JPS58134064A - Novel substance mg245-cf2-a, its metal complex, and their preparation - Google Patents

Abstract

NEW MATERIAL:The compound of formulaI. USE:A carcinostatic agent. PREPARATION:A microorganism in Saccharopolyspora, capable of producing MG245-CF2-A, (FERM-5734) is cultured in a medium to accumulate MG-245-CF2- A in the culture mixture and the substance is collected from the mixture. The reaction of the compound of formulaIwith a transition metal in the fourth period in the periodic table gives a metal complex of MG245-CF2-A of formula II (Me is a transition metal in the fourth period in the periodic table; m, n are integers).

Description

[Detailed description of the invention] The present invention is a new substance MG245-CF2-A.

This invention relates to metal complexes and methods for producing them.

The present invention is based on the knowledge that microorganisms of species belonging to the genus Satucharopolyspora produce a substance that inhibits thymidylate synthase prepared from Maus-Ehrlich ascites carcinoma cells in the presence of vanadium salts. It is. This antibiotic (MG 245-CF 2-
The microorganism capable of producing A) was collected and isolated by the present inventor from the soil within the premises of the Microbial Chemistry Research Institute, Tokyo Parts Yo-ku.

New substance MG of the present invention 2. , j 5-CF 2-A has the following physical and chemical properties.

(1) Elemental analysis value = 1°C48.80% N5.80% N9.37% 088.87% (2)
Molecular weight: Estimated to be 427 from the mass spectrum peak.

Calculated from [1(i) of C-C-13N spectrum and elemental analysis to be 427, supporting (l(i) by mass spectrometry.

(3) Molecular formula: C1C13H25N309C-13
cis H'25 from N spectrum and elemental analysis
Estimated to be N309. From analysis of acid hydrolyzate 2.
3-dihydroxybenzoic acid, threonine, and N-hydroxyornithine were confirmed, and from the H-I NMR spectrum, a structure in which these substances were bonded to peptides at one apex was deduced, and from this structure, the molecular formula was Cl8H25N3.
09, which supports the molecular formula obtained from the c-18 NMR spectrum and elemental analysis.

(4) Structural formula: As a result of comparing the acid hydrolyzate with the standard product, MG245-CF
2,3-dihydroxybenzoic acid, L-threonine, and N-hydroxyornithine were confirmed as constituent components of 2-A. From the H-INMR spectrum, these substances are 1
A structure in which each molecule is bonded to a peptide is estimated, and a structure with connections as shown in the structural formula is estimated by mass spectroscopy.

[5] Melting point: 105°C to 109°C (6) Specific optical rotation: [α) D-1-6,1° (c = 1 methanol) 47+ Ultraviolet absorption spectrum: As shown in Figure 1 .

(8) Infrared absorption spectrum: As shown in FIG.

(9) Solubility in solvents: Well soluble in water and methanol, slightly soluble in ethanol and acetone, but almost insoluble in chloroform, ethyl acetate and hexane.

(101 color reaction: positive for Lydon-Smith reaction, triphenyltetrazolium chloride reaction, permanganic acid reaction, negative for ninhydrin reaction, Sakaguchi reaction, and Anthrone reaction.

0υ Distinguishing between basic, acidic, and neutral: i Togami shows the behavior of acidic substances as a result of electropulsion.

Bi-white powder of O2 substance 03 Cellulose thin layer chromatography [Use Avicel SF (manufactured by Funakoshi Yakuhinjiru) as cellulose]
Rf value: n-butanol:methanol:water (4:1:
2) 0.76 MG245-CF2-A and its vanadium complex were used to culture mouse leukemia L-1210 for 48 hours in IBM medium at a concentration of 105 cells/mf. In case, 50μg/dragon\45μVm respectively
1 inhibits cell growth by 50%. MG245-CF
2-A vanadium complex is mouse leukemia L-1210,
Treatment experiments for Ehritzchi's cancer showed a life-prolonging effect as shown below.

MG 245-CF 2-A vanadium complex (hereinafter referred to as:,
MG245-CF2-VA)! 11 Cancer activity (a) Tori prefecture (il') against mouse Shiromen disease L-1210
-iP strain) 1 group of 8 female CDF I mice (withdrawal 1 at 6-7 weeks old)
05 cells 10.25 mJ/mouse of L-1210 leukemia cells were inoculated intraperitoneally, immediately dissolved in physiological saline, and MG 245-CF 2-V, A was administered intraperitoneally 181 times in total for 10 days. The life extension rate was calculated. The results are shown in Table 1.

Table 1 (b) Effect on Ehrlich ascites 2 x 106 cells 10.25m were added to 4 female ICR mice (6 withdrawals) per group.
f/ MG245-CF2-V dissolved in physiological saline immediately after intraperitoneal inoculation of mouse Ehrlich cancer cells.
A was administered intraperitoneally 101 times for 10 consecutive days to examine the survival rate.

The results are shown in Table 2. MG 245-CF 2-V
A also controls Ehrlich ascites cancer Mii '2h
The prefecture was shown.

Table 2 Average age of control group: 14 days, beginning of OD MG 245-CF 2-VA has low toxicity, and as a result of testing using mice, intraperitoneal injection 5. T-p.

is 200 m! It was A or higher. Therefore, this substance is expected to be used as an anticancer agent.

As mentioned above, the physical and chemical properties and biological properties of MG 245-CF 2-A and its metal complex have been described in detail. In terms of containing
Enterobactin produced by N. coli, N-
Although it is similar to albomycin produced by Streptomyces subtropicus in that it contains hydroxyornithine, the chemical structure of MG245-CF2-A is as shown above, and it is clearly different from them.
-CF2-A can be determined to be a new substance.

Next, a method for producing MG 245-CF 2-A and its metal complex will be explained.

MG 245-CF 2-A is produced by culturing a microorganism belonging to the genus Satucharopolyspora and having the ability to produce MG 245-CF 2-A in a medium, and adding MG245-CF 2-A to the culture medium.
MG245-CF2-A was accumulated from the culture.
It can be obtained by harvesting. Also MG
The metal complex of 245-CF 2-A is MG 245-C
It can be easily obtained by adding a transition metal belonging to the fourth period of the j'i1 period rule to an aqueous solution of F 2-A and causing it to be anti-IS. The details will be explained in 1. below.

An example of a bacterial strain that can be used in the present invention is Saccharopolyspora (Saccharopolyspora) described in Examples.
~・-5pora) MG 245-CF 2 (Microorganism accession number υ: Kaikoken Kanyori No. 5734) or a mutant strain of this strain can also be used.

This producing bacterium was discovered in June 1978 by microorganisms (e-science (11[
This actinomycete was isolated from the soil within the laboratory premises at a sudden point, and was deposited with the Institute of Fine Technology and given the strain number MG245-CF2.

Mycological properties of the MG245-CF2 strain 1. Morphology The MG245-CF2 strain has relatively long substratum hyphae, which are irregularly branched, and there are 1 to 2 hyphae at the tips of the branched substratum hyphae.
Individual spores are observed.

Particularly, something like this is observed in starch/inorganic salt agar medium (15P-medium 1) and malate lime agar medium. Aerial hyphae are short, hook-shaped or spiral-shaped, and a chain of spores is observed at the tip. Spores are oval to oblong;
The size of the spider is 0.5μ x 0.8μ, and when it is mature, it is 1
It becomes a chain of 5 to 20 or more. The surface of the spore exhibits a unique hair-like structure.

2. Regarding the description of growth state colors in various media, the standards shown in brackets are based on the Color Harmony Manual of Container Corporation of America.
on of America's C01o1-harm
onemanual) was used.

(1) Sucrose/nitrate agar medium (27°C culture) Growth of shoots C8ni; C1ove Brown
, :], white aerial mycelium grows sparsely and tinglingly.

Soluble pigments are observed from around the 7th day of culture and exhibit a brown color.

(2) Glucose/Aspa) Gin agar medium (2γC
Culture) The growth of the bacteria is poor, and colorless growth can be observed around the 100th culture. The growth around the 144th eye is pale yellow (2
ca; Lt Ivory) to pale yellow (2gc; Bamboo), but no aerial mycelium is attached. Also, no production of soluble pigment was observed.

(3) Glycerin-asparagine agar medium (I5P-medium 5.27°C culture) Slightly aching white aerial mycelia are grown on growing pale yellow to pale shoots (2gc; Bamboo).

In addition, a brownish soluble pigment was observed from around 71111 after culturing.

(4) Starch/Inorganic Salt Agar Medium (ISP-Medium 4.2
7°C culture) Growth is light yellow to light yellow (2 gc; Bam)
-boo], aerial mycelia grow in places 6 and are white to brownish white. Soluble dyes cannot be used.

(5) Tyrosine agar medium (ISP-medium 7.27°CI
Wk) Light yellow [2gc; Bamboo] to thin shoots [2ee:01d Gold], with thin white aerial mycelia attached to the growth. In addition, soluble pigment with a brownish tint is cultured 7
It was widely recognized from around [''■.

(6) Nutrient agar medium (27°C culture) Growth is light yellow to light shoots (8ie; Maple)
Sugarl, it grows sparsely tingling white aerial mycelia. Also, no soluble dye was observed.

(7) Yeast/malt agar medium (-ISP-Medium 2.2
7°C culture) Thin shoot C3ie; Maple Sugar)
～Golden Bro (3pg;, Golden Bro
A thin layer of white to brown aerial mycelium grows on the growing wnl.

Soluble pigments are observed from around the 7th day of culture and exhibit a brown color.

(8) Oatmeal agar medium (ISP-medium 3.27°
C culture) The growth was light yellow (2ec; B15cuit) to light yellow (2ea; Lt Wheat:), and the attachment of aerial mycelium was poor, and a slight brownish-white aerial mycelium was observed in the IC filter on the 22nd day of culture. That's about it. No production of soluble pigment is observed.

(9) Glycerin/nitrate agar medium (27°C culture) Light yellow ~ Light yellow (2gc; Bamboo) ~
Growth of thin shoots (2 μg, Mustard Gold) is accompanied by white to brownish aerial mycelium.

The soluble pigment becomes slightly brownish from around day 1 of culture and takes on a brownish color.

(10) Starch agar medium (27°C culture) On the growth of pale yellow to light yellow Daidai [2gc; BambOo], white to grayish-white aerial mycelia were formed as stones, and a slightly brownish solubility was observed. T' the dye? arise.

0]) Malate lime agar medium (cultured at 27°C) Growth is good with pale yellow to pale shoots [3j? e; Cinnamon
), with sparsely growing white aerial mycelium. Soluble pigments are observed around the 70th day of culture, and are brownish at first, but later become light reddish brown.

02 Cellulose (cultured at 27°C) Growth is colorless, aerial mycelia are attached, and no soluble pigments are observed.

03 Gelatin Puncture] In pure gelatin medium (20°C, J8 culture), small numbers of bacteria were observed to grow from around 10 days after culture, the growth was colorless, no aerial mycelia were formed, and soluble pigments were also observed. I can't do it.

In the case of glucose-peptone-gelatin medium (cultured at 27°C), colorless development and f\ are observed from around March of the culture, but no aerial mycelia are attached and no soluble pigment is observed.

04) Skimmed milk (cultured at 37°C) Smooth white aerial mycelium grows on the growing thin shoots. A small amount of soluble pigment was observed from around July in the culture.
It takes on a brownish tinge.

3. Physiological properties (1) Growth temperature range MY agar medium (maltose 1.0%, yeast extract 7.0
．． 4%, thread agar 3.5%, pH 6.0), 20
°C124°C127°C, aO°c133°c, 37
As a result of culturing at temperatures of 11'C, 40°C, 142°C, 145°C, and 50°C, it grew at temperatures of 20 to 42°C, but the optimum temperature was 30 to 37°C.

(2) Liquefaction of Seratin (15% simple gelatin, 2°C
Culture and Glucose, Peptone, Gelatin, 27°C Culture) Simple gelatin (20"C culture) begins to liquefy slowly from around 10 days after culture, and its effect is weak even after 200 days.Glucose, peptone, gelatin, In the case of gelatin (cultured at 27° C.), liquefaction begins around 10 days of culture, and the liquefaction is somewhat strong or moderate compared to simple gelatin.

(3) Starch hydrolysis (starch/inorganic salt) (culture medium and starch agar medium, both cultured at 27°C) In both cases, ice-melting was observed from around July after the start of culture. The effect is moderate to strong.

(4) Coagulation and peptonization of skimmed milk (skimmed milk, 37°
C culture) ' No coagulation effect was observed. Peptonization was completed around July after the start of culture, and was completed by 10 days (1). (5) Production of melanin-like pigment (tryptone yeast broth, I
(SP medium 1: peptone/yeast/iron agar, 15P medium 6: large amount of tyrosine, ■ST medium 7: both cultured at 27°C) No production of melanin-like pigments was observed in any of the media.

(6) Availability of carbon sources (Pritham-Gotlieb agar medium, l5P-medium 9.27°C culture) glucose, D-
Grow using fructose, sucrose, raffinose, D-mannitol, L-arabinose, D
-Does not utilize xylose, inositol or L-rhamnose.

(7) Dissolution of malate lime (malate lime agar, 27°
C culture:! #) From around July of culture, the malic acid lime on the growth 1a1 side is dissolved, and its effect is moderate to strong.

(8) Nitrate reduction reaction (1.0% potassium nitrate-containing peptone water, l5P-medium 8.27°C culture) is positive.

To summarize the above characteristics, the MG245-CF2 strain has relatively long basal hyphae that are irregularly branched under a microscope, and one or two spore-like objects may be observed at the tips of the hyphae. Hook-shaped or spiral-shaped? A spore button is formed at the tip of the aerial hyphae, and the surface of the spore has a unique hair-like structure. On various agar media, white to brownish-white aerial mycelia are attached to the growth of light shoots, Nibucha daisies, and yellowish browns, and the soluble pigments are light reddish brown to light reddish brown. The formation of melanin-like pigment 'L' was negative in both media. Raw ash is peptonized without coagulating, and its effect is moderate to strong. The liquefaction effect of gelatin is recognized to be weak. In addition, the ice-melting properties of starch are moderate to strong. The growth temperature range is 20-42°C, and the optimal growth temperature is 30-37°C.

MG245-CF2 strain Chivalier et al. (Lechr-v
Alier et al.; International Journal of Swissmatic Batteriology, Volume 20, 1
It shows the n-type cell wall type proposed by 970) and contains 2,6-diaminopimelic acid, arabinose, and galactose. Furthermore, LCN-A (Lipid Characteristics of Nocardia) was detected as a lipid by thin layer chromatography, and Nocardiac acid (Nocardia) was detected by gas chromatography.
It was confirmed that it does not contain ard'omyco-1ic acid).

Based on the above results, when searching for known bacterial genus, MG24
Micropolyspora (M
icropolyspora genus (Birshi's Manual of Determinative Bacteriology,
8th edition, p. 861) and the genus Satucalopolisvora (J. Lacey, Lacey et al.: Journal of General Microbiology, Vol. 88, p. 75, 1)
975). The spore surface of Micropolyspora does not show a hair-like structure, and is clearly different from the MG245-CF2 strain. On the other hand, the genus Satucharopolisvora has a unique king structure on the spore surface and is extremely similar to the MG245-CF2 strain. Next, actually Saccharopolyspora hyspora.

hirsuta) KCCA-017 strain was obtained, and M
The following table summarizes the results of a comparative study with the G245-CF2 strain.

Literature ill 5taneck, J, L, etal, Appl
iedMicro-biology, 28.226
(1974) (2) Mopdarska, H, et.
al, J, Gen, Microbio-1ogy;
71.77 (1972) f3) Goodfellow, hLetal, J
, Gen, Micro-biology, 74.18
5 (1978) (410 KAMI, Y, etal,
Proceedings of the First I
international conference o
nCulture Co11ections (Pu'
blised by University of To
kyo Press, PP457゜1975) (5) Lixcey, J, etal, J, Gen,
Microbiology.

88, 75 (1975) (frE) Large Reference (5) states that substratum hyphae with spores attached are observed, but this is either aerial hyphae or images of collapsed colonies on the surface. It states that it will be.

X, <according to literature (5) 25~50'c※※※ According to literature (5) 37~40°C※※￥※According to literature (5) + As shown in the table, MG245-CF2 The strain and Satsukalopolisbora hirsuta KCCA-0170j Vermilion are morphologically similar, such as the shapes of aerial and ground hyphae and the hair-like structure on the spore surface. In the MG245-CF2 strain, basal spores are rarely observed, but as described in Lacey et al.'s description of Saccharopolyspora hirsuta, these are either aerial hyphae with spores attached or on the colony surface. It may be a collapsed statue (Reference 5). In addition, the rainforest is the color of aerial mycelia,
The growth colors and soluble pigments are also almost identical.

The growth temperature range is 20-42°C for the former, but
The latter is characterized by a slightly higher temperature of 24 to 45°C, and the same can be said about the optimum temperature for growth. Other physiological properties': ``Ni, the il properties of nitrate show different results, but the properties of rainforests include production of melanin pigment, hydrolysis of starch, coagulation of milk, peptone C, liquefaction of gelatin, etc. Regarding the availability of carbon sources, the MG245-CF2 strain does not use inositol and L-rhamnose, whereas the KCCA-0170 strain uses them to grow.It also agrees with D-xylose. However, the utilization of other carbon sources is shown to be the same as that of the rainforest.

Furthermore, analysis of cell wall composition reveals that both rainforests are of the ■ type proposed by Lysivalier et al., and Nocardo is the lipid.
It was found that it did not contain mycoliCacid and LCN-A.

Based on the above results, strain MG245-CF2 is considered to be a bacterium belonging to the genus Satucalopolyspora, and a species extremely closely related to Satucalopolyspora hirsuta. Therefore, MG
245-CF2 strain was transformed into Satsukalopolisbora hirsuta (S
accharopolysporahirsuta)
MG 245-CF 2. In addition, MG245-
CF2 strain was transferred to the Institute of Microbial Technology, Agency of Industrial Science and Technology in 1972.
I applied for storage entrustment on October 3, 2017, and the application receipt number is No. 5734. “MG24 using the above strain
A method for producing 5-CF2-A and MG245-CF2-VA will be explained.

Six types of media commonly used for fermentation of microorganisms are used for culturing the MG245-CF2-A producing bacteria according to the present invention. For example, carbon sources include glucose, glycerin,
Sucrose, etc., and as a nitrogen source, daisu flour,
Yeast m extract, asparagine, etc. are used. Inorganic salts such as common salt and calcium carbonate may be used together, and an antifoaming agent may be added if necessary. As a culture method, methods using a liquid medium such as a vigorous culture method and a deep aeration agitation culture are suitable. The culture temperature is 15-40°C, preferably 20-85°C. The appropriate number of culture cells is 2 to 5 days. MG 245-CF 2-A is accumulated in the culture medium as a substance.

MG 245-CF 2-A was determined by grinding Maus-Ehritzg ascites cancer cells and centrifuging them at 10,5000 g. MG 245-CF 2-A reacts in the presence of a vanadium compound such as vanadyl sulfate, taking advantage of its harmful properties.

Since MG 245-CF 2-A has the above-mentioned physical and chemical properties, it can be extracted and purified by conventional methods according to its properties, and for example, the method shown below is efficient.

When the culture solution containing the active ingredient is passed through a column filled with Diaion HP-20 (manufactured by Mitsubishi Kasei), the active ingredient is adsorbed. After washing the column with deionized water, the active ingredient is eluted with 20% acetone.

After concentrating the eluate under reduced pressure, the pH is adjusted to 2 and the active ingredient is extracted with n-butanol. After drying the extract under reduced pressure, water was added to make an aqueous solution, and pEAE-Sephadex C
The active ingredients are adsorbed when the column is passed through a column packed with -25 (Pharmacia Inc.).After washing the column with deionized water, a concentration gradient elution of 0 to 1 mol of common salt is carried out.The active ingredients are The aqueous solution containing the active ingredient was passed through a column packed with Diaion H, P-20, eluted with 20% acetone, and the solution containing the active ingredient, excluding salt, was dried under reduced pressure and then dissolved in a small amount of methanol.Then, it was dissolved in a small amount of methanol. A purified product is obtained by gel filtration using a column packed with LH-20 (manufactured by Pharmacia) using methanol as an eluent.

Also MG 245-CF 2-VAfi, MG245-
It can be produced by conventional methods depending on the properties of CF2-A, and for example, the method shown below is effective.

Titanium, vanadium, iron, etc. are added to a 0.1-1061D, preferably 1-5% aqueous solution of MG 245-CF 2-A.
A 0.1 to 10% aqueous solution of a transition metal belonging to the fourth period of the periodic law is added and mixed to form a complex. Then, D E A
E-5ephadex-A-25, Dowex-1
or anion exchange resins such as and Diaio
The complex is adsorbed on an adsorption resin such as n-HP-20,
Wash the resin with water to remove unreacted substances. Then, a”'+
When using a desorbing agent such as an anion exchange resin □゛.

In the case of adsorption resin, it is eluted with saline solution or salt cape, and in the case of adsorption resin, it is eluted with water-containing methanol or water-containing acetone. Then,
When eluting with saline, desalt using a 111 degree adsorption resin, and when eluting with hydrochloric acid-containing methanol or aqueous acetone, directly concentrate under reduced pressure and dry to obtain MG245-CF2-A. Obtain a metal complex.

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

Example 1 1% glucose, 1% glycerin, 1% sucrose, 0.5% oatmeal, 2% soybean flour, 1% dry yeast, 0.5% casamino acid, 0.0% calcium carbonate were added to 10 500mf Yosaka flasks. Pour two 100 ml portions of a liquid medium containing 1% and 0.01% antifoaming silicone into a container, apply a cotton plug, and
Autoclave sterilized at 1520 min at °C, and then
ccharopolyspora) M G245-C
Inoculated with F2 (Feikoken Bacteria No. 5734), 80"C,
Culture with shaking for 40 minutes. Next, this culture solution was grown for 500 m
/100 Yosaka flasks with 1.5% glycerin, 1.5% cottonseed flour, 0.3% salt, and 0.2% L-asparagine.
110ml of liquid medium containing! Dispense 8 mJ each into a medium that has been autoclaved with 120" of Cl2O and add cotton plugs. Culture with shaking at 27°C for 1411 hours to obtain a culture solution of 91.
I got it. Transfer this culture solution (pH 7, 8) to Diaion HP.
When the resin is passed through a column filled with If-20 (manufactured by Mitsubishi Kasei), the Q-containing components are adsorbed by the resin. After washing this tower with water,
Elute the active ingredient with 80% acetone water, concentrate under reduced pressure, and concentrate II! Make into an aqueous solution. When the pIl is set to 2 with 1N hydrochloric acid and 1 liter of n-butanol is added and transferred, the active ingredient is extracted into n-butanol. The aqueous layer is further extracted twice with 1 liter of n-butanol, and the n-butanol extracts are combined and concentrated to dryness under reduced pressure.

When the active ingredient is dissolved in 500 mJ of water and passed through a column filled with 200 ml of DEAE-Sephadex C-25 (manufactured by Pharmacia), the active ingredient is adsorbed by the resin. After washing this column with water, a concentration gradient elution of 0 to 1 mol of common salt (
21). The components having Q are eluted at a salt concentration of around 0.8 M.

Next, Diaion HP-20 (manufactured by Mitsubishi Chemical) 20
When this eluate is passed through a column packed with 0 ml/g of i&+1, the active ingredient is adsorbed by the resin. After washing this tower with water, the active ingredient was eluted with 2,017 tons, and the pressure was reduced to 11 akr.
i Dry. The active ingredient was dissolved in 5 m/m of methanol, gel filtration was performed using methanol as an eluent in a tower packed with 400 mJ of Sephadex LH-20, and the active fraction was concentrated to dryness under reduced pressure to obtain 86 m/m of pure MG245-CF2-A.
got '.

Add 20 mg of this MG 245-CF 2-A to 5 m of water.
l! Dissolve 27.6 mg of vanadyl sulfate in 5 mj?
Add the solution dissolved in water and stir and mix for 5 minutes.

Next, this mixed solution is passed through a tower filled with 50 mf of DEAE-Sephadex C-25 (manufactured by Pharmacia), and the active ingredients are adsorbed by the resin. After washing the column with water, elute with 0.5M brine.

Collect the active fraction and use Diaion HP-20 (manufactured by Mitsubishi Kasei)
When passed through a column packed with 100 mf, the active ingredients are adsorbed by the resin. After washing the column with water, the active ingredient was eluted with 50% acetone water and concentrated to dryness under reduced pressure.
CF2-VA(7) pure product 9:1 m7 was obtained. The vanadium complex of MG245-CF2-A contained 2.18% vanadium and was estimated to have a molecular weight of 2500-3000 on Sephadex G-25. Therefore, V (MG 245 CF 2 A)
It is estimated that it has the structure of a. Melting point is 185-190
°C, it is a green powder.

Example 2 Glucose 1% in 500 ml Yosaka flask IO volume
, glycerin 1%, sucrose 1%, oatmeal 0.5%
, soybean flour 0.2%, dry yeast 1 curse, casamino acid 0.5%
, calcium carbonate 0.1%, antifoaming silicon 0.01%
A liquid medium containing 100 m/each was dispensed, a cotton plug was applied, and the temperature was kept at 120°C for 20 minutes. The cells were sterilized, inoculated with Saccharopolysbora MG245-CF2 (Feikoken Bibori No. 5734), and incubated at 30° C. for 411 hours.

Next, this culture solution was placed in a 201-volume stainless steel culture tank containing 1.5% glycerin, 0.5% cottonseed powder, and 0.0% salt.
1 zl of liquid medium containing 3% and 0.5% L-glutamic acid
Prepare, sterilize at 120°C for 20 minutes, and cool.

240 mj after dismissal? 4ii! j Bacteria. Culture with aeration and stirring at 27°C. Aeration: 1 ton, 12 ff/min, stirring number: 25 Orp.
m) Culture solution 10.51 after 4 days! The obtained product was subjected to HP-20 column chromatography, n-butanol extraction, DEAE-Sephadex column chromatography, and Sephadex LH-20 gel filtration in the same manner as in Example 1, and then dried to dryness under reduced pressure. MG245-CF2-A pure product 250m
I got a 9. This was dissolved in 50m of water, 100m5i' of vanadium trichloride was added and stirred for 5 minutes, followed by DEAE-Sephadex column chromatography and IP-20 column chromatography in the same manner as in Example 1. Dry to dryness, MG 245-CF 2-VA
247 mg was obtained.

[Brief explanation of drawings]

Figure 1 shows the ultraviolet absorption spectrum of MG 245-CF 2-A.
It was measured by dissolving it in water at a concentration of e. FIG. 2 shows an infrared absorption spectrum of MG 245-CF 2-A, measured as a potassium bromide tablet. Patent applicant: Makoto Asano, patent attorney representing the Microbial Chemistry Research Foundation

Claims (4)

[Claims] (1) Structural formula %formula% (2) A complex of MG 245-CF 2-A represented by the structural formula and a transition metal belonging to the 4th period of the periodic law, general formula % formula %) (where Me is a transition metal belonging to the 4th period of the periodic law) MG2452C, where m and n are integers)
Metal complex of F2-A. (3) Belongs to the genus Satucharopolyspora and MG245-
A microorganism capable of producing CF2-A is cultured in a medium, MG245-CF2-A is accumulated in the culture, and MG245. - A method for producing neonatal substance MG 245-CF 2-A, which comprises recovering CF 2-A. (4) Belongs to the genus Satucalopolisvora and is NG-2
A microorganism capable of producing 45-CF2-A is cultivated in a medium, MG-245-CF2-A is accumulated in the culture, MG245-CF2-A is collected from the culture, and then MG-245-CF2-A is collected from the culture.
245-CF2-A MG characterized by reacting with a transition metal belonging to the fourth period of the periodic law in an aqueous solution
245-CF2-A metal complex production method.