CH617227A5 - Process for the preparation of the antibiotic B-98891 - Google Patents

Description

The invention relates to a method for producing a new antibiotic with the designation B-98891, which is obtained by cultivating a strain of the genus Streptomyces which forms the antibiotic B-98891, and a method for producing salts of this antibiotic.

In the search for new antibiotic substances, the owner isolated a large number of soil microorganisms and examined their biologically active metabolites. The research led to the finding that certain soil organisms are able to form a new antibiotic called B-98891, that these microorganisms belong to the genus Streptomyces, and that it is possible to cultivate these antibiotics in such a manner that the antibiotic is accumulated in the culture medium to gain.

The invention accordingly relates to a process which is characterized in that a microorganism which forms the antibiotic B-98891 and belongs to the genus Streptomyces is cultivated under aerobic conditions in a culture medium which contains an assimilable carbon source and a usable nitrogen source. until the antibiotic is accumulated in the fermentation medium and the antibiotic is isolated from the fermentation medium.

Furthermore, it was surprisingly found that the new antibiotic B-98891 is excellent for combating plant diseases, e.g. powdery mildew.

The invention accordingly also encompasses its use in a crop protection agent used to control plant diseases which does not show any significant phytotoxicity and is essentially non-toxic to animals, including the fish. Particularly suitable is a concentrate of the plant protection product which, after simple dilution, can be applied to plants for the abovementioned purposes and is more stable and easier and more convenient to store or transport than the ready-to-use diluted preparation.

The antibiotic B-98891 is thus produced by cultivating a strain belonging to the genus Streptomyces and forming the antibiotic B-98891, preferably as long as it is able to form the antibiotic. For example, the strain classified as Streptomyces rimofaciens strain isolated from a soil sample taken by the owner in New Guinea and designated Streptomyces rimofaciens No. B-98891 and its related strain can be used with particular advantage. : o The following are some microbiological and cultural features of Streptomyces rimofaciens No. B-98891.

1) Morphological properties j5 This strain forms an aerial mycelium on various culture media, and the branching type of spore-bearing hyphae is double-bubbly. On glucose-asparagine agar, loops and hooks (not on the verticals) are found, albeit rarely. The spores are oval to cylindrical. Their m size is in the range of 0.6 to 0.8 X 0.7 to 1.3 [i ,. The spores usually appear in chains from 3 to 16. The surface of each spore is smooth or warty. Neither sporangia nor goat nor sclerotium etc. can be found on various culture media.

15

2) Cultural characteristics

The cultural characteristics of the strain used according to the invention are listed in Table 1 below. Unless otherwise stated, the characteristics were determined as a result of cultivation at 28 ° C for 21 days. The color names in the tables were taken from the Color Harmony Manual, 4th edition, Container Corporation of America.

Table 1

45

Cultural characteristics of Sterptomyces rimofaciens No. B-98891 on various media Medium characteristics *

Sucrose- W: Colorless, penetrates the medium. 50 stepsagar R: White to light ivory (Light Ivory)

(2ca)

LM: Sparse, fluffy, shell (3 ca) LP: Not available

Glucoseni-stiragar

Glycerinni-step agar

65

W: Colorless, penetrates the medium. R: Yellow Maple (3 ng) LM: Moderate, pearl-colored (Pearl) (2 ba) to light ivory (2 ca)

LP: Brown

W: Colorless, penetrates the medium. R: honey gold (2 ic) to mustard gold (mu

stard gold) (2 pg)

LM: moderate, cream-colored (l'A ca) LP: pale yellowish brown (Pale yellowish brown)

3rd

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Glucose-W: Colorless, penetrates the medium.

Asparagine Agar R: Light ivory (2 ca) to Chamois (2 gc) to cinnamon (3 le)

LM: Thin, white with a yellow tinge or moderate, woolly shell (3 ca) to pearl-colored ben (3 ba)

LP: Not present or very weak, culture yellowish brown.

Glycerin W: Chamois (2 gc) to tan (tan) (3rd

Asparagine agar ie), penetrates into the medium. Nutrient agar

R: cinnamon colors (3 le)

LM: thin, white to light ivory (2 ca)

or moderate, fluffy, white with a tinge of yellowish pink, (withe with a tinge of pinkish yellow)

LP: pale brown nutrient agar

Calcium maleate- W: cream (IV2 ca), later cha-agar mois (2 gc)

R: honey gold (2 ic) to cinnamon (3 le)

LM: Not present or thin, light- glucose-ivory (2 ca) peptone-

LP: Pale brown gelatin

Inorganic salts

Starch agar

W: Colorless, penetrates the medium

R: cream (1 'h ca) to light ivory (2 ca)

LM: Abundant, fluffy, white to pearl-colored (2 ba), later pearl-colored (3 ba) to mother-of-pearl (Shell) (3 ca) to light beige (3 ec)

LP: Pale yellow

Yeast extract- W: colorless, wrinkled agar R: honey gold (2 ic) to yellow maple (yel low maple) (3 ng)

LM: moderate, dull yellow (dusty yellow) (1 'A gc) or fluffy, white to pearl (white to pearl) (3 ba) LP: light brown (3 lg)

Yeast malt agar

Oat flour agar

î5 skimmed milk

Abbreviations:

cb), partly mother-of-pearl (Shell) (3 ca)

LP: The color of the stitch changes to grayish brown

W: Good, mother-of-pearl (3 ca) LM: Abundant, fluffy, white to pearl-colored (3 ba) to mother-of-pearl (3 ca) LP: The color of the stitch hardly changes

W: Colorless

R: light maize (2 ea)

LM: Thin, white

LP: Pale brown

W: Good, colorless, wrinkled R: Honey gold (2 ic) to topaz (3 ne) LM: Moderate, pearl-colored (3 ba) to light ivory (2 ca)

LP: Pale brown

W: surface growth good, colorless, growth in the medium sparse, colorless

LM: Moderate, cream-colored (11 h approx) LP: Brown pigment, just for growth. After incubation for about 10 days, bluish green pigment appeared near the surface, later transitioning to dark bluish green and diffusing downward. Gelatin not liquefied.

W: ring-shaped surface, light ivory

(2 ca) to light maize (2 ea) LM: not available LP: maple (maple) (4 le), coagulation and peptonization were observed.

W: growth R: back LM: aerial mycelium LP: soluble pigment

W: Good, colorless, penetrates the medium;

R: light ivory (2 ca) to golden brown

(goldenbrown) (3 pi)

LM: Moderate, light ivory (2 ca) to pastel yellow (pastel yellow) (1 db) to chamois (2 gc) or fluffy, white to pearl-colored (3 ba)

LP: Pale brown

W: Good, colorless

R: yellow maple (3 ng) LM: abundant, fluffy, white to pearly (3 ba) to light beige (3 ec) LP: pale yellowish brown

Potato W: Corn colors (Maize) (2 ga) to cha-

Stitch culture mois (2 gc)

LM: Abundant, fluffy, white to pearl-colored (3 ba) to sand-colored (sand) (3

3) Physiological properties

1) Growth temperature range Growth takes place at 15 to 38 ° C; better growth and formation of aerial mycelium is observed at 28 to 36 ° C. 50 2) Liquefaction of gelatin (cultivation 28 days at 24 ° C:

Not liquefied.

3) Hydrolysis of starch: positive

4) Reduction of nitrates: negative

55 Bactonitrate broth (ISP No. 8: Method: Manual of International Cooperative Project for Description and Déposition of Type Cultures of Streptomycetes, 1964) and Czapek solution

5) Coagulation of skim milk: positive Peptonization of skim milk: positive

(, o 6) Formation of black pigment:

positive (peptone-yeast-iron agar)

negative (tyrosine agar)

7) Assimilation of carbon sources (method of Pridham and Gottlieb): Well assimilated carbon sources: ss inositol, D-galactose, D-glucose, maltose, D-mannose, starch, glycerin, sodium acetate, sodium succinate and sodium citrate. Fairly well assimilated carbon sources: D-fructose, Tre-halose;

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4th

unassimilated carbon sources:

Erythritol, adonite, D-sorbitol, D-mannitol, Dulcit, D-xylose, L-arabinose, L-sorbose, rhamnose, melibiose, sucrose, lactose, raffinose, salicin, esculin, inulin.

As already mentioned, the vegetative mycelium of this strain has a colorless to pale yellow surface. The back is pale yellow to yellowish brown to brown. The aerial mycelium is either yellowish white or in the fluffy part yellowish orange to yellowish with a tinge of gray. Light yellowish brown to brown soluble pigment is formed on certain synthetic agar.

The color formation of this strain on protein-containing media is low, but the formation of a blackish-brown soluble pigment in peptone-yeast-iron agar is clearly visible, a sign that the strain is melanin-positive. A soluble brown pigment and a soluble bluish green pigment are formed on glucose-peptone gelatin.

On the basis of the cultural characteristics observed in this way, the strain used according to the invention was compared with the known species which are described in the literature, for example by S.A. Waksman "The Actinomyce-tes" II (The William and Wilkins Co., 1962), Cooperative Description of Type Cultures of Streptomyces II (International Journal af Systematic Bacteriology 18, p. 69), dto. III (loc.cit. 18, P. 279), dto. IV (loc.cit. 22, p. 265), Rocci et al "The Genus Streptoverticillium: A Taxonomic Study" (Giornale di Microbiologia 17 (1969) 1) and others. As a result, it was found that the strain used in the present invention is similar to Streptomyces rimofaciens (Japanese Patent Publication No. 7598/1967).

The differences between S.rimofaciens and the strain used according to the invention are listed in Table 2. As for the other characteristics that are not mentioned in the table, most of the morphological characteristics, cultural characteristics and physiological characteristics of both strains are similar.

The differences mentioned in the table are not sufficient to consider the strain used according to the invention as a new species. In view of the fact that the strain used according to the invention does not form Destomycin A and B, but the new antibiotic B-98891 according to the invention, the holder assumes, however, that this microorganism and Streptomyces rimofaciens are different strains. The new strain is hereinafter referred to as Streptomyces rimofaciens No. B-98891 and was registered on March 21, 1974 under No. IFO-13592 at the Institute for Fermentation, 17-85 Juso-honmachi 2-chome, Yodogawa-ku, Osaka 532 ( Japan), on March 26, 1974 under No. FERM-P 2549 at the Fermentation Research Institute, Agency of Industriai Science and Technology, 8-1 Inage-Higashi 5-chome, Chiba City 281 (Japan) and on January 20, 1975 No. ATCC-31120 with the American Type Culture Collection, 12301 Parklawn Drive, Rockville, Maryland 20852 (USA).

Table 2

Comparison of S. sp. B-98891 with S.rimofaciens Medium S.rimofaciens S. sp.Nr.B-98891

Antibiotic formed

Destomycin A & B B-98891

Bouillon agar and glucose bouillon agar

Carrot stitch culture

Cracks formed in growth; soluble pigment dark brown no growth no cracks; soluble pigment pale brown good growth with aerial mycelium

Assimilation of sorbitol and mannitol neither sorbitol nor carbon sources are assimilated. Mannitol is assimilated

As common features, the species of Strepto-5 myces show a willingness to change certain cultural features and physiological properties, and this also applies to Streptomyces rimofaciens No. B-98891. Thus, the above-mentioned characteristics of the new strain are not constant, but different mutants can easily be generated. However, as long as the ability to form antibiotic B-98891 is not lost, these mutants can also be used for the purposes of the invention. It does not matter, of course, whether this mutation occurred spontaneously or was artificially caused by mutagenic treatment.

In the method according to the invention, the cultivation is carried out using a medium which, in addition to an assimilable carbon source and a utilizable nitrogen source, generally contains inorganic salts etc. 2ii Trace nutrients, growth-promoting agents (e.g. vitamins, amino acids, animal oil, vegetable oil and mineral oil), precursors and other materials that are effective in small quantities can be added to the medium as required. Examples of suitable assimilable carbon sources are 25 glucose, sucrose, molasses, starch, dextrin, glycerin. Suitable nitrogen sources which can be used are, for example, meat extract, soybean meal, corn steep liquor, peptone, casein and cottonseed meal as well as inorganic nitrogen compounds, e.g. Nitrates and ammonium compounds, î »In addition to these components, anti-foaming agents etc. can be added to the medium as required. All of these materials can be used to advantage. Although the surface culture can be carried out, the aerobic submerged culture is generally more advantageous. When carrying out i5 the aerobic submerged culture, the pH of the medium is preferably in the vicinity of the neutral point) i.e. pH 6.0-8.0) and the incubation temperature is preferably kept at about 20 ° to 37 ° C. These cultivation conditions such as composition and pH of the medium, cultivation temperature, degree of movement or aeration, etc. can of course be regulated and selected depending on the strain of the microorganism used, the external conditions and other factors in such a way that good results are obtained will.

45 In each case, however, the highest concentration of antibiotic B-98891 is generally found in the medium 96 to 144 hours after the start of cultivation.

The culture medium obtained contains the antibiotic B-98891 in high concentration. For the isolation of the antibiotics in optimum purity from this culture medium, the separation methods usually used for the isolation of microbial metabolites from cultures can be used with advantage. For example, since the antibiotic B-98891 is largely present as a water-soluble base in the filtered culture medium, a filter aid is first added to the culture medium, which is then filtered off to remove the cells. The filtrate is then treated with a suitable adsorbent, on which the active fraction is adsorbed and then desorbed with a suitable solvent. Such a fractionation process can be used advantageously.

Suitable as adsorbents are, for example, activated carbon, as adsorbents effective resins, cation exchange resins, activated aluminum oxide and silica gel. In addition, separation and purification processes are also suitable which take advantage of the differences in molecular weight, e.g. a process carried out using molecular sieves. The type of desorbing solvent depends on the type

5

617 227

and the properties of the absorbent, however, for example, aqueous acetone, aqueous methanol, aqueous ethanol, aqueous propanol, aqueous butanol, etc., and acidic and alkaline buffers and aqueous solutions of inorganic or organic salts can be used.

For example, when activated carbon or an adsorbable resin is used as the adsorbent, the active substance is adsorbed in the filtered culture medium under neutral or weakly basic conditions and, for example, with water containing a suitable amount of acetone, methanol, propanol or the like, water containing a salt or contains an acid, or desorbs a buffer solution.

Since the antibiotic is a basic substance, it can be adsorbed on a cation exchange resin and eluted with a suitable acidic or alkaline solution or buffer solution. The product “Amberlite IRC-50” or “Amberlite CG-50” (manufacturer Rohm and Haas Co., USA) is suitable as a cation exchange resin. For example, the antibiotic B-98891 can be adsorbed on a column of the resin and then, for example, eluted with dilute aqueous ammonia or a buffer solution. In addition to the above-mentioned methods, adsorption chromatography on silica gel (manufacturer Merck, West Germany) is suitable for cleaning the antibiotic B-98891. For example, the active eluate can be developed by developing the chromatogram with a suitable solvent system, e.g. a mixture of methanol, methylamine and water. The desired purification is also possible by adsorbing the antibiotic on a carrier material which has the property of a molecular sieve, e.g. on the product “Sephadex G-15” (manufacturer Pharmacia, Sweden) and elution, for example with water or a buffer solution. The cleaning can also be carried out using such an adsorbent and a molecular sieve in a suitable combination.

The antibiotic B-98891 purified in this way can then be made from methanol, acetone or the like. can be obtained as a colorless powder.

The properties of the powdered free antibiotic B-98891, which was prepared in the manner described in Example 2, are mentioned below.

physical and chemical properties

1) Melting point: gradual discoloration begins at around 220 ° C; the powder shows no sharp melting point.

2) Specific rotation:

[oc] 20 = + 91.8 ° ± 10 ° (c = 0.5 in water);

+ 68.5 ° ± 10 ° (c = 0.5, 0, In HCl).

3) pKa: 4.2 ± 0.2.7.0 ± 0.2 (determined using the titration method)

4) Molecular weight: 529 ± 100 (determined according to the titration method)

5) Composition according to elements and elemental analysis: consisting of carbon, hydrogen, nitrogen and oxygen without halogen, phosphorus and sulfur.

C: 42.73 ± 1.5 H: 6.01 ± 0.5 N: 20.48 ± 1.0 O: 30.05 ± 2.0

6) UV absorption spectrum:

The UV absorption spectrum is shown in Fig. 1. In the diagram, the solid line represents the spectrum measured in aqueous solution at pH 7, the dotted line the spectrum O.ln-NaOH and the dashed line the spectrum in 0.1n-HC1.

The absorption maxima are as follows:

\ pH7 271 ± 2nm (E]% m = 157)

Max

, 0, lN-NaOH 271 ± 2 nm (eI ^ L = 154)

I fi A »* ■ cm

II max

X.0, ln-HCl 280 ± 2nm (E \ fm = 228)

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7) IR absorption spectrum

The IR absorption spectrum, which was recorded using the KBr disk method, is shown in FIG. 2. The significant absorptions (wavenumbers) are as follows: 1 (1 3330, 3160 (shoulder), 2900, 2850, 1660, 1600, 1505, 1485, 1400, 1375, 1295, 1230, 1180, 1130, 1065, 910, 780, 700, 580 cm -1.

8) Thin layer chromatography (silica gel F2S4, manufacturer Merck)

25th

Solvent system Rf value

Chloroform-methanol 17% aqueous

Ammonia (2: 2: 1 top layer) 0.34

.m

Propanol-pyridine-acetic acid-water 0.15 (15: 10: 3: 10)

Ethyl acetate-acetone-acetic acid water 0.37 15 (45: 5: 5: 45 lower layer)

9) Paper chromatography (Whatman # 1 filter paper)

Solvent system Rf value

4 (| butanol-acetic acid-water (2: 2: 1) 0.13

Propanol-water (7: 3) 0.27 isopropanol-5% aqueous ammonia (6: 4) 0.26

10) Whatman filter paper No. 1, 500 -V paper electrophoresis; completely migrated to the cathode side in 2 hours)

cm -0.7

-1.1

-3.3 11) Solubility

Easily soluble in water, but slightly soluble in organic solvents (e.g. methanol, butanol, pyridine, acetic acid, dimethyl sulfoxide and tetrahydrofuran).

12) Color reactions

Positive reactions with Greig Leaback, Sakaguchi and potassium permanganate reagents; uncertain positive reactions with aniline phthalate and ninhydrin; negative reactions with Pauly, Ehrlich, Dragendorff, Barton, benzidine periodate and iron (III) chloride sulfosalicylic acid reagents.

(.5

13) Stability

Slightly unstable in acidic aqueous solution, but stable in neutral or basic aqueous solution.

Buffer solution

1/10 molar glycine-NaCl-NaOH (pH 9.32) 5H 1/15 molar phosphate buffer (pH 7.05) 1/10 molar citrate-HCl buffer (pH 3.62)

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6

Biological properties 1) Antimicrobial spectrum (in vitro)

The antimicrobial spectrum of antibiotic B-98891, determined by the agar dilution method and diffusion method, is shown in Table 3. The table 3 given in the table

Antimicrobial spectrum

Test microorganism Test conditions bene "diameter of the inhibitory zones" is the diameter of the inhibitory zones, which are formed around a paper disk (7 mm diameter), which are immersed in a solution containing 5000 (xg / ml of the antibiotic produced according to the invention.

Minimum inhibitory concentration

Diameter of the inhibition zone

medium

temperature

time

([ig / ml.)

(mm)

(° C)

Aspergillus niger IFO-4066

A

28

40

> 500

+

Pénicillium chrysogenum IFO-4626

A

28

40

> 500

13

Penicillium expansum IFO-7813

A

28

40

100

25th

Pyricularia oryzae IFO-5279

A

28

40

> 500

-

Cochliobolus miyabeanus IFO-5277

A

28

40

> 500

15

Sclerotinia sclerotiorum IFO-9395

A

28

40

500

20th

Botrytis cinerea TKF-12

A

20th

88

250

15

Guignardia laricina IFO-7888

A

28

88

100

18th

Trichophyton mentagrophytes IFO-5809

D

28

88

500

+

Candida albicans IFO-0583

A

28

40

> 500

-

Saccharomyces cerevisiae IFO-0209

A

28

40

> 500

12

Rhodotorula rubra IFO-0870

A

28

40

50

80

Bacillus subtilis IFO-3513

B

37

20th

> 500

14

Staphylococcus aureus IFO-3061

B

37

20th

> 500

10th

Escherichia coli IFO-3044

B

37

20th

500

14

Proteus vulgaris IFO-3045

B

37

20th

> 500

11

Pseudomonas aeruginosa IFO-3080

B

37

20th

> 500

-

Mycobacterium phlei IFO-3158

C.

37

40

50

27th

Mycobacterium smegmatics ATCC-607

C.

37

40

250

24th

Test media, see

A 3.0% sucrose, 0.2% L-asparagine, 0.3% NH4N03,

0.1% KH2P04.0.1% MgS04.7 H20.0.001% iron gelate compound «Versenol» (manufacturer Dow Chemical Co.; content of iron-sodium-ethanol-ethylenediamine triacetate 57.04%) 1.5% agar (pH 7). Before use, the following vitamins (4) are added to the medium up to the stated final concentrations in (ig / ml): thiamine 1, riboflavin 1, calcium pantothenate

1, niacin 1, biotin 0.005, folic acid 0.5, pyridoxine hydrochloride

2, p-aminobenzoic acid 0.5, cyanocobalamin 0.0002

B 0.5% Ehrlich meat extract, 0.5%, polypeptone, 0.5% 4S NaCl, 1.5% agar (pH 7)

C3.0% glycerin, 0.5% Ehrlich meat extract, 0.5% polypeptone, 0.5% NaCl, 1.5% agar (pH 7.0)

D 1.0% glucose, 0.4% (NH4) 2HP04, 0.07% MgS04.7 H20.0.1% KH2P04, 0.1% NaCl, 0.003% FeS04.1.5% agar (pH 7, 0)

The values in Table 3 show that antibiotic B-98891 is weakly active against certain gram-positive bacteria, gram-negative bacteria, phythogenic fungi and certain yeasts. 55

The antibiotic B-98891 was tested by the diffusion method using potato-sucrose agar (3% sucrose, pH 5) as the test medium and Rhodotorula rubra IFO-0907 as the test microorganism.

net to determine the effect of the test preparation. The results are shown in Table 4 below.

Table 4

Effect against powdery mildew of barley

concentration

Lesion Area,%

50 ppm

0

25th

0

12.5

0

6.25

26

3.12

40

62.5 **

14

100

Test connection

B-98891

Wetting powder «Benomyl» *

Control untreated

♦ Agricultural chemical preparation containing methyl 1- (butylcarbamoyl) -2-benzimidazole carbamate as an active ingredient.

♦♦ Concentration as methyl l- (butyIcarbamoyl) -2-benzimi-dazol carbamate.

The results show that antibiotic B-98891 is more effective than the commercially available preparation.

2) Efficacy against powdery mildew of barley

Seven day old barley seedlings were used. The roots were covered with small pieces of cotton wool moistened with a predetermined concentration of antibiotic B-98891. The unfolded leaves were inoculated with conidia from Erysiphe graminis with a small brush. Each seedling was kept in a test tube at 18 ° C for 7 days. The percentage area of the lesion formed was calculated

3) Toxicity

The determination of the acute toxicity in mice and rats had the following results:

administration

Mice

Rats

LD5 () (mg / kg) intravenously orally

> 500> 2000

> 500> 2000

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When determining the effect of the preparation on the cornea and skin of rabbits, no change was observed at a concentration of 1000 ppm after 10 days of observation.

In the toxicity test on fish, no toxic effect on the Japanese kerping (killifish) was observed after 7 days of observation at a concentration of 20 ppm.

Comparison with known antibiotics

The above properties show that antibiotic B-98891 is a nucleoside antibiotic. Of the known antibiotics of this type, gougerotin, anthelmycin, blasticidin S, aspiculamycin, hikijimycin, moroyamycin A, B (gougerotin analogs) and C and cytovirin in the UV absorption spectrum are somewhat similar to antibiotic B-98891. The UV absorption spectra of these known antibiotics all show maxima at 268 nm (in alkaline aqueous solution) and 276 nm (in acidic aqueous solution), while the antibiotic B-98891 maxima at 271 ± 2 nm (in alkaline aqueous solution) and 280 ± 2 mm (in acidic aqueous solution) shows. Furthermore, of the known antibiotics mentioned, all compounds whose chemical structure has been elucidated contain cytosine as chromophore, while 5-hydroxymethylcytosine was determined as the chromophore in the antibiotic B-98891. According to the above results, the antibiotic B-98891 can certainly be regarded as a new antibiotic.

As its biological properties show, this new antibiotic B-98991 is weakly effective against gram-positive and gram-negative bacteria.

Its toxicity in mice and rats is also low and there is practically no irritating effect on the cornea and skin of rabbits. The antibiotic is therefore considered to be weakly toxic.

On the other hand, the antibiotic has a characteristic strong effect against the cause of the mildew of the barley (Erysiphe graminis) and also the extremely advantageous property that it is essentially harmless to fish.

Powdery mildew is widespread and there are various problems with the known powdery mildew control agents, e.g. Efficacy (including the problem of resistance), damage to the plant, residues, the problems of contamination and risk associated with its production, etc. The antibiotic does a lot to solve these problems.

The antibiotic B-98891 is contained as an active ingredient in fungicidal preparations intended for plant protection.

Antibiotic B-98891 can be used either alone or in combination with other chemicals whose applicability to plants is known. The antibiotic B-98891 can be the fermentation medium itself or any concentrate which contains the antibiotic B-98891 in a concentration of more than about 50%. The concentrates are the filtrate, the condensate of the filtrate or any material which is available as an intermediate product of the purification.

All forms of application in which conventional fungicides are usually used are suitable for the antibiotic. For example, the antibiotic can be dissolved or dispersed in a liquid carrier (e.g. in a solvent) or mixed or adsorbed with a suitable carrier (e.g. extender, excipient, etc.). If necessary, emulsifiers, dispersants, suspending agents, scattering agents, penetration aids, wetting agents, thickeners, stabilizers and other additives can be incorporated into the preparations so that they are used in application forms such as oily suspensions, emulsions, wettable powders, dusts, tablets, granules, aerosols, etc. can be.

The preferred concentration range of the main active ingredient in preparations such as emulsions and wettable powders is in the range from about 1 to 70% by weight and in the case of oils, dusts and other forms of use in the range from about 0.01 to 10% by weight. In the case of emulsions and wettable powders, the preparations are diluted or stretched to a suitable concentration (e.g. 100 to 10,000 times) with water or other extenders before use.

Examples of the abovementioned solvents which are suitable in the abovementioned preparations are: water, alcohols (for example methanol, ethanol and ethylene glycol), ethers (for example dioxane, tetrahydrofuran and cellosolve), aliphatic hydrocarbons (for example gasoline, kerosene, heating oil) and machine oil), aromatic hydrocarbons (e.g. benzene, toluene, xylene, mineral spirits and methylnaphthalene), organic bases (e.g. pyridine and aldehyde-collidine), halogenated hydrocarbons (e.g. chloroform and carbon tetrachloride), acid amides (e.g. dimethylformamide), esters (e.g. ethyl acetate, butyl acetate and fatty acid glyceride ester) and nitriles (eg acetonitrile). These solvents can be used alone or in combination.

Vegetable powders (e.g. soybean flour, tobacco flour, wheat flour and sawdust), mineral powders (e.g. kaolin, ben-tonite, acid clay and other clay minerals, talcum powder, pagodite and other types of talc, diatomaceous earth, mica powder and other silicon dioxide) are, for example, suitable as extenders and / or auxiliaries ), Aluminum oxide, sulfur powder and activated carbon. These materials can be used alone or in combination.

As the surfactants used as emulsifiers, spreading agents, penetration aids, dispersants, etc., various soaps, esters of sulfuric acid with higher alcohols, alkylsulfonic acids, alkylarylsulfonic acids, quaternary ammonium salts, oxyalkylamines, fatty acid esters, surface-active polyalkylene oxides, surface-active agents can be used as required Base of anhydrosorbitol etc. can be added. Casein, gelatin, starch, alginic acid, agar, polyvinyl alcohol, wood turpentine oil, rice bran oil, bentonite, cresol soap, etc. can be used for the same purposes.

The preparations produced in this way can be further enhanced by other types of fungicides and pesticides (for example copper fungicides, mercury fungicides, organic sulfur fungicides, phenol-type fungicides, etc.), herbicides, insecticides (organic chlorine insecticides, organophosphorus insecticides and natural insecticides), other miticides, plants, nematocides , Synergists, insect-attracting or repellent agents, fragrances, plant nutrients, fertilizers and the like. be supplemented.

The correct amount of fungicidal preparations, the types of other chemicals to be mixed with the preparations and the proportions of active ingredients in the final preparation depend, for example, on the type of plants to which the preparation is to be applied, the growth phase and the condition of the plant, the cultivation method of the plant, the type of disease, the condition of the infestation, the time at which the preparation is used and certain factors of the external environment, the type of application, the cost-effectiveness of the application and other conditions. In general, however, an application rate of 1 to 300 g per 100 ares, calculated as compound B-98891, is sufficient. With regard to the effective concentration, it is expedient that the final concentration of the compound is in the range from 1 to 10 mg / ml. In practice, the fungicidal preparations can be applied directly to the accrued part of the plant or to the soil.

S

10th

15

20th

25th

30th

35

40

45

50

55

60

() 5

617 227

8th

It is only necessary to ensure that the preparations are used safely, reliably and effectively on the plants.

The fungicidal preparations are effective against a wide variety of plant diseases and also as miticides. For example, the preparations are particularly effective against the mildew of field crops (e.g. barley), industrially used plants (e.g. tobacco and Japanese mulberry), fruit trees (e.g. apple and grape), forest plants (e.g. oak), vegetables (e.g. cucumber, oriental melon, pepper , Tomatoes, peas and gooseberries), ornamental plants (e.g. rose) etc. and the brown rot of tomatoes, potatoes and other plants. The preparations are practically non-toxic (e.g. low acute toxicity, no irritation to the cornea and skin, no toxicity in fish etc.) and are extremely safe to use.

Some test data and examples are given below, which illustrate the advantageous effects of the preparations in comparison with various chemicals which are commercially available as crop protection agents for combating the abovementioned plant diseases.

The various chemicals used in the tests for comparison are commercial products, the active ingredients of which are listed below. The concentrations given in the tables are the concentrations of these active substances.

Table 5

Effect of antibiotic B-98891 against the mildew of the oriental melon test compound

B-98891

Karathane EM control (untreated; treated)

Concentrate

Area of the lesion,

%

tration

7 days 14 days

21 days ppm

40

0

19.0

40.0

80

1.5

7.0

24.0

160

0.6

0.6

5.0

148

0.5

5.0

14.5

-

65.0

97.0

100

Phytotoxicity

Karathane EM

(Manufacturer Sanyo Boeki Co; Japan) Dinitromethylheptylphenylcrotonat 37%

Takeda-Mycin Conc.

(manufactured by the applicant) Dihydrostreptomycin sulfate Benlate WP (manufacturer duPont, USA)

Methyl 1 - (butylcarbamoyl) -2-benzimidazole carbamate polyoxin AL WP

(Manufacturer Kumiai Chemical Co., Japan) as polyoxine complex B Morestan WP

(Manufacturer Nihon Tokushunoyaku Co., Japan) 6-methylquinoxaline-2,3-dithiocarbonate 25%

Trial 1

Fighting the powdery mildew of the oriental melon

The unfolded leaves of oriental melon plants (Cucumis melo L-var.makuwa Makino, f.Ginsen), which were grown in pots (phase with 6 to 7 leaves), were pollinated with Sphaerotheca fuliginea (Schlechtendahl) by dusting the infected leaves with the spores infected with the fungus. Antibiotic B-98891 was diluted with water to a certain concentration. A spreading agent ("Dyne", manufactured by the applicant) was added in such an amount that a 3000-fold final dilution resulted. The comparative product was diluted with water in the same way.

A sufficient amount of all the fungicidal solutions were sprayed onto the plants of the oriental melon mentioned 2 days after the infection. The plants were then grown and cared for in the usual way in a greenhouse. At certain intervals, the percentage area of fungal growth on the leaves that had already developed at the time of spraying was determined. The experiment was carried out twice for each group.

Trial 2

Control of powdery mildew from pepper

The backs of unfolded leaves of potted pepper plants (9-10 leaf phase, Capsi-cum annuum L.f. California Wonder) were infected by pollination with the pathogenic fungi from infected leaves. 2 days later, the leaves were sprayed with a sufficient amount of the test solutions as in experiment 1. The plants 25 were then cultivated further in the greenhouse and cared for. The percentage area of fungal growth was determined at certain time intervals. This test was carried out twice for each group.

, d Table 6

Effect of antibiotic B-98891 against powdery mildew of pepper

Test preparation

concentration

Lesion Area,%

Phytotoxicity

12.5%

ppm

7 days

10 days

B-98891

80

7.9

10.3

-

50%

160

5.0

4.6

-

Karathane EM

148

7.2

18.1

-

4o control

10%

(untreated)

-

24.6

76.2

-

Trial 3

Control of brown rot of tomato 45 Tomato plants (Lycopersicon esculentum Mill, f.Ohgata Fukuju), which had been grown in 12 cm pots for 30 days, were used for the experiment. The test was carried out twice per group of 4 pots. The fungicidal solutions 5o prepared in the same way as in test 1 were sprayed on in sufficient quantity. Two days later, a suspension of Phytophthora infestans (Montagne) de Bary spores was also sprayed over the plants. The pots were kept in the wet room for 4 days, after which the percentage area of the lesions (on the third and fourth sheets) 55 was determined.

Table 7

Effect of antibiotic B-98891 against the brown rot of the tomato

Test preparation conc. Lesion

area, %

B-98891

,, 5 Takeda-Mycin Conc. Control (untreated)

Concentration, ppm 250 500 500

100 0 28

100

Degree of spore formation moderately zero sparsely good

Phyto

toxic

zitat

9

617 227

Trial 4

Control of barley powdery mildew

Barley seedlings with 5 or 6 leaves (Hordeum vulgare Lf Shiga Hakkoku No. 5) were cultivated in pots in a box provided with lamps (Plant-Lux, manufacturer Tokyo Shibaura Electric Co. Ltd., Japan) at 20 ° C. . The pots were placed next to the pots of infested barley plants so that the seedlings took over the disease. The fungicidal solutions prepared in the same way as in Experiment 1 were sprayed on the seedlings in sufficient quantities at intervals of 10 days. After the third application, the plants were cared for in the manner described above until day 10. To determine the degree of disease incidence, the percentage area of the lesions was determined in accordance with official Japanese regulations for determining the percentage area of mildew lesions in wheat, oats, barley and rye (available from the Ministry of Agriculture and Forestry Office, Japan) determined.

Table 8

Effect of antibiotic B-98891 against the powdery mildew of barley

Trial 6

Control of powdery mildew

For this experiment, rose plants (Rosa spp.) Were cultivated in pots in the greenhouse. The test plants (species: s Super Star and Arlene Francis) were used in groups of 3 pots for each treatment. The fungicidal solutions prepared in the same way as in Experiment 1 were sprayed on in sufficient quantities at intervals of 7 days. The plants were kept under the same conditions until spontaneous infection with the disease took place. The number of infected leaves on three branches per pot was examined 7 days after the third application. The percentage of infected leaves in relation to the total number of examined leaves was calculated.

15

Table 10

Phytotoxicity

test

Concentrate

Lesion area,%

preparation tration,

6th sheet

9th sheet

ppm

Investigative

Investigative

Day

Day

A

B C

B C

B-98891

15.6

0.4

2.1 0.8

2.3 10.5

31.2

0.3

2.0 1.3

0.3 '1.3

62.5

0.1

0.1 0

0.5 0.4

Benlate WP

125

0.3

0.4 0.2

1.3 1.6

control

-

8.4

100 mildew

68.5 mildew

(uninhibited

delt)

Examination day:

Test-

Concentrate

Infested leaves, ° A

0

Preparation tration

Super

Phyto

Arlene

2 (1st

(ppm)

Star toxicity

Francis

B-98891

40

5.6

_

14.9

80

3.2

-

6.7

Karathane EM 160

2.6

-

2.2

25th

148

52.8

-

59.9

control

-

100

88.8

A: 10 days after the first application B: 10 days after the second application C: 10 days after the third application The first application was made in the six-blade phase.

Trial 5

Control of the mildew of the tobacco plant

Potted tobacco plants (Nicotiana tabacum L.f. MC), the fourth mature leaf of which was sufficiently developed, were prepared in a greenhouse in groups of 2 pots for this experiment. To promote the disease, pots with infected plants were placed next to the test pots for self-infection. The fungicidal solutions were prepared in the same manner as in Experiment 1 and sprayed onto the test plants in sufficient amounts at two intervals in two divided doses. The percentage areas of the lesion were determined 9 days and 15 days after the last application.

Table 9

Effect of antibiotic B-98891 against tobacco mildew

45

(untreated)

Trial 7

Combating apple mildew

Apple seedlings (Malus pumila Miller var.domestica Schneider f.Rall's Janet) in the five to eight leaf phase were used. Pathogenic fungi from infected leaves of an apple plant were dusted over the seedlings, which were then kept in the greenhouse. After 2 days, the fungicidal solutions prepared in the same manner as in Experiment 1 were sprayed on in sufficient quantities. After the application, the seedlings were further cultivated in the greenhouse. The percentage area of the lesion was determined after 12 days. The test was carried out 6 times for each group.

Table 11

Effect of antibiotic B-98891 against apple mildew test preparation

B-98891

Karathane EM

control

(untreated)

Concentrate

Lesions

Phyto

tration,

area, %

toxicity ppm

12 days

40

0.9

-

80

0.6

-

160

0

-

148

0.9

-

-

27.9

-

test

Concentrate

Lesion area,%

Phyto

Trial 8

preparation tration,

toxicity

Control of powdery mildew ppm

This trial was done with saplings of about 10 years

9 days

15 days

M) old naturally grown oak trees (Quercus glauca

B-98891

40

11.6

20.4

-

Thunb.) Performed. The same way as with

80

10.0

13.2

-

Experiment 1, applied fungicidal solutions were in fog

160

6.0

8.5

-

form applied. The old adult leaves were

Polyoxin

160

61.5

90.0

-

already heavily infested, so that self-infection of the young leaves

AL WP

65 ter was expected on the saplings. The number of treated

control

-

95.0

100

The number of leaves was 50 per group. The percentage area of the

(uninhibited

Lesion on the treated leaves was 7 days after the delt)

Treatment determined.

617 227

10th

Table 12

Effect of antibiotic B-98891 against powdery mildew

Test preparation conc. Lesion area,% phyto-

tration toxicity ppm

7 days 14 days

B-98891 40 8.3 26.4

80 1.1 1.5

160 0.6 0.5

Karathane EM 148 1.8 3.4 -

Morestan WP 83 7.5 22.1

Control - 16.7 35.8 (untreated)

Trial 9

Action against a pest-resistant strain

The leaves of a mildew-infested cucumber plant (Cucumis sativus L.f.Suyo) were lightly beaten in a pot over a one-month-old cucumber plant, so that the plant was infected with Sphaerotheca fuliginea pollacci.

After one day, the fungicidal solutions prepared in the same manner as in Experiment 1 were sprayed onto the plant in sufficient quantities. After about 14 days, the fungal growth formed was used to infect healthy cucumber plants. The same fungicidal solution was sprayed on one day after infection and the percentage area of the lesion was determined after 7 days.

Table 13

Effect of antibiotic B-98891 against mildew which is resistant to pesticides.

Lesion area,%

Benlate WP

Lesions after application of Benlate WP (250 ppm) 78.3

Lesions after application of B-98891 0.8 (80 ppm)

Control (untreated) 5.0

B-98891 control

(untreated)

21.7

18.3

15.8

100

66.7 76.7

The fungicidal solutions prepared in the same way as in Experiment 1 were sprayed in sufficient quantity at intervals of 7 days on strawberry plants (var.Hogyoku) which were cultivated in the greenhouse at the flowering stage. The percentage area of the lesions was determined 7 days after the second application. In the group treated with the preparation containing the antibiotic B-98891 in a concentration of 20 ppm, a remarkable effect was found in comparison with the control group. The effect was stronger than when using Morestan WP in a concentration of 80 ppm and Polyoxine AL WP in a concentration of 100 ppm on the test plants in the same way.

It has also been found that the above-mentioned preparation shows excellent effects against mildew of garden peas, grapes and other plants.

The invention is further illustrated by the following examples. In these examples, parts are by weight unless otherwise specified. Parts by weight are related to parts of the room like grams to cubic centimeters.

example 1

In a 31-Sakaguchi flask were filled 500 ml of a medium containing 1.0% glucose, 0.3% yeast extract and

0.5% Bacto-tryptone (pancreatically digested casein, which is used as a nutrient in microbiology, manufacturer Difco Laboratories, USA) (pH 7), after which the medium was sterilized. The flask was then inoculated with an inoculum of Streptomyces rimofaciens No. B-98891 (IFO-13592) from a slant culture and incubated on a reciprocating shaker (120 cycles / min) for 48 hours at 28 ° C.

Separately, a medium containing 3.0% glucose, 2.2% soybean meal, 0.3% polypeptone, 0.4% precipitated calcium carbonate and 0.05% one was placed in a 501 tank made of stainless steel 301 stainless steel Antifoam ("Antifroth F 102", manufacturer Daiichi Kogyo Seiyaku KK, Japan) contained. After sterilization, the medium with the i? the entire amount (500 ml) of the previously prepared culture from the Sakaguchi flask.

Submerged culture with aeration at 301 air / min. was carried out at 100 rpm and 28 ° C for 48 hours, whereby a seed culture was obtained.

2 (i) In a 2001 stainless steel tank, 100 liters of a medium having the same composition as the medium used in the tank culture mentioned above was added. After sterilization, 10 liters of the above-mentioned inoculum culture was added. 25 mers culture carried out with aeration at 501 / min at 100 rpm and 28 ° C. 114 hours

1.6 kg of filter aid (“Hyflo-supercel”, manufacturer Johnes-Manville, USA) were added to 80 l of the fermentation medium obtained in this way, and the mixture was then filtered. 30 60 l of filtrate were obtained. The filtrate obtained was effective against Erysiphe graminis in a concentration corresponding to a 32-fold dilution. This experiment was carried out in the manner described above.

The filtrate obtained in the manner described was adjusted to "pH 8 and passed through a column which was filled with 7 1 ion exchange resin" Amberlite IRC-50 "(manufacturer Rohm & Haas Co., USA). The column was filled with 40 1 water and then washed with 40 10.5% aqueous ammonia. Then the active component was eluted with 4012% aqueous ammonia. The obtained eulate was passed through a column filled with 3 liters of chromatographic activated carbon (manufactured by the applicant). The column was washed with 20 l of water and then with 2015% acetone. The active component was eluted with 15 130% acetone and 15 1 45 50% acetone. The active fractions were poured together and concentrated Addition of 0.51 methanol gave 38 g of a brown, raw powder.

The crude powder thus obtained completely inhibited the growth of Erysiphe graminis in a concentration of 25 to 50 ppm. This experiment was carried out in the manner described above. The crude powder was dissolved in water and the aqueous solution passed through a column filled with 351 Amberlite CG-50 ion exchange resin. The column was washed with 2.51 water and then with 1.510.5% aqueous 55 ammonia. The active component was then eluted with 1.5 1 pyridine-acetic acid-water buffer (2: 1: 97) and then with 1.5 1 pyridine-acetic acid-water buffer (4: 2: 94). The eluate obtained was concentrated and methanol was added to the concentrate. 16 g of B-98891 acetate were obtained as a colorless 60 powder.

Example 2

In a 2001 stainless steel tank, 1001 f, 5 of a medium (pH 7) were made up containing 3.0% glucose, 2.2% soybean meal, 0.3% polypeptone, 0.4% precipitated calcium carbonate and 0.05 % Antifoam ("Antifroth F102", manufacturer Daiichi Kogyo Seiyaku, KK, Japan) contained. To

11

617 227

Sterilization was inoculated with the entire amount of a culture prepared in the manner described in Example 1 in a Sakaguchi flask. The submerged culture was carried out with aeration at 50 l air / minute and at 100 rpm for 48 hours at 28 ° C., an inoculum culture 5 being obtained.

1000 l of a medium (pH 7) containing 5.0% glucose, 3.5% soybean meal, 1.0% "Pharmamedia" (protein nutrient obtained from cottonseed, which is used in microbiology used, manufacturer Trader's Oil Mill Company, USA), contained 0.5% NaCl and 0.5% precipitated calcium carbonate. After sterilization, 100 l of the inoculum described above were added, whereupon the submerged culture was aerated with 500 l air / min. was carried out at 100 rpm for 114 hours at 28 ° C. 15.

The fermentation medium thus obtained was treated in the manner described in Example 1 to obtain 7501 of filtrate. 225 g of a crude powder were obtained from this filtrate. 211

This crude powder was dissolved in water and the aqueous solution passed through a column filled with 5 l of "Amberlite CG-50" ion exchange resin. The column was washed with 25 1 water and then with 25 10.5% aqueous ammonia and then eluted with 17 11% aqueous 25 ammonia. The eluate obtained was passed through a column of 10 l of chromatographic activated carbon which had been pretreated with 2% aqueous ammonia. The column was washed with 50 12% aqueous ammonia and then with 5015% acetone-2% aqueous ammonia and finally with 50 15% acetone. Then it was eluted with 80 120% acetone. The eluate obtained was concentrated. By adding methanol to the concentrate, the free antibiotic B-98891 was obtained as a colorless powder (150 g). «

Example 3

A crude powder was obtained from 700 liters of a filtered fermentation medium, which had been prepared by cultivation and filtration in the manner described in Example 2. The raw powder was dissolved in water and the aqueous solution was cleaned with the ion exchange resin «Amberlite CG-50». The product was then passed through a column of 5 l of chromatographic activated carbon. The column was washed with 25 liters of water and then with 25 15% acetone. The active substance was then eluted with 35% 120% acetone-0.5% formic acid. The eluate obtained was concentrated. By adding methanol to the concentrate, B-98891 formate was obtained as a colorless powder (115 g). This format had the following key figures:

Melting point: 228 ° C (dec.)

[«] £ 4- 88.4 ° ± 10 ° (c = 0.5 in water)

+ 66.9 ° ± 10 ° (c = 0.5 in 0, in HCl)

UVXmax: nm (E} c%

pH 7 271 (142)

0, ln-NaC) H271 (139)

0.1 HCl 280 (210)

Elemental analysis:

C40.50 ± 1.5; H 5.77 ± 0.5; N 18.55 ± 1.0 The corresponding hydrochloride was eluted with 20% acetone-0.05N hydrochloric acid instead of 20% acetone-0.5%

Get formic acid. This hydrochloride had the following key figures:

Melting point: 224 ° C (dec.)

[ex] ^ + 81.3 ° ± 10 ° (c = 0.5 in water)

+ 63.3 ° ± 10 ° (c = 0.5 in 0.1 n-HCl)

UV? W: nm (e}%

pH 7 272 (142)

0, ln-HaOH 272 (147)

0.1 HCl 280 (213)

Elemental analysis:

C 38.81 ± 1.5; H 5.75 ± 0.5; N 18.25 ± 1.0.

These salts had essentially the same biological activity as the free B-98891.

Example 4

A wettable powder was prepared by mixing 10 parts of antibiotic B-98891.2 parts of sodium lignin sulfonate, 3 parts of polyoxyethylene alkylaryl ether and 85 parts of clay. This powder was diluted 500- to 1000-fold with water and the dilution was sprayed uniformly onto agricultural plants or horticultural plants in an application amount of 100 to 200 1/10 ares.

Example 5

A dusting agent was prepared by mixing 0.5 part of antibiotic B-98891 and 99.5 parts of clay. This product is used directly in an amount of 3 to 5 kg / 10 ares.

Example 6

A water-soluble preparation was prepared by mixing 10 parts of antibiotic B-98891.5 parts of polyoxyethylene alkyl aryl ether and 85 parts of lactose with stirring. The preparation was diluted with water to a suitable concentration and sprayed uniformly onto agricultural plants and horticultural plants in an amount of 100 1/10 ares.

Example 7

An aqueous solution was prepared by mixing 50 parts of antibiotic B-98891, 5 parts of methanol, 5 parts of amine stearate and 40 parts of water. The solution was diluted with water in the manner described in Example 4 and sprayed onto the plants.

Example 8

Granules were prepared by mixing 0.5 part of antibiotic B-98891, 5 parts of gum arabic, 30 parts of bentonite and 64.5 parts of talc and granulating the resulting mixture. The product is used directly in an amount of 3 to 5 kg / 10 ares.

Like the product used in the experiments described above, the preparations described in the above exemplary embodiments showed excellent effects as pesticides against powdery mildew of barley, rose powdery mildew and tobacco mildew, the cucumber mildew resistant to benlate, the brown rot of the tomato and other plant diseases.

1 sheet of drawings

Claims (10)

617 227 1. Process for the preparation of the new antibiotic B-98891, characterized in that a microorganism, which forms the antibiotic B-98891 and belongs to the genus Streptomyces, in a culture medium which contains an assimilable carbon source and a usable nitrogen source, under aerobic conditions cultured, the antibiotic is accumulated in the fermentation medium, and the antibiotic is isolated from the fermentation medium. 2. The method according to claim 1, characterized in that one uses a microorganism belonging to Streptomyces rimofaciens. 2nd PATENT CLAIMS 3. Process according to Claims 1 and 2, characterized in that Streptomyces rimofaciens No. B-98891, IFO 13592, is used as the microorganism. 4. A process for the preparation of salts of the new antibiotic B-98891, characterized in that said antibiotic is prepared by the process according to claim 1 and the latter is converted into the corresponding salt by reaction with an inorganic or organic acid. 5. The method according to claim 4, characterized in that one produces the hydrochloride, formate or acetate of antibiotic B-98891. 6. Use of the new antibiotic B-98891 as a component in a crop protection agent used to combat plant diseases. 7. Use according to claim 6, characterized in that the crop protection agent contains raw antibiotic B-98891 in a purity of more than 50%. 8. Use according to claim 6, characterized in that the crop protection agent is in liquid form and contains the active ingredient in an amount of 1 to 70 wt .-%. 9. Use according to claim 6, characterized in that the crop protection agent is in powder form and contains the active ingredient in an amount of 0.01 to 10% by weight. 10. Use according to claim 6, characterized in that the crop protection agent is in the form of an oil and contains the active ingredient in an amount of 0.01 to 10 wt .-%.