JPS63264599A - Antibiotic substance tan-865 and production thereof - Google Patents

Abstract

NEW MATERIAL:A basic peptide based antibiotic substance TAN-865-A, B or C or salt thereof, having 2,3-diaminopropionic acid (4mol.), L-phenylalanine (1mol.) and threonine (1mol.) as constituent amino acids and capable of exhibiting the following physico-chemical properties. TAN-865A; Appearance; white solid. Molecular formula; C35H55N11O8. TAN-865B; Appearance; white solid. Molecular formula; C36H57N11O8. TAN-865C; Appearance; white solid. Molecular formula; C36H57N11O9, etc. USE:An antibacterial agent against Gram-negative bacteria. PREPARATION:A microorganism, belonging to the genus Cytophaga and having the ability to produce the antibiotic substance TAN-865A, B or C [e.g. Cytophaga sp. MK-30 strain (FERM P-9345)] is cultivated, preferably at 10-28 deg.C and pH 5-9 for 24-144hr.

Description

DETAILED DESCRIPTION OF THE INVENTION Field of Industrial Application The present invention relates to a novel basic peptide antibiotic and a method for producing the same.

BACKGROUND OF THE INVENTION To date, various antibiotics have been created and are being used in clinical settings as therapeutic agents for infectious diseases.

Various peptide antibiotics with various properties are known, and TAN-536A and B, for example, have been reported [JP-A-60-94088].

Problems to be Solved by the Invention Diseases caused by bacteria have been largely overcome by the development of therapeutic methods by administering antibiotics.

However, due to the long-term or large-dose administration of conventional antibiotics, there is an increase in diseases due to changes in the number of bacteria (microbial replacement phenomenon) and the emergence of resistant bacteria (tolerance phenomenon), or opportunistic bacterial infections due to weakened immunity. The increase in the number of infectious diseases has become a major problem in the current medical field of infectious disease treatment. To overcome this problem, the field is constantly developing new frameworks,
There is a need for substances that exhibit new biological activities or intermediate raw materials for synthesizing them.

Means for Solving the Problems The present inventors isolated a large number of microorganisms from soil for the purpose of searching for new substances, and conducted a separate search for the substances they produced. The microorganism is a species belonging to the genus Cytophaga; By culturing the microorganism in an appropriate medium, a substance exhibiting antibacterial activity mainly against Durham-negative bacteria is accumulated in the medium. TA11-865A, TA11-865A, TA11-865A,
I decided to call them B and C.

The present inventors completed the present invention as a result of further research based on these findings.

The present invention uses 2,3-diaminopropionic acid (4 mol), L-phenylalanine (1 mol) as constituent amino acids.
and L-threonine (1 mol), a basic peptide antibiotic TAN with the following physicochemical properties:
865 A, B or C and salts thereof, and belongs to the genus Zytophaga, and the antibiotic TAN-865A
, culturing a microorganism capable of producing B or C in a medium,
producing and accumulating antibiotic TAN-865 in the culture,
Antibiotic TAN-8 characterized by collecting this
65A.

The present invention provides a method for producing B or C and a salt thereof.

In addition, in this specification, antibiotics T, AN-
865A, B and C collectively as antibiotic TAN-86
5 or simply TAN-865.

In addition, in this specification, antibiotic TAN-865A
simply 'I' A N 865A and antibiotic T A
N 865B is sometimes simply referred to as TAN-865B, and antibiotic TAN-865C is sometimes simply referred to as TAN-865C.

The TAN-866 producing bacteria used in the present invention include:
Any microorganism may be used as long as it belongs to the genus Cytophaga and has the ability to produce the antibiotic TAN-865.

An example of a TAN-865 producing bacterium is Zytophaga sp, strain MK-30 (hereinafter referred to as r
It is sometimes abbreviated as "MK-30 strain". ) can be given.

The mycological properties of Cytophaga sp, MK-30 strain are as follows.

(a) Morphology Observation of cells cultured on broth agar slants at 24° C. for 5 days revealed that the cells were rod-shaped with a diameter of 0.5 to 1.0 μm and a length of 1.5 to 5.0 μm, and filamentation was often observed. There are no flagella. Demonstrates motility through briding. It does not form spores, Durham staining is negative, and it does not show acid fasting properties.

(b) Growth status on various media The cells were cultured at 24° C. and observed for 1 to 14 days.

■Meat juice agar plate culture Forms bicircular, convex circular, golden-edged colonies. No diffusible dye is produced.

■Juice agar slant culture: Shows medium spread-like growth, opaque, and pale yellow in color.

■ Broth liquid culture: Grows well in a turbid state and produces precipitates.

Forms a thinly closed ring.

■Meat juice gelatin puncture culture: Liquefies in layers.

■ Lidmus milk: Lidmus is not reduced and peptonization is not observed, but some coagulation is observed.

(c) Physiological properties ■ Nitrate reduction: negative ■ Denitrification reaction: negative ■ MR (methyl red) test: negative ■ VP (Voges Proscalaer) test: negative ■ Indole production: negative ■ Hydrogen sulfide production (TSI agar): Negative ■ Hydrolysis of starch: Positive ■ Utilization of citric acid (Christenzen's medium): Positive (Kosel's medium and Simmons' medium): Negative ■ Utilization of inorganic nitrogen sources i) Potassium nitrate: Negative ii) Ammonium sulfate: Negative [strain] Pigment production (King A, B and Mannitol yeast extract agar): Negative ■ Urease: Negative ■ Oxidase: Positive [Strain] Catalase: Negative ■ Range of production i) pH: pH 5, 1 to 9.3 It grows at a temperature of 9.5 to 27.5°C, but the optimum pH is 5.3 to 8.6°C.ii) The optimum temperature is 19 to 27.5°C.

■Attitude towards oxygen: Aerobic [stock] 0-F (oxidative fermentative)
Test [Hugh Leifso
n) Method]: Generation of acid and gas from non-degradable O-sugar: Acid Gas Availability (Peptone water) (Davis medium) L-arabinose --- D-xylose --- Shiri-glucose --- KiD- Mannose − −+ D-fructose − −+ D-galactose − −+ Malt sugar −−+ Milk sugar −−↓ Trehalose −−+ D-Sorvit −−± D-Mannitol − −+ Inosit − − − Glycerin − −+ Starch − − Decomposition of Tween 80: Positive ■ Decomposition of polysaccharide: Carboxymethyl cellulose: Negative, Colloidal chitin: Positive, Alginic acid: Negative, Agar, Negative 3D N, G of A C content (%, Tm method): 38.
The MK-30 strain having mycological properties of 8±1.0 or more was subjected to Bergey's Manual of Determinative + Hacteriolonow (Bergey's Manual of
termi++ativc Bacteriology
y) Comparing with the description in the 81st edition, it is a Durham-negative bacillus that does not have flagella and does not show motility using flagella, but it does show motility by briding and decomposes colloidal chitin aerobically, so it is a site. Cytophaga
) and identified the IK-30 strain as Cytophaga s.
p.

MK-30 (Cytophaga sp, MK-
30) and named it f2.

Cytophaga sp, stock was deposited with the Fermentation Research Institute (IFO) on April 9, 1986, with accession number IFO1460.
It has been deposited as 2. In addition, this microorganism was transferred to the Microbial Research Institute (FRI), Agency of Industrial Science and Technology, Ministry of International Trade and Industry on April 18, 1986, with the accession number FERM Pq.
3(+”;).

The properties of the Cytophaga bacteria used in the present invention are generally susceptible to change, for example, when exposed to ultraviolet light.

TAN-865, which is the subject of the present invention, can be easily mutated by artificial mutation methods using X-rays, chemicals (e.g., nitrosoguanidine, ethyl methanesulfonic acid), etc. Anything that has production capacity can be used in the present invention.

When culturing TAN-865 producing bacteria, carbon sources such as glucose, sucrose, maltose, blackstrap molasses, glycerol, oils and fats (e.g., soybean oil, olive oil, etc.), organic acids (e.g., citric acid, succinic acid, etc.) are recommended. acid, gluconic acid, etc.) are used as appropriate. Examples of nitrogen sources include organic nitrogen compounds such as soybean flour, cottonseed flour, corn steep liquor, dried yeast, yeast extract, meat extract, pep, ammonium urea disulfate, ammonium nitrate, ammonium chloride, and ammonium phosphate. and inorganic nitrogen compounds can be used. In addition, as the inorganic salt, for example, inorganic salts normally required for culturing bacteria such as sodium chloride, potassium chloride, calcium carbonate, magnesium sulfate, potassium phosphate, and disodium phosphate are used alone or in appropriate combinations. .

In addition, heavy metals such as ferrous sulfate and copper sulfate, and vitamin B
1. Vitamins such as hyotine are also added as necessary. Furthermore, antifoaming agents and surfactants such as silicone oil and polyalkylene glycol ether may be added to the medium. Helps the growth of bacteria, TAN 865
Organic or inorganic substances that promote the production of may be added as appropriate.

The culturing method may be the same as a general antibiotic production method, and solid culture or liquid culture may be used. For liquid culture, static culture and agitation culture are used.

Although any method such as shaking culture or aerated culture may be used, aerated agitation culture is particularly preferred. Also, the culture temperature is approximately 1
The range of 0°C to 28°C is preferable, and 1) l(
is cultured for about 8 hours to 168 hours, preferably 24 hours to 144 hours.

In order to collect the target antibiotic TAN-865 from the culture, separation means that are normally used to collect metabolites produced by microorganisms from the culture of the microorganisms are appropriately used. Antibiotic TAN-865 exhibits properties intermediate between water-soluble and fat-soluble, and since it is a basic substance, it is purified using this property. Furthermore, since TAN 865 is mainly contained in the culture filtrate, first, a filter aid is added to the culture solution and the culture solution is filtered or centrifuged to remove bacterial cells.

Advantageously, a method is used in which the obtained culture filtrate is brought into contact with a suitable carrier to avoid adsorption of the active ingredient, and then the effective substance is desorbed with a suitable solvent, and then fractionated and collected.

Supports include ion exchange resin, adsorption resin, and silica gel. Adsorbents such as activated carbon and cellulose, or carriers that utilize differences in the molecular weight of compounds such as molecular sieves, etc., are used. The elution solvent is the type of carrier.

Depending on the nature, for example, organic solvents or aqueous solutions of water-soluble organic solvents, such as aqueous acetone, aqueous methanol, aqueous butanol, or acids.

Aqueous solutions of alkalis, inorganic salts, organic salts, etc. alone or mixtures of these with organic solvents that are miscible with water are advantageously used.

Further, TAN-865 can be extracted from a neutral to weakly alkaline aqueous solution with an alcoholic organic solvent that can be separated from water.

Next, the mixture of TAN-865 purified by a combination of ion exchange chromatography, adsorption chromatography, organic solvent extraction, etc.
A, B and C, respectively.

More specifically, as a carrier, a cation exchange resin such as Amberlite IRC-50 or CG-
50 (manufactured by Rohm and Haas, USA), the antibiotic in the solution is adsorbed to the resin and eluted with an aqueous solution containing acid or salt. In addition, adsorbent resins such as Amberlite XAD-I (manufactured by Rohm and Haas, USA), Diamond Ion
P-20 (manufactured by Mitsubishi Chemical Industries, Ltd.) is used to adsorb the active substances in the solution, and a water pot containing an appropriate amount of acetone, methanol, etc., is used to absorb salts or It is eluted with a mixture with an acid, an aqueous solution containing it, or a buffer solution. When a silica gel such as Kieselgel 60 (manufactured by Merck & Co., West Germany) or a molecular sieve carrier such as Sephadex LI (-20 (manufactured by Farmana, Sweden)) is used, the active substance is adsorbed, and when an appropriate solvent such as ethyl acetate, Elute with chloroform, acetone, methanol or a mixture thereof.

TAN-865 can also be used in organic solvents that can be separated from water in neutral to alkaline aqueous solutions (1) 86-9), such as n-butanol, 1so-butanol, 1so-
Extracted with amyl alcohol.

Examples of carriers used in reverse phase preparative high performance liquid chromatography include YMc gel (manufactured by Yamamura Kagaku Kenkyusho).
, TSK gel (manufactured by Toyo Soda Co., Ltd.), etc.
A mixed solution of methanol, acetonitrile, or the like and a buffer solution is used as the moving phase.

Since TAN-865 is a weakly basic substance, it may form a pharmacologically acceptable salt with an acid. Examples of the acid used to form the salt include hydrochloric acid, sulfuric acid, etc. as mineral acids. Organic acids such as phosphoric acid, taste acid, acetic acid,
Examples include oxalic acid. The salt formation takes place according to methods known per se.

TAN-865A obtained in Example 2 described below.

The physicochemical properties of B and C (educts) are as shown.

TAN-8 6 5 A B C (1) Exterior
White solid White solid White solid (2) Specific rotation [α]2'-33.5° [α]2'-31
．． 3° [α]2'-38.8.

D D D (c=0.5Q
,H. O) (c=(1.53,11tO) (c-
=0.52. Ito) (3) Molecular weight measurement value 774 (M
! ()” 78g(λlt()” 78K
M+I)” (SI-MS method) TAN-865 ABC-(4)
Molecular formula C35H,,NllO8・CjgH,7NllO
．．・C381157NI,o,・(4)1.0)
(3H!0) (2HtO) (5) Elemental analysis value: Actual value (calculated value) (%) C, 49.57 (4
9.69) 51.37 (51.36) 52.3
2 (52.48)H, 7.50 (7.51) 7
．． 69 (7.54) 7.72 (7.46: iN
．． 18.22 (18.21) 18.53 (18JO
) 18.57 (18.70) 0, (24.5
9) (22.80) (21.35)(
6) UV spectrum - Underwater terminal absorption Terminal absorption Terminal absorption Shoulder near 24hm) (Shoulder near 24Onm x Shoulder near 240) (7
) IR Speccil. K B rat (am-'): 33
20, 2950 3330.2950, 3
320.2950.

1640, 1520.1640,152 (1, 1
640,152 (1.

1370, 1260, 1380.1260,
+360.1260.

1110, 700. +110.700. 1
110,700(8) H PLC : carrier, ODS
．． YMC-Pack A-312 (manufactured by Yamamura Chemical Research Institute), f
Hyperactivity (old 25% acetonitrile 10.01M phosphate buffer, 2 flow cycles/min TAN-865, ~ B C
RL=2.8 Rt=4.0
Rt = 4.4 (9) amino acids (to PA' (4)
mol) to DPA" (4 mol) DAPA" (4 mol)"'2,3-noaminopropionic acid (10)" CNNIR spectrum (loOMIiz,
in Dta, δppm) 180.4 (s)
180.3 (s) 180.0 (s) 178
．． 1 (S) 178.1 (s> ty
g, t (S) 177.4 (s) 177.5
(s) 178.1 (s) 17! i, 9 (
s) 176.7 (s) 176.7
(s)! 75.5 (s) 175.5 (s)
175.5 (s) 172.6 (s)
172.5 (s) 172.5 (s) 17
1.2 (s) 171.1 (s) 1
71.1 (s) 139.3 (s) 139.
4 (s) 139.4 (s) 131.8 (
s) 131.9 (s) 131.9
(s) 131.8 (d) 131.7 (d)
131.7 (d) T, 8 N-865 A B C
131.8(d) 131.7(d)
131.7 (d) 130.2 (d)
130.0 (d) 130.0 (d)1
24.5 (d> 124.2 (d)
124.2 (d) 69.8 (d) 6
9.8 (d) 69.8 (d) 62.7
(d) 62.6 (d) 62.6
(d) 59.1 (d) 59.1 (d)
59.1 (d) 57.0 (d)
57.0 (d) 57.0 (d) 56.
8 (d) 56.7 (d) 56
．． 7 (d) 56.1 (d) 56.0
(d) 56.0 (d) 55.4+ (d)
55.6 (d) 55.6 (d
)46.0 (t) 46.0 (t)
46.1 (t) 45.7 (t) 4
5.7 (t) 45.7 (t) 42.6
(t) 42.6 (t) 42.6
(t) 40.5 (t) 40.5 (t
)39.1 (t) 39.1 (t)
39.1 (t) TAN-865 A B C
38,6 (t) 36.4 (t)
38.8 (t) 33.4 (t) 36.
3 (d) 29.9 (d) 27.9 (t
) 34.8 (t) 26.1 (
t) 24.5 (t) 31.4 (t)
24.7 (q)21.9 (li 21.
9 (q) 21.9 (Q) 16.1 (Q
) 21.0 (Q) 14.5 (
Q) i45 (q) 14.5 (q)13.
5 (q) (11) Color reaction, same positive for all three components, ninhydrin, Brave Reback.

Ehrlich (acidic), dimethylaminohenzaldehyde, phosphomolybdic acid reaction (12) Distinction of substances - All three components are basic substances (i3) soluble, three components (same soluble, water, methanol, dimethyl sulfogizide) mm: Ethyl acetate, chloroform From the comparison of the above results and the physicochemical properties of known compounds, the present antibiotics TAN-865A, B, and C are considered to be new compounds.

Next, describe the biological properties of TAN-865. T
The antibacterial properties of AN-865A, B and C against various bacteria are shown in Table 1.

Table 1 Minimum inhibitory concentration (q+c) for antibacterial activity of TAN-865, (Note) Test bacteria
μg/enrichia coli CPC202512
,,56,25 Klebnoella pneumoniae
> 100 > 100 > 1001PO33
17 7 Trobacter freundii > 100
> 100 > l [101F012681 Acinetobacter calcoaceticus 25
25 25IFO13006 Proteus mirabilis ATCC21+00 >
100 > 100 > 100 Proteus morganii + FO3963 > 100 > too >
100 Neudomonas aeruginosa
25 2525IF0 3080 Staphylococcus aureus > 100
> 100 > IaOFDA 209P - (Note): Medium: YA medium (Bacto Antibiotic Medium 3.+7.5g, Bacto Yeast Extract: 5g, Bacto Agar 20g.

Distilled water 1000 ml, pH: 7.0) Inoculum amount: 10'CFU/J of bacterial solution was used.

Furthermore, TAN-865A, B, and C have an antibacterial activity-enhancing effect on various antibiotics and antibacterial substances, as shown in Table 2. That is, TAN-865A, B and C contain lankacidin C, novobiocin, micamycin,
It enhances the antibacterial activity of antibiotics such as rifampicin, chlorampheniconil, rifamycin Sv, actinomycin D6 and penicillin G, and antibacterial substances such as nalidixic acid.

Table 2 Antibacterial activity enhancement effect of TAN-865A, B and C on various antibiotics and antibacterial substances Growth inhibition circle diameter (mm) (Note) Additive agent (μg/-) TAN-865A TAN-86, lB TAN-86
5C antibiotic (μg/d) 0 100
100 100 Lancasidin C1000-+-2
12422 Novobionn 100 - (
15) (17) (1g) Mikamycin
1000 - 13.5' 17.5
17 Nalidixonic acid 100 15 21
22 22 riff 1 pinon 100
10 19.5 21.5・ 21.5 Chloramphenicol 100 + 1617.5 1
7.5 Lihuamynon too -1517
17,5 actinomain D 1000 −
13 13 13 Penicillin G
1000 12.515.5 15 17.5
(Note) Medium: 10 g of polypebuton, 10 g of meat extract.

1g of table salt, 15g of agar, 1000ml of distilled water, pH 7.0 Test bacteria, Escherichia coli
colt) NIHJ JC-2 Test method: Paper disc method (diameter 8 mm) Judgment Knee indicates no growth inhibition, ± indicates inhibition circle of 10 mm or less, () indicates incomplete inhibition.

Also, Pseudomonas aeruginosa using mice
In the P-9 infection experiment, TA,N-865C showed efficacy (EDs: 160 mg/kg) when administered subcutaneously.

As is clear from these facts, TAN-365 is effective against Durham-negative bacteria either alone or in combination with other antibiotics.

The toxicity of TAN-865 is low, e.g. TAN-8650
The acute toxicity of L
D 50400 mg/kg or more (mouse).

Therefore, TAN-865 can be administered parenterally (as an injection, suppository, etc.) as a single agent or as a combination agent to mammals (
It can be administered to (cows, pigs, dogs, monkeys, humans) as a preventive/therapeutic agent for infectious diseases. r Kotoba TA
N-865 was dissolved in physiological saline and administered as TAN-865 to patients infected with Pseudomonas aerogyno'.
Administer 100 mg/kg.

Similarly, it can be used as a sterilizing and disinfecting agent. In this case, for example, a mixture of TAN-865 or fanpicin is prepared as an aqueous solution at a concentration of 10 to 100 μg/d, for example, in a birdcage, laboratory equipment, in the hands of two people, etc.
It can be used by spraying or applying it for the purpose of sterilizing and disinfecting the feet.

Example 1 Cytophaga sp grown on nutrient agar slopes,
strain MK-30 (I FOI 4620) with 2% glucose, 3% soluble starch, and 1% raw soybean flour.

Corn steep liquor 1%, polypeptone 0.5
%1 salt 0.3%, precipitated calcium carbonate 0.5% (p
2Q containing 500% of a medium (pH 7.0) containing H7.0) was inoculated into a Yosaka flask and cultured in a reciprocating tray at 24°C for 48 hours. The entire amount of this culture solution was inoculated into a Z50Q, C''1 tank containing 30C of the above medium supplemented with 0.05% Actocol (manufactured by Takeda Pharmaceutical Co., Ltd.), and the aeration rate was 30Q/min at 24°C. Culture was carried out for 48 hours under the conditions of 200 revolutions/min.The entire volume of the culture solution was mixed with a medium containing 2.0% dry mother paste (Kanebuchi (1=manufactured by Gakukogyo Co., Ltd.) and 0.1% glucose (pI). Volume 220 (6,0) containing medium TOOC supplemented with 0.05% Actocol
0 (2 tank was inoculated and the aeration rate + oO (/
Minutes.

Cultivate for 66 hours at 170 rpm.

Example 2 Hyflo Super Cell (manufactured by Noyon's Manville, USA) was added to the culture solution (I20ρ) obtained in Example 1 to obtain a filtrate of filtration 1 (100υ).
Amber light IFt (, -50 (H+ type,
2, 5 (2) was subjected to column chromatography. Elute the antibacterial active ingredient with 0.5N hydrochloric acid (15ρ),
The eluate was extracted with 1so-butanol (10i2), and the extract was concentrated.

The concentrated liquid (lρ) was added to Diaion HP-20 (100-2
00 mesonyu) column chromatography.

50% methanol water and 50% methanol10. OI
Among the eluted fractions of N-hydrochloric acid, the fractions with strong antibacterial activity were collected,
It was concentrated and lyophilized to obtain a powder (18.4 g). This powder was subjected to preparative high performance liquid chromatography using YMC gel as a carrier to obtain a 16% acetonitrile 70.02M phosphoric acid solution (pH 3.0). It was eluted. After removing acetonitrile, each of the obtained four fractions was extracted with 1so-butanol, and the extract was concentrated and dried.
Purified powders of (585 mg), B (432 mg), B and CCI complex (4.78 g) and c (3.1 g) were obtained.

Effects of the Invention The antibiotics TAN-865A, B and C of the present invention have antibacterial action and the action of enhancing the antibacterial activity of various antibiotics,
It is useful as a preventive and therapeutic agent for infectious diseases.

[Brief explanation of the drawing]

Claims (1)

[Scope of Claims] [1] Contains 2,3-diaminopropionic acid (4 mol), L-phenylalanine (1 mol) and L-threonine (1 mol) as constituent amino acids, and has the following physicochemical properties Basic peptide antibiotic TAN-865A, which shows
B or C or a salt thereof (1) Antibiotic TAN-865A (a) Appearance: White solid (b) Molecular formula: C_3_5H_5_5N_1_1O_9
(c) Infrared absorption spectrum (cm^-^1, KBr tablet): 3320, 2950, 1640, 1520, 136
Absorption is shown at 0, 1260, 1110, and 700. (d)^1^3C-NMR spectrum (δppm, heavy water): 180.4 (s) 178.1 (s) 177.
4 (s) 176.9 (s) 175.5 (s) 172
．． 6 (s) 171.2 (s9 139.3 (s) 13
1.8(s) 131.8(d) 131.8(d)
130.2(d) 124.5(d) 69.8(d)
62.7(d) 59.1(d) 57.0(d) 56
．． 8(d) 56.1(d) 55.6(d) 46.0
(t) 45.7(t) 42.6(t) 39.1(
t) 38.6 (t) 33.4 (t) 27.9 (t)
24.5(t) 21.9(q) 16.1(q) 1
The signal is shown in 4.5(q). (2) Antibiotic TAN-865B (a) Appearance: White solid (b) Molecular formula: C_3_6H_5_7N_1_1O_9
(c) Infrared absorption spectrum (cm^-^1, KBr tablet): 3330, 2950, 1640, 1520, 138
0, 1260, 1110, 700 (d)^1^3C-NMR spectrum (δppm, in heavy water): 180.3 (s) 178.0 (s) 177.
5 (s) 176.7 (s) 175.5 (s) 172
．． 5 (s) 171.1 (s) 139.4 (s) 13
1.9(d) 131.7(d) 131.7(d)
130.0(d) 124.2(d) 69.8(d)
62.8(d) 62.6(d) 59.1(d) 57
．． 0(d) 56.7(d) 56.0(d) 55.6
(d) 46.0(t) 45.7(t) 42.6(
t) 40.5 (t) 39.1 (t) 36.4 (t)
36.2(d) 34.8(t) 31.8(t) 2
1.9 (q) 21.0 (q) 14.5 (q) 13.
5(q) shows the signal. (3) Antibiotic TAN-865C (a) Appearance: White solid (b) Molecular formula: C_3_6H_5_7N_1_1O_9
(c) Infrared absorption spectrum (cm^-^1, KBr tablet): 3330, 2950, 1640, 1520, 136
0, 1260, 1110, 700 (d)^1^3C-NMR spectrum (δppm. in heavy water): 180.0 (s) 178.1 (s) 178.
1 (s) 176.7 (s) 175.5 (s) 172
．． 5 (s) 171.1 (s) 139.4 (s) 13
1.9(d) 131.7(d) 131.7(d)
130.0(d) 124.2(d) 69.8(d)
62.6(d) 59.1(d) 57.0(d) 56
．． 7(d) 56.0(d) 55.6(d) 46.1
(t) 45.7(t) 42.6(t) 40.5(
t) 39.1 (t) 38.8 (t) 29.9 (d)
26.1(t) 24.7(q) 21.9(q) 1
The signal is shown in 4.5(q). [2] Belongs to the genus Cytophaga and is an antibiotic TAN-86
Antibiotic TAN-865 is produced and accumulated in the culture by culturing a microorganism capable of producing 5A, B or C in a medium, and the antibiotic TAN-865 is collected.
865A, B or C or a salt thereof.