JPH02240007A - Antifungous agent - Google Patents

Abstract

PURPOSE:To provide an antifungal agent capable of being added or sprayed to soils or plants to inhibit the growth of plant disease fungi by containing alpha-1,4-galactosamine, a decomposed product thereof or a fraction of the decomposed product as an active ingredient. CONSTITUTION:An antifungal agent contains as an active ingredient an alpha-1,4- galactosamine having a mol.wt. of >=50000, 10000-50000 or <10000 and prepared by culturing an imperfect fungus Paecilomyces I-1 (FERM BP-1180) as PF-101 or PF-102, by hydrolyzing the PF-101 or PF-102 with a polygalactosamine- decomposing enzyme produced by Pseudomonas sp H881 (FERM P-8935) or by fractionating the decomposed products of the acid hydrolysis with an ultrafiltration membrane.

Description

DETAILED DESCRIPTION OF THE INVENTION (Industrial Application Field) The present invention is directed to the use of filamentous fungi, particularly the plant pathogenic fungus Fusari.
This invention relates to an antifungal agent against bacteria of the genus Um and genus Botrytis. More specifically, the present invention relates to an antifungal agent containing α-1,4-linked polygalactosamine as an active ingredient.

The antifungal agent of the present invention can inhibit the growth of plant pathogenic bacteria by adding or spraying it to soil or plants.
It can be used very effectively as a growth inhibitor for bacteria of the genus Ytis. (Prior art and problems) Conventional antifungal agents include copper preparations such as Bordeaux solution, heterocyclic compounds used for powdery mildew, and imidazole agents used for scab. It has been known. However, in recent years, there has been interest in the acute toxicity, chronic toxicity, carcinogenicity, and genotoxicity of drugs, and there is a demand for safer antifungal agents. 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 polygalactosamine is also a natural polysaccharide produced by microorganisms, and is structurally similar to chitosan, which has glucosamine as a constituent sugar, and is expected to have similar physiological activities.

However, polygalactosamine (α-1,4 galactosaminogalactan) is extremely rare in the natural world, and only PF-101 and PF-102 derived from Bacillus deuteromycetes are known (Japanese Patent Publication No. 12639/1983). Unexamined Japanese Patent Publication 1986
-294093). In addition, it is generally known that polysaccharides change their physicochemical properties when they are reduced in molecular weight, making them easier to handle than in a polymer state, and making them more physiologically active. Regarding this polygalactosamine, methods for reducing 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 clarified (particularly 1986-164884, Japanese Patent Publication No. 63-164884
3-164885). (Means for Solving the Problems) An object of the present invention is to provide a safer antifungal agent.

The present invention relates to an antifungal agent containing α-1,4-linked galactosamine as an active ingredient. α-1,4-linked galactosamine, which is the active ingredient of the antifungal agent in the present invention, is
Incomplete bacterium Baecilomycesto 1 (Feikoken Joyori No. 1180
By culturing F[ERM BP-1180). It can be obtained as PF-101 and PF-102.

Also, PF-101. The decomposition product obtained by hydrolyzing PF-1.02 with a polygalactosamine-degrading enzyme produced by Pseudomonas sp H881 (Feikoken Bacteria No. 8955) or by acid hydrolysis was filtered using an ultrafiltration membrane. By fractionation, α-1,4-linked polygalactosamines having a molecular weight cut-off of 50,000 or more, 10,000 or more to less than 50,000, and less than 10,000 can be obtained, respectively.

The antifungal agent of the present invention includes PF-101. PF-10
2. All α-1,4-linked galactosamines such as these decomposed products and fractionated products can be used alone or in combination.

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

Deuteromyces I-1 isolated from a humus layer in Wakayama Prefecture was recognized as belonging to the genus Paecilomyces and was named Paecilomyces I-1.
This strain has been deposited with the Microtech Institute as FERN BP, 1180. Next, Peecilomyces I-1. (Paacylomy
ces I-1).

(a) Observation under a microscope This fungus lacks conidiophores, and conidia are individual phialides that grow directly from vegetative hyphae or vegetative hyphal bundles.
) is derived in a long chain at the tip. The phialide has a length of ≠ 20 to 45μ at the shade, the base is slightly thick (1.0 to 1.5μ), and the tip is slightly tapered (0.5 to 1.5μ).
i. oμ), and some have a straight or slightly curved tip. The conidia are cigar-shaped (
or rod type), and its size is 4 to 6 x 1.0
~1.4μ. Conidia usually form a chain of 25 to 35 conidia, but in rare cases long chains are also observed. This chain of conidia is extremely fragile and can be easily broken by the slightest shock.

(b) Growth status fi in each medium (25°C flat culture)
(1) The colony grows 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 area around the colony. Water drops/
No 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) Malt agar medium The colony grew well, with a diameter of about 54 mm on the 14th day, and the colony periphery was 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 bacterial flora is medium, and the central part is slightly concave. No water droplets or wrinkles were observed, and the entire back surface of the colony was pale yellow. A pale yellow pigment was produced on the agar medium.

(3) Potato dextrose agar medium Colonies grow very well and reach a diameter of about 60 mm on the 14th day. Forms a white, velvety to woolly, fairly thick bacterial flora; the central part is slightly swollen, the subcentral part is a pale yellow, slightly thinner flora, and the peripheral area is a relatively thick white bacterial flora. ．． 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 is observed. There is diffusion of pale yellow pigment into the agar.

(4) YpSs agar medium (composition starch 1.5%, yeast extract 0.4%, K, IIPO 40.1%.M
(gSO40.05%, agar 2%) Colony growth was good and reached a diameter of about 50 mm on the 14th day. 'A fluffy, fluffy, thick flora of flora all over the white body, with no water droplets or wrinkles. There are no noteworthy features on the back side of the colony. No pigment production. (5) MY. . 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 m+a. Aerial mycelium does not form much and there are many fine wrinkles, and 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 using a conventional liquid culture method for filamentous fungi. Spores or hyphae of Baecilomyces 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 nitrogen sources, inorganic nitrogen such as ammonium sulfate and sodium nitrate, and organic nitrogen such as peptone and yeast extract can be used. The culture temperature may be changed as appropriate within the range that allows the flocculating active substance producing bacteria to produce the flocculating substance, but 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. You can estimate the time it takes for the active substance to reach its maximum level and finish at an appropriate time.
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. 12639/1988 can be obtained.
01 is obtained.

In addition, PF-lot producing bacterium Paecilomyces I
-1 is cultured and filtered, various salts are added to the obtained culture filtrate or filtrate concentrate, and if no precipitation occurs, alkali is added as necessary to adjust the pH to 7 to 9 to precipitate the new product. Separate the product, wash it with water, dissolve it in a dilute acid aqueous solution, and add salt again or add an alkali etc.
Precipitating with H of 7 to 9, JP-A-62-2940
PF-102 described in Publication No. 93 can be obtained. The physical and chemical properties of PF-101 are as follows. (1)
ii) Collecting activity: Agglomerates suspended fine particles in extremely small amounts. (2) Aggregation activity p}ll[! Circle: Shows stable aggregation activity at pH 2-9. (3) Temperature range for flocculation activity; flocculation activity is observed between 0 and 100°C. (4) Coagulation active ionic strength; carbonate ions and Fe,
The aggregation activity is inhibited by (so4), but the aggregation activity is not affected by other various ions and ionic strength. NaCl, K, and So4 have no effect on IM. (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 - Decaphenol sulfuric acid reaction - Tenrerosen test - (9) Hydrolysis with acid 16N hydrochloric acid, 110°, decomposition for 20 hours produces galactosamine and ammonia. obtained, 4
Decomposition with N-hydrochloric acid at 100°C for 16 hours yields 74% of the standard galactosamine and a small amount of ammonia. (10) Electrophoresis: Confirmed as a single substance by density gradient isoelectric focusing, and the isoelectric point (p!) is 8.5. (1l) Color of substance: Pale yellowish white (12) Distinction between basic, acidic, and neutral; Slightly basic with an isoelectric point of 8.5. (The pH of 0.1% aqueous solution is 6.2,
PL of deionized water is 5.8) (13) Solubility in solvents: Soluble in hot water, does not deteriorate even if cooled after dissolution. Slightly soluble in cold water.

Slightly soluble in dilute acids and dilute alkalis. 7 Insoluble in alcohols, acetone, chloroform, benzene, ethyl acetate, and n-pentane.

The physical and chemical properties of PF-102 are as follows. (1)
Agglomeration activity: Agglomerates suspended fine particles in extremely small amounts. (2) Ii aggregation activity pF1 range; shows stable aggregation activity at ρH2-9. (3) Temperature range for flocculation activity; flocculation activity is observed between 0 and 100°C. (4) Ill collection activity ionic strength dicarbonate and Fe2(S
Although the aggregation activity is inhibited by O4)j, the aggregation activity is not affected by other various ions and ionic strength.
Naσ1, K. So. It does not affect LM at all.

(5) Elemental analysis; Nitrogen 8.64% & Carbon 42.80%
, hydrogen 6.87% General 2: (C.H,,No,・xll.O)n(6)
Ultraviolet absorption spectrum; as shown in Figure 3. (7) Infrared absorption spectrum; as shown in Figure 4. (8) Color reaction lininhydrin reaction The point (pI) is 8.5. (10) Color of substance; pale yellow (11) Distinction between basic, acidic, and neutral When suspended in water at 0.5% -/V, pl1 is 7.5 (
The pH of deionized water is 5.8). (B) Soluble in fR agent, slightly soluble in hot water, slightly soluble in cold water, easily soluble in dilute acid, slightly soluble in dilute alkali, alcohols, acetone, chloroform, benzene,
Insoluble in n-bentane.

(l3) Average molecular weight 160,000 or more The above PF-101 and PF-102 are both α-1,
4-polygalactosamine, and in the present invention, all of them were found to have antifungal properties, and their decomposed products and fractions of the degraded products were also found to have antifungal properties, and the present invention was completed. be. When α-1.4 polygalactosamine 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 a polygalactosamine degrading enzyme will be explained.

This polygalactosamine-degrading enzyme is produced by Pseudomonas, q
pH881 FERM P-8955.

(1) Action and substrate specificity This enzyme acts on oligo and polygalactosamines (α-1,4 polygalactosamine) having a degree of polymerization n; 4 (Te1-lagalac I-samine) or higher to produce oligogalactosamine.

Other polysaccharides, starch (α-1,4 glucan), glycogen (α-1,4 glucan), pullulan (α-1,4
glucan), dextran (α-1.6 glucan), laminaran (β-1,3 glucan), carboxylcellulose (β-1,4-glucan), xyl-san (β-1.
4 glucosaminoglucan), ethylene glycol chitin (β-1.4N-acetylglucosaminoglucan), P
poly N-acetylgalactosaminogalactan (poly β-1
, 3N-acetylgalactosaminogalactan) (Y.
Akiyama. , at. al. , Agric.
Biol. ChelI. , 50(3)747, 1
986) etc. has no effect at all. Furthermore, it does not act on α-1,4 galactosaminooligosaccharides having a degree of polymerization of n=3 (trigalactosamine) or less.

(2) Optimal pH and stable pH range When using citrate phosphate buffer, the optimal pi is 4.5
~7.0. In addition, the stable pu range is p}14.5~
It is 8.0. 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 measured using PF-101 or PF-102 produced by Paecilomyces I-1 bacteria (the main constituent sugar being α-1.4 galactosaminogalactan). Add 0.5IQ of the enzyme solution to this 0.5% 70.1M acetate buffer PH6.0 solution 0.5+an, and store at 37°C.
React for 0 minutes, and reduce the resulting reducing power with Somogyi-Ne
Measured using the lson method. The enzyme unit is 1 per minute.
The activity that increases the reducing power equivalent to μmol of galactosamine was defined as one unit. (4) Range of optimal temperature and temperature stability The optimal 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 decomposed 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 decomposition product of α-1,4 polygalactosamine has anti-fungal properties, it can be used as an anti-mold agent as is, but the decomposed product of α-1,4 polygalactosamine can be separated using an ultrafiltration membrane, etc. By fractionating, it is possible to obtain fractions separated into various molecular weight units. The degradation product of α-1.4 polygalactosamine is 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? However, all fractions have antifungal properties and can be used as the antifungal agent of the present invention. The present invention is an antifungal 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 additive as it is, or formulated into various liquids and powders as an antifungal agent. 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. , Ca
125 g of Cl, 6211■0 was dissolved in 17Q of tap water, adjusted to pH 7.0 with concentrated NaOH solution, and then transferred to a 30Q jar fermenter. This medium solution was sterilized under pressure and heat by injecting steam ( 121°C for 20 minutes). After cooling, add 150% to the medium (final liquid volume 20Q) to a 500°■Q Erlenmeyer flask.
mΩ Medium with the same composition (glucose 3%, polypeptone 0.
3%, CaC1, 0.5%, PH7.0) at 26℃
, Baecilomyces I-1., cultured with shaking for 4 days. FE
RM OP-1180 (FIIERM P-3928)
Approximately 10% of the volume was inoculated aseptically. 27℃ after inoculation,
Air flow rate 0.5VVM. The cells were cultured for 5 days at a stirring rate of 20 ORPM. After the culture is completed, culture filtrate 1 is obtained by filtering the culture with a filter cloth.
I got 712. While heating this culture filtrate to 50°C to 60°C, an ultrafiltration membrane (Mitsubishi Rayon,
The low molecular weight fraction was removed by passing through a UF membrane tubular module F type manufactured by Engineering Co., Ltd., and concentrated until the liquid volume was approximately 3Q. Furthermore, about 14,000
Cell residue and heat-denatured proteins were removed by centrifugation at XG. After centrifugation, approximately 750 g of salt was added to supernatant fraction 3Q (approximately 2
5% concentration) was added and stirred, and after dissolving, the pH was adjusted to 7.0 to 8.5 with concentrated NaOH. After leaving it for one night to extract ten samples of salted-out material, the salted-out material was collected on Saran cloth (a copolymer of vinylidene chloride and vinyl chloride). Furthermore, a large amount of slightly alkaline water (pH 7.0 or higher) is poured over this salted-out product.
By removing the water, excess salt, neutral sugars, and other impurities mixed in the culture solution were washed away. Next, 0. Approximately three times the volume of IM hydrochloric acid solution was added and dissolved. A concentrated NaOH solution was added to this solution to adjust the pH to 8.5, which is the isoelectric point of polygalactosamine. After leaving it overnight to precipitate ten analytical precipitates, the precipitates were collected on Saran cloth in the same manner as above, and washed with a large amount of tap water. Repeat this washing with water one more time. After dissolving in IM hydrochloric acid, it was purified by isoelectric precipitation and repeated washing with water.

This purified precipitate was sterilized at 121°C for 15 minutes and then freeze-dried to obtain 7 g of purified powder of PF-102 (approximately 99% purity as α-1.4 polygalactosamine) containing polygalactosamine as the main component. Obtained. Production Example 2 Partial purification of α-1.4 polygalactosamine α-1.
4-polygalactosamine (PF-102) IOOJC was dispersed in 2Q of 4N hydrochloric acid, and hydrochloric acid hydrolysis was performed at 80° C. for 4 hours in an Erlenmeyer flask equipped with a cooling tube.

After decomposition, this hydrochloric acid hydrolysis solution was neutralized to pH 7 with 10N sodium hydroxide. This solution was first treated with an ultrafiltration membrane with a molecular weight cutoff of 50,000 (manufactured by Grace Japan), and then further treated with an ultrafiltration membrane with a molecular weight cutoff of 10,000. In this way, α-1,4-linked polygalactosamines having molecular weights of 50,000 or more, 10,000 to 50,000, and less than 10,000, respectively, were obtained. A powder sample was obtained by freeze-drying both parts of each sample. Production Example 3 Production of polygalactosamine degrading enzyme Pseudomonas sp H881, FERM P-895
5 in a 500 mQ Erlenmeyer flask with 0.5% glucose
, yeast extract 0.05%, and polybebutone 0.05% in 100v2 of the seed medium and cultured at 30°C for 20 hours.

The obtained seed culture solution was placed in a 30Q jar fermenter with 0.25% polygalactosamine (PF-102),
Inoculate 181 m of enzyme production medium containing 0.25% glucose, 0.05% yeast extract, and 0.05% polypeptone, and
Aeration (18 Q/min) and stirring (20 Or
pm) Cultivate.

The obtained culture solution was centrifuged (14,000rρI1) to remove the bacterial cells, and the obtained culture filtrate (enzyme activity: 0.00
Proteins are precipitated by adding chilled ethanol to 60% concentration (35 Ll/mQ, total strength 631 J/18+2), and the precipitated proteins are separated from the solution by centrifugation. The obtained protein was dissolved in 0.1 molar acetic acid buffer (P
Adsorb onto a C tosephadex C-25 column (2.5 x 60 cm) equilibrated with H5.0) and elute using the same buffer with a concentration gradient of 0 to 0.5 molar saline. Collect the eluted enzyme activity fraction and concentrate using an ultrafiltration device (maintaining a molecular weight of 10,000). Next, a Sephadex G-50 column (s
x 90cm) chromatography. Next,
A Phenyl-Sepharose CL-4B column equilibrated with the same solution with the salt concentration in the active section increased to 4 molar (
2.5 x 20 cm) and eluted with a 0.1 molar acetate buffer with an inverse concentration gradient of sodium chloride to degrade purified polygalactosamine! 150 1g (yield 23.1%, specific activity S2
pg galN/win/mg protein) was obtained. Production Example 4 α-1,4 Polygalactosamine fractional method 25 g of purified polygalactosamine was dissolved in 0.1 mol of acetic acid of 4.8 12, and then adjusted to ρ of 116.0 with sodium hydroxide.
The total liquid material was 5Q. This polygalactosamine solution was used as a substrate, and the purified polygalactosamine degrading enzyme 10+B 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 cutoff of 50,000 (Grace Japan Co., Ltd. 1l). Further. Molecular weight cut-off 1
It was treated with 10,000 ultrafiltration membranes. In this way, each molecule!
Polygalactosamines with α-1,4 bonds of 50,000 or more, 10,000 to 50,000, and less than 10,000 were obtained. Furthermore, both parts of each sample were freeze-dried to obtain a powder sample. The physical and chemical properties of the α-1,4-linked polygalactosamine with a molecular weight cut-off of 50,000 or more are as follows. 1. Elemental analysis: Nitrogen 8.6%, Carbon 42.8%, Hydrogen 6
．． 9%, oxygen 41.7% 2. Specific optical rotation; [α] also' = + 215 to 2253
．． Melting point: Browning at 160℃ or higher, carbonization at 210℃, 230℃
Ashing at ℃4. Color of substance: light yellow5. Color reaction; ninhydrin reaction, dexanthate protein 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 Figure 5. 8. Red external absorption spectrum; shown in Figure 6. The physical and chemical properties of α-1,4-linked polygalactosamine with a molecular weight cut-off of 10,000 to 50,000 are as follows. 1. Elemental analysis: Nitrogen 8.7%, Carbon 45.7%, Hydrogen 3
．． 2%, oxygen 42.4% 2. Specific optical rotation; [α]i)' = +210 to 2203.
Melting point: Does not have a specific melting point, turns brown at 160℃, 210℃
Carbonization at 230℃, ashing at 230℃4. Color of substance: light yellow5. Color reaction; Ninhydrin reaction Decaxanto protein 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 Figure 8. Also,
The physicochemical properties of α-1,4-linked polygalactosamine with a molecular weight cut-off of less than 10,000 are as follows. 1. Elemental analysis;
Nitrogen 8.6%, Carbon 44.3%, Hydrogen 6.9%, Oxygen 4
0.2% 2. Specific optical rotation: [α]B″=+206.83. Melting point: Does not exhibit a particular 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 reaction 16. 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 Figure 10. Example 1 α-1,4 polygalactosamine (P
F-102), the hydrochloric acid decomposition product of α-1,4 polygalactosamine prepared in Production Example 2 and the fractions of the hydrochloric acid decomposition product prepared in Production Example 2 with each molecular weight, each α-1
．． 4 Polygalactosamine in potodextrose medium (potato dextrose agar medium: Eiken Chemical Co., Ltd.) 30
After heating and dissolving 1000@ll of purified water, 0.1 g of each
% and autoclaved at 121°C for 15 minutes, an agar plate medium was prepared.

One platinum loop was inoculated from a slant of pre-cultured mold onto the plate medium containing polygalactosamine of each molecular weight, and after culturing at 27°C for 4 days, the diameter of the colony was measured.
The antibacterial activity was expressed as the ratio of colonies in each test plot, with the diameter of mold colonies inoculated in polygalactosamine-free medium taken as 100. The mold tested was ^itarnaria kikuchiana I
FO 8414, Botrytis cinarea
．． Cochlibolus miyabeanus
IFO 5277, Diaporths citr
i IFO 9170, Elsinoa fav
cetti IFO 8417, Fusarium
Oxysporum capaa, Fusari
um solni, Gibbarslla zeae IFO 71
60, Guignardia larisina
IF0 7888,lalminthosporiu
m sigmoidium var. irrBular
e IFO 5273. Pasealotia f.
unarea, Piricularia oryzaa
IFO 5279, Sclarotinia
scleotiorum IFO 9395. Ve
nturla prina IFO 6189.
The results are shown in Table 1. Example 2 The molecular weight cutoff of less than 10,000 obtained in Production Example 2 was further fractionated to obtain galactosaminooligo-hexasaccharide. The properties of galactosaminoligo 611f are as follows. 1) Galactosamine hexasaccharide GalN, which is composed of only α1→4 bonds, 4. GalN1-+. GalN,→4Ga
1N,→4GalNt→4GalN (GalN represents α1D-galactopyranosyl group.) 2) Color and shape: Pale yellow amorphous powder.

3) Taste: Has a weak sweet taste.

4) Solubility: Soluble in dilute acids, salt solutions, and water. Poorly soluble in common organic solvents except dimethyl sulfoxide.

5) Indicate the following elemental analysis values. C: 43.90. H: 6.91, N: 8.
54. O: 40.65 Expected molecular formula: C*a
HiaOasNa6) The molecular weight and structural formula are as follows. Molecular weight: 984.6 7) Color reaction: Positive for indole-hydrochloric acid reaction, Elson-Morgan reaction, and Somogyy-Nelson reaction. 8) Optical rotation [α) 5”: +190.2 9) I!! point: Carbonized at 160°C or above. 10) Figure 11 shows the ultraviolet absorption spectrum. 11》’12 @ shows the infrared absorption spectrum The obtained galactosaminoligo-hexasaccharide was added to a potato dextrose agar medium as in Example 1 at a concentration of 0.1%. Contrast classification 10
The antifungal activity of each strain is shown in the table. Table 2

[Brief explanation of drawings]

Figure 1 shows the ultraviolet absorption spectrum of PF-101, Figure 2 shows the infrared absorption spectrum of PF-i 01, and Figure 3 shows the ultraviolet absorption spectrum of PF-102. So, Figure 4 shows the infrared absorption spectrum of PF-102, Figure 5 shows the ultraviolet absorption spectrum of the fraction of the decomposed product of PF-102 with a molecular weight of 50,000 or more, and Figure 6 shows its ultraviolet absorption spectrum. Figure 7 shows the infrared absorption spectrum of the decomposed product of PF-102 with a molecular weight of 10,000 to 50,000, and Figure 8 shows the infrared absorption spectrum. Figure 9 shows the ultraviolet absorption spectrum of the decomposed product of PF-102 with a molecular weight of less than 10,000.
Figure 10 is a diagram showing the infrared absorption spectrum.
Figure 1 shows the ultraviolet absorption spectrum of galactosaminooligo-hexasaccharide, and Figure 12 shows its infrared absorption spectrum. Agent Patent Attorney Toda Chika Male U1 d N Said - Hokai Diagram Wavelength

Claims (4)

[Claims] (1) An antifungal agent containing α-1,4 polygalactosamine, a decomposition product thereof, or a fraction of the decomposition product as an active ingredient. (2) The antifungal 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 antifungal 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 antifungal 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.