acetate buffer pH 6.0 solution. Add 5mQ and heat to 37℃
, 10 minutes, and the resulting reducing power is
Measured by Nelson method. The enzyme unit was defined as the activity that increases the reducing power equivalent to 1 μmol of galactosamine per minute. (4) Range of optimum temperature and temperature stability for this enzyme The optimum temperature for this enzyme is 55°C, and the temperature decreases rapidly above this temperature. 70% activity remains after 1 hour at 50°C. A decomposition product of α-1,4 polygalactosamine can be obtained by adding a polygalactosamine degrading enzyme to a solution of α-1,4 polygalactosamine and enzymatically decomposing it at about 35 to 40°C. Since the decomposed product of α-1,4 polygalactosamine has antibacterial properties, it can be used as an antibacterial agent as it is, but the decomposed product of α-1,4 polygalactosamine can be fractionated using an ultrafiltration membrane, etc. By doing this, fractions separated into various molecular weight units can be obtained.

The decomposition product of α-1,4 polygalactosamine can be obtained by using an ultrafiltration membrane to obtain fractions with a molecular weight of 50,000 or more, a molecular weight of 10,000 or more and less than 50,000, and a molecular weight of less than 10,000. The painting also has antibacterial properties and serves as the antibacterial agent of the present invention.

The present invention is an antibacterial agent containing α-1,4 polygalactosamine, a decomposition product thereof, or a fraction of the decomposition product as an active ingredient.

In the present invention, the active ingredient can be effectively used as an antibacterial agent as it is as an additive, or formulated into various liquids and powders.

Next, production examples and examples of the present invention will be shown.

Production Example 1 Production of α-1,4 polygalactosamine (PF-102) 600 g of glucose, 60 g of polypeptone. CaCl2
・Dissolve 125g of 2H,0 in 11i of tap water. '7
3 After adjusting the pH to 7.0 with NaOH solution, 30Q. Transferred to jar fermenter.

Pressure and heat sterilization (121° C., 20 minutes) was performed by injecting steam into this medium solution. After cooling, add 150ml to the medium (final volume: 20a) in a 500ml Erlenmeyer flask.
1IIQ with a medium of the same composition (glucose 3%, polypeptone 0.3%, CaC1, 0.5%, pH 7.0).
Baecilomyces I-1, FE cultured with shaking at ℃ for 4 days
RN OP-1180 (FERM P-3928) was aseptically inoculated at a volume ratio of about 10%. After inoculation, 27℃, aeration rate 0.5VVM, stirring number 20ORPM (7) under the following conditions:
Cultured for 1 day.

After the culture is completed, culture filtrate i is obtained by filtering the culture with a filter cloth.
I got ll. While heating this culture filtrate to 50°C to 60°C, an ultrafiltration membrane (Mitsubishi Rayon Engineering Co., Ltd. IBUF membrane tubular module
The low molecular weight fraction was removed by passing the solution through a liquid (type) and concentrated until the liquid volume was approximately 3Q. Furthermore, about 14000
Cell residue and heat-denatured proteins were removed by centrifugation at G. After centrifugation, approximately 750 g of salt was added to supernatant fraction 3Q (approximately 2
5% concentration), stirred, and after dissolving, diluted with concentrated NaOH.
was adjusted to 7.0 to 8.5. After leaving it for one night to extract ten samples of salted-out material, the salted-out material was collected on Saran cloth (copolymer of vinylidene chloride and vinyl chloride).

Furthermore, a large amount of slightly alkaline water (PH
7.0 or higher) to wash away excess table salt, neutral sugars and other impurities mixed in the culture solution at the same time.6 Next, a 0.1M hydrochloric acid solution was added to the salted out product after washing with water. About 3 times the volume was added and dissolved. 9 Add concentrated Nail solution to this dissolved material and Polygalac! - Adjusted to pH 8.5, which is the isocenter of samin. After leaving it overnight to precipitate the analyte, the precipitate was collected on a saran cloth in the same manner as above and washed with a large amount of tap water. I'm going to wash this dish again. After dissolving in 1M hydrochloric acid, it was purified by isoelectric precipitation and repeated washing with water. This purified precipitate was sterilized at 121°C for 515 minutes and then freeze-dried to obtain a purified powder of PF-102 (α-1,4 polygalac l-
7 g of samin (purity of about 99%) was obtained.

Production Example 2 Partial purification of α-1,4 polygalactosamine α-1.
4 polygalactosamine (PF-102) 100g
The mixture was dispersed in 2Q normal salt anesthesia and hydrochloric acid hydrolyzed at 80° C. for 4 hours in an Erlenmeyer flask equipped with a cooling tube.

After decomposition, this hydrochloric acid hydrolyzed solution was diluted with 1-10N sodium hydroxide.
Neutralize with Rium p! {7. This solution was first treated with an ultrafiltration membrane with a molecular weight of 50,000 (manufactured by Grace Japan) and then further treated with an ultrafiltration membrane with a molecular weight of 17,000.

In this way, polygalactosamines with α-1,4 bonds each having a molecular weight of 50,000 or more, 10,000 to 50,000, and less than 10,000 were obtained.

Both portions of each were freeze-dried to obtain a powder sample.

Production Example 3 Borigalac}-Samine Decomposition #Production of Element Pseudomonas Sp H881, FERM P-895
5 in a 500 mQ Erlenmeyer flask with 0.5% glucose
, yeast extract 0.05%, polypeptone O. 100 ml of seed medium with a composition of 0.05% was inoculated and grown at 30'C for 2
Incubate for 0 hours.

The obtained seed culture solution was added to 0.25% polygalactosamine (PFl02) in a 30Q Jarfer Menta.
Inoculated into enzyme production medium 18Q containing 0.25% glucose, 0.05% yeast extract, and 0.05% polypebutone, and
Aeration (18 Q/min) stirring (200 rpm) at ℃ for 48 hours
Cultivate.

The obtained culture solution was centrifuged (14.00 Orpm) L
, the culture filtrate obtained (enzyme activity 0.0
0351J/rrrQ. Total activity 631J/18f'! )
The proteins were precipitated by adding ethanol cooled to 60% concentration, the precipitated proteins were separated from the solution by centrifugation, and the resulting proteins were dissolved in 0.1 molar acetate buffer (pH 5.0). Adsorb onto an equilibrated C tocefadesox C-25 column (2.5 x 60 cm) and elute using the same buffer with a concentration gradient of 0 to 0.5 molar saline.

The eluted enzyme activity fraction was collected and filtered using an ultrafiltration device (molecular weight 1
Concentrate using 1,000,000 ml of water. Next, 0 containing 2 molar salt
．． A Sephadex G-50 column (5X
90 cm) chromatography. The salt concentration in the active fraction was then increased to 4 molar and a Phenyl-Sepharose CL-4B column equilibrated with a similar solution (2.
5 x 20e+i) and eluted with 0.1M acetate buffer with an inverse concentration gradient of sodium chloride to obtain purified polygalactosamine degrading enzyme 50g (yield 23.1%, specific activity 52μ).
g gal. N/min/mg protein) was obtained.

Production Example 4 α-1.4 Polygalactosamine fractional method Purification Polygalactosamine 25K was dissolved in 0.1 mol of 4,8Q acetic acid, and then the pH was adjusted to 6.0 with sodium hydroxide. This polygalactosamine solution containing 5Q liquid was used as a substrate, and 10 mg of the purified polygalactosamine degrading enzyme obtained in Production Example 3 was added thereto and enzymatically decomposed at 37°C. This decomposed solution was first treated with an ultrafiltration membrane with a molecular weight of 50,000 (manufactured by Grace Japan), and then further treated with an ultrafiltration membrane with a molecular weight of 10,000. In this way, α-1,4-linked polygalactosamines having molecular weights of 50,000 or more, 150,000, and 1-10,000, respectively, were obtained.

Furthermore, both parts of each sample were freeze-dried to obtain a powder sample.

The physical and chemical properties of the α-74-linked polygalactosamine with a molecular weight cut-off of 50,000 or more obtained in 22 are as follows.

1. Elemental analysis: Nitrogen 8.6%, Carbon 42.8%, Hydrogen 6
. 9%, oxygen 41.7% 2. Specific optical rotation; Cα]6°=. +215~2253. Melting point: Browning at 160℃ or higher, 210'CTF carbonization, 230
Ashing at ℃4. Color of substance: light yellow5. Color reaction; Ninhydrin reaction Decaxantoprotein reaction Ehrlich reaction Molisch reaction Phenol sulfuric acid reaction 6. Solubility in solvents: Slightly soluble in water, soluble in dilute acids, methanol, ethanol, acetone, chloroform,
Poorly soluble in hexane7. Ultraviolet absorption spectrum; shown in FIG.

8. Infrared absorption spectrum; shown in FIG.

Further, the physical and chemical properties of α-1,4-linked polygalactosamine having a molecular weight cut-off of 10,000 or more and less than 50,000 are as follows.

■． Elemental analysis: Nitrogen 8.7%, Carbon 45.7%, Hydrogen 3
．． 2%, oxygen 42.4% 2. Specific optical rotation; [α] et al' = +210 to 2203. Melting point: Does not have a specific melting point, browns at 160°C, carbonizes at 210°C, ashes at 230°C4. Color of substance: light yellow5. Color reaction; Ninhydrin reaction Decaxantoprotein reaction Ehrlich reaction Molisch reaction Phenol sulfuric acid method 6. Solubility in solvents: Soluble in water, slightly soluble in methanol, slightly soluble in dimethyl sulfoxide, slightly soluble in ethanol, acetone, chloroform, hexane7. Ultraviolet absorption spectrum; shown in FIG.

8. Infrared absorption spectrum; shown in FIG.

The physical and chemical properties of α-1,4-linked polygalactosamine with a molecular weight cut-off of less than 10,000 are as follows.

■． Elemental analysis: Nitrogen 8.6%, Carbon 44.3%, Hydrogen 6
．． 9%, oxygen 40.2% 2. Specific optical rotation; [α]♂': +206.83. Melting point: Does not show a specific melting point and begins to carbonize at temperatures above 160°C.

4. Color of substance: light yellow5. Color reaction; ninhydrin reaction, ten indole hydrochloric acid reaction, ten Somogyi-Nelson reaction, ten phenol sulfuric acid reaction, iodine reaction6. Solubility in solvents; Soluble in water, Soluble in dilute acids, Slightly soluble in methanol, Slightly soluble in dimethyl sulfoxide, Slightly soluble in ethanol, acetone, and chloroform7. Ultraviolet absorption spectrum: Shown in Figure 9. 8. Infrared absorption spectrum; shown in FIG.

Example 1 Culture medium; 10 g of meat extract, peptone Log, and 5 g of sodium chloride were dissolved in deionized water. After adjusting the pH to 6.0, it was determined as IQ. Separately, α-1.4 polygalactosamine (PF-102) produced in Production Example 1, the hydrochloric acid decomposition product of α-1,4 polygalactosamine produced in Production Example 2, and the hydrochloric acid decomposition product prepared in Production Example 2. For each molecular weight in the image, each α-1.4 polygalactosamine was dissolved in 0.1 N acetic acid, and the final concentration was 0.1, 0.05, and 0.0125, respectively.
, 0.006, and 0.003% to the culture medium.

Furthermore, add agar to a final concentration of 1.5% and heat at 121℃.
After autoclaving for 15 minutes, a polygalactosamine-containing plate medium was prepared.

The antibacterial effect was determined by the minimum concentration of polygalactosamine in the medium that was confirmed to inhibit the growth of the inoculated bacterial strain. The strain was cultured at 30°C for 2 days, and the results were evaluated.

The strain used in the test was Esehcrichia eoH. IFO 1273
4, Bacill. us subtjlis IFO
3513, Pseudomonas aerugi. n
osa IFO 3449, Mierococcus
luteus IFO 12708, Mycobact
erium phlej. IFO 3158, Sta.
phyloeoccus aureus IFO 22
It is 732.

The results are shown in Table 1.

[Brief explanation of drawings]

Figure] shows the ultraviolet absorption spectrum of the old PF-1, Figure 2 shows the infrared absorption spectrum of PF-101, and Figure 3 shows the ultraviolet absorption spectrum of PF-102. Figure 4 shows the infrared absorption spectrum of PF-102, and Figure 5 shows the ultraviolet absorption spectrum of a fraction of the decomposition product of PF-102 with a molecular weight of 50,000 or more. 6th
The figure shows its infrared absorption spectrum, Figure 7 shows the ultraviolet absorption spectrum of the fraction of the decomposed product of PF-102 with a molecular weight of 10,000 to 50,000, and Figure 8 shows its ultraviolet absorption spectrum. Figure 9 shows the ultraviolet absorption spectrum of a fraction of 10,000 drops of molecular weight of the decomposed product of PF-102, and Figure 10 shows its infrared absorption spectrum. It is. Agent Patent Attorney Chika Toda Diagram Wavelength nm Diagram Wavelength nm

Claims (4)

[Claims] (1) An antibacterial agent containing α-1,4 polygalactosamine, a decomposition product thereof, or a fraction of the decomposition product as an active ingredient. (2) The antibacterial agent according to item 1, wherein the fraction of the decomposition product of α-1,4 polygalactosamine is α-1,4 polygalactosamine having a molecular weight of 50,000 or more. (3) The antibacterial agent according to item 1, wherein the fraction of the decomposition product of α-1,4 polygalactosamine is α-1,4 polygalactosamine having a molecular weight of 10,000 or more and less than 50,000. (4) The antibacterial agent according to item 1, wherein the fraction of the decomposition product of α-1,4 polygalactosamine is α-1,4 polygalactosamine with a molecular weight of less than 10,000.