JPS6053597B2 - New antibiotic N-461 substance, its manufacturing method, and agricultural and horticultural fungicides containing it as an active ingredient - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a new antibiotic substance N-461, its production method, and an agricultural and horticultural fungicide containing the substance as an active ingredient.

The present inventors have discovered that a newly isolated microorganism belonging to the genus Streptomyces produces and accumulates in its culture a substance that has antibacterial activity and is effective against various diseases of plants such as vegetables and fruit trees. , as a result of isolating and analyzing this substance,
It was confirmed that it is a new substance.

This new substance is called N-461 substance. The new antibiotic N-461 substance is produced by culturing Streptomyces bacteria that have the ability to produce N-461 substance, and then producing N-461 from the culture.
It can be manufactured by collecting 461 substances.

Microorganisms used in the production of N-461 substances include N-461.
All microorganisms belonging to the genus Streptomyces can be used as long as they have the ability to produce the -461 substance, but for example, Streptomyces SP. N
Bacterium No.-461 (Feikoken Bacteria No. 4513) is particularly preferred.

Furthermore, mutant strains obtained from this bacterium by artificial mutagenesis using ultraviolet rays, X-rays, radiation, chemical agents, etc. can also be used. The actinomycete N-461 strain used in the present invention has true vegetative hyphae and aerial hyphae, produces long chains of conidia (conidia) on conidiophores (sporophytes), and produces sporangia and zoospores. It is aerobic and mesophilic, and LL-dianomipimelic acid and glycine were characteristically detected in the hydrolyzate of whole cells, and based on the diameter of hyphae and conidia, it is classified as a member of the order Actinobacteria, family Streptomycetes, and family Streptomycetes. It is a strain belonging to the genus.

The mycological properties of strain N-461 are shown below. (D Morphological properties: The vegetative hyphae exhibit the usual characteristics of the genus Streptomyces, are well developed and branched, but usually do not produce septa and do not produce spores.

The diameter of vegetative hyphae is approximately 0.6 to 0.87 TLP,
It does not exhibit zigzag branching. In addition, in a liquid medium such as tryptone yeast extract medium, 28'Cl2
Even if it is left to stand for a week, the hyphae will not divide into rod-like or cocci-like forms and will not make the medium cloudy. Aerial mycelium grows well on, for example, sucrose/nitrate agar, glycerin/asparagine agar, tyrosine agar 5 agar, yeast/malt agar, etc., and has a gray powder-like appearance, with a wavy main axis under the microscope. It is flexible, usually has alternating branches, no true whorls, and uniaxial branching. The tips of the conidiophores form 11 long chains of spores as they mature, typically in the form of a spiral with 5 to 6 turns, with an average diameter of about 2.6 TrL, μ. In addition to the typical spiral shape, it may take on a wavy shape, an incomplete spiral shape, a hook shape, an annular shape, etc., or it may simply remain in a straight shape.
The shape of the conidia is oval to oblong, forming long chains of one or more caps, often 20 to 3 or more caps, and according to scanning electron micrographs, the size is 0.4 to 0.6 × 0.7
It is about 1.3μ. The surface of conidia is smooth or slightly rough. No sclerotia, sporangia or zoospores are observed.
(Properties of co-cultivation: Experimental methods are described in the paper by E.B. Scherling et al. (International Journal of Systematics)
Bacteriology, 162. Volume, pp. 313-340, 1
96), and in addition, known culture media and experimental methods were used in combination.

The color tone was determined under the standard light source of a xenon lamp, using the Color Harmony Manual, 4th edition (Container Corporation 3 Yon of Chicago, USA, 2019) as the standard color chart. If a color chart was available, the common name was indicated first, and if it was repeated, the common name was omitted. Next, I also wrote the color chart code from the Color Harmony Manual in parentheses. The following is the state of growth in each medium after culture at 28° C. for 3 weeks, unless otherwise specified. (1) Growth on sucrose/nitrate agar medium (Difco): Vigorous growth with raised ridges, color tone unknown due to strong soluble pigment (black) (first week of culture: head-like yellowish brown (21 g).

Back: Unknown due to dark soluble pigment (black).

Aerial mycelium: Powder-like, vigorously epiphytic.

Beaver color (411).

Soluble pigment: dark brown or black.

Others: Soluble dyes do not change color due to pH.

). ) Growth on glycose-asparagine agar medium: Moderate growth with small colonies.

Bamboo color: dull yellow; Straw color: wheat color).

Back: 2fd0 Aerial mycelium: good powdery, natural color (3dc) to silvery gray (3fe).

Soluble dye: Not produced.

3) Glycerol-asparagine agar medium (ISP-5
, Difco) Growth: Good, raised growth that does not spread much.

The color cannot be observed because it is a dark brown soluble pigment. In the first week (21 g), it gradually becomes blackish brown due to the soluble pigment.

Back: Unknown due to soluble dye.

Aerial mycelium: powder-like, adheres well, 3fe or soluble pigment:
Dark brown.

(4) Growth on starch agar medium (ISP-4, Difco): Good growth with slightly spreading and ridges.

Camel yellow, maple brown, tan (31e) to sepia brown, seal brown, dark brown, coffee brown (3pe).

Back side: 31e to 3pn (depending on soluble dye).

Aerial mycelium: Adheres well in powder form, dark gray (2fe to 3f)
e). Soluble pigment: blackish brown. Others: Starch is hydrolyzed.

(5) Growth on tyrosine agar medium (ISP-7, Difco): Good growth with raised ridges that do not spread too much, chocolate brown, dark brown (4pn) color tone is due to soluble pigment.

Back: Unknown due to soluble dye.

Aerial mycelium: very well attached, powdery, ? .

Soluble pigment: Dark brown (positive for melanin production).

(6) Nutrient agar medium (Difco) Growth: Moderate growth, spreading and slightly raised.

Ivory color (2db). Back side: 2db.

Aerial mycelium: Adheres well in powder form, ? .

Soluble pigment: dark yellowish brown.

(7) Yeast/barley agar medium (■SP-2, Difco) Growth: Uptight and slightly spreading, vigorous growth, color cannot be determined due to soluble pigment.

Back: Dark brown due to soluble pigment.

Aerial mycelium: powdery and vigorously epiphytic (3fe).

Soluble pigment: blackish brown. (8) Oatmeal agar medium (I
SP-8) Growth: Moderate growth that is raised and does not spread (
2dp).

Back: 2db0 Aerial mycelium: powdery, well attached (2fb).

Soluble dye: yellow (does not change color depending on pH). (■Physical and biochemical properties (1) Growth temperature range (yeast/malt agar medium, pH 7. Temperature gradient device, results on day 10 of culture) Suitable temperature: around 26°C.

Non-growth temperature: Will not grow above 40°C and below 17°C.

(2) Liquefaction of glucose ● peptone ● gelatin medium: Liquefy.

(3) Hydrolysis of starch: Hydrolyze.

(4) Coagulation and peptonization of skim milk: Peptonization.
It does not grow at 37°C.

(5) Melanin production: Positive in tryptone yeast extract liquid medium (ISP-1), peptone yeast iron agar medium (ISP-6), and tyrosine agar medium (ISP-7).

(■) Assimilation of carbon sources (indicated by growth after 2 weeks of culture on Pridham-Gosesorb agar medium, 28°C) Positive utilization: L-arabinose, D-xylose, D-glucose, D-fructose, Use of sucrose, L-rhamnose, D-mannitol Negative: inosyte, raffinose (■) Salt tolerance: Bennett's agar medium, +0, 4, 7, 10 or 13% salt addition, 28°C, results after 2 weeks of culture, It grows up to 7%, but not at 10%.

To summarize the above mycological properties, Streptomyces N.
In strain No. -461, the tip of the conidiophore is spiral and consists of a chain of w or more long conidia, the surface of the spore is smooth or slightly rough, and the color of the aerial mycelium is grayish.

The color of vegetative hyphae is yellowish when melanin is not produced, and is not a pH-indicating color. It strongly produces melanin in various synthetic and natural media. It shows extremely good growth on sucrose/nitrate agar medium (Tsapek medium). Species similar to the above-mentioned mycological properties from known literature include Streptomyces Diastatochromogenes (Kleinski) Waxman and Henrich, 19
4 & instep can be raised.

There is no type strain for Strepfutomyces diastatochromogenes, and the unofficially proposed new type strain is ATCCl.
23O9 (IFO3337 and 13389, CBS69
O. 72, RIAl35O, ISP5449).

Its mycological properties were described in a report by E.B. Scherling and D.G.Sorb (Intersional Systematic Bacteriology, Zamaki, No. 4, 290-291)
When compared with strain N-461, the morphology of spore chains, the surface of spores, the color tone of aerial hyphae, and the ability to form melanin are in good agreement. The ISP54 phase strain differs from strain N-461 in that it does not produce melanin-like pigments on glycerol/asparagine agar and yeast/malt agar, and uses inosyte and raffinose. Streptomyces huaeochromogenes varaetus icarganensis sakai, the producing bacterium of icargamycin whose chemical structure is very similar to the new antibiotic N-461 substance, 197 reviews (Journal of Antibiotics, Vol. 25, 5) No., pp. 271-280, 19n 5
When compared with strain N-461, the morphology of aerial mycelia, spore surface, melanin production ability in synthetic and natural media, and color tone of aerial mycelium are in good agreement. It grows poorly on nitrate agar) and tyrosine agar, and does not form melanin on the latter medium, does not form aerial mycelium or melanin at starch cold temperature, and only slightly grows on fructose, xylose, arabinose, and sucrose. They are clearly different in that they do not grow.As mentioned above, strain N-461 is different from both Streptomyces diastatochromogenes and Streptomyces huaeochromogenes Varaetus icarganensis. S.A. Waxman's book: The Actinomycetes, Volume 2, 2
01-202, 1961, (The Williams and Wilkins Kamper, Baltimore) and Ralph Hützter: Distemateik Streptomitzeren, pp. 12-14, 1967, (S. Kagel, Basel). When referring to the search table, it was found that it was very close to Stretoptomyces diastatochromogenes and was identified as a new subspecies thereof.

Cultivation can be carried out by a known method for culturing actinomycetes or a modified method thereof, but from an industrial perspective, it is preferable to culture with aeration in a fermenter.

The culture temperature can be changed as appropriate within a range where microorganisms can grow and produce the N-461 substance, but 25 to 30°C is generally preferred. As the culture medium, the usual raw materials used for culturing actinomycetes are used, that is, various carbon sources, nitrogen sources, inorganic salts, and optionally growth promoters or growth inhibitors, antifoaming agents, etc. are appropriately combined. Can be used.

As the medium carbon source, assimilable carbon compounds such as glucose, maltose, lactose, sucrose, starch, dextrin,
Glycerin, molasses, soybean oil, linseed oil, etc. can be used.

As a nitrogen source, available nitrogen compounds or those containing the same, such as soybean flour, corn steep liquor, cottonseed flour, peptone, meat processing scratches, yeast processing scratches, yeast, chilitin hydrolysates, ammonium salts, Nitrates and the like can be used. Other inorganic salts such as phosphates, potassium salts, sodium salts, calcium salts, magnesium salts, iron salts, manganese salts, cobalt salts, etc. may also be used as necessary. Although the culture time varies depending on various conditions, the amount of N-461 substance produced and accumulated usually reaches its maximum within about 48 to 1201 days.

Most of the N-461 substance is usually contained in the culture P solution and the rest in the bacterial cells, but depending on the culture method, it may be contained in quite large amounts in the bacterial cells. N-46 from culture
Isolation and purification of one substance can be carried out by appropriately selecting and combining known means.

For example, you can operate as follows. Since the N-461 substance is soluble in ethyl acetate and other organic solvents under acidic conditions, N-461 can be dissolved by mixing the culture well with these solvents.
-461 substance can be transferred to a solvent and extracted. Alternatively, it is also possible to extract the N-461 substance from the solution from which the bacterial cells have been separated by overnight aging, etc., using the above-mentioned solvent, and then extract the N-461 substance from the bacterial cells using a hydrophilic solvent such as acetone. In the latter case, the extract from the bacterial cells is concentrated under reduced pressure to form a concentrated aqueous solution, which is then cultured. It can be extracted using ethyl acetate or the like in an acidic state in the same way as the liquid, and the extracts are combined. When the extract thus obtained is made alkaline, water is added, and the mixture is shaken and stirred, the N-461 substance is transferred to the aqueous layer. When the aqueous layer is made acidic again and extracted with chloroform, ethyl acetate, etc., the N-461 substance is transferred to the organic solvent layer. After washing the organic solvent layer with water, the solvent is evaporated under reduced pressure. The N-461 substance is precipitated by adding a solvent such as n-hexane that does not dissolve the N-461 substance to the resulting syrup, and the precipitate is filtered and dried under reduced pressure to form a coarse powder of the N-461 substance. is obtained. Crude N-461 material can be purified preferably by column chromatography.

The crude powder is dissolved in a small amount of chloroform, and column chromatography is performed using a column packed with silica gel. In this case, it is effective to use chloroform and a chloroform-methanol mixture in sequence. The active fractions are collected and evaporated to dryness under reduced pressure, the residue is dissolved in hot ethanol, and the solution is left to cool to precipitate crude crystals of the target substance. When this is collected and recrystallized, pure crystals of N-461 are obtained. Preferred recrystallization solvents are alcohols such as methanol or ethanol. The properties of the N-461 substance thus obtained are as follows.

(1) Acidic substance in the form of colorless needle-like crystals (crystallized from ethanol).

(2) From the melting point of 186°C, it begins to turn brown and does not show a clear melting point.

7(3) Molecular formula (molecular weight) C3OFl4ON2O6 (5
24.64) (4) Elemental analysis value') (5) Solubility Easily soluble: Chloroform, pyridine, alkaline water Slightly soluble:
Ethyl acetate, acetone, methanol, ethanol Insoluble: ether, benzene, n-hexane, water (6) Thermal stability and ultraviolet stability (a) 1 in 50% methanol water with pH 4-11
Stable for 5 minutes at 00°C, but slightly unstable at pH 4 or lower.

(b) In the state of 50% methanol water with pH 2 to 11, 4
Stable for 5 hours at 0°C.

(c) Chemical lamp (Toshiba GLl5 type germicidal lamp) 45
) When ultraviolet rays were irradiated from a distance of Cm, almost no decrease in antibacterial activity was observed even after 4 hours of continuous irradiation.

(7) Positive color reaction: Remieux reaction, 2●4-dinitropherhydrazine reaction Negative: ninhydrin reaction, Moritzhu reaction, Anthrone reaction (8) Thin layer chromatography thin layer: Merck Kieselgel GF2,l detection: 10%
Sulfuric acid water (9) Specific rotation [α]? +196 (C1, chloroform) (1Cj/
The ultraviolet absorption spectrum W absorption spectrum is shown in FIG.

In this figure, curve a is methanol and curve b is 0.1N.
-NCl/methanol, curve c is 0.1N-NaOH/
This is a value measured using methanol.

The maximum absorption wavelength 2 is as follows. K. K? H32
5Tnμ(E(?500)230~2407T1,μ(
S)K. K Shou-HOllCH3OH325TrLμ(E
)? 594) 220mμ(E(?918)
1λ Akira-NaOHlOH3C'H32Omμ(
E(?567)220~240TrL,μ(S)(11
) Infrared absorption spectrum The IR absorption spectrum measured by the KBr tablet method is shown in FIG.

The characteristic absorption is as follows. 308012 to 338
95 to 290 287011705, 1640, 158
to 1435, 1370s134 to 1300, 1250,
1240, 1170, 1140, 1090, 1040,
995, 9801960, 940, 870, 840, 8
20, 800, 780, 760, 730167 to 62 480C77! -1(12) Nuclear Magnetic Resonance Spectrum Tetramethylsilane (TMS) in Deuterochloroform Dissolution
FIG. 3 shows the NMR spectrum measured using the internal standard.

The presence of two C1CH2, one O-CH3, trans double bond (H>C=C<H), -NH and -0H in the molecule is considered. (13) Nuclear magnetic resonance spectrum by 13C-NMR From the 13C-NMR spectrum measured in CDCl3 using TMS as an internal reference, the presence of a total of 30 carbon atoms in the molecule is recognized.

(a) There are three primary carbon methyl groups, 17.3, 1
7.6 and 55.0 ppm.

(b) There are 6 secondary carbons, 20.9, 25.5, 2
7. λ36. It is 38.96 ppm.

(c) There are 16 tertiary carbon atoms, 33.7, 40.
5, 41.1, 45.8, 47.1, 47.5.48.
8, 50.1, 53.7, 58. 61.5 sleep 77.3
s122. Snake 124. Next 139.8 and 150.8pp
It is m. (d) There are 5 quaternary carbon atoms, 100.
6, 166.2, 173.5, 175.7 and 196.
It is 1 ppm.

(14) The molecular ion (M+) in the mass spectrum is 5>4. This substance is recognized as a new substance because the above-mentioned physical and chemical properties do not match any of the known substances.

The present invention has antibacterial activity and is effective against various diseases of vegetables, fruit trees, and other agricultural and horticultural plants, such as various downy mildews, late blights, damping-off diseases,
It is effective in controlling soft rot, charcoal stain, powdery mildew, rice blast, sheath blight, etc.

Accordingly, the present invention further provides an agricultural and horticultural fungicide that contains the N-461 substance as an effective ingredient.

When this substance is used as an active ingredient in agricultural and horticultural fungicides, it may be used as it is, but it is usually used in the form of a preparation mixed with an appropriate carrier and used with common adjuvants if desired. .

Crude extracts containing the substance can also be used. As the carrier, both solid and liquid carriers can be used. Examples of solid carriers include vegetable carriers, fibers, clays,
For example, water, alcohols, ketones, ethers, aliphatic or aromatic hydrocarbons, esters, acid amides, etc. are used as liquid carriers for powders and granules of talc and other inorganic minerals, fertilizers, etc. It will be done. Preparations suitable for use include, for example, powders, granules, wettable powders, emulsions, oils, etc., and these preparations can be produced by conventional methods. The amount of this fungicide to be used will vary depending on the dosage form, type of crop, season, etc., but for example, when using a hydrating powder, it is generally 1 to
The objective can be achieved by spraying 50% hydrating powder in an amount of 50-300e/10a.

The agricultural and horticultural fungicide of the present invention can also be used in combination with other fungicides, insecticides, herbicides, and the like.

Example 1 Glucose 1%, sucrose 3.5%, germ 3.5%, granular yeast 0.5%, ammonium sulfate 0.3%, ammonium chloride 0.2%, calcium carbonate 0.9%, magnesium sulfate 0. 2%, ferrous sulfate 0.004%, manganese sulfate 0
．． A liquid medium having a composition of 0.05% zinc sulfate and 0.01% zinc sulfate is adjusted to pH 7.6, dispensed into flasks, and sterilized.

In addition, Streptomyces Sp. Bacterium No. N-461 (Feikoken Bacteria No. 4513) was inoculated, cultured with shaking at 29-30°C for 2 days, and treated with a pre-cultured bacterial solution. Separately, 100 mL of a medium with the same composition as above was placed in a 200 e capacity stainless steel fermenter, inoculated with pre-cultured bacterial solution 1 e, and heated to 29-30°C.
Culture with aeration and agitation.

The ventilation rate was 100 e/min, and the rotation speed was 250 r'Pm. Antibacterial activity reaches its maximum after 9 hours.

After culturing, 4% diatomaceous earth was added to the culture mixture and filtered, and the Oki liquid was adjusted to pH 4. O and extracted with 50' of ethyl acetate, and the solvent layer was separated.

Wet bacterial cells (filtered residue) were extracted by thoroughly mixing with acetone 30e, filtered, and the solution was concentrated under reduced pressure to 7'.
4. O and extracted with the same amount of ethyl acetate. The extracts obtained from the cultured cells and the bacterial cells were combined and heated for about 5' under reduced pressure.
Concentrate to

The same amount of water was added to the mixture to adjust the pH to 8.5, and after stirring for about 1 hour, the ethyl acetate layer and the aqueous layer were separated, and the aqueous layer was collected. The aqueous layer was adjusted to pH4. The mixture is extracted with ethyl acetate 3e, and the ethyl acetate layer is washed with water and concentrated under reduced pressure to obtain a syrup-like substance. When one dose of n-hexane is added to this, substances insoluble in n-hexane precipitate. When the precipitate is collected and dried under reduced pressure, approximately 30% of the N-461 substance is removed.
15 g of containing powder are obtained. The crude powder of N-461 material can be further purified as follows.

The crude powder 5y dissolved in a small amount of chloroform was passed through a packed tower filled with silica gel (Wako C-200) 150y filled with chloroform, and the chloroform was allowed to flow down to remove impurities. : 1) and fractionate the eluate. Using Phytophythora capsici (gray blight fungus) as the test bacterium, parts showing antibacterial activity are collected, combined, and evaporated to dryness under reduced pressure.
When the residue is dissolved in ethanol heated to 70°C and left in a refrigerator, crude crystals of the N-461 substance are precipitated. When the crude crystals are further recrystallized from hot ethanol, pure N-4 is obtained.
61 substance 770WL9 is obtained as needle-shaped crystals. The N-461 substance thus obtained has the properties described above.

Example 2 95 parts by weight of clay is uniformly mixed with 5 parts by weight of the crude extract containing about 30% N-461 substance obtained in the same manner as in Example 1 to prepare a powder.

Test Example 1 (Pot Test) (a) Gray Phytophthora: Zoosporangia of cucumber Gray Phytophthora (Phytophytora capsiki) were immersed in water for 30 to 60 minutes (adjusted to 4 to 5 per field at 15@). , which was used as the inoculation source.

The chemical was sprayed on cucumber seedlings at the two-leaf stage, air-dried, and then the above-mentioned inoculum was sprayed. 30℃ and 100% humidity after inoculation
After 3 days, the degree of disease onset was investigated. 7(b
) Seedling damping-off: The cucumber damping-off fungus (Pythium debayanum) was grown in advance on a potato-sugar agar medium and then mixed with 300% water.
It was ground together with m1 and used as an inoculum.

The chemical was sprayed on cucumber seedlings at the two-leaf stage, air-dried, and then the above-mentioned inoculum was sprayed. 30℃ and 100% humidity after inoculation
After 3 days, the degree of disease onset was investigated. The following drugs were used for comparison.

Difortan (manufactured by B Honyaku): N-(1,12,2-tetrachloroethylthio)-4-cyclohexene-1,2
-Decarboximide daconyl (manufactured by Takeda Pharmaceutical): Tetrachloroisophthalnitrile control value was calculated using the following formula.

The results obtained are shown in the table below.

Test Example 2 (Acute Toxicity) ICR/JCL female mice (5 weeks old, body weight 22.0±
LD, O by oral administration of N-461 substance determined by up-down method using 1.0f) and whether it was alive or dead after 1 week.
was 600-700 mg/kg.

[Brief explanation of drawings]

Figure 1 is the ultraviolet absorption spectrum of N-461 substance, Figure 2 is
The figure shows the infrared absorption spectrum of the N-461 substance by the KBr tablet method, and Figure 3 shows the nuclear magnetic resonance spectrum of the N-461 substance in deuterated chloroform.

Claims (1)

[Claims] 1. An acidic substance in the form of colorless needle crystals (crystallized from ethanol), which begins to turn brown at 186°C and does not show a clear melting point, and whose molecular formula is C_3_0H_4_0N_2O_6 (molecular weight: 52
4.64), elemental analysis values are C: 68.66%, H: 7.
66%, N: 5.30%, O: 18.34% (theoretical value C
:68.68%, H:7.69%, N:5.34%O:
18.30%), the specific rotation is ▲ There are mathematical formulas, chemical formulas, tables, etc. ▼ +196 (C1, chloroform), and the ultraviolet absorption spectrum is ▲ ▲ There are mathematical formulas, chemical formulas, tables, etc. ▼: 325 mμ (▲ Mathematical formulas, There are chemical formulas, tables, etc.▼
500), 230-240mμ (S), ▲There are mathematical formulas, chemical formulas, tables, etc.▼: 325mμ (▲There are mathematical formulas, chemical formulas, tables, etc.▼594), 220mμ (▲There are mathematical formulas, chemical formulas, tables, etc.▼918) , ▲ There are mathematical formulas, chemical formulas, tables, etc. ▼: 320 mμ (▲ There are mathematical formulas, chemical formulas, tables, etc. ▼ 567), shows maximum absorption of 220 to 240 mμ (S), infrared absorption spectrum (potassium bromide tablet method)
in 3380, 3080, 2950, 2900, 2
870, 1705, 1640, 1580, 1435, 1
370, 1340, 1300, 1250, 1240, 1
170, 1140, 1090, 1040, 995, 98
0, 960, 940, 870, 840, 820, 800
, 780, 760, 730, 670, 620, 480c
It exhibits a characteristic absorption of m^-^1, is easily soluble in chloroform, pyridine, and alkaline water, slightly soluble in ethyl acetate, acetone, methanol, and ethanol, and insoluble in ether, benzene, n-hexane, and water, and has antibacterial properties. N-461 substance. 2. Cultivate Streptomyces bacteria that have the ability to produce the N-461 substance described below, and extract N-461 from the culture.
It is an acidic substance in the form of colorless needle-like crystals (crystallized from ethanol), which is characterized by the fact that it collects a substance.It starts to turn brown from 186℃ and shows a clear melting point.The molecular formula is C_3_0H_4.
_0N_2O_6 (molecular weight 524.64), elemental analysis values are C: 68.66%, H: 7.66%, N: 5.30%
, O: 18.34% (theoretical value C: 68.68%, H: 7
．． 69%, N: 5.34%, O: 18.30%), the specific rotation is ▲ There are mathematical formulas, chemical formulas, tables, etc. ▼ +196 (C
1 chloroform), and in the ultraviolet absorption spectrum there are ▲ mathematical formulas, chemical formulas, tables, etc. ▼: 325 mμ (▲
There are mathematical formulas, chemical formulas, tables, etc.▼500), 230-2
40mμ(S), ▲There are mathematical formulas, chemical formulas, tables, etc.▼:
325 mμ (▲There are mathematical formulas, chemical formulas, tables, etc.▼594
), 220 mμ (▲There are mathematical formulas, chemical formulas, tables, etc.▼9
18), ▲There are mathematical formulas, chemical formulas, tables, etc.▼: 320m
μ (▲There are mathematical formulas, chemical formulas, tables, etc.▼567), 22
It shows a maximum absorption of 0 to 240 mμ (S), and in the infrared absorption spectrum (potassium bromide tablet method), it has a maximum absorption of 3380, 3
080, 2950, 2900, 2870, 1705, 1
640, 1580, 1435, 1370, 1340, 1
300, 1250, 1240, 1170, 1140, 1
090, 1040, 995, 980, 960, 940,
870, 840, 820, 800, 780, 760, 7
It exhibits characteristic absorption at 30, 670, 620, and 480 cm^-^1, and is easily soluble in chloroform, pyridine, and alkaline water.
A method for producing N-461 substance, which has antibacterial properties and is sparingly soluble in ethyl acetate, acetone, methanol and ethanol, and insoluble in ether, benzene, n-hexane and water. 3 An acidic substance in the form of colorless needle-like crystals (crystallized from ethanol), which begins to turn brown at 186°C and does not show a clear melting point, and whose molecular formula is C_3_0H_4_0N_2O_6 (molecular weight: 52
4.64), elemental analysis values are C: 68.66%, H: 7.
66%, N: 5.30%, O: 18.34% (theoretical value C
:68.68%, H:7.69%, N:5.34%, O
: 18.30%), the specific rotation is ▲ There are mathematical formulas, chemical formulas, tables, etc. ▼ +196 (C1 chloroform), and the ultraviolet absorption spectrum is ▲ ▲ There are mathematical formulas, chemical formulas, tables, etc. ▼: 325 mμ (▲ Mathematical formulas, There are chemical formulas, tables, etc. ▼ 500), 230-240 mμ (S), ▲ There are mathematical formulas, chemical formulas, tables, etc. ▼: 325 mμ (▲ mathematical formulas, chemical formulas,
There are tables, etc. ▼594), 220 mμ (▲ There are mathematical formulas, chemical formulas, tables, etc. ▼ 918), ▲ There are mathematical formulas, chemical formulas, tables, etc. ▼: 320 mμ (▲ There are mathematical formulas, chemical formulas, tables, etc. ▼ 567), 220 It shows a maximum absorption of ~240 mμ (S), and in the infrared absorption spectrum (potassium bromide tablet method), it has a maximum absorption of 3380, 3080, 2950, 2900,
2870, 1705, 1640, 1580, 1435,
1370, 1340, 1300, 1250, 1240,
1170, 1140, 1090, 1040, 995, 9
80, 960, 940, 870, 840, 820, 80
0, 780, 760, 730, 670, 620, 480
It exhibits a characteristic absorption of cm^-^1, easily soluble in chloroform, pyridine, and alkaline water, slightly soluble in ethyl acetate, acetone, methanol, and ethanol, ether, benzene, n
- N-461 with antibacterial properties, insoluble in hexane and water
A fungicide for agriculture and horticulture that contains a substance as an active ingredient.