JPS6054691A - Novel antibiotic sf-1130-x2, its preparation and immunoactivating agent - Google Patents

Abstract

PURPOSE:To produce a novel antibiotic SF-1130-X2 substance exhibiting the growth inhibiting action to Gram-negative bacteria and useful also as an immunoactivator, by culturing a microbial strain belonging to Streptomyces genus. CONSTITUTION:A microbial strain belonging to Streptomyces genus and capable of producing an antibiotic SF-1130-X2 substance, e.g. Streptomyces myxogenes SF-1130 strain (FERM-P NO.676), is cultured at 25-38 deg.C for 2-5 days under aerobic condition, and the objective antibiotic SF-1130-X2 substance is collected from the filtrate of the cultured product.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention provides a novel antibiotic 8F"-l 1 isolated from a culture of a microorganism selected from the genus Streptomyces.
30 X2 substances, their production methods, and uses as immunostimulants.

The present inventors discovered that an antibiotic that exhibits a growth inhibiting effect on Gram-negative bacteria is produced in the culture solution of an organism belonging to the genus Streptomyces, isolated the effective substance, and produced 8F I It was named I 30 X 2 substance.

Furthermore, it was discovered that this substance has the effect of suppressing the decline in immunity caused by cancer carriers or the administration of anticancer drugs, and the present invention was completed. Elemental analysis values: 43.84% carbon, 6.41% hydrogen, nitrogen +
, og-, oxygen 49.67 cm (difference); specific rotation +155
° (Water), has an absorption maximum in the ultraviolet region, has an infrared absorption spectrum shown in the ＠+ figure of the attached drawing, a hydrogen nuclear magnetic resonance absorption spectrum shown in Figure 2, and has an absorption maximum in the ultraviolet region. Easily soluble in sulfonate, tS in alcohol, acetone,
It is insoluble in ethyl acetate, chloroform, and benzene, and is positive in the tests for reed, silver nitrate, red tetrazolium 1 anthrone, ninhydrin, and flake saw/4 ivy.
-・G- When a developing solvent of ethyl acetate/pyridine/water (lO: 4:3) is used in chromatography, R
Rough Ino, is 0.57. n-butanol pyridine
When using a developing solvent of acetic acid/water (6:4:1:3)
The substance is 8F'-1130-12, which is characterized by being a white weakly basic powder with 74 lotus of 0.33. In the second invention, 5F'-1130-12, which belongs to the genus Streptomyces, is -1130-X2 Substance-producing bacteria were cultured under aerobic conditions, and SF-11 was added to the culture solution.
The gist of the invention is a method for producing a new antibiotic 8F-1130-X2 substance, which is characterized by accumulating and collecting the 30-X2 substance.
-An immunostimulant characterized by containing an X2 substance.

An example of the 8F-1130-xx substance-producing bacteria belonging to the genus Streptomyces used in the method of the present invention is the 8F-1130 strain isolated from soil by the present inventors. This bacterium has been stored at the Institute of Microbial Technology, Agency of Industrial Science and Technology since September 4, 1971, with the microbial accession number 6.
The mycological properties of strain 08F-1130 deposited as No. 76 are as follows: @1
Morphobase hyphae grow well on many media, but formation of aerial hyphae is generally poor. ・Elongation was good on Scooch agar and starch yeast extract (or starch/yeast extract) agar, where aerial hyphae are observed. Short, dense aerial hyphae are formed from basal hyphae. The branch is a simple branch and there are no axle branches.

The tip of the aerial hyphae is mostly composed of closed spiral threads, but incomplete spiral threads and open spiral threads are also observed.・No nucleation is observed.0 Spore formation by electron microscopy Table 1 τth 1q structure is smooth.0 Spores are coffin-circular or cylindrical in shape and have a size of 0.6-0.7 x 0.9-1.0 microns.

8. Physiological properties Liquefaction of gelatin: Positive Hydrolysis of starch: Positive Generation of tyrosinase: Positive Generation of hydrogen sulfide: Positive Chromodynic action: Positive Decomposition of fibers: Negative Reduction of nitrate: Negative Peptonization of skim milk : Negative Coagulation of skim milk : Negative Liquefaction of Loeffler's coagulated serum: Negative In addition to the above physiological properties, strain 8F-1130 has the property of producing mucilage i in agar and liquid media. Yes, this is a large 4I microorganism of this strain.◎Starch yeast extract (or starch/yeast extract) becomes sticky from the 10th day of culture on agar media such as agar, starch agar, glucose/asparaquine agar, etc. It is also observed that it is produced on the bacterial body.

In addition, a liquid medium containing appropriate carbon sources (glucose, starch, etc.) and nitrogen sources (yeast extract, soybean flour, wheat germ, etc.) is used at 8F.
When strain -1+30 is cultured with shaking, the culture solution gradually becomes viscous and the production of mucilage is observed.

4. Utilization of carbon sources: xylose, glucose, galactose, malto-sose, sutucarose, lactose, raffinose, dextrin.

Starch, glycerol, wild boar Thor, Sodium Acetate, Citric acid soda, mannose 2. Near-ubinose and fructose whose use is questionable.

Salicin 3, not used: rhamnose, inulin, dalgit, mannitol, sorbitol, sodium succinate, cellulose, etc. To summarize the mycological characteristics of the Sv-1130 strain, (1) The tips of aerial hyphae are mainly spiral. The spore surface is smooth (2) The aerial hyphae are grayish brown or gray, but the epiphytic hyphae are extremely poor.

(3) Growth on synthetic media is brown to gray-brown (4) Growth on organic media is chromogenic.

(5) Producing mucilage in agar medium and liquid medium.

Based on the above mycological properties, Streptomyces pheochromolnes is a closely related bacterial species to the 5F-1130 strain.

Examples include Streptomyces purpureochromogenes and Streptomyces novolidoensis, but as shown below, strain 5F-1130 does not match any of the bacterial species.

Streptomyces 7eochromogenes grows abundantly on aerial mycelium and produces brown soluble pigment on sucrose/nitrate agar and calcium malate agar, whereas strain 8F-1+30 has no ability to grow aerial mycelium. The two are clearly distinguishable in that they have poor pigmentation and do not produce soluble pigments in the above-mentioned medium.

Streptomyces purpureochromogenes grows orange to orange-red on potatoes, does not produce hydrogen sulfide, and coagulates skim milk, whereas strain 8F-1130 grows on potato pieces brown and produces hydrogen sulfide. There are clear differences, such as the absence of skim milk coagulation.

In addition, Streptomyces novolidoensis does not form spiral threads and produces green soluble pigment in starch agar (although the literature does not mention the production of green soluble pigment,
This is noticeable in type strains). On the other hand, SF-11
Strain 30 forms spiral threads and does not produce soluble pigments on starch agar.The two strains also differ in the availability of mannitol and sucrose, and strain 8F-1130 can be distinguished from Streptomyces novolidoensis. Ru.

Furthermore, although the 8F-1130 strain has the unique property of producing mucilage, there are no reports that any bacterial species of the genus Streptomyces, including the three bacterial species mentioned above, produce mucilage.
In this respect as well, strain SF', -1130 has no match among known bacterial species.

Therefore, at the time the present inventors isolated strain 8F'-1130, we considered this strain to be a new bacterial species of the genus Streptomyces, and believed that this strain was a new strain of Streptomyces.
As seen in the case of other Streptomyces species, the O8F-1130 strain, named ces myxogenes 8P, nov, ), is susceptible to changes in its properties, such as UV rays, X-rays, radio frequencies, and radiation.

Bacterial strains belonging to the genus Streptomyces that have the ability to produce the SF l 130-X2 substance, including such mutant strains, which can be mutated by artificial means such as drugs, , all can be used in the method of the invention.

In the method of the present invention, the above-mentioned bacterial strain is cultured in a medium containing nutrients that can be used by ordinary microorganisms. As the nutrient source, any known nutrient source conventionally used for culturing Streptomyces can be used. For example, as a carbon source, starch, starch syrup, molasses, etc. can be used @Also, as a nitrogen source, soybean flour, wheat germ, dried yeast, pezotone, meat extract, cornstarch liquor, ammonium sulfate, sodium nitrate, etc. can be used .

Calcium carbonate, sodium chloride, etc. as necessary.
In addition to adding inorganic salts such as potassium chloride and phosphates, organic and inorganic substances that aid the growth of bacteria and promote the production of SF-1130-1 and other substances may be appropriately added.

As for the culture method, the liquid culture method, especially the deep culture method, is most suitable, as is the case with general antibiotic production methods. Cultivation is carried out under aerobic conditions, and the appropriate temperature for cultivation is 2 s D.
c~3goC, but in most cases the culture is carried out at around 26°C.Thus, the production of the 8F'-1130,-x2 substance reaches its maximum in 2-5 days in both shaking and tank cultures.

8F'-l 130-12 The following method is used for assaying substances.・As a medium for assay・0.12
5-mycin assay agar medium containing maltotriose (peptone O, SS, 0.3% in Mitoe,
Using agar 1.5, pH 6), test bacteria Kid, Escherichia coli, and Escherichia buri (geeherle) were prepared.
For the 6-assay method using K-12R, the conventional page/arm disc method is used.

Since the EsV-1130-12 substance is a weakly basic oligosaccharide that exhibits the physical and chemical properties described below, it can be collected from the culture solution and purified according to its properties. For example, carbon adsorption, hydroalcohol elution, or water-
It can be made blue by the ethanol reprecipitation method.

That is, by acid-filtering the culture solution of %SF-1130 strain at pH 3, the bacterial cells and P were immediately adsorbed onto a carbon column, eluted with 50% thiacetone water, the eluate was concentrated to dryness, and then dissolved in water. Then, it was adsorbed on a carbon column again, and the first elution was carried out with 5 to 25 ml of ethanol water.
A coarse powder can be easily obtained by concentrating the fraction containing the F-1130-x2 substance and precipitating it with ethanol.The coarse powder thus obtained usually contains neutral oligosaccharides, In particular, the following method is preferable in order to obtain a highly pure product of the SF'-I13O-X2 substance, which is contaminated with a large amount of maltodextrin. That is, the fermentation liquid is passed through a strongly acidic ion exchange resin such as DOWEX 50wx2 (H), and the 5F-I 13O-X2 substance is adsorbed onto the resin. After washing thoroughly with water, elute with O, IN aqueous ammonia. ◎ After drying the eluent, dissolve in water and adsorb on a carbon column at pH 3.5. After washing with water, bathe with 30-35% ethanol water. , concentrate to dryness. Next, this was again adsorbed onto DOWEX 50W x 2 (NIP) resin, washed with water, swamped with O,IN ammonia water, and dried. ), developed with 0.1M pyridine-formate buffer (pH 3, 1), and concentrated to dryness of the antibacterial f8 fraction. -1130-x Substance (patent application 1973)
1-99757) can be obtained as a white powder, which can be used as is for physiological activity tests. Developed with pro-gnol-ethyl acetate-water = 6:l:3), and subjected to benochlorite atomography (acid 1jll = c-tyl-pyridine water + = IO:4
:3), if the fraction showing a dark spot of R,,i,-X: 0.39 (assuming Yf of raffinose is 1.00) is concentrated to dryness, the by-produced SF-30-X ,
The substance is obtained as a colorless powder. At the same time, R5 I, -X
The 8F-1130-12 substance of the present invention is obtained as a colorless powder by concentrating the fraction of = 0.57 to dryness. In addition to the 5F-1130 =
9756) are also produced and accumulated. 5F-1
130-x, --"2 The substance belongs to oligosaccharides that exhibit antibacterial properties and weak basicity, and is easily soluble in water, sparingly soluble in methanol and ethanol, and insoluble in acetone. In contrast, the above-mentioned malt Pentaose.

Since maltohexaose is a neutral oligosaccharide that is easily soluble in water and insoluble in ethanol and acetone, it can be separated from the other by utilizing the above-mentioned differences in properties, i.e., 5
F-113 (When the P solution obtained by acidic filtration of the Zelkova culture solution is passed through a strongly acidic ion exchange resin, 8t' l I 3
0-x + ,-]C2 substance is adsorbed to the resin, but
The middle saose passes through maltopentaose and malt without being adsorbed. The resin was added to 0. When treated with IN ammonia water, the 8F-30-11 s-X2 substance is eluted.

The physical and chemical properties of the SF-1130-12 substance of the present invention are shown in Table 2. For reference, SF
The properties of l 30 x + substances are also listed in Table-2.
S F l l 30 x + and -x2 The substance is
It is an oligosaccharide because it adsorbs to acidic resin and migrates to the cathode in -1.9 F paper electrophoresis, it is a weakly basic substance, and it gives a large amount of glucose by acid hydrolysis. It is represented by ◎, which is considered to be 5F-
1l 30-X+ For substances, (m+n)=5 and 5
For F-l 130-X2 substance (m+n)=
It turned out to be 4.

8F-11307X2 The substance is ineffective against acid-fast bacteria against various microorganisms.
It was discovered that the antibacterial activity of C2 substances increases significantly when maltodextrins such as maltose, maltotriose, maltotetraose, maltopentaose, and maltotetraose coexist.However,
In this case, as an exception, there was no Enhancement Roca The test examples are shown below.
The toxicity of the S' I l 30-X 2 substance is extremely weak, and in acute tests using mice,
In the case of subcutaneous injection, all cases survived even at 400 rug/9. SF-1130-x2 substance is Sarcom
a In an immunostimulation test (delayed skin reaction method) in mice transplanted with Igo ascites tumors, it was discovered that it has an effect of preventing immune decline. It has also been found that this effect is also effective in preventing the immunological decline that occurs when anticancer drugs are administered. In this case, as in the case of antibacterial activity, the addition of maltodextrin enhanced immunostimulation. One test example is shown in Table 4. The test method was to inject Sarcoma into ICR mice (6 mice per group). 180 ascites tumors were transplanted, and 24 hours later, each sample (drug) and anticancer drug (g
ndoxan) were administered subcutaneously and intraperitoneally, respectively, and transplantation 2
On the fourth day, 6% pyrrolyl chloride ethanol solution was applied to the abdomen for sensitization, and then the specimen was administered once a day for 4 days.On the 4th day, Endoxan was readministered, and on the 9th day, 1% Apply chlorinated picryl olive oil solution to the front and back of both ears for secondary sensitization. The degree of increase in ear thickness was measured 24 hours later, and the degree of delayed skin reaction was determined based on the rate of increase in ear thickness.

Table 5 shows the immunostimulation test using Sarcoma 180 solid gun.The test method is Sarcoma C80.
to ICR mice (5 mice per group) - 8F-l 130- obtained in Example 2 on the 6th and 5th day before subcutaneous transplantation.
A mixture of the x2 substance and M1-<ntaose was administered intraperitoneally, and 100 days after transplantation, the tumors were excised and their sizes and *m were measured. As seen in Table 5, significant regression of solid tumors was observed by repeated administration of the compound of the present invention.

On the other hand, the results of a therapeutic test in which 8arcoma 180 was previously implanted into the abdominal cavity of a mouse and then using 8F-I 130 )C2 substance were completely ineffective. Therefore, it is clear that the 8Fl 130-C2 substance indirectly shows a curative effect by activating the immune resistance of the host. It has been found that there is a significant decrease in the number of patients, and so-called immunotherapy has begun to be used clinically to prevent this decrease and increase the therapeutic effect. As an immunostimulant for this purpose, B
Although CG bacterial cells and various polysaccharides are well known, they are all polymers.1) There have been no reports to date that oligosaccharides with a molecular weight of less than 10,000 yen as shown in the present invention have an immunostimulatory effect. This is truly unexpected. It is clear that such low-molecular-weight oligosaccharides are practically superior to polysaccharides in terms of absorption and excretion, tuberculosis infectivity, frequency of allergic reactions, etc.

Furthermore, it is clear from the following test examples that the SF I 130 C2 substance exhibits a protective effect against bacterial infections and is effective as an immunostimulant. That is, J.C.
L: ICR male mice were used in groups of 8F.
I j30-X2 substance was dissolved in phosphate buffer at pH 7.2 and administered intraperitoneally three times at 48 hour intervals at a daily dose of J50v/kIi or 10 my/kg. Final administration route 7
After 72 hours, the culture solution of Staphylococcus aureus Smith S-424 strain was diluted with physiological saline, and a bacterial suspension containing 5 mucins was inoculated intraperitoneally.@The bacterial inoculation volume was 0. 5rnt/mouse.

Observe the number of surviving mice for 5 days after inoculation, and compare the numbers in the table below.
The results shown in 6 were obtained.

5F-I 130-X2 is an untreated control group (ContCI
When administered at 50ty/kg, the LD50 (which is a measure of the effectiveness in preventing infection) of the inoculated bacteria increased by approximately 11.4 times, and when administered at 9 loq/kg, it increased approximately 8.1 times. afx SF l 130-! z As shown in Table 4, the substance itself has no antibacterial activity against Staphylococcus aureus @ It is also known that immunostimulants are effective in treating autoimmune diseases such as arthritis (rheumatism). This time, according to the test examples shown below, 5F-1130 of the present invention
The -x2 substance was found to be effective in preventing the development of experimental arthritis, which is said to be closely related to rheumatism.

That is, in this test, the bacterial cell mass of Mifpacterium butyricum suspended in physiological saline was 5.8η/m.
8D rats (male, average weight 1
Inflammation was caused by subcutaneous injection into the right foot of 10 animals per group. SF − 1 day after bacterial injection for this inflammation
1130-x substance (SF -1130-X+) and 8F
-l 130- X2 in a 1:2 weight ratio)
A solution prepared by dissolving the
, subcutaneously injected for 10 consecutive days.
It was set to 50 wJ/kg. Signs of inflammation that occurred in the legs, ears, nose, and eyes of rats on day 14 after bacterial inoculation (degree of onset)
Investigated and evaluated on a scale of 0 to 304 (score) and calculated the average value.Furthermore, the left foot pedal volume*fi- was measured and SF
The anti-inflammatory efficacy of the -1130-x substance was determined.For comparison, phenylbutacin tm was orally administered once a day for 11 days at a dose of 20/97 days in place of the SF-1130-x substance. A control test was also conducted in exactly the same manner except that administration of SF-1130-x and phenylbutycin was omitted. The results are shown in Table 7 below.

This test showed that the SF I' 30 X 2 substance was effective in preventing experimental arthritis.

SF 1130! 2. When administering the substance, intravenous drip, local injection, intramuscular, thoracic or intraperitoneal injection is possible. 0, daily or 2 to 3 times a week, 50 to 4 ooη/kg.

Preferably, around iQO++/kg is administered. Next, examples of the present invention will be shown. Example 1 Streptomyces myxogenes SF'-1130 strain C
ffA Koken Bokuyori No. 676), 5.0 g of starch syrup, 2.5% of soybean flour, 1.0% of wheat germ, and 0.25 of sodium chloride.
The cells were inoculated into 200 L of a liquid medium (pH 7.0) of 100 ml of chloride, and cultured with aeration and agitation for 64 hours at 28'C in a jar fermenter.

After culturing, the culture solution was acidified with p)I 3, and P solution 1
Obtained 40t. Add this P solution to DOWEX 5owX2 (H
) strongly acidic ion exchange resin ft:, ti
5. After adsorbing on A and thoroughly washing with water, it was eluted with O, 1N and 7 liters of water. Of the eluate, Eeoli
The fraction with antibacterial activity against K-,12EC was concentrated to dryness to obtain 250 tons of brown powder. 150 tons of pure liquid
Adjust L to 3.0 again with hydrochloric acid.

It was adsorbed onto a column packed with 20 t'i of DOWEX 50WX2 (H) resin, and after washing with water, 0. IN ammonia water,
A total of 80 tons were drained, and the antibacterial active fraction was concentrated to dryness to obtain brown powder 511.

Firstly obtained powder 47f was dissolved in 250 td of water, p
After adjusting to H30, it was adsorbed on a column (5,5 x 26 c1n) packed with Wako activated carbon 600-, and after washing with water,
Bathed with 30% ethanol water and then 35% ethanol water.Of each fraction, 4t of the fraction with antibacterial activity against Escherichia coli and 12R was concentrated and dried to give 5.6% yellowish brown powder.
I got t.

Add this powder again to water + 00. d and pH 3 with hydrochloric acid.
After adjusting to DOWEX s OWX 2 (NH
4+) in a column filled with resin 600-,
After washing thoroughly with water, 0. The antibacterial active fraction was eluted with IN ammonia water and concentrated to dryness to obtain 1.5 t of yellowish brown powder.
The formic acid bath solution... = 3.1) was dissolved in 70-, adsorbed on a column (3.5 x 60 m) packed with pyridinium-type DOWEX 50WX2 (200-400 mesh), and eluted with a buffer solution. The eluate was fractionated into 10-2 fractions, and each fraction was subjected to high-pressure P paper electrophoresis and separated with silver nitrate reagent.

Antibacterial activity fraction ≦63 to 79 showing a single sulfur concentration of 0.53 (as Rf of alanine is 1.Oo)
Concentrate to dryness to obtain 8F-1130-x, substance and 5F-
380 da of white powder was obtained as a mixture of 1130x2 substances.

Next, this powder is dissolved in a small amount of water, and mixed solvent (n
- Add pro/4'nol, ethyl acetate, water; 6:1=3) and cellulose 2ooIl! Column (3
x30cIII) and developed with a mixed solvent.The eluate was separated into 7 fractions, and each fraction was purified with a mixed solvent of ethyl acetate, pyridine, and water (10:4:3). R, i, -10,57 (raffinose 1.0
Fractions 351 to 445 were concentrated to dryness at -8F' 11.
30 The substance '70iaf was obtained. Furthermore, 8F l l 3O-X2 substance powder 150q
was dissolved in water and adsorbed on a column (2 x 5.5 crn) packed with activated carbon.
The mixture was washed out with ethanol and concentrated to dryness to obtain white powder 12011f. Melting point 195
°C (decomposition) -8F-I 130-It The powder 70■ of the substance was treated in the same way to obtain 55 cape as a white powder 0 melting point 203DC (decomposition) 02nd Dowex 50 WX 2 (H+) Resin eluate 51f was treated in the same manner and 8F -1130-X+
Substance 210 W, 8F -1130-x2 Substance 5
Example 2 Streptomyces mikingenes 5r-II 30 strain l Koken Bacteria 676-1'q) was inoculated into 20 L of liquid medium having the same composition as in Example 1, and placed in a jar 7 armer. te2
Aerated agitation culture was carried out for 66 hours in g'c.
This P solution was applied to a column (6 x 3
6crn), washed thoroughly with water, and then
Drain with 5 tons of thiacetone water, -1 for Escherichia coli in the eluate
The fraction having antibacterial activity against 28 was concentrated to dryness to obtain yellowish brown powder 67.52.

Of these, 45F was dissolved in 200 d of water, adsorbed on a column (5,6 x 42 vice) filled with activated carbon IL manufactured by Wako, and after washing with water, 5, 10. The first elution was carried out with +5, 20, and 25 ml of ethanol water, and the eluate was fractionated into 15 ml of eluate. Antibacterial active fractions 366-510 were concentrated to dryness to obtain a white powder 5.72. This white powder 5f' was dissolved in +5- water and after filtration, p
Gradually pour 180-g of ethanol into the armpit and re-precipitate to obtain 4.6 f of a mixture of 5F-1130-x2 substance (+ol) and maltopentaose (99 g)◎

[Brief explanation of drawings]

Figures 1 and 2 are from the original publication 013F-1130, respectively.
-x A curve diagram of an infrared absorption spectrum (in KBr tablet) and an OOMHz hydrogen nuclear magnetic resonance spectrum (in heavy water) is shown. Figures 30 and 4 show the infrared absorption spectrum of 5F-1130X+ substance (in KBr tablet) and 100MHz of water, respectively.
O showing the curve diagram of hydrogen nuclear magnetic resonance absorption spectrum (in heavy water)

Claims (1)

[Claims] ■ Showing the following physical and chemical properties, that is, elemental analysis values: carbon 43.84, hydrogen 6.41, nitrogen 1.08, oxygen 49.67 (difference); ratio Optical rotation +155° (water)
It has an absorption maximum in the ultraviolet region, an infrared absorption spectrum shown in Figure 1 of the attached drawings, and a hydrogen nuclear magnetic resonance absorption spectrum shown in Figure 2. Easily soluble in alcohol, slightly soluble in alcohol, insoluble in acetone, ethyl acetate, chloroform, benzene, silver nitrate, red tetrazolium, anthrone. It was positive for each reagent of ninhydrin and Blake Lee. Ethyl acetate/pyridine/water (10:1) was detected by paper chromatography.
4: 3) + 7) Illut R57 Ia'-x for expansion W3s 0.57. When a developing solvent of n-butanol, pyridine, acetic acid, and water (6:4:I:3) is used, F(,
A S FI I 30 X 2 substance characterized by being a white weakly basic powder having a phynose content of 0.33. 2. f3kj'-1130 belonging to the genus Streptomyces
-x2 A new antibiotic SF-l 1 characterized by culturing substance-producing bacteria under aerobic conditions to accumulate SF-1130-X2 substance in the culture solution and collecting it.
30 x Method for producing 2 substances. 8. An immunostimulant characterized by containing SF I I 30 X 2 substance as an active ingredient.