JP2002315592A - Mb5747 substance, salt thereof, production method thereof, microbicidal agent including mb5747 substance or salt thereof in the fields of agriculture and horticulture - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a novel substance useful as an active ingredient for bactericidal agents in the fields of agriculture and horticulture and a bactericidal agent for the agricultural and horticultural purposes containing the same. SOLUTION: A microorganism belonging to Nigrospora genus is cultured in a medium including the nutritives available for usual microorganisms and the resultant culture mixture is extracted with a solvent, or subjected to the ion exchange resin or the gel filtration column chromatography to isolate the MB5747 substance in the state of sodium salt represented by the molecular formula of C25 H33 O6 Na or the salt thereof. The MB5747 substance and salt thereof can be expected to be useful as an active ingredient for bactericidal agent for agricultural and horticultural purposes.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The present invention relates to a novel MB574.
The present invention relates to a fungicide for agricultural and horticultural use containing at least one of the seven substances and salts thereof, a production method thereof, and at least one of the MB5747 substance and salts thereof.

[0002]

2. Description of the Related Art Conventionally, various antibiotics produced by microorganisms are known as antibiotics, and these antibiotics have been put to practical use in the fields of pharmaceuticals, cosmetics, veterinary drugs, agricultural chemicals and the like.

In the field of agriculture and horticulture, various fungicides are used for controlling diseases. However, the use of existing chemicals is restricted due to the emergence of resistant bacteria, and there is an important problem in controlling.

[0004]

Accordingly, there is always a need in these fields for the emergence of new substances as active ingredients in fungicides. An object of the present invention is to provide a novel fungicide active ingredient in the agricultural and horticultural field and a novel agricultural and horticultural fungicide containing the same.

[0005]

Means for Solving the Problems The present inventors have produced a substance having an antibacterial activity against phytopathogenic fungi such as cucumber gray mold in a culture of a microorganism belonging to the genus Nigrospora. Thus, the present invention was completed by isolating the active ingredient and determining its physicochemical properties.

That is, the present invention provides an MB5747 substance or a salt thereof having the following physicochemical properties in a sodium salt state, a method for producing the same, and a fungicide for agricultural and horticultural use containing at least one of the MB5747 substance or a salt thereof as an active ingredient. It is. (1) Color and shape: white powder (2) Molecular formula: C ₂₅ H ₃₃ O ₆ Na (3) Specific rotation: [α] _D ²² = + 122.6 ° (c = 0.31, MeO
H) (4) Mass spectrum (TSP-MS): m / z 430 (M) ^- , (FAB-MS): m / z 453 (M + Na) ⁺ , (HR-MS): m / z 453.2251 ( (C ₂₅ H ₃₃ O ₆ Na) (5) UV absorption spectrum: λ _max (MeOH) nm = 257 (6) Red absorption spectrum: ν _max (KBr) cm ⁻¹ = 340
0, 2930, 1650, 1570, 1405, 1255 (7) Solubility: Easily soluble in methanol, poorly soluble in ethyl acetate, chloroform, water, and insoluble in hexane. (8) Basic, acidic, and neutral: basic substance (9) Color reaction: positive for molybdic acid reagent and iodine vapor, negative for ninhydrin (10) Thin-layer chromatography on silica gel: chloroform: methanol: acetic acid ( 10: 1: 0.5)
The Rf value under the developing solvent condition is 0.59. Hereinafter, the present invention will be described in detail.

[0007]

BEST MODE FOR CARRYING OUT THE INVENTION The MB5747 substance or a salt thereof of the present invention has the above-mentioned physicochemical properties in a sodium salt state. The above sodium salt, ie MB5747
The physicochemical properties of the sodium salt of a substance can be measured by known methods. Regarding the physicochemical properties shown above, the state of the sodium salt of MB5747 substance to be measured, the purification conditions, the state of the reagent used for measurement,
The values may slightly vary depending on the conditions of the employed measuring method, measuring environment, used measuring instrument, and the like, but such numerical values are included in the above-mentioned physicochemical properties specified in the present invention as errors relating to measurement.

The MB5747 substance of the present invention can form various salts. Examples of the salt of the MB5747 substance include metal salts such as lithium salt, sodium salt, potassium salt, magnesium salt and calcium salt, and ammonium salts. Methyl ammonium salt, dimethyl ammonium salt,
Examples thereof include ammonium salts such as trimethylammonium salt and dicyclohexylammonium salt.

In order to form a sodium salt, a sodium salt can be formed from a base such as sodium hydroxide or a basic salt such as sodium carbonate and the MB5747 substance. Also, a salt of MB5747 substance and a weak base can be formed like an ammonium salt of MB5747 substance, and a sodium salt can be formed from this salt by a commonly used salt exchange reaction.

According to the present invention, the MB5747 substance or a salt thereof can be obtained by culturing a microorganism capable of producing the MB5747 substance and collecting the compound from the obtained culture.

[0011] Examples of the microorganisms having the ability to produce the MB5747 substance include molds classified into the phylum Incomplete Fungi Omentum Dementia spp. It is a microorganism belonging to the genus Nigrospora of the reticulum Moniliidae, and more preferably Nigrospora oryzae.
A microorganism belonging to the genus, particularly preferably nigrospora
Oryzae (Nigrospora oryzae) strain D7886.

[0012] Nigrospora oryza used for the production of the compound MB5747 of the present invention.
e) The strain D7886 (hereinafter sometimes abbreviated as "strain D7886") is a strain belonging to the order of the Impermatophyte incomplete fungi Omentia moniliria isolated from natural soil. Nigrospora oryzae strain D7886 was deposited on March 22, 2001 with the National Institute of Advanced Industrial Science and Technology, Biotechnology and Industrial Technology Research Institute, and FERM P-1827.
A deposit number of 0 is assigned.

Hereinafter, Nigrospo oryzae (Nigrospo)
ra oryzae) The details about strain D7886 are shown.

[Morphological properties] Colony growth is rapid,
25 ° C. on a potato dextrose agar medium (PDA)
The culture reaches a diameter of 94 mm after 3 days of culture. The colonies are fluffy, light brownish white. The back surface of the colony has a dark olive color, and when the agar medium is used, a soluble pigment is produced in the medium and the agar has an olive color. When cultured on Miura medium agar medium (LCA) at 25 ° C. for 3 days, the colony reaches a diameter of 72 mm, spreads fluffy and thin, and exhibits a colorless to light olive green color. When the above agar medium is used, production of a soluble pigment is not recognized.

[0015] The basal hypha has a dark olive green color. The conidiophore is formed laterally or at the tip of the aerial mycelium, has a pot-like shape, is colorless to pale green, and has a smooth surface. The size of the conidiophore is 10 to 28 × 4 to 9 μm.

[0016] Conidia formation is observed from day 14 of the culture. The mode of conidium formation is aleuro-type, conidia are subglobular to elliptical, unicellular with no septum, blackish brown to black, and smooth in surface. Conidia size is 1
1 to 14 × 9 to 12 μm. No teleomorph was observed.

[Physiological properties] Growth temperature range 10-35 ° C Optimal growth temperature 25 ° C (Miura agar medium, cultured for 3 days) Growth pH range 3-10 Optimal growth pH 6 (Miura liquid medium, 25 ° C, cultured for 3 days) )

[Taxonomic considerations] This strain (D7886 strain)
Is characterized by the following features: 1) forming a subcellular to elliptical unicellular conidia in black, 2) conidia formation mode is aleuro-type, 3) conidia stalk is short-pot-shaped.

Based on the above features, HL Barnett and B.
B. Hunter`s Illustrated generaof imperfect fung
i '(1972) As a result of searching for genera using the search table on pages 35 to 49, the strain was found to be incomplete mycota (Deuteromycota)
Hyphomycetes (Monilial)
es) Dematiumaceae (Dematiaceae) Nigrospora (Nigro
spora). So, MB Ellis
As a result of searching for species using the species search table of the genus of the genus, `Dematiaceous Hyphomycetes', pages 319 to 320, the strain (D7886) was found to be Nigrospora oryzae.
ra oryzae).

Microorganisms which are classified as incomplete fungal phylum nets and have the ability to produce the compound of the present invention can be cultured in a medium containing nutrients such as carbon sources and nitrogen sources which can be used by ordinary microorganisms. can do. Rice, glucose, syrup, dextrin, sucrose, starch, molasses, animal and vegetable oils and the like can be used as the carbon source.

As the nitrogen source, soybean flour, wheat germ, corn steep liquor, cottonseed meal, meat extract, peptone, yeast extract, ammonium sulfate, sodium nitrate, urea and the like can be used. In addition, if necessary, sodium, potassium, calcium, magnesium, cobalt, chlorine,
It may be advantageous to add to the medium inorganic salts capable of forming phosphate, nitric acid and other ions.
In addition, an appropriate organic or inorganic substance that assists the growth of bacteria and promotes the production of the compound of the present invention can be appropriately added to the medium.

The most suitable culture method for the microorganisms is a culture method under aerobic conditions, particularly a solid surface culture method using an agar medium or a rice medium, and a submerged culture method using a liquid medium. .

The temperature suitable for culturing is from 15 to 30 ° C, preferably around 20 to 27 ° C. Although the production of the compound of the present invention varies depending on the culture medium and culture conditions, the solid surface culture method in a flask usually maximizes its accumulation in 10 to 15 days. It is preferable to stop the culture when the accumulation of the compound in the culture is maximized, and to separate and purify the compound from the culture.

[0024] The MB5747 substance obtained by the present invention can be obtained from a culture of a bacterium producing MB5747 substance by using a usual separation means utilizing its properties, for example, a solvent extraction method, an ion exchange resin method, an adsorption or distribution column chromatography method, or a gel method. Extraction and purification can be performed by a filtration method, a dialysis method, a precipitation method, or the like, alone or in an appropriate combination.

For example, the MB5747 substance is extracted from the cultured cells with an organic solvent such as acetone water and methanol water. After distilling off the organic solvent components such as acetone and methanol contained in the extract, the aqueous layer was washed with Diaion H.
Adsorbed on a synthetic adsorbent such as P-20 (manufactured by Mitsubishi Chemical Corporation), washed with a 50% acetone solution, eluted with an 80% acetone solution, and furthermore, pH of an aqueous layer from which an organic solvent component such as acetone was distilled off. Is adjusted to be basic with an alkali or alkaline earth metal salt such as NaOH, or an ammonium salt such as ammonium carbonate, etc., washed with a low-polarity organic solvent such as ethyl acetate, and then with a medium-polarity solvent such as n-butanol. Extract. The sample thus obtained was subjected to silica gel column chromatography and then preparative thin layer chromatography to purify the MB5747 substance as a salt,
Can be taken out.

Further, since the MB5747 substance of the present invention is an acidic substance, metal salts such as lithium salt, sodium salt, potassium salt, magnesium salt and calcium salt, or ammonium salt, methyl ammonium salt, dimethyl ammonium salt, trimethyl Ammonium salts such as ammonium salts and dicyclohexylammonium salts can be formed.

These may be obtained by changing the type of the basic substance used in the above-mentioned purification / extraction method, or by subjecting a salt of MB5747 substance obtained by the above-mentioned purification / extraction method to a salt exchange reaction usually used. Alternatively, another salt may be produced. Alternatively, the MB5747 substance may be obtained as an MB5747 substance by introducing a salt of the substance into an acidic system.

The agricultural and horticultural fungicide of the present invention comprises the novel M
It contains at least one of the B5747 substance or a salt thereof as an active ingredient, and has a remarkable plant disease controlling effect.

The agricultural and horticultural fungicide of the present invention can take any form or use form that can be employed as an agricultural or horticultural medicine, particularly as a fungicide.

Specifically, for example, MB5747 of the present invention
At least one of the substance or a salt thereof, as it is, or diluted with a liquid carrier, a solid carrier, a gas carrier, or another suitable carrier, or used with an auxiliary agent such as a spreading agent if necessary, Alternatively, it is mixed with various liquid, solid or gaseous carriers by a method generally used in the production of agricultural chemicals, and if necessary, aids such as wetting agents, spreading agents, dispersing agents, emulsifiers, fixing agents, lubricants, etc. Hydrating agents, solutions, emulsions,
It can be used in various formulations such as powders, granules, fine granules, and aerosols.

In producing these preparations, as the liquid carrier, a liquid carrier which is a solvent for at least one of the MB5747 substance of the present invention or a salt thereof, or a liquid carrier which can be dispersed or dissolved with an auxiliary agent is used. For example, water, aromatic hydrocarbons, aliphatic hydrocarbons, chlorinated hydrocarbons, alcohols, ethers, esters, ketones, polar solvents such as dimethylformamide, dimethylsulfoxide and the like can be mentioned. Examples of the solid carrier include clay, kaolin, talc, diatomaceous earth, bentonite, calcium carbonate, silica, vermiculite, mineral powders such as white carbon, gravel, wood powder and other organic powders and fine powders, urea, ammonium sulfate, and the like. Used. For example, as the gas carrier, air, carbon dioxide gas, chlorofluorocarbon gas, nitrogen gas or the like is used.

The adjuvants can be used for further improving properties such as dispersion, emulsification, spreadability of the drug, adhesion to plants, absorption and migration, and examples of such adjuvants. As the non-ionic, cationic, anionic, amphoteric surfactants, lignin sulfonic acid or its salt,
Examples include gums, fatty acid salts, and pastes such as methylcellulose.

In the production of the fungicide for agricultural and horticultural use of the present invention, the above-mentioned carriers, auxiliaries and the like can be used alone or in an appropriate combination according to the respective purposes. When the MB5747 substance of the present invention or at least one of its salts is formulated as a fungicide for agricultural and horticultural use, the concentration of the active ingredient is usually 1 to 90% by weight for an emulsion, usually 1 to 90% for a wettable powder, and a flowable agent. Then usually 1-50
% By weight, usually 0.1 to 50% by weight for powder, 0.1 to 50% by weight for granules, 0.1 to 50% by weight for oils, and 0.1 to 30% by weight for aerosols. However, it is desirable for exhibiting effective bactericidal activity and maintaining a stable dosage form. Preferably, the content is 10 to 50% by weight for the emulsion and 10 to 50% by weight for the wettable powder.

The fungicide for agricultural and horticultural use of the present invention can be used by spraying it on the foliage of a plant according to the purpose, and can also be used by applying it to the water surface, water or the soil surface or soil. It can also be used by directly applying to seeds and fruits. In this case, the agricultural and horticultural fungicide of the present invention may contain other additives such as a compatible agricultural and horticultural drug or fertilizer that does not inhibit the efficacy of at least one of the MB5747 substance or a salt thereof. it can. Such agricultural and horticultural agents include, for example, fungicides, insecticides, herbicides, plant growth regulators, and the like.

The application rate of the fungicide for agricultural and horticultural use of the present invention varies depending on the kind and degree of disease of the target disease, the kind and target site of the target crop, the form of application, and the like. For example, 50-500 liters for an emulsion per 10 ares, 50-500 liters for a wettable powder, 2-6 kg for a powder, for example.
A moderate application rate is desirable. In addition, for example, when granules are applied to soil, for example, 1 to 6 k per 10 ares
g application rates are desirable.

Examples of the present invention will be described below.
Since the properties of the 47 substances have been elucidated by the present invention, various methods for producing an MB5747 substance or a salt thereof, and a method for producing an agricultural and horticultural fungicide containing at least one of them as an active ingredient based on those properties have been devised. can do.

Therefore, the present invention is not limited to the following examples. Not only the modifying means of the examples but also the known means based on the properties of the MB5747 material revealed by the present invention are applied to the MB5747 material etc. It encompasses all methods for producing, concentrating, extracting, and refining, and fungicides for agricultural and horticultural use containing an MB5747 substance or the like as an active ingredient.

[0038]

EXAMPLES Example 1 <Example of Preparation of Sodium Salt of MB5747 Substance> Cultivation of strain D7886 Twenty 500-mL Erlenmeyer flasks were prepared, 60 g of rice and 20 mL of tap water were dispensed into each flask, lightly stirred, and then subjected to high-pressure steam sterilization at 121 ° C. for 20 minutes. Three platinum loops of the D7886 strain were inoculated into this, and cultured at 27 ° C. for 14 days.

2. Purification of MB5747 substance produced by cultivation
0 mL of a 50% acetone aqueous solution was added and stirred, and the mixture was left overnight at 4 ° C. This aqueous acetone solution is filtered,
About 2.2 L of the filtrate was obtained as a bacterial cell extract.

The aqueous layer from which the acetone in the cell extract was distilled off was adsorbed on Diaion HP-20 (manufactured by Mitsubishi Chemical Corporation), washed with a 50% acetone solution, and the adsorbed components were eluted with an 80% acetone solution. Was. Further, the pH of the aqueous layer from which acetone was distilled off was adjusted to basic with NaOH, washed with ethyl acetate, and extracted with n-butanol. The sample thus obtained subjected to silica gel (silica gel 60, manufactured by Merck) column chromatography, followed by preparative thin layer chromatography (silica gel 60 F _254, manufactured by Merck), MB57
106 mg of 47 sodium salts were obtained.

The physicochemical properties of the sodium salt of MB5747 thus obtained are shown below. (1) Color and shape: white powder (2) Molecular formula: C ₂₅ H ₃₃ O ₆ Na (3) Specific rotation: [α] _D ²² = + 122.6 ° (c = 0.31, MeO
H) (4) Mass spectrum (TSP-MS): m / z 430 (M) ^- , (FAB-MS): m / z 453 (M + Na) ⁺ , (HR-MS): m / z 453.2251 ( (C ₂₅ H ₃₃ O ₆ Na) (5) UV absorption spectrum: λ _max (MeOH) nm = 257 (6) Red absorption spectrum: ν _max (KBr) cm ⁻¹ = 340
0, 2930, 1650, 1570, 1405, 1255 (7) Solubility: Easily soluble in methanol, poorly soluble in ethyl acetate, chloroform, water, and insoluble in hexane. (8) Basic, acidic, and neutral: basic substance (9) Color reaction: positive for molybdic acid reagent and iodine vapor, negative for ninhydrin (10) Thin-layer silica gel chromatography: chloroform: methanol: acetic acid ( 10: 1: 0.5)
The Rf value under the developing solvent condition is 0.59.

In this example, the above physicochemical properties were measured as follows. (1) The color and shape were judged from the appearance of the sodium salt of the obtained MB5747 substance.

(2) The molecular formula was calculated from the molecular weight of HR-MS.

(3) About the specific rotation, JASCO D
It measured using the IP-370 type polarimeter.

(4) TSP-MS (Th
HEWLE for ermospray mass spectrometry)
Using HP-5989A manufactured by TT PACKERD, F
AB-MS (fast atom bombardment mass spectrometr
y: fast atom collision mass spectrometry)
HR-MS (high resolutionmass) using MS-700
s spectrometry (high-resolution mass spectrometry)
The measurement was performed using JMS-700 manufactured by L.

(5) Regarding the ultraviolet absorption spectrum,
It measured using Shimadzu self-recording spectrophotometer UV-260.
FIG. 1 shows the obtained spectrum.

(6) Regarding the infrared absorption spectrum,
It measured using Shimadzu Fourier transform infrared spectrophotometer FTIR-8100. FIG. 2 shows the obtained spectrum.

(7) Solubility was determined from the amount of solvent required until the sodium salt of MB5747 substance was suspended in various solvents and completely dissolved.

(8) The distinction between basic, acidic and neutral was determined by measuring with a pH meter and pH test paper.

(9) With regard to the color reaction, as the molybdic acid reagent, 12.1 g of Na ₂ MoO ₄ .2H ₂ O,
Of H ₃ PO ₄ 7.5mL (85%), and 25mL of H ₂
Using a reagent prepared by making SO ₄ constant volume to 500 mL with H ₂ O, a color reaction using a molybdic acid reagent was performed by spraying the above reagent on a sample and then heating at 110 ° C. for 15 minutes. The color reaction using iodine vapor was performed by exposing the sample to a container containing iodine vapor for 15 minutes. The color reaction using ninhydrin is
The test was performed by spraying a ninhydrin spray manufactured by Tokyo Kasei on the sample and heating it at 110 ° C. for 15 minutes.

(10) Regarding silica gel thin layer chromatography, Merck S
Ilicagel60F ₂₅₄ used (thickness 0.25 mm), using chloroform, methanol, each of the acid reagent chemicals, those dehydrated by mixing a volume ratio of the as a developing solvent, to measure the Rf value according to a conventional method .

Further, the chemical shift value and the binding constant of 400 MHz ¹ H-NMR (FIG. 3) of sodium salt of MB5747 substance in deuterated methanol, and 100 MHz ¹³ C
The chemical shift values of -NMR (FIG. 4) are shown below. In addition,
The peak of heavy methanol was used as a reference peak.

¹ H-NMR δ (ppm): 7.13 (1H, d, J = 16.1 H
z), 6.54 (1H, s), 6.24 (1H, s), 5.90 (1H, s), 5.73 (1H,
d, J = 16.1 Hz), 4.72 (1H, d, J = 12.3 Hz), 4.66 (1H,
d, J = 12.3 Hz), 4.40 (1H, dd, J = 6.8, 7.1 Hz), 3.84 (1
H, dd, J = 2.2, 8.3 Hz), 3.52 (1H, d, J = 10.5), 2.87 (1
H, brs), 1.93-1.77 (5H, m), 1.56-1.19 (11H, m), 0.
90 (1H, m), 0.76 (3H, d, J = 6.6 Hz)

¹³ C-NMR δ (ppm): 208.5, 175.1, 157.6, 14
2.9,138.9,137.7,133.6,132.4,129.6,121.8,95.0,76.1,
75.1,74.2,44.1,43.1,34.6,32.8,31.6,30.9,23.5,20.8,
20.5,19.9,19.8

The above physicochemical properties were compared with known antibiotics, but no corresponding substance was found. Thus, it was concluded that this antibiotic is novel.

Example 2 <Preparation of preparation> Preparation of Emulsion The sodium salt of MB5747 substance was uniformly mixed with 20% by weight of MB5747 substance itself, 20% by weight of cyclohexanone, 45% by weight of xylene, 12% by weight of polyoxyethylene alkylphenyl ether and 3% by weight of calcium alkylbenzenesulfonate, Upon dissolution, an emulsion was obtained.

2. Preparation of wettable powder The sodium salt of MB5747 substance was uniformly converted into 20% by weight of the MB5747 substance itself, 20% by weight of clay, 55% by weight of diatomaceous earth, 3% by weight of ligninsulfonic acid, and 2% by weight of polyoxyethylene alkyl allyl ether. The wettable powder was obtained by crushing and mixing.

Example 3 <Antibacterial test against phytopathogenic fungi> The sodium salt of substance MB5747 was 1, 10, or 1
On a potato decoction agar medium containing 00 ppm, a mycelium fragment of a phytopathogenic bacterium shown in Table 1 which had been separately cultured in advance was placed. After culturing at 28 ° C. for 5 days, the diameter of the resulting hyphal colony was measured, and the growth inhibition rate represented by the following formula was determined in comparison with a non-treated section containing no sodium salt of the MB5747 substance. Table 1 shows the test results.

[0059]

## EQU1 ## Growth inhibition rate = (1-hyphae colony diameter of treated section / hyphae colony diameter of untreated section) × 100

[0060]

[Table 1]

Example 4 <Efficacy Test> Cucumber gray mold control test After seeding in a plastic pot containing culture soil, 1
A cucumber seedling (variety: four-leaf) that had been raised for about 0 days and had completely developed cotyledons was used as a test plant. The cucumber seedlings were sprayed with a chemical solution prepared at a predetermined concentration at a concentration of 5 mL per 3 pots using a spray gun and air-dried.
mm paper disc was placed. On the other hand, a conidiospore suspension (1 to 1) collected from a petri dish in which the gray mold fungus was cultured in advance and prepared using a potato decoction medium
5 × 10 ⁵ / mL) was dropped on a paper disc placed on cotyledons, 30 μL of which was then allowed to stand in a moist chamber at 21 ° C. to cause disease. Three days after inoculation, the lesion diameter was measured, and the lesion was measured by the following formula. The control value was calculated. Table 2 shows the test results.

[0062]

## EQU2 ## Control value = (1−average lesion diameter in treated section / average lesion diameter in untreated section) × 100

[0063]

[Table 2]

[0064]

Industrial Applicability The MB5747 substance or a salt thereof of the present invention is expected to be used as a fungicide for agricultural and horticultural use.

More specifically, according to the present invention, MB57 which is a novel substance expected to be used as an agricultural and horticultural fungicide
A novel agricultural / horticultural fungicide containing 47 substances or salts thereof, and at least one of them as an active ingredient can be provided.

According to the present invention, a microorganism of the genus Nigrospora that produces the above-mentioned MB5747 substance is cultured, and the MB5747 substance is collected from the culture to obtain MB574.
Seven substances or salts thereof can be produced. In the production of the substance, the use of a microorganism belonging to Nigrospora oryzae is more effective in producing the substance.

[Brief description of the drawings]

FIG. 1 is a drawing showing a UV spectrum of a sodium salt of MB5747 substance in methanol.

FIG. 2 is a drawing showing an IR spectrum of sodium salt of MB5747 substance by a KBr method.

FIG. 3 is a drawing showing a 400 MHz ¹ H-NMR spectrum of sodium salt of MB5747 substance in deuterated methanol.

FIG. 4 is a drawing showing 100 MHz ¹³ C-NMR spectrum of sodium salt of MB5747 substance in deuterated methanol.

 ──────────────────────────────────────────────────続 き Continuing on the front page (72) Inventor Masato Sugiyama 1000-san, Kamoshida-cho, Aoba-ku, Yokohama-shi, Kanagawa Prefecture Inside the Yokohama Research Laboratory (72) Inventor Kazuhiko Kikutake 1000, Kamoshita-cho, Aoba-ku, Yokohama-shi, Kanagawa (No. 72) Inventor Nozomi Nozomi Nakanishi 760, Shioka-cho, Kohoku-ku, Yokohama, Kanagawa Prefecture Meiji Seika Co., Ltd. (72) Inventor Go, Teraoka 760, Shioka-cho, Kohoku-ku, Yokohama, Kanagawa Meiji Seika Pharmaceutical Research Institute Co., Ltd. CE08 CE10 DA02 4H011 AA01 BA01 BB21 BC01 BC05 BC07 BC19 BC20 DA15 DA16 DC01 DC05 DD03 DH03 DH10 4H055 AA02 A A03 AB03 AC60 AD22 AD32 CA40 DA06 DA79 DA82

Claims (5)

[Claims] 1. An MB5747 substance or a salt thereof having the following physicochemical properties in the form of a sodium salt. (1) Color and shape: white powder (2) Molecular formula: C ₂₅ H ₃₃ O ₆ Na (3) Specific rotation: [α] _D ²² = + 122.6 ° (c = 0.31, MeO
H) (4) Mass spectrum (TSP-MS): m / z 430 (M) ^- , (FAB-MS): m / z 453 (M + Na) ⁺ , (HR-MS): m / z 453.2251 ( (C ₂₅ H ₃₃ O ₆ Na) (5) UV absorption spectrum: λ _max (MeOH) nm = 257 (6) Red absorption spectrum: ν _max (KBr) cm ⁻¹ = 340
0, 2930, 1650, 1570, 1405, 1255 (7) Solubility: Easily soluble in methanol, poorly soluble in ethyl acetate, chloroform, water, and insoluble in hexane. (8) Basic, acidic, and neutral: basic substance (9) Color reaction: positive for molybdic acid reagent and iodine vapor, negative for ninhydrin (10) Thin-layer silica gel chromatography: chloroform: methanol: acetic acid ( 10: 1: 0.5)
The Rf value under the developing solvent condition is 0.59. 2. A method for producing an MB5747 substance or a salt thereof, comprising culturing a microorganism belonging to the genus Nigrospora and capable of producing the MB5747 substance according to claim 1, and collecting the MB5747 substance from the culture. . 3. The MB according to claim 2, wherein the microorganism is a microorganism belonging to Nigrospora oryzae.
A method for producing a 5747 substance or a salt thereof. 4. The method for producing an MB5747 substance or a salt thereof according to claim 2, wherein the MB5747 substance produced by the microorganism is taken out of the culture in a salt state. 5. A fungicide for agricultural and horticultural use comprising at least one of the MB5747 substance according to claim 1 or a salt thereof as an active ingredient.