JPS6094088A - Antibiotic substance tan-536a,b, its production and microorganism - Google Patents

Abstract

PURPOSE:To produce novel antibiotic substance TAN-536A and/or B, by culturing a microbial strain belonging to Cytophaga genus and capable of producing said antibiotic substance, and collecting the antibiotic substance produced and accumulated in the culture product. CONSTITUTION:A microbial strain capable of producing antibiotic substance TAN-536A and/or B, e.g. Cytophaga sp. YK-33 strain, is cultured in a medium containing a C source such as glucose, an N source such as soybean flour, an inorganic salt such as NaCl, etc. The cultivation is carried out preferably under aeration at 15-35 deg.C and about 4-8pH. The objective antibiotic substance TAN- 536A and/or B can be separated from the cultured product by conventional separation means. These antibiotic substances are purified taking advantage of their intermediate properties between water-soluble and fat-soluble properties, and weak basicity.

Description

[Detailed description of the invention] The present invention is novel antibiotics TAN-536A and B.

Regarding its manufacturing method and microorganisms.

The present inventors isolated a large number of microorganisms from soil for the purpose of searching for new antibiotics, and conducted a separate search for the antibiotics produced by the microorganisms. It is known that the microorganism belongs to the genus Cytophaga, and that by culturing the microorganism in an appropriate medium, an antibiotic that exhibits antibacterial activity and exhibits a synergistic effect when used in combination with other antibiotics is accumulated in the medium, Based on its physicochemical properties, it has been recognized that the antibiotic can be used as a new antibiotic, and has been designated as Antibiotic I! t'I''AM-536. Since TAN-536 consists of at least two components, we named them TAN-536A and B. Based on these findings, the present inventors conducted further research. As a result,
The invention has been completed.

The present invention provides (1) antibiotic TAN-536A, B or its L; (2) antibiotic TAH belonging to the genus Cytophaga;
- Antibiotic ITAN-536A and/or B characterized by culturing 536A and/or B producing bacteria in a medium, producing and accumulating antibiotic TAN-536A and/or B in the culture, and collecting the antibiotic TAN-536A and/or B. and (3) Cytophaga sp, YK-33, which is positive for the availability of citric acid and D-mannitol.

In addition, in this specification, the antibiotic lαTAN-536
A and B may be collectively referred to as antibiotic TAN-536 or simply TAN-536.

In addition, in this specification, antibiotic TAN-536A
simply TAN-536A and antibiotic TAN-536B
are sometimes simply referred to as TAN-53 (3B).

Antibiotics used in the present invention'rAN-536A
and/or B-producing bacteria include Cytophaga (
Any microorganism may be used as long as it belongs to Cytophaga ffjl and has the ability to produce the antibiotic TAN-536A and/or B.

An example of a producing bacterium for the antibiotic TAN-536A and/or B is Hiro, for example, Cytophaga sp.
3 strain (hereinafter sometimes abbreviated as "YK-3.3 strain").

The mycological properties of Cytophaga PJ's SP YK-33 strain are as follows.

(a) Morphology Observation after culturing for 2413.5 days on a broth agar slant shows that
#l cells are 0.7-1.0 pm in diameter and 1.5-4 pm in length.
．． It has a rod shape of 5 μm and shows no polymorphism. lacks mobility,
-Has no hair. 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.

■Juice agar plate culture: Forms circular, convex, or golden-edged colonies. The surface is smooth, opaque, and white to gray-yellow in color. No diffusible dye is produced.

■ Broth agar slant culture g1: Shows moderate wart-like growth;
Opaque, pale gray-yellow color.

■ Broth liquid culture: Grows in a thin turbid state and produces a small amount of precipitate. Does not form bacterial membrane.

■ Meat juice gelatin puncture culture Liquefies into two layers.

■Lidmus filtration: Lidmus is not reduced, and neither peptonization nor coagulation is observed.

(c) Physiological properties ■ Nitrate reduction: Positive ■ Denitrification reaction: @ ■ MR (Methyl Red) test: Negative ■ vP (Voges-Proskauer) test: Negative ■ Indo μ formation: Negative ■ Hydrogen sulfide Formation (TSI agar): Negative ■ Starch hydrolysis M, Niyang + Cob ■ Utilization of citric acid (Kose μ, Christensen and Simmons media): Positive ■ Utilization of inorganic nitrogen sources 1) Potassium nitrate: Negative i) Ammonium sulfate: Negative ■ Pigment production (King A, B and Mannitol yeast extract Makuten): Negative ■ Urease: Negative ■ Oxidase: Positive ■ η Crase: Amphoteric ■ Growth range 1) pH: pll 5.0 ~ It grows at 8.5, but
The optimum pH is 5.6-7.1°■) It grows at a temperature of 26.0-28.5 t, but the optimum temperature is 17-23°C.

0 Attitude towards oxygen: aerobic or facultative anaerobic 00-1
'(Oxidate Fufu Armentate Tape) Test [Hugh Leifson Method]
: Production of acid and ganu from fermentative sugars: L-arabinose -- D-xylose -- D-glucose 10 -- + D-mannose 10 --+ D-hookdose + -- D-galactose ± --+ Malt Sugar −+ Sucrose −− Lactose −−+ Trehalose −−± D-Sorvit − − D-Mannitol −−+ Inoshift −− Glycerin −− Starch − − More than 10 mycological properties The XK-33 strain having
erminative Bacteriology)
In comparison with the description in the 758th edition, it is a Guhum-negative bacillus, has no flagella, does not exhibit flagellar motility, but exhibits motility by briding, is aerobic or facultatively anaerobic, and is 〇-F.
The test is fermentation, decomposes chitin, and the optimum temperature for growth is 3.
Because it grows below 0°C and in the absence of salt,
The YK-33 strain was identified as Cytophaga sp.
33 (Cytophaga sp, YK-33).

Furthermore, no Cytophaga bacteria were found to be positive for citric acid and D-mannitol utilization, and therefore, this property is characteristic of the YK-33 strain.

The YK-33 strain was deposited with the Fermentation Research Institute (IFO) on October 5, 1988 under accession number IF014289. In addition, this microorganism
Ministry of International Trade and Industry, Agency of Industrial Science and Technology, Microbial Technology Research Institute (F'
RI) 412: Deposited under accession number F'ERMp-7#da.

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

It can be easily mutated by artificial mutation means using X-rays, chemical substances (e.g., nitrosoguanidine, ethyl methanesulfonic acid), etc., and the TAN that is the subject of the present invention, regardless of the paternal strain. Anything capable of producing -53OA and/i or B can be used in the present invention.

When culturing the bacteria producing the antibiotic TAN-536A and/or B, the carbon source was
course, sucrose, maμdose.

Wild goose molasses, glycerol, fats and oils 4A1 (e.g., soybean oil, olive oil, etc.), organic acids (e.g., citric acid, succinic acid,
Glucon M, etc.) are used as appropriate.

Examples of sources include soybean flour, cotton seeds, corn, etc.
Nuteig liquor, dry fholfj+, (1)1
．． Organic and inorganic nitrogen compounds such as u extract, meat extract, peptone, urea, ammonium sulfate, ammonium nitrate, ammonium chloride, and ammonium phosphate can be used. Examples of inorganic salts include sodium chloride, potassium chloride, calcium suboxide, magnesium sulfate, potassium phosphate, and disodium phosphate, which are normally necessary for culturing bacteria. i! Salts may be used alone or in appropriate combinations.

In addition, heavy metals such as ferrous sulfate and copper sulfate, and vitamin B
Vitamins such as T and biotin are also added as necessary. Furthermore, antifoaming agents and surfactants such as silicone oil μ and boria μ xylene glycol μ ether may be added to the medium. Helps the growth of bacteria, TAN-536A
And/or organic or inorganic substances that promote the production of B 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.

Any method such as shaking culture or aerated culture may be used, but aerated agitation culture is particularly preferred. Also, the culture temperature is approximately 15C.
The temperature is preferably in the range of ~35°C, and the pH of the medium is in the range of approximately 4 to 8 for approximately 8 to 168 hours, preferably 24 to 8 hours.
Incubate for 144 hours.

In order to collect the target antibiotic WTAN-536A and/or B from the culture, separation means that are normally used to collect metabolites produced by microorganisms from the culture of the microorganisms are appropriately used. Antibiotics TAN-536A and B exhibit properties intermediate between water solubility and fat solubility, and are weakly basic substances, so they are purified using this property. Also TA
Since N-536A and B are mainly contained in the culture p solution, first, a temporary 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 adsorb the active ingredient, and then the effective substance is de-responsed with a suitable solvent and then fractionated and collected.

As the carrier, an adsorbent such as an adsorbent resin, silica gel, activated carbon, or cellulose, or a carrier utilizing the difference in molecular weight of compounds such as a molecular sieve may be used. The elution solvent varies depending on the type and properties of the carrier, and includes, for example, an organic solvent or an aqueous solution of a water-soluble organic solvent, such as aqueous acetone, aqueous methanol, aqueous butanol, acid, or alkali.

S-free mounds, mixtures of aqueous solutions of organic salts, etc., and organic solvents that are miscible with water, and the like are advantageously used.

Further, TAN-536A and B are extracted from a neutral to weakly alkaline aqueous solution with an alcoholic organic solvent that can be separated from water.

Next, TAN-53 (3A,
The mixture of B is isolated into T'AN-A and B, respectively, by reverse phase preparative high performance liquid chromatography.

In more detail, adsorbent resins such as Amberphyto The active substance in the solution is adsorbed and eluted with a mixture of an ammonium medium and an aqueous solution, i.e., water containing acetone, methanol, etc., or a mixture of an aqueous solution or buffer containing salts, acids, etc. In addition, silica gel such as Kieserge/I/60 (manufactured by Melli AG, West Germany) or molecular sieving carrier such as Sephadex LH-2
0 (manufactured by Fapman, Sweden), the active substance is adsorbed, and a suitable solvent such as ethyl acetate μ,
Elutes with chloroform, acetone, methanol, or a mixed solution of these.

In addition, TAN-536A and B are organic solvents that can be separated from water in neutral to alkaline aqueous solutions (pH 6 to 9), such as n-butano-μ, 1sO-butano-μ, 180-
It is extracted by AmiμAμCoμ, n-AmiμAμCoμ, etc.

Examples of carriers used in reverse phase preparative high performance liquid chromatography include TSK Ge/I/ (manufactured by Toyo Soda Co., Ltd.), MMC GeLV (Yamamura Kagaku Kenkyujo! 11), etc.; A mixture of methanol, acetonitrile μ, etc. and FWk liquid is used.

Since TAN-536A and B are weakly basic substances, they may form pharmacologically acceptable salts with acids. Examples of vinegar used to form the salt include mineral acids such as hydrochloric acid, sulfuric acid, and phosphoric acid, and organic acids such as formic acid, acetic acid, and oxalic acid. The salt formation takes place according to methods known per se.

TAN-536A and B obtained in Example 2 (described later)
The physical and chemical properties of the free form are as follows.

TAN-53 (5 A 73 (1) White powder White powder (2) Specific rotation cα] Weight-6MQlt=20' (a
) Static-70,5120' (in methanol) (c-1,
o) (c-t, o) (3) Elemental analysis value C:, 55.
32: t2.0 55.28: t2.0 (Real 111 [)
H, 7, 73shi 1.0 7.69f: 1. ON, 17
．． 76±1.5 17.90 1.5 (4) Molecular role measurement it 915 (MH+) 901 (Mu+) (SI
MS method) 953 (MK”) 939 (MK+)A
B (6N-ncl Threonine Threonine 105t
:, 15 o'clock iL At least the following valine valine (7) ultraviolet absorption smooth terminal absorption terminal absorption
(8) Infrared absorption) # (KB
r), main absorption (c'suA; 3350,3060
,2960,2940,2870゜1660.1530
．． 1460, 1380.1270゜1230.1180
．． 1150,1120.1080゜930.840,7
00,650.490 (see Figure 1) B; 3350
,3060,2960,2940.2870°1660
．． 1530, 1440.1380.1270゜1230
．． 1180, 1110, 1080.930, 840°7
40.700,650,580,490 (See Figure 2) (9) 13C-Nuclear Magnetic Resonance Spectrum (100MH
2, DMSO-46) At least the following signals are observed.

δppm A; 173.70, 173.55, 173.42,
172.25°171.55, 171.09.170.
64,169.39゜165.31.13&21,13
1.44, 128.93° 128.12, 126.27
, 117.22. 66.01°60.28, 57.
93, 56.35, 54.84, 54.57.

53.41, 52.35.43.93, 43.08
．． 42.56.

38.33, 36.14, 35.08, 30.05.2
8.85°27.27, 26.44, 25.31
, 22.41, 20.51.

19.19.18.15, 11.92°B i 173
．． 67.173.54, 173.41, 172.27°171.57, 171.11, 170.66.169.
39°165.33,138.21.131.42,1
28.95°12EL13.126.27,117.2
4. 66.01゜60.27, 57.93, 56
．． 38, 54.83, 54.56.

53.38, 52.34, 43.91, 43.06.4
2.55゜40.23.36.13, 35.09.31
．． 14.30.05゜28.55, 28.37, 2
5.30, 21.97, 20.51.

19.20, 18.16, 13.83, 11.9
3゜ (Incorporated Foundation) color reaction, AB both positive; ninhydrin, Brave Revac, dimethymu-aminobenzadehyde, phosphomolybdic acid reaction I thin layer chromatography, spot film.

Cellulose (manufactured by Tokyo Kasei Kogyo Co., Ltd.) f 1# Medium thread AB Acetonitrile/L/:Water (4:1) 0.3 0.3
(2) Liquid chromatography (manufactured by Waters).

USA), -111 salary; YMCA-312 (Yamamura Chemical J
Nf Institute a), Mobile Layer i 759jjri/-IL'10
．． 01M phosphate buffer (PH7,3), 2wt
/ mtn.

Rt (min, ) A B 6, 6 5.0 Stability: Both AB are stable in aqueous solution with pU of 3.7.9.

Classification of Substances: Weak salts;
(Both) TAH-536A and B are new compounds. it is conceivable that.

Next, the biological properties of TAN-536A and B will be described. The antibacterial properties of TAN-536A and B against various bacteria are 9 as shown in Table 1.

Table 1 Antibacterial activity of TAN-536A and B
Near: 1 Re MXII with JC-225100V Gusiewo Pneumoniae IFO331725100V Trobacter Proindi Dt) 12G81 25 10
0ffi4 Neph Typhimurium IPF 12529
50 50 Comamonas terrigena nη13299
25 1007V net/9/-・force fr7-qf ixyo 13006 3.13 3aa alcaligenes-
7m Charis IFD l3111 12.5 125 pu"C'Ttsu-t?biliX A!21100 >100
>100 Propius Prius is a squirrel IK) 3988
＞100 ＞100 ν Theus Morga Tomi-Fuki υ396
8 100 >100'-L-V-etIZ・xmetsu4
1m3080 50 50 (c7f'7・YAs?/l
/J If012648 100 100 SIFIA:
x Cas-1tvtxFDA 209P 100 10
0 Micrococcus lutecus EFO12708505
0/<f/Li-zuffU7. PCI 219 1
00 100 (Note): wI base: YA medium (Bacto Antibiotic Medium 3 i 17.59,
Pact yeast extract; 5g, Pact agar 20□, distilled water 100 (LH/, pH 7,0) Amount of inoculated bacteria: l 0
A bacterial solution of 6 cFu/ptl was used.

As is clear from Table 1, TAN-536A and B are effective against gram-negative bacteria and gram-negative bacteria.

The acute productivity of TAN-536A or B was 2% in T, D5Q 400 Iv/in both daily administration.
(h (mouse) is 6 ka, every sex is low.

Therefore, TAN-536At or B can be used as a disinfectant or disinfectant. For example, TAN-5
36A or B as an aqueous solution with a concentration of about 10 to 100 μ9/ml, for example, a birdcage, a fruit wtn tool, 9 people's hands 1
It can be used by spraying or applying it for the purpose of sterilizing and disinfecting the feet.

Furthermore, TAN-536A and B have an antibacterial activity-enhancing effect on various antibiotics, as shown in Table 2.

namely, TAN-536A and Bii erythromycin, oleandomycin.

maridomycin, leucomycin, tylosin.

Macrocyclic lactone antibiotics such as Funkacidin C.

Ansamycin antibiotics such as amycin SV, rifampicin, tribomycin, and streptovaricin, caryomycin, and fusidic acid.

actinomycin D, chromomycin A3, novobiocin, nalideixic acid, pleomycin.

It enhances the antibacterial activity of antibiotics such as chloramphoenicol μ, micamycin, and chlortetofcycline.

Table 2 Erythromycin 1001-20 Oleandomycin 100 1-10 Maridomycin c xooi-15 0 Icomycin 100 1-21 Tylosin 1
oo i 13 Funcasidin C10024 Rifuamycin BY Zoo □ -26 Rifuampicin 1oo: 11 32 Tribomycin y loo
-265 Streptovaricin 1ooo H-1
9 Carry My 7in 1000! xas fucidin 1
4!12too l -17,5 actinomycin D1
00 1-19 (Note 1) TAN-536A and B
Mixing ratio (2,6:1) (World 0 medium 2 polypeptone 10g, meat extract 10q9 salt If, agar 15g, distilled water 1000I++/, pH 7
,0 Test bacteria: Escherichia coli (Escheri-chi)
acoli) NIHJ JC-2 test method; Paper disc (diameter 8 mm) Method judgment: Diameter of growth inhibition circle (
Expressed in mm).

(-) indicates no growth inhibition. means.

Table 3 shows the results of investigating whether this antibacterial activity enhancing effect is additive or synergistic.

Table 3 Synergistic effects of TAN-536A and B with various antibiotics Tylosin i >400>400>400 25 1
25 113'<0313□ Funkacidin C] 200 100 50 1.56
(χ78 039, <D, 258)! j；j纬4yy
Y; 12,5 12,5 12.5 1% 039
02 4) 375 Caryomycin, >400>400
400 12510.054) D5, <0.281
Oninovobiocin, >100 25 3.13 1,56
α39 0f125 1 <0.156 (Note 1) TA
Mixing ratio of N-536A and B (+, 8:1) (Song) medium; YA medium (see footnotes in Table 1), test bacteria: Engineering 7 Erichia coli I) TJ JC-2 bacterial quantity: 1Q6
A bacterial solution of CFU/We was used.

P' I C index: The single MIC of A is a.

B's independent MIC.

When used together, the MIC of OA is a The MIC of B when used together is b be.

It is said that there is moderate synergy when the F engineering C index is 0.5 to 0.25, and strong synergy when it is 0.25 or less [
Antimicrobial Agent and Chemotherapy
and Chemotherapy), 15XJ
e KS684 page.

[1979], it is clear that TAN-536A and B exhibit strong synergy with both of the antibiotics used.

Next, the therapeutic effects of TAN-536 and B on experimental mouse I8 infection are shown in Table 4.

Table 4: Therapeutic effect of TAN-536A and B on actual maternity infections Infecting bacteria Administered drug Administration method ``D50, lIVkg (
Note) Mixing ratio of TAN-536A and B (L, S:
1) As is clear from Table 4, the combined use of both TAN-536A and B and erythromycin shows a significant therapeutic effect.

Therefore, the present antibiotic TAH-536A or B is
It can be used as an antibiotic action enhancer. For example, antibiotics such as erythromycin can be
Pi
n milk'ah! i Of course ((>11, mouse.

It can be administered to rats, dogs, rabbits, and humans as a bacterial infection therapeutic agent for the purpose of treating I51 bacterial infections.

For the above-mentioned administration, both antibiotics are formed into tablets, capsules, powders, etc., using a method known per se, together with pharmacologically acceptable carriers, excipients, and diluents, and administered orally. It can be formulated into an injection and administered parenterally.

When using the antibiotics TAN-536A and B of the present invention, a mixture thereof may be used, and the mixing ratio of TAN-536A to B is preferably about 1 to 3 to 1.

Next, the present invention will be explained in more detail with reference to Examples.

Percentages are weight/volume % unless otherwise specified.
shows.

Example 1 A strain of Cytophaga sp. YK-33 (IP'0 14289.FERM P-n3/g) grown on a nutrient agar slant was grown on 2% glucose.

3% soluble starch, 1% raw soybean flour, 1% corn steep liquor, 0.5% polypeptone (Daigo Eikan Kagaku Kogyo Co., Ltd. 111I), 0.3% salt, 0.5% precipitated calcium carbonate. Medium containing (pH 7,0) 40
Inoculate two Erlenmeyer flasks containing 200 seeds/containing
Shaking culture was carried out at 4tE for 48 hours, and the culture was used as an inoculum.

Next, corn starch 196. A medium (pH 7,0) 2500*/ containing 2 ppm of Corn Gluten Meal/L/3% and Coba Chloride/LZ) was dispensed into 200 gt triangular Fufusco vessels in an amount of 40 tnt each, and heated at 120°C.
The seeds were sterilized by autoclaving for 20 minutes, inoculated with 1 rtte of seeds, and incubated at 24° C. for 90 hours at 200 rpm.

Example 2 The culture solution obtained in Example 1 was separated by centrifugation, and the supernatant ('1
01. p11B, 5) was obtained. After adjusting the p solution to p) 17.0, it was subjected to column chromatography using 7 mm/(-fi)XAD-I [(200 ff/) and eluted with 80% methanol-μ water (0.6 J). After methanol-μt-distillation in the eluate, the aqueous layer was extracted with n-butanol (300 ml?), the extract was concentrated, ether was added to the concentrated solution, and a crude powder (x
, 14y) were obtained. Coarse powder/l/(22g)
Column chromatography with 60% Mell/-
)V/ethylacetate/L/(270ml). Rom the eluate, add Eteμ to the concentrate, and add powder (466
4) was obtained. This powder was subjected to high performance liquid chromatography using TSK gel, and 65% methanol 10.01
Elution was performed with M phosphate buffer (1) H7,4). The two peak fractions were extracted with n-butanol, the concentrate was extracted with n-butanol, the concentrate was triturated with ether, and the concentrate was triturated with TIN-536A (118W) and B
Obtain (4311v).

Example 3 Cytophaga sp. YK-33 (IFO14289, F'ERM P-7) grown on nutrient agar slant
3/da) strain was added to the microorganisms of Cornu Cornu 296, 3% soluble starch, 1% raw soybean flour, corn steep liquor ig.
6. Polypeptone 0.5%.

Salt 0.3*, precipitated carbonate, 5% (pH 7
, 0) was inoculated into a 2J-capacity slope flask containing 500 g of a medium (pH 7, 0) and subjected to reciprocating shaking w5qI at 24°C for 48 hours. Add the entire amount of this culture solution to the above medium and add 0.05g/1/(manufactured by Takeda Pharmaceutical Co., Ltd.)
was inoculated into a 1ic5os tank containing medium 301 supplemented with 301, and aerated at 24°C [301j min.

The cells were cultured for 48 hours at 200 revolutions/min. ) Total amount of culture solution, 1% corn starch, corn gluten,
A medium (pH 7,0) containing 2 ppm of miR396 m chloride (tz<μ) was inoculated into an 8 Ja2 o□j tank containing 100 J of a medium supplemented with Actocol 0.054, and incubated at 24°C with an aeration rate of 10017 min. The cells were cultured for 90 hours at 170 revolutions/min.

Example 4 Hyflo Super 7/l/(manufactured by Johns Manville, USA) (
as#) was added and filtered to obtain a solution (786). After adjusting the p solution to p) I 7, add Amberlite XAD-II (s
p) was subjected to column chromatography. 809
The active M active ingredient was eluted with 6 methanol water (20 J), and the eluate was concentrated by 1'g4. ikA condensate (2,8j) as n-
Extraction was carried out with butano-/I/(2j), the extract was concentrated, and ether was added to precipitate a crude powder (66.2 g). The crude powder was subjected to column chromatography using silica gel/l/(241) and eluted with 60-80% methano/I//acetic acid. The eluate was concentrated to dryness, and the dried product was 4096
Dissolve in methanol water and add lf earion HP-20 (1
The resulting product was subjected to column chromatography using 00-200/1 VYU).

Among the elution fractions of 70 to 8096 methanol μ water, the fractions with strong antibacterial activity were collected, concentrated, frozen and transferred to a powder (19
, 79) were obtained. 1v of the powder was subjected to preparative high performance liquid chromatography using YMC gel as a carrier, and 709
6 methanol/L'10.01M phosphate buffer (pFf7
．． 4) was eluted. The obtained two fractions were each treated with methanol.
After removing p, it was extracted with n-butanol, the extract was concentrated and dried, and TAN-536A (341) and B (1
A purified powder of 95Iv) was obtained.

[Brief explanation of drawings]

Figures 1 and 2 represent the infrared absorption spectra of antibiotics TAN-536A and B, respectively)/l/(in KBr).

Claims (1)

[Claims] Ci) Veptide antibiotic TAN-536A, B or a salt thereof having the following physical and chemical properties = (1) Antibiotic TAN-536A (free form): (a) Elemental analysis value (l
C,55,32±2.0°H7,73±1.0. N
17476±1,5 (b) Infrared absorption spectrum, potassium bromide tablet, principal wave number showing maximum absorption (cm-1): 3350.3060.2960.2940.2870,
1660°1530.1460,1380.1270.
1230.11B0゜1150, 1120, 1080,
930,840,700,650゜90 (0) Molecule ff1 (by S engineering M8 method): 914 (d)
Color reaction: Positive: Ninhydrin, Brave Reback. J4 and II7--; l A# 1--
2-j Coub 1 Item 1-4--1-monodenic acid reaction (2) Antibiotic TAN-s36B (free form): (a)
Elemental analysis i*(m: c 55.28±2.0°H7,
69±1.0. N 17.90±1.5 (b) Infrared absorption spectrum μ, potassium bromide tablet, principal wave number showing maximum absorption (CIll-1): 3350.3060, 2960.
2940.2B70.1660°1530.1440.
1380, 1270, 1230, 1180°1110.
1080,930.840,740,700,650°580.490 (0) Molecule 1 (by sxus method): 900 (d) Color reaction: Positive: Ninhydrin, Brave Reback. Dimethylaminobenza μdehyde, phosphomolybdic acid reaction. [2] Antibiotic Tjl-5 belonging to the genus Cytophaga
36A and/or B-producing bacteria are cultured in a medium to produce and accumulate antibiotics TAli-536A and/or B in the culture, which is then collected. Method for producing W-ζ 16 molecule/or B. [3] Cytophaga sp, YK-33°, which is positive for citric acid and D-mannitol utilization