JPH0316115B2 - - Google Patents

Description

[Detailed description of the invention]

[] Summary of the Invention The present invention relates to a new antibiotic K-51A substance, a method for producing the same, and a plant disease control agent containing the same as an active ingredient. More specifically, the new antibiotic K-51A substance obtained by culturing K-51A substance-producing bacteria belonging to the genus Streptomyces in a culture medium, its production method, and containing it as an active ingredient. The present invention relates to a plant disease control agent characterized by: The present inventors discovered that a substance exhibiting control activity against rice sheath blight was produced in a culture of a certain strain belonging to the genus Streptomyces, and isolated the effective substance purely from the culture material. As a result of investigating its properties, it was confirmed that it was a new substance different from known substances, and the present invention was completed by naming this effective substance K-51A substance. The new antibiotic K-51A substance according to the present invention exhibits antibacterial activity against a wide range of bacteria and fungi, and also exhibits control activity against a wide range of agricultural and horticultural crop diseases.
In particular, it has excellent control effects on rice sheath blight, tomato and cucumber seedling blight, and cucumber anthracnose. [] Detailed Description of the Invention 1) Description of the K-51A substance (1) Physical and chemical properties of the K-51A substance 1 Appearance: yellow amorphous powder 2 Melting point: 194-195°C (decomposition) 3 Specific optical rotation: [α ] ²⁴ _D −29.7° (C1, CHCl ₃ :
MeOH=1:1) 4 Ultraviolet absorption spectrum: λ _nax nm (E1% 1cm) [MeOH]: 310 (65) The spectrum is shown in FIG. 5 Infrared absorption spectrum: (KBr tablet method
cm ^-1 ) 3400, 1720, 1700, 1600, 1280,
1060, its spectrum is shown in Figure 2. 6 Nuclear magnetic resonance spectrum (NMR) (400M
Hz, 70% heavy acetone-heavy water mixed solution) δ (ppm): 7.72 (1H, d), 7.68 (1H,
s), 7.08 (1H, m), 6.66 (1H, d), 6.02
(1H, d), 5.34 (1H, d), 5.25 (1H,
m), 5.18 (1H, d), 5.06 (1H, m), 4.78
(1H, d), 4.65 (1H, m), 4.17 (1H,
m), 4.02 (1H, m), 3.96 (1H, m), 3.88
(1H, d), 3.83 (2H, m), 3.74 (1H,
m), 3.67 (1H, d), 3.65 (2H, m), 3.55
(1H, m), 3.54 (3H, s), 3.52 (1H,
m), 3.50 (1H, m), 3.43 (1H, dd), 3.10
(1H, t), 2.90 (3H, s), 2.82 (2H,
m), 2.52 (1H, m), 2.43 (1H, m), 2.31
(2H, m), 2.10~1.40 (25H), 1.34 (3H,
d), 1.26 (3H, d), 1.09 (3H, d), 0.96
(3H, d), 0.95 (3H, t), 0.92 (3H,
d), 0.87 (3H, d), 0.78 (3H, d) The spectra are shown in Figure 3. 7 Molecular weight: 1491 (FAB mass spectrum) 8 Elemental analysis values: carbon 56.54%, hydrogen 8.04%,
Nitrogen 1.05% 9 Color reaction Positive for Remieux, sulfuric acid, vanillin sulfate, negative for ninhydrin, Sakaguchi reaction 10 Rf value on silica gel plate (Merck & Co., Kieselgel) Developing solvent Rf value CHCl ₃ :MeOH:H ₂ O = 65:25 :4 0.21 BuOH:AcOH: _H2O =2:1:1 0.56 BuOH:MeOH: _H2O =4:1:1 0.41 11 Stability When treated at 100℃ for 10 minutes at PH2, 7, and 9, PH7, It was stable at PH 9, but at PH 2
The activity was slightly reduced in the treated area. 12 Solubility in solvents Soluble in dimethyl sulfoxide, aqueous methanol, aqueous ethanol, aqueous n-butanol, and aqueous acetone. Insoluble in water, chloroform, and ethyl acetate. (2) Antibacterial activity 1 Antibacterial test against various microorganisms The antibacterial activity of substance K-51A was investigated by the Agarhole method using the microorganisms shown in the table below as test bacteria. 5 per hole for exam
A solution of K-51A substance with a concentration of mg/ml is added to 100μ
This was done using both. The results are shown in the table below.

[Table] 2 Antibacterial test against plant pathogenic bacteria Using the plant pathogenic bacteria shown in the table below as test bacteria, the presence or absence of mycelial growth on an agar plate was investigated, and the minimum growth inhibitory concentration (MIC) was determined. That is, a dilution series was prepared by mixing the K-51A substance of the present invention into a potato decoction agar medium, and the mixture was poured into a shear dish and solidified to prepare an agar plate. The test bacteria were inoculated onto the agar plate, and after culturing at 25°C for 48 hours for bacteria and 72 hours for filamentous fungi, the presence or absence of growth of the test bacteria was observed. The results are shown in the table below.

[Table] (2) Use as an antibiotic As mentioned above, substance K-51A has antibacterial activity against bacteria and filamentous fungi, so it is used as a drug for the prevention and treatment of infectious diseases caused by these microorganisms. Alternatively, it can be used as a veterinary drug. When the antibiotic K-51A substance of the present invention is used as a medicine for the prevention and treatment of infectious diseases, the dosage and administration route should be determined in consideration of the above-mentioned antibacterial properties and in accordance with the patient's condition. It may be determined as appropriate in the same manner as in the case of antibiotics based on bacteria of the genus. 2) Explanation regarding the method for producing K-51A substance (1) K-51A substance-producing bacteria The K-51A substance-producing bacteria used in the method of the present invention must contain a sufficient amount of K to be collected into the culture. Any strain may be used as long as it has the ability to produce the -51A substance, but an example of such a strain is the strain newly isolated by the present inventors from the soil of Tamaki Village, Shimajiri District, Okinawa Prefecture. There is K-51A strain. The mycological properties of the K-51A strain are as follows. 1. Morphology Aerial hyphae have long main axes, branch irregularly into uniaxial branches, and their tips exhibit a spiral shape (coiled diameter 1.3-1.5 μm, 2-4 turns) or an incomplete spiral shape. At least 10 spore chains are attached, and the surface structure of the spores is smooth and 0.3 to 1.4 μ in diameter.
m, oval-shaped with a length of 0.6-0.7 μm. sclerotia,
Special forms such as flagellated spores and sporangia are not observed. Contains LL type diaminopimelic acid during whole bacterial cell hydrolysis. 2. Growth status on various media The following observation of growth status on various media:
ISP Methods Manual 1941
I followed it. The observation results are summarized in Table 3.

【table】

[Table] 3 Physiological properties and assimilability of carbon sources Physiological properties are shown in Table 4, and assimilability of carbon sources is shown in Table 5. Table 4 Growth temperature range 20-30℃ Optimum growth temperature 20-30℃ Liquefaction of gelatin ± Hydrolysis of starch + Coagulation of skim milk ± Peptonization of skim milk ± Production of melanin-like pigment Tyrosine agar − Peptone yeast iron agar - Tryptone yeast liquid medium - (Note) +: Positive ±: Slightly positive -: Negative Table 5 L-arabinose D-xylose D-glucose D-fructose + Sucrose - Inositol - L-Rhamnose Raffinose - D-Mannitol Pridham Gottlieb agar medium was used (Note): Good growth +: Growth -: No growth To summarize the above results, strain K-51A has LL type diaminopimelic acid and forms 10 or more spore chains. , is included in the genus Strepmyces and is characterized as follows. (1)
Spore chain is spiral, (2) Spore surface structure is smooth, (3)
(4) The color of the underside of the colony is pale yellow to pale yellow-brown, light brown-gray, and light red-orange.
(5) Production of melanin-like pigment was negative. Based on the above-mentioned characteristics, a search using "Mr. Bergier's Bacterial Identification Handbook, 8th edition (1974)" and Mr. Nonomura's search table for ISP-listed species revealed that K.
Strain -51A was closely related to the described species of Streptomyces aminophilus. When strain K-51A was cultured at the same time as the standard strain of Streptomyces aminophilus and comparative observation was made, it was found that Streptomyces
Aminophilus differs from the K-51A strain in that it has good aerial hyphae on various media, exhibits a pale yellow color, and does not exhibit reddish-orange or pink underside color on glycerin-asparagine agar and tyrosine agar. Other properties match well. Based on the above characteristics, strain K-51A was identified as a subspecies of Streptomyces aminophilus subsp.
notonesogenes) 647-AV ₁ (K-51) strain. This strain was submitted to the Microtech Institute as Streptomyces sp. 647-AV ₁ (K-51) strain.
It was entrusted to us on December 20, 1981, and its FERM acceptance number is FERM P-6843. (2) Culture The K-51A strain is susceptible to changes in its properties, as seen in the case of other actinomycetes, and can be mutated by artificial mutagenesis methods using, for example, ultraviolet rays, X-rays, radiation, chemicals, etc. All Streptomyces bacteria capable of producing the K-51A substance, even such mutant strains, can be used in the method of the present invention. In the method of the present invention, the K-51A strain is usually cultured in a medium containing nutrients that can be utilized by the microorganism. For example, glucose, sucrose, dextrin, starch, starch syrup, molasses, vegetable oil, animal oil, etc. can be used as the carbon source. In addition, soybean flour, wheat germ,
Peptone, meat extract, yeast extract, cornstarch liquor, sodium nitrate, ammonium sulfate, etc. may be used. In addition, inorganic salts such as calcium carbonate, potassium chloride, and phosphates may be added as necessary, and K may help the growth of bacteria.
Organic and inorganic substances that promote the production of -51A substances can be added as appropriate. As for the culture method, if it is a culture method under aerobic conditions like the general antibiotic production method,
Although any method may be applied, deep culture is the most suitable. The temperature suitable for culturing is 20 to 35°C, but in most cases it is preferable to culture at around 26 to 32°C. Production of K-51A substance reaches its maximum accumulation in 2 to 7 days in both shaking culture and tank culture. (3) Assay For assaying the K-51A substance, a method is used in which the growth of the bacterial color of the assay bacterium Rhizoctonia solani on a slide glass is observed. That is, a solution containing K-51A substance is placed on a hole slide glass together with Rhizoctonia solani, and the degree of inhibition of hyphal elongation of Rhizoctonia solani is observed. According to this method, K-51A substance is
Even at a concentration of 20 mcg/ml, the mycelial elongation of Rhizoctonia solani can be inhibited. (4) Collection The K-51A substance obtained by the present invention is a neutral fat-soluble substance and has the above-mentioned physical and chemical properties. It can be extracted and purified using its properties. In other words, the K-51A substance accumulated in the culture solution can be extracted with an organic solvent that does not mix with water, such as n-butanol.
The 51A substance is extracted into the organic solvent layer. Also,
It is extracted from the cultured bacterial cells with acetone-water or methanol-water. In order to further purify the K-51A substance, chromatography using an adsorbent such as silica gel or alumina or Sephadex LH20 (Pharmacia) may be performed. By using the methods described above or by appropriately combining these methods, a highly pure K-51A substance can be obtained as a yellow amorphous powder. 3) Description of the K-51A substance as a plant disease control agent The K-51A substance according to the present invention has a high control effect on plant diseases, so it can be used as a plant disease control agent such as an agricultural and horticultural fungicide using the K-51A substance. It is possible to use it as The plant disease control agent according to the present invention can be in any form or usage mode that can be adopted as an agricultural and horticultural agent, especially a fungicide, except that the active ingredient is the above-mentioned K-51A substance. can be taken. Specifically, for example, the K-51A substance of the present invention may be used as it is, or it may be diluted with water, solid powder, or other suitable carrier, and if necessary, an auxiliary agent such as a spreading agent may be added. Alternatively, various liquid or solid carriers may be mixed using methods commonly used in agricultural chemical manufacturing, and if necessary, wetting agents, spreading agents, dispersing agents, emulsifiers, fixing agents, lubricants, etc. may be added. By adding adjuvants, it can be used in various formulations such as wettable powders, solutions, emulsions, powders, granules, and fine granules. In producing these preparations, the liquid carrier used is one that serves as a solvent for the K-51A substance of the present invention, or one that can be dispersed or dissolved with the aid of an auxiliary agent. For example, water
Aromatic hydrocarbons, aliphatic hydrocarbons, alcohols, esters, ketones, highly polar dimethylformamide, dimethyl sulfoxide, etc. Solid carriers include clay, kaolin, talc, diatomaceous earth, bentonite, calcium carbonate,
Mineral powders such as silica, sand and gravel, wood flour and other organic powders and granules can be used. Examples of auxiliary agents include nonionic, anionic, cationic, and amphoteric surfactants, ligninsulfonic acid or its salts, gums, fatty acid salts, and thickeners such as methylcellulose. The plant disease control agent of the present invention can be used by spraying on the foliage of crops, and can also be applied to water surfaces, water, soil surfaces, and soil. In this case, compatible agricultural and horticultural chemicals or fertilizers may be used. Such agricultural and horticultural chemicals include, for example, fungicides, insecticides, herbicides, and plant growth regulators. When using the plant disease control agent of the present invention as a liquid, the K-51A of the present invention is usually added to the spray solution.
It is desirable that the substance be contained in a concentration of 10 to 1000 ppm (in the case of concentrated small-volume spraying, aircraft spraying, etc., a more concentrated spray liquid can be used as necessary), and powders, granules,
0.1 to 30% when used as fine granules, etc.
It is desirable that it be included. The amount of application varies depending on the type and severity of the target disease, the type of target crop, the mode of application, etc., but for example, when applying to soil, if the hydrating agent (20% active ingredient) is applied per 10 ares. 50～
20 liters, for example 50 to 200 liters for aqueous solution (10% active ingredient), granules (5% active ingredient)
For example, 2 to 6 kg, powder (active ingredient 2)
%), an application amount of, for example, about 2 to 6 kg is generally appropriate. 4) Explanation regarding Experimental Examples It goes without saying that the present invention is not limited to the following examples, and that many modifications and modification means not exemplified here can be adopted. (1) Production of K-51A substance Streptomyces sp. 647-AV ₁
(K-51) stock (FERM accession number FERM P-
6843) spores with 1% starch and 3% soybean flour.
(PH7) liquid medium 1 (500ml Erlenmeyer flask)
The seeds were inoculated into 10 seedlings (used) and cultured with shaking at 28°C for 43 hours. Liquid medium 35 consisting of 1.5% glucose, 1.0% glycerin, 1.0% meat extract, 1.0% polypeptone, 0.4% calcium carbonate (PH7.2)
The above-mentioned seed mother was inoculated and cultured with aeration and stirring at 28°C for 76 hours (50 jar fermenter). A filter aid (Hyflo Super Cell) was added to this culture and filtered, resulting in a culture solution of 16
Extracted twice with half the amount of n-butanol,
A brown syrup was obtained by concentrating under reduced pressure. Add 300ml of methanol to this syrup to remove insoluble matter, and then add 50ml of this methanol solution.
Cefdex filled with methanol
It was placed on a LH20 (500 ml) column and developed with methanol for chromatography.
It was fractionated into 10 g fractions, Nos. 14 to 25 having activity against Zoctonia solani were collected, and concentrated under reduced pressure to obtain 465 mg of light brown powder with a purity of 10 to 15%. This operation was repeated to obtain about 2 g of powder. Silica gel (2) where 1.7g of this powder was sprinkled on silica gel and filled with chloroform.
After washing with chloroform:methanol (5:1) 2, chromatography was performed using chloroform:methanol (2:1) as a developing solvent. Nos. 120 to 130 having activity against Rhizoctonia solani were collected in 15 g fractions and concentrated under reduced pressure to obtain 400 mg of yellow powder with a purity of about 70%. Toyo Pearl was prepared by dissolving 350mg of this yellow powder in a small amount of methanol and filling it with methanol.
Chromatography was performed by placing it on a HW-40f (1.5) column and developing with methanol. When fractionated in 10 g fractions, fractions No. 67 to 74 showed a single spot of K-51A substance in silica gel thin layer chromatography (chloroform:methanol:water=65:25:4). This fraction was concentrated to dryness to obtain 135 mg of yellow powder of substance K-51A. (2) Formulation example 1 (Wettable powder) Part by weight K-51A substance 20 Clay 10 Diatomaceous earth 65 Lignosulfonic acid 3 Polyoxyethylene alkyl allyl ether 2 It is effective if the above components are uniformly ground and mixed. A hydrating agent containing 20% of the ingredients is obtained. Formulation Example 2 (Granules) Parts by Weight K-51A Substance 5 Clay 92 Carboxymethyl Cellulose 3 If the above ingredients are mixed with the above ingredients, an appropriate amount of water is added, kneaded into a mold, and then dried, the product contains 5% of the active ingredient. Obtain granules. Formulation example 3 (Powder) Part by weight K-51A substance 2 Calcium stearate 1 Silicic anhydride powder 1 Clay 48 Talc 48 If the above ingredients are uniformly ground, the active ingredient will be 2%
Obtain a powder containing. (3) Medicinal efficacy test test example 1 (rice sheath blight control effect test) The above formulation example 1 was applied to paddy rice (cultivar "Jukoku") at the heading stage grown in a 1/5000 are Wagner pot.
The wettable powder produced by
cm ² ) at a rate of 70 ml/3 pots. Immediately after air-drying, the sheath blight bacteria obtained by plating on a peptone-added potato decoction agar medium for 48 hours were grown.
Inoculation was performed by punching out a 0.5 cm cork borer and inserting a piece of mycelium-containing agar into the center of the plant, 15 cm from the ground surface. After inoculation, each pot was covered with a vinyl cylinder to encourage the invasion of the sheath blight fungus and left in a glass greenhouse at 30°C during the day and 24°C at night to develop the disease. Ten days after inoculation, the infected stems were examined. The lesion length was measured and the control value was calculated according to the following formula. At the same time, the occurrence of drug damage was also observed. Control value = (1 - average lesion length in treated area/average lesion length in untreated area) x 100 The results were as shown in the table below.

[Table] Test Example 2 (Cucumber seedling damping-off control test) The cucumber seedling damping-off bacterium was cultured on a potato decoction agar medium and mixed and ground with three times the amount of rice bran to prepare an inoculum. 1/50 using cucumber (variety "Tokiwa Jiho") as a joint trial crop and filling with sterilized field soil
20 germinated seeds per pot were sown in a 00 are Wagner pot and covered with soil, and a mixture of equal amounts of the above inoculum and sterilized field soil was uniformly spread over the pot for inoculation. After inoculation, store in a constant temperature room at 28℃ for 48 hours.
After standing for a period of time, the wettable powder produced according to Formulation Example 1 was adjusted to a specified concentration and used as an irrigation solution.
100 ml of this chemical solution was poured uniformly onto the ground surface using a pipette. Thereafter, in order to facilitate the progress of invasion of the inoculum, the plants were transported to a glass greenhouse at a temperature of 30°C to 28°C, and the soil in the pot was kept in a slightly dry state to develop the disease. The test was conducted using a 3-pot system for each section. In the investigation, the number of germination and the number of healthy seedlings up to 3 weeks after sowing were investigated, and the germination rate relative to the number of sown grains and the percentage of healthy seedlings relative to the number of germination were calculated. The results were as shown in the table below.

【table】

[Table] Test Example 3 (Test on the effect of controlling cucumber anthracnose disease) Produced according to Formulation Example 1 using cucumber seedlings (cultivar "Tokiwa Jigo") at the second true leaf development stage grown in three seedlings in a No. 3 clay pot. Prepare a chemical solution of the wettable powder to a specified concentration, spray 40 ml per 3 pots using a spray gun, air dry, enter a moist room at 24°C, and collect conidia of the melon anthracnose fungus. The suspension was uniformly sprayed for inoculation, kept in a humid room overnight, and then transferred to an artificial climate room to induce disease. The degree of disease onset was determined 5 days after vaccination.
0 for those without any disease, 1 or more lesions per leaf
5 items have an index of 1, 6 to 15 items have an index of 2, 16 to 30 items have an index of 3, 31 to 50 items have an index of 4, and 51 or more items have an index of 4. An index of 5 was given to each species, and the pesticidal value was calculated using the following formula. Control value (%) = (1 - average disease index of sprayed plots/average disease index of non-sprayed plots) ×100 The results were as shown in the table below.

【table】 [Brief explanation of drawings]

Figure 1 is the ultraviolet absorption spectrum of the K-51A substance of the present invention, Figure 2 is the infrared absorption spectrum of the K-51A substance (potassium bromide tablet method), and Figure 3 is the K-51A substance.
Proton NMR of substance 51A (70% heavy acetone in heavy water).

Claims (1)

[Claims] 1. A new antibiotic K- having the following physical and chemical properties.
51A substance. (a) Elemental analysis values Carbon: 56.54%, Hydrogen: 8.04%,
Nitrogen 1.05% (b) Molecular weight (by FAB mass spectrum) 1491 (c) Ultraviolet absorption spectrum λ _nax nm (E1% 1cm) [MeOH]: 310 (65) (d) Infrared absorption spectrum (KBr tablet method cm
^-1 ) 3400, 1720, 1700, 1600, 1280, 1060 (e) Nuclear magnetic resonance spectrum (400MHz, 70% heavy acetone-heavy water mixed solution) δ (ppm): 7.72 (1H, d), 7.68 (1H, s ),
7.08 (1H, m), 6.66 (1H, d), 6.02 (1H, d),
5.34 (1H, d), 5.25 (1H, m), 5.18 (1H, d),
5.06 (1H, m), 4.78 (1H, d), 4.65 (1H, m),
4.17 (1H, m), 4.02 (1H, m), 3.96 (1H, m),
3.88 (1H, d), 3.83 (2H, m), 3.74 (1H, m),
3.67 (1H, d), 3.65 (2H, m), 3.55 (1H, m),
3.54 (3H, s), 3.52 (1H, m), 3.50 (1H, m),
3.43 (1H, dd), 3.10 (1H, t), 2.90 (3H,
s), 2.82 (2H, m), 2.52 (1H, m), 2.43 (1H,
m), 2.31 (2H, m), 2.10-1.40 (25H), 1.34
(3H, d), 1.26 (3H, d), 1.09 (3H, d),
0.96 (3H, d), 0.95 (3H, t), 0.92 (3H, d),
0.87 (3H, d), 0.78 (3H, d) (F) Melting point 194-195℃ (decomposition) (G) Fat-soluble (H) Neutral (L) Yellow amorphous powder (N) Solubility Dimethyl sulfoxide, water content methanol, aqueous ethanol, aqueous n-butanol,
Soluble in aqueous acetone, insoluble in water, chloroform, and ethyl acetate (R) Color reaction Positive for Remieux, sulfuric acid, and vanillin sulfuric acid reactions, negative for ninhydrin and Sakaguchi reactions (W) Rf value Developing solvent in silica gel thin layer chromatography Chloroform-methanol-water (65:
25:4) and 0.21; n-butanol-
0.56; n when developed with acetic acid-water (2:1:1)
-Butanol-methanol-water (4:1:1)
When expanded with , it is 0.41. 2. A new antibiotic K-51A substance that belongs to the genus Streptomyces and has the following physical and chemical properties. (a) Elemental analysis values Carbon: 56.54%, Hydrogen: 8.04%,
Nitrogen 1.05% (b) Molecular weight (by FAB mass spectrum) 1491 (c) Ultraviolet absorption spectrum λ _nax nm (E1% 1cm) [MeOH]: 310 (65) (d) Infrared absorption spectrum (KBr tablet method cm
^-1 ) 3400, 1720, 1700, 1600, 1280, 1060 (e) Nuclear magnetic resonance spectrum (400MHz, 70% heavy acetone-heavy water mixed solution) δ (ppm): 7.72 (1H, d), 7.68 (1H, s ),
7.08 (1H, m), 6.66 (1H, d), 6.02 (1H, d),
5.34 (1H, d), 5.25 (1H, m), 5.18 (1H, d),
5.06 (1H, m), 4.78 (1H, d), 4.65 (1H, m),
4.17 (1H, m), 4.02 (1H, m), 3.96 (1H, m),
3.88 (1H, d), 3.83 (2H, m), 3.74 (1H, m),
3.67 (1H, d), 3.65 (2H, m), 3.55 (1H, m),
3.54 (3H, s), 3.52 (1H, m), 3.50 (1H, m),
3.43 (1H, dd), 3.10 (1H, t), 2.90 (3H,
s), 2.82 (2H, m), 2.52 (1H, m), 2.43 (1H,
m), 2.31 (2H, m), 2.10-1.40 (25H), 1.34
(3H, d), 1.26 (3H, d), 1.09 (3H, d),
0.96 (3H, d), 0.95 (3H, t), 0.92 (3H, d),
0.87 (3H, d), 0.78 (3H, d) (F) Melting point 194-195℃ (decomposition) (G) Fat-soluble (H) Neutral (L) Yellow amorphous powder (N) Solubility Dimethyl sulfoxide, water content methanol, aqueous ethanol, aqueous n-butanol,
Soluble in aqueous acetone, insoluble in water, chloroform, and ethyl acetate (R) Color reaction Positive for Remieux, sulfuric acid, and vanillin sulfuric acid reactions, negative for ninhydrin and Sakaguchi reactions (W) Rf value Developing solvent in silica gel thin layer chromatography Chloroform-methanol-water (65:
25:4) and 0.21; n-butanol-
0.56; n when developed with acetic acid-water (2:1:1)
-Butanol-methanol-water (4:1:1)
1. A method for producing a new antibiotic K-51A substance, which comprises culturing a microorganism capable of producing 0.41 when expanded as follows, and collecting the K-51A substance from the culture. 3 Belongs to the genus Streptomyces and is a new antibiotic K-
51A The ability of the microorganism to produce the substance Streptomyces sp. 647-AV ₁ (K-51) (FERM P
-6843) The method according to claim 2. 4 New antibiotic K- with the following physical and chemical properties
51A substance (a) Elemental analysis value Carbon: 56.54%, Hydrogen: 8.04%,
Nitrogen 1.05% (b) Molecular weight (by FAB mass spectrum) 1491 (c) Ultraviolet absorption spectrum λ _nax nm (E1% 1cm) [MeOH]: 310 (65) (d) Infrared absorption spectrum (KBr tablet method cm
^-1 ) 3400, 1720, 1700, 1600, 1280, 1060 (e) Nuclear magnetic resonance spectrum (400MHz, 70% heavy acetone-heavy water mixed solution) δ (ppm): 7.72 (1H, d), 7.68 (1H, s ),
7.08 (1H, m), 6.66 (1H, d), 6.02 (1H, d),
5.34 (1H, d), 5.25 (1H, m), 5.18 (1H, d),
5.06 (1H, m), 4.78 (1H, d), 4.65 (1H, m),
4.17 (1H, m), 4.02 (1H, m), 3.96 (1H, m),
3.88 (1H, d), 3.83 (2H, m), 3.74 (1H, m),
3.67 (1H, d), 3.65 (2H, m), 3.55 (1H, m),
3.54 (3H, s), 3.52 (1H, m), 3.50 (1H, m),
3.43 (1H, dd), 3.10 (1H, t), 2.90 (3H,
s), 2.82 (2H, m), 2.52 (1H, m), 2.43 (1H,
m), 2.31 (2H, m), 2.10-1.40 (25H), 1.34
(3H, d), 1.26 (3H, d), 1.09 (3H, d),
0.96 (3H, d), 0.95 (3H, t), 0.92 (3H, d),
0.87 (3H, d), 0.78 (3H, d) (F) Melting point 194-195℃ (decomposition) (G) Fat-soluble (H) Neutral (L) Yellow amorphous powder (N) Solubility Dimethyl sulfoxide, water content methanol, aqueous ethanol, aqueous n-butanol,
Soluble in aqueous acetone, insoluble in water, chloroform, and ethyl acetate (R) Color reaction Positive for Remieux, sulfuric acid, and vanillin sulfuric acid reactions, negative for ninhydrin and Sakaguchi reactions (W) Rf value Developing solvent in silica gel thin layer chromatography Chloroform-methanol-water (65:
25:4) and 0.21; n-butanol-
0.56; n when developed with acetic acid-water (2:1:1)
-Butanol-methanol-water (4:1:1)
A plant disease control agent containing 0.41 as an active ingredient. 5. The plant disease control agent according to claim 4, which is an agricultural and horticultural fungicide.