WO1985000607A1

WO1985000607A1 - Antibiotic tan-542 and process for its preparation

Info

Publication number: WO1985000607A1
Application number: PCT/JP1983/000229
Authority: WO
Inventors: Setsuo Harada; Yukimasa Nozaki; Hideo Ono
Original assignee: Takeda Chemical Industries, Ltd.
Priority date: 1983-07-15
Filing date: 1983-07-15
Publication date: 1985-02-14
Also published as: JPS6041490A

Abstract

Antibiotic TAN-542 produced by microorganisms belonging to genus Flexibacter and its salts possess an antibacterial effect against Gram-positive and Gram-negative bacteria, and can be used as agents for treating bacterial infection of mammals and poultry.

Description

i Akira ^ Antibiotics ^ 赏 T AN-542 and its production

The present invention relates to a novel antibiotic A-542, a salt thereof, and a method for producing the same. Background art

Conventionally, sulfazesine and isosulfazecin are known as monocyclic and lactam anti-producing substances produced by patella (Nature, Vol. 289, 590-591, 1998). 80, 26700, 26823, 26875, 26970, 26812, etc. (Nature, 291, 489-491, 1981). Known as an antibiotic with weak antibacterial properties.

The present inventors have separated a large number of organisms from soil for the purpose of searching for new antibiotics, and have separated and searched for the antibiotics produced. Producing the substance, that the organism belongs to the genus Flexibacter, and culturing the organism in an appropriate medium to produce an antibiotic that exhibits antibacterial activity against Gram-positive bacteria and Gram-negative bacteria. He knew that it could accumulate in the medium, isolated this antibiotic, and from its physicochemical and biological properties, confirmed that the antibiotic was a novel antibiotic. This was designated as the antibiotic TAN-542.

The present inventors have further studied based on these findings, and have completed the present invention.

The present invention relates to (1) an antibiotic TAN-542 and a salt thereof (2) an antibiotic TAN-542-producing bacterium belonging to the genus Flexipactor is cultured in a medium, and the antibiotic TAN-542 is contained in the culture. This is a method for producing TAN-542 and its salts, which is characterized by producing, accumulating, and collecting N-542.

In this book, the antibiotic TA-54 2 is simply referred to as ΤΑ 54 — 54

O PI

"''!: ■ O 2 ".

The antibiotic TAN-542 producing bacterium used in the present invention may be any bacterium which belongs to the genus Flexiactor <Flexibacter> and has the ability to produce the antibiotic TAN-542. .

As an example of an antibiotic TAN-542 producing bacterium, for example, a Flexipactor genus YK-114 strain (hereinafter referred to as “YK-114 strain”) collected from soil of Nabari City, Mie Prefecture by the present inventors. ).)). The bacteriological properties of the YK-114 strain are as follows.

(a) Form

On gravy agar 241 ;, 5 observations after cultivation revealed that the cells were rod-shaped or 0,7-0,9 long and 2.4-9 in length.

It has a filament shape of 35-75〃ffl or more. Has mobility due to gliding. No hair is found. No spores are formed, and Gram stain is negative, showing no acid resistance.

(b) Growth on various media

Cultured at 24 and observed for 1-4 days.

1. Gravy agar plate culture: colonies are opaque, orange, irregularly irregular, the surface is flat raised, and the periphery is dissected. No diffuse dye is formed. *

2. Gravy agar slant culture: Shows medium-sized or spread-like growth, opaque and orange in color.

3. Broth liquid culture: Grows in a thin turbid state, produces a small amount of sediment, and forms a pellicle.

4. Broth gelatin stab culture: shows weak growth on the surface and liquefies in a bag form.

5. Litmus, milk: Does not reduce litmus, does not convert into peptone, and does not condense.

(c) Physiological properties

1. Reduction of nitrate: +

2. Denitrification reaction: one

3. MR (Methyl Red) Test: Soil (thickening)

4, VP (Forges Proscowell) Test: 1

5. Production of Indole: One

OMPI '' Production of hydrogen sulfide (TS I agar and lead paper): ー

7. Hydrolysis of starch: one

8. Utilization of citrate (Kosel 'Christensen and Simmons's media):

9. Use of inorganic nitrogen sources

I) Potassium nitrate: one

II) Ammonium sulfate: +

IE) Sodium glutamate: +

10. Pigment formation: No diffusible pigment formation is observed in each medium of King A, B and Mannitol yeast extract agar.

11. Perease: Negative in the early stage of culture, but positive by the 15th observation.

12. Oxidase: +

13. Power Cod: One

14. Range of growth

I) pH: Grows at pH 4.9 to 7.1, but the optimal pH is 5.2 to 6,6.

I) Temperature: Grows at 13-35¾, but the optimal temperature is 23-29.

15. Attitude to oxygen: aerobic

16. 0-F (Hyti-Leifson method) Test (Hugh ♦ Leifson method): Fermentative.

17. Generation and utilization of acid and gas from sugar:

Acid gas availability

(Peptone water) (Davis medium)

D—Xylose

D-glucose + +

D—Mannose +

D—Practice Soil

D—galactose soil

+

Sucrose +

Lactose +

Trehalose +

D—Sorbit D—Mannit

Inoshit

Glycerin

Starch + — earth

18. Utilization of agar, cellulose, and chitin: None of them is assimilated or decomposed.

19. Sensitivity to actinonomycin D: III.

YK-114 strains having the above mycological properties were compared with those described in the 8th edition of Bergey's Manual of Deterinative Bacteriology (Purges Manual). . It is a gram-negative bacterium of rod 扰 or filament 、, and motility by gliding is recognized only in filamentous bacterium, does not utilize agar, cellulose, and chitin, and orange cells that are carotenoid pigments Since it produces a pigment, it was identified as a bacterium belonging to the genus Flexipacter, and the YK-114 strain was designated as Flexipactor sp. YK-14 strain (Flexibacter sp. YK-4).

The strain YK-114 was reported on July 15, 1983 by the Ministry of International Trade and Industry, Ministry of Industry and Industry's Industrial Technology Research Institute (FRI, 1-3-1 Higashi Yatabe-cho, Tsukuba-gun, Ibaraki, Japan) Accession number FERM-P 71 55, and from July 6, 1983, the Fermentation Research Institute

(IFO, 2-17-85, Jusanhoncho, Yodogawa-ku, Japan, Japan).

The properties of the bacteria of the genus Flexipactor used in the present invention are generally easily changed, and can be easily changed by artificial mutation using, for example, ultraviolet rays, X-rays, chemicals (eg, nitrosoguanidine, ethyl methanesulfonic acid) and the like. Mutants that can be mutated can be used in the present invention, regardless of any mutant strain that has the ability to produce TAN-542, which is the subject of the present invention.

When cultivating TAN-542-producing bacteria, the carbon sources include, for example, glucose, sucrose, maltose, waste condensate, glycerol, oils and fats (eg, soybean oil, liana oil, etc.), and organic substances. Acids

Those that can be utilized by bacteria, such as (down, quen, kohaku, glucon, etc.) are used as appropriate. Nitrogen sources include, for example, large flour, cottonseed flour, corn * steep ♦ liquor, dried yeast, yeast extract, meat

ΟΜΡΓ Organic nitrogen compounds such as extract, peptone, nitrogen, SI ammonium, nitrite ammonium, ammonium chloride, ammonium phosphate and inorganic nitrogen compounds can be used. Examples of the inorganic salt include inorganic salts necessary for culturing normal bacteria, such as sodium chloride, potassium chloride, calcium carbonate, magnesium sulfate, monopotassium phosphate, and sodium phosphate, alone or in combination. Used. Also, an antibiotic belonging to the genus Flexipactor TAN-542 Sulfur compound which can be used by bacteria producing t! Inorganic sulfur compounds such as sulfates (eg, ammonium sulfate, etc.), sulphates (eg, ammonium sulphate, etc.), sulphite (eg, ammonium sulphite), sulfur-containing amino acids (down, cystine) , Cysteine, L-thiazolidine-41-carponic acid), hypotaurine, sulfur-containing peptide (inverted, glutathione) or other organic sulfur compounds, or when adding these to the culture medium increases the amount of 100% There is.

In addition, heavy metals such as ferrous sulfate and sulfuric acid steel, and vitamins such as vitamin B 1 and bistin are added as necessary. Further, a defoaming agent such as silicone oil or polyalkylene glycol ether or a surfactant may be added to the medium. In addition, organic and inorganic substances that promote the growth of bacteria and promote the production of TAN-542 may be added as appropriate.

The culturing method may be the same as that of a general antibiotic production method, and may be a solid culture or a liquid culture. In the case of liquid culture, any of static culture, stirring culture, shake culture, and aeration culture may be performed, but aeration and agitation culture is particularly preferred. The culture temperature is about

The culture is preferably performed in the range of 5 to 35, and the pH of the medium is in the range of about 4 to 8 for about 8 hours to 168 hours, preferably 24 hours to 144 hours.

In order to collect the desired antibiotic TAN-542 from the culture, the fractionation means normally used to collect the metabolites produced by the freshwater culture from the fresh culture is appropriately used. You. For example, the antibiotic AN-542 exhibits the properties of a water-soluble acidic substance and is mainly contained in the culture broth, so first add an auxiliary agent to the culture broth or centrifuge. Remove the cells. A means for contacting the obtained culture solution with a suitable carrier to adsorb the effective component in the solution, then adhering the effective material with a suitable solvent and separating and collecting the fraction is advantageously used.

OMPI

r WIPO You. Use the difference in the adsorptive properties of compounds such as activated carbon, silica gel, powdered cellulose, and adhesive resin as the carrier for chromatography, or use the difference in the functional groups of compounds such as anion exchange resin and anion exchange cellulose. Alternatively, those utilizing the difference of the compound molecules a such as molecular sieve carriers are advantageously used. In order to elute the target compound from these carriers, the combination varies depending on the type and properties of the carrier.For example, aqueous solutions of water-soluble organic solvents, that is, aqueous acetone, aqueous alcohols, etc. A buffer solution or an aqueous solution containing an inorganic or organic salt is used in an appropriate combination.

In addition, the crude substance of the present antibiotic obtained by these chromatography can be subjected to preparative high-performance liquid chromatography for further purification.

More specifically, as the carrier, an anion exchange resin such as Dowex-1 (manufactured by Dow ♦ and ♦ Chemical Co., USA), Amberlite IRA-68, 400, 402, 410 (manufactured by Rohm and Haas Co., Ltd. When U.S.A. and Diaion SA-21 A and C (Mitsubishi Kasei, Japan) are used, the antibiotic in the solution is adsorbed and eluted with an aqueous solution or buffer containing salts or acids. Anion exchange cellulose such as DE-32

(Wattman, Fukui), DEA E—cellulose (Brown, West Germany), etc., or anion-exchange molecular sieve resin such as DEA E— or QA E—Sephadex (Falmasha, Sweden) ) Can be adsorbed onto a carrier such as) and eluted with an aqueous solution or buffer containing salts "or 羧. To remove salts, colored substances, etc. in these eluates, activated carbon for chromatography (Takeda Pharmaceutical Co., Japan) or adsorbent resin such as Diaion HP-20 (Mitsubishi Kasei Co., Japan), Amberlight XADH (Roam ♦ and ♦ Haas Co., USA) The fractionated elution fraction is converted into powder through processes such as concentration, freezing, and drying, etc. The powder thus obtained has poor purity. A high-performance liquid chromatography method is advantageously used for purification to a carrier such as TSK gel (manufactured by Toyo Soda Co., Ltd., Japan) YMC gel (manufactured by Yamamura Chemical Laboratory, Japan) ) And the mobile bed is methanol or

OMPI A mixture of acetonitrile, etc. with a neutral aqueous solution or buffer

Is used.

The TAN-542 thus obtained is usually a salt used for purification.

, Ions in the buffer, such as sodium, potassium, lithium, lithium

, Calcium, ammonium ion, etc.

It is simply done as The obtained TAN-542 salt as a free form

It is advantageous to use activated carbon chromatography to obtain

is there. That is, dissolve T A — 54 2 salt in water and dilute hydrochloric acid to PH

Adjust to 5-4 and chromatograph on activated carbon. Washing

After that, the active part is eluted with aqueous alcohol, etc.

Drying gives the free form of TAN-542.

Antibiotic TA N-542 forms metal and ammonium salts

I do. Metal salts include, for example, sodium salts, potassium salts,

Zium salt, calcium salt and the like.

Treatment of TAN-542 sodium salt obtained in Example 4 described later

The chemical properties were as follows:

(1) Appearance; white powder

(2) Elemental prayer value (96);

(Sample dried at 4 CTC on phosphorus pentoxide at 6 o'clock)

C 35 26 ± 20

H 4 9 7 Sat Ί 0

N 1 4 36 ± 1 5

0 35 87

S 5 74 ± Ί 0

Na 3 8 Sat 1 0

(However, oxygen is calculated by subtracting other elements.)

(3) Molecular weight; The molecular ion peaks by SIMS are as follows.

It is.

571, 549, 527, 469, 447, 426.

(4) Estimated molecular formula (molecule 蟁); C / ra H zs N 0

S «Na (548.45)

(5) Ultraviolet-absorbing spectrum (in water); unique to 210 nm or more

Does not show income.

(6) Infrared absorption (IR) spectrum;

The main absorption spectrum of potassium bromide absorption spectrum (Fig. 1)

,, Is

3000, 2950 1 765, 1730,

¾O C 11 1 540, 1 460 410, 1 350,

(

Wave o 77 250, 200 100, 1 050,

9 0 ¾ o, oo 780, 650, 570 (cm, " ^-1 1)

7) ¹³ C—nuclear magnetic resonance (CMR) spectrum (100MH:

Heavy water>; at least the following signals are observed.

1 77 8 (s, 77.6 (s 174.4 (s)

1 726 (s, 168.9 (s), 161.4 (s)

75 .. 2 (d) 66.2 (t), 64.1 (t)

60 .. 7 (d) 57.9 (d), 52.4 (d)

50 .. 6 (t) 44.0 (t), 35.4 (a)

19 .. (q) (PPffl), (where s singlet

d: doublet, t: triplet, q: quartet ₀ )

8) solubility;

Soluble; water, dimethyl sulfoxide.

Poorly soluble; ethyl acetate, chloroform,

Getyl ether.

(9) Color reaction

Positive ninhydrin reaction,

Negative Greig, ♦ Reapak reaction, Sakaguchi reaction,

Ehrlich reagent, parton reaction,

Dragendorff reaction.

(10) Amino acid analysis; in 6 N-hydrochloric acid, 20 o'clock

Glycine and alanine are detected as known amino acids when hydrolyzed.

(11) Stability: Stable at pH3 and PH5 in aqueous solution, slightly at pH7

Unstable, unstable at PH9.

(12) Thin-layer chromatography (cellulose f, manufactured by Tokyo Chemical Industry Co., Ltd.)

Japan);

Rf value of solvent system

Case 2: water (4: 1) 0.33

Butanol:: water (1: 1: 1) 0.58

Case two: 3% ammonia 0.52

(4: 1)

O PI (13) High performance liquid chromatography (Hitachi, Japan); YM CA-303 (Yamamura Chemical Laboratory, Japan), transfer layer: 896 methanol ZO.01M phosphoric acid buffer (PH6.3)

-. 1 Zsiin.

R ΐ (min) = 7.0

(14) Specific rotation: [〗 ² - 33. ± 10. (C = 0.5,

Dimethyl sulfide>

{15) Distinguishing between acidic, neutral, and basic substances; neutral substances (TAN-542 in the arrested body is a dark substance). Next, the biological properties of TAN-542 sodium salt are described. Table 1 shows the antibacterial spectrum of this compound against various symptoms.

Table 1

Test bacteria Minimum growth inhibition

Strain (jug X) Staphylococcus aureus FDA 209 P 3.13 Staphylococcus epidermides FS 5010 1.56 Bacillus subtilis NIHJPC 219 219 0.78 Bacillus cereus FDA 53.1 3 Bacillus * Pumils IF 03813 0.78 Micrococcus' Luteus IFO 12708 0.78 Escheri-Hia Coli LD— 2 25

Escheri Hir 'Kori C P-13 0.39 Klepsiella * Numonier IF 03317 1 00

Proteus '' Mirabilis AT C G 21 100 1 00

Pseudomonas 'Earginosa ί F O3030 12.5 Alcaligenes' Feucaris IF 0 13111 12.5

I Salmonella '' Cifimurium I F O 12529 1 00

(Note 1) Medium: Antibiotic medium 3

1 7.59, Pactagar 2 Og, distilled water;

PH 7.0.

(Note 2) About 1 O ⁶ ^ was used as the inoculum.

The therapeutic effect of TAN-542 sodium salt on practical mouse infections is shown in Table 2.

ΟΜΡΙ Table 2

Furthermore, subcutaneous administration of TAN-542 sodium salt to mice at 1 g Zkg did not show any acute toxicity.

As is clear from these data, TAN-542 is an antibiotic that shows antibacterial activity against Gram-positive bacteria and Gram-negative bacteria, and does not show toxicity for dairy animals. Therefore, TAN-542 can be used to treat bacterial infections in humans, livestock, and poultry.

To use TA-542, for example, as a therapeutic agent for B. aureus infection, for example, dissolve TAN-542 in physiological saline and inject parenterally subcutaneously or intramuscularly with 1- 5 days OigXkg, preferably SSO gZl ^ Z. As an oral preparation, the antibiotic TAN-542 is mixed with lactose to form a capsule, and TAN-542 is administered at 1 to 10 OmgZkgZ days, preferably 5 to 50 mgZkgZ days.

Further, TAN-542 obtained by the present invention can be used as a bactericide. For example, ぱ TAN-542 was dissolved in distilled water at a concentration of 0.2 Ί Ί ~ 0, 1 WZV6. Based on a liquid formulation or vaseline or lanolin, the ratio of TAN-542 per gram was 0.2. As a plaster containing lag of 209, preferably 1 to 10 lag, it can be used for sterilization and disinfection of human and human hands, feet, eyes, ears and the like.

Antibiotic TAN-542 is also a very promising compound as a synthetic intermediate for new pharmaceuticals.

From the properties described above, the antibiotic TAN-542 is considered to be a monocyclic β-lactam antibiotic. However, among the known antibiotics belonging to these, the physicochemical properties are relatively similar. Sulfazecin (su azecin). However, TAN-542 is a standard monocyclic product because the infrared, CMR, and antibacterial spectra of the antibiotic TA-542 are clearly different from those of sulfazecin. JS—Lactam antibiotic.

OMPI

Non

· BRIEF DESCRIPTION OF THE DRAWINGS

Fig. 1 shows the infrared absorption spectrum (KBr method) of antibiotic J-Jing AN-542 sodium salt. BEST MODE FOR CARRYING OUT THE INVENTION Next, the contents of the present invention will be described in more detail with reference to embodiments, but the present invention is not limited thereto. Percentages indicate weight capacity 9 unless otherwise noted.

Example 1

FlexiPactor grown on nutrient agar slope * SP YK-4 (FERP-7155; ίF014263) strain, glucose 2%, Soluble starch 396, raw soy flour 1% waste sugar Fine 1%, polypeptone (manufactured by Daigo Kunitachi Kagaku Co., Ltd., Japan) 0.5% salt containing 0.3% aqueous solution (食塩 7.0) plus sedimentable calcium carbonate 0.59 A 200-volume triangular flask was inoculated and cultured with shaking at 24 for 24 hours at 24 hours, and the culture was used as a bacterium.

Next, glycerol 3%, glucose 0.196, meat meat (manufactured by Wako Pharmaceutical Co., Japan) 0.596, and polypton (manufactured by ^ ¾, Japan) 0.596, salt A medium 600 containing 0.5%, sodium thiosulfate 0.0596, and cobalt chloride (PH 7,0) was dispensed 40 times each into 20 Erlenmeyer flasks. The inoculum was inoculated one by one, and cultured at 17 ° C with shaking at 230 rpm for Z minutes for 72 hours. Defeat 2

The pH of the culture solution (5.5 g) obtained in Example 1 was adjusted to pH 6, and acetone (22) was added thereto, followed by stirring, and Hyflo Supercell (John Manville, USA) was added. The filtrate (27.5i) was concentrated under reduced pressure, the filtrate (5.5i) was adjusted to PH6, and subjected to Dow Ix 1 x 2 (CI type) chromatography. After washing the column with water, the active portion was eluted with 49 saline (175). The eluted product was adjusted to PH6 and then subjected to chromatography on activated carbon (100 ^). Antibiotics were eluted with aqueous butanol (500 ^). After the solvent was distilled off, the solution was subjected to chromatography on QA E-Sephadex A-25 (CI type) (5 Oi). The column was washed with 0.02M saline (250) and then eluted and fractionated with 0.1M saline. The antibiotic-containing fraction (100e) was chromatographed on activated carbon (10 ^). After the column was washed with water, 7% isoptanol antibiotic was eluted. The eluted solution was concentrated, and the concentrated solution was subjected to preparative high performance liquid chromatography using TSK gel, and antibiotics were added with 296 methanol ZO, 01 M disodium phosphate monophosphate (PH 6.3). The substance was eluted and fractionated. Effective fraction (1 20

The inorganic salts in) were analyzed by chromatography on activated carbon (5j?), And the eluate was concentrated and freeze-dried to obtain white powder (2 Oag) of TAN-542 sodium 馑. Example 3

Flexi-pactor grown on nutrient agar slopes ♦ SP YK-114 strain (FERP-71 55; IFO 14263) 2% glucose, sorbul starch 396, raw soybean flour 1%, abolished 196, 0.5 g of polybeptone, 0.3% of sodium chloride in an aqueous solution (pH 7.0) containing 0.5% of sedimentable calcium carbonate The cells were inoculated and cultured at 241; The entire amount of the culture was inoculated into a 50-capacity tank containing 30 g of the above medium supplemented with 0.05% of Actol, and aerated at 30 with aeration of 30 β min and 200 rpm. For 48 hours. Glycerol 396, Darcos 0.Ί96, Meat extract (Wako Pure Chemical Industries, Japan) 0.596, Polypeptone 0.596, salt 0,5%, sodium salt 0.05% Baba Coppart 2 ppia, Actol 0 5% Inoculate a 200 g tank containing medium 1 containing 20% (PH 7.0), and aerate at 17 ^ G 60 Cultivation was carried out for 66 hours under the conditions of Z minute and 170 rpm. Actual £ 1 defeat 4

The medium (111) obtained in step 3 is passed using a high-flow super-cell (7 * 5 ^). The solution (110) was adjusted to pH 6.5, and adsorbed to Dowexoo 1 X 2 (7) by the patch method.

OMPI Antibiotics were eluted with 4% food water (35). The eluate was chromatographed on activated carbon (3 £), and the effective portion was eluted with 7% isopeptanol water. The eluate was poured, and the condensed solution was subjected to chromatography using Amberlite IRA-68 (CI type) (1) and fractionated with 0.1 M saline (6). The effective fraction (2.5i) was chromatographed on activated carbon (1SO) and eluted with 796 aqueous isopropanol. Reduce the eluate and concentrate (1

Was applied to the chromatograph of QAE-Sephadex A-25 (CI type). The column was washed with 0.02M saline (500) and fractionated with 0.1M saline. The active fraction was desalted with activated charcoal, concentrated and freeze-dried to obtain TAN-542 sodium salt (64 Omg). Industrial applicability

Antibiotic TAN-542 and its salts have antibacterial activity against Gram-negative bacteria and negative bacteria, and these bacteria can be administered to humans, livestock, poultry, etc. by parenteral or parenteral administration. It can be used to treat infectious diseases.

Claims

Slave_

Antibiotics with physicochemical properties of ^ as a sodium salt

Quality TAN—54 2 and its salts

(1) Appearance: White powder

(2) Elemental analysis (96), on phosphorus pentoxide at 40 and 6 o'clock

Dried sample:

C, 35.26 ± 2.0

H, 4.97 ± 1.0

N, 1 4.36 ± 1.5

S, 5.74 ± 1.0

Na, 3.8 ± 1.0

(3) Molecular weight: 548. 45 (SIMS method)

(4) Ultraviolet absorption spectrum (in water): Unique to 21 Onss or more.

Does not show yield.

(5) Infrared absorption tank: absorption tank using potassium bromide tablets

The main absorption (wave number) of the tool is as follows.

3400, 3000, 2950, 17, 6, 452-5, 1 730,

1 670, 1 540, 1 460, 1 4 10, 1 350,

1 2 70, 1 250, 1 200, 1 1 00, 1 050,

900, 780, 650, 570 (on-re.

(6) Solubility: Soluble in water and dimethyl sulfoxide.

Insoluble in ethyl acetate, chloroform, and getyl ether

(75 Color reaction: Positive for ninhydrin reaction.

Negative to the reaction, the slope q reaction, the Ehrlich reagent, the Barton reaction, and the Dragendorff reaction.

)-Nuclear magnetic resonance (CMR) spectrum (100MHz

Heavy water>: At least the following signals are observed.

77.8 (s, 17 7, 6 (s 1 74.4 (s

1 72.6 (s, 16.8.9 (s 161.4 (s

75.2 (d) 66.2 (t),. 1 (t)

60.7 (d) 57.9 (d).. 4 (d)

50.6 (t) 44.0 (t),. 4 (q)

19.1 (q) (PPi3). (However, s si ng let

d: doublet, t: triplet, q: quartet ₀ )

2. Antibiotics belonging to the genus Flexibacta TA N—542

,

' Is cultured in a medium, and the antibiotic TAN-54.2 is produced and accumulated in the culture medium, and then collected and collected.

54 2 or the manufacturing method of the idea.

O-M-PI ノ).-