JPS6035112B2 - New antibiotic SF-1130-X↓2 substances, their manufacturing method and immunostimulant - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention provides a new antibiotic SF-1130 isolated from a culture of a microorganism selected from the genus Streptomyces.
-x2 substances, their production methods, and uses as immunostimulants.

The present inventors discovered that an antibiotic that exhibits a growth-inhibitory effect on Gram-negative bacteria is produced in the culture solution of a microorganism belonging to the genus Streptomyces, isolated the effective substance, and It was named -11301x2 substance. Furthermore, the present invention was completed based on the discovery that this substance has the effect of suppressing the decline in immunity caused by cancer carriers or the administration of anticancer drugs.

That is, the first invention exhibits the following physical and chemical properties, namely, elemental analysis values: carbon 43.84%, hydrogen 6.41%, nitrogen 1.08%, oxygen 49.67% (difference) : has a specific optical rotation of +155o (water), does not have an absorption maximum in the ultraviolet region, has 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 water and dimethylsulfoxy, sparingly soluble in alcohol, insoluble in acetone, ethyl acetate, chloroform, and benzene, positive in silver nitrate, red tetrazolium, Anthrone, ninhydrin, Gleak, and Reapak tests, and paper In chromatography, use a developing solvent of ethyl acetate, pyridine, and water (10:4:3). The substance is characterized in that it is a white weakly basic powder having R-raffinose of 0.33 when used. In the second invention, SF-1130- belonging to the genus Streptomyces is cultured under aerobic conditions, and SF-1130-×
The gist of this invention is a method for producing a new antibiotic SF-1130, which is characterized by accumulating and collecting two substances. Further, the third invention has SF-1130 as an active ingredient.
The gist of the present invention is an immunostimulant characterized by containing a uniform substance.

An example of the SF-1130 substance-producing bacteria belonging to the genus Streptomyces used in the method of the present invention is SF-113 Gongju, which was isolated from soil by the present inventors.

This bacterium was transferred to the Institute of Microbial Technology, Agency of Industrial Science and Technology in 1968.
Since September 4th, it has been deposited under the microbial accession number 676. The mycological properties of SF-113 beads are as follows. Although morphobase hyphae grow well on many media, aerial hyphae formation is generally poor.

Starch agar with aerial mycelium, starch yeast extract (
(or starch, yeast extract) agar, etc., short, dense aerial hyphae are formed from well-elongated basal hyphae. The branch is a simple branch and there are no axle branches. The tip of the aerial hyphae consists mostly of compact, closed spiral threads, but incomplete spiral threads and open spiral threads are also observed. No sclerotia formation is observed. The surface structure of the spores observed by electron microscopy is smooth. Spores are oval or cylindrical, 0.6-0.7 x 0.
It has a size of 9 to 1.0 microns. 2 Properties on various media -- 1 3 Physiological properties Liquefaction of gelatin: Positive Hydrolysis of starch: Generation of positive tyrosinase: Generation of positive hydrogen sulfide: Positive chromogenic action: Degradation of positive and negative cellulose: Negative nitrate In addition to the above physiological properties, strain SF-1130 has the ability to absorb viscous substances in agar and liquid media. This is a major feature of this strain.

When using starch yeast extract (or starch/yeast extract) agar, starch agar, glucose/asparagine agar, etc., sticky substances have been observed to climb onto the bacterial bodies and be produced from around the 10th day of culture. Ru. In addition, when SF-113 Koshu is cultured in a liquid medium containing an appropriate carbon source (glucose, starch, etc.) and nitrogen source (yeast extract, soybean flour, wheat bud, etc.), the culture solution gradually becomes a viscous bowl and becomes viscous. Purchase generation is recognized.

4 Utilization of carbon sources 1 Used: xylose, glucose, galactose, maltose, sutucarose, lactose, raffinose, dextrin, starch, glycerol, inositol, sodium acetate, sodium citrate, mannose 2 Difficult to use: arabinose , fructose, salicin 3
Not used: Rhamnose, inulin, dal..., mannitrate, sorbitol, sodium succinate, cellulose From the above, the mycological characteristics of SF-113 vermilion can be summarized as follows: '1} The tips of aerial hyphae are mainly spiral. (closed type) and the spore surface is smooth.

{2) Aerial hyphae are grayish brown or gray in color, but their ability to settle is extremely poor.

(3) Growth in the synthesis tower area is brown to grayish brown.

'4) In organic substrates, it has chromogenic properties. (
5) Produce mucilage on agar medium and liquid column.

Based on the above mycological properties, the near green bacteria species of SF-1130 strain include Streptomyces phaeochromogenes, Streptomyces purpureochromogenes, and Streptomyces novolitoensis, but as shown below, S
Strain F-1130 does not match any bacterial species.

Namely, Streff.

Tomyces pheochromogenes grows abundantly on aerial mycelium and produces brown soluble pigment on sucrose-nitrate agar and calcium malate agar, whereas SF-1
The strain 130 has a poor ability to attach to aerial mycelium, and the two strains are clearly distinguished from each other in that they do not produce soluble pigments on the above-mentioned soil. Streptomyces purpureochromoges grows on potato pieces from light-red to light-red, does not produce hydrogen sulfide, and coagulates skim milk, whereas strain SF-1130 grows on potato pieces as brown.
It has clear differences such as producing hydrogen sulfide and no coagulation of skim milk.

In addition, Streptomyces novolitoensis does not form spiral threads and produces a green soluble pigment in starch agar (although the production of green soluble pigment is not discussed in the literature, it is clearly observed in the type strain). .

On the other hand, SF-113 forms porridge spiral threads and does not produce soluble pigments in starch agar. Furthermore, the two also differ in the availability of mannitol and saccharose.
Distinguished from SF-113 powder Streptomyces novolitoensis.

Furthermore, although the SF-1130 strain has the unique property of producing sticky substances, there are no reports of Streptomyces species, including the three cocoon species mentioned above, producing slimy substances.
In this respect, strain SF-1130 has no match among known bacterial species.

Therefore, at the time the present inventors isolated the SF-1130 strain, we considered this strain to be a new bacterial species of the genus Streptomyces,
smisoogenesSP. nov. ) was named.

As seen in the rudder of SF-113 and other cocoon species of the genus Streptomyces, the properties are easily changeable and can be mutated by artificial mutagenic means such as ultraviolet rays, X-rays, radio frequency, radiation, and chemicals. and SF, including such mutant strains, belong to the genus Streptomyces.
-1130-Things that have the capacity to produce substances are:
All can be used in the method of the invention.

In the method of the present invention, the strain is cultured in a medium containing nutrients that can be used by common microorganisms.

As the nutrient source, known nutrient sources conventionally used for culturing Streptomyces can be used. For example, as a carbon source,
Starch, starch syrup, molasses, etc. can be used. Further, as a nitrogen source, soybean flour, wheat germ, dried yeast, veptone, meat extract, corn staple liquor, ammonium sulfate, sodium nitrate, etc. can be used. In addition to adding other inorganic salts such as calcium carbonate, sodium chloride, potassium chloride, and phosphates as necessary, organic and inorganic substances that support the growth of bacteria and promote the production of SF-1130 Ichiume substances are added as appropriate. Can be 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.
500 to 38 oo, but in most cases it is cultured around 28 o'clock. Thus, the production of SF-1130-x2 substance reaches its maximum in 2 to 5 days in both shaking moat culture and tank culture. SF-1130-For the verification of private substances, the following method is used.

The assay medium was Mysin Assay Agar (Beptone 0) containing 0.125% maltotriose.
．． 5%, meat extract 0.3%, agar 1.5%, pH 6
), and the test bacteria include Escherichia coli and Escherichia coli (
Echerichia coli) K-12R is used. The normal paper disk method is used for the assay.
Since SF-1130-cedar 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, it can be purified by carbon adsorption, hydrous alcohol elution, or water-ethanol reprecipitation. That is, SF
-113 Gongzhu culture solution was treated in an acidic oven at pH 3 to separate it from bacterial cells, and then immediately adsorbed on a carbon column.
Elute with 50% acetone water, evaporate the eluent to dryness, dissolve in water, adsorb onto a carbon column again,
SF-1130-
A coarse powder can be easily obtained by concentrating the fraction containing the x2 substance and precipitating it with ethanol.

The coarse powder thus obtained usually contains a large amount of neutral oligosaccharides, especially maltodextrin, and S
In order to obtain a highly pure F-1130 Hitosugi substance, the following method is preferred. That is, the fermentation liquid was mixed with DOWEX 50W×
SF-113 through a strongly acidic ion exchange resin of 2 (H+)
0-Let the substance be adsorbed onto the resin. After washing thoroughly with water,
Elute with 0.1N aqueous ammonia. After drying the eluent, it is dissolved in water, adsorbed on a carbon column at pH 3.5, washed with water, eluted with 30-35% ethanol water, and concentrated to dryness. Next, this was reused with DOWEX 50W x 2 (N
After adsorption on H40) resin, and washing with water, elution was carried out with 0.1N aqueous ammonia, and the mixture was dried. Furthermore, this was adsorbed onto Dowex 50W x 2 (pyridinium salt type) resin,
It was developed with 0.1M pyridine-formate buffer (pH 3.1), the antibacterial active fraction was concentrated to dryness, and SF-1130-×2
SF-11301x produced at the same time as the substance. A mixture with a substance (see specification of Tokuiso Sho 51-99757) is obtained as a white powder. This powder can be used as is for physiological activity tests, but this mixture was adsorbed onto a cellulose column, developed with a developing solvent (n-propanol/ethyl acetate/water = 6:1:3), and then subjected to paper chromatography. (Ethyl acetate/pyridine/water = 10:4:3), R roughinose = 0.39 (Rf of raffinose
By drying the fraction showing a single spot of 1.00, the by-product SF-11301x is obtained as a colorless powder. At the same time, 0.57 fractions of R-raffinose were concentrated to dryness, and the SF-1 of the present invention was converted into a colorless powder.
1301×2 material is obtained. In addition, when SF-113 vermilion is cultured in the method of the present invention, in addition to the substances SF-1130- (See Samurai Gan No. 51-99756 filed on the same day) has also been produced and accumulated.

The SF-1130-x,,-ike substance belongs to oligosaccharides that exhibit antibacterial properties and weak basicity, and is easily soluble in water, slightly soluble in methanol and ethanol, and insoluble in acetone. On the other hand, maltobentaose and maltohexaose are neutral oligosaccharides that are easily soluble in water and insoluble in ethanol and acetone, so they can be separated from each other by utilizing the above-mentioned difference in properties. That is, for example, when the culture solution of the SF-1130 strain is exposed to an acidic environment and the obtained solution is passed through a strongly acidic ion exchange resin, the SF-1130-x,,-& substances are absorbed by the resin due to the force of the reaction. , maltobentaose, and maltohexaose pass through without being adsorbed. When the resin is treated with 0.1N aqueous ammonia, the SF-1130-x2, 1x2 substance is eluted. The physical and chemical properties of the SF-1130-x2 substance of the present invention are shown in Table 2.

For reference, SF-1130-x. The properties of the substances are also listed in Table-2. SF-1130-x and -x2 substances have the ability to adsorb acidic resin and have pHI. In the furnace paper electrophoresis method of No. 9, it is a weakly basic substance because it migrates to the cathode, and it is considered to be an oligosaccharide because it gives a large amount of glucose by acid hydrolysis. Table-2 In addition, SF-1130-x, and SF-1130-x2
The substance is represented by the following general formula, SF-1130-×
, for the substance (m+n)=5 and SF-11301×
It was found that (m+n)=4 for the two substances.

The antibacterial spectrum of SF-1130-private substance against various microorganisms is as shown in Table 2. It has activity against Gram enterobacteria, but is ineffective against Gram negative bacteria and acid-fast bacteria.

Furthermore, the present inventors have found that the antibacterial activity of SF-1130-& substance increases significantly when maltodextrins such as maltose, maltotriose, maltotetraose, maltobentaose, and maltohexaose coexist. discovered. However, in this case, Klebzilla New*
*Exceptionally, no increase was observed for Mony. The test examples are shown in Table-3. Table 3 * Maltotriose was added to the plate upper layer medium (Mycin Assey agar) at a rate of 1.25 c/cc.

Under these conditions, maltotriose itself has no antibacterial activity. The toxicity of SF-1130 is extremely low, and acute toxicity tests using mice showed that when subcutaneously injected, the toxicity of SF-1130 is extremely low.
Even at 9/kg of 0, all cases survived. SF-1130-×
In an immunostimulation test (delayed skin reaction method) in mice transplanted with Sarcoma 180 ascites, it was discovered that the two substances had 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, addition of maltodextrin enhanced immunostimulation. One test example is shown in Table 4. The test method was to inject Sarcoma1 into ICR mice (6 mice per group).
80 ascitic fluids were transplanted, and 24 hours later, each specimen (drug) and anticancer drug (Endoxan) were administered subcutaneously and intraperitoneally, respectively, and on the second day of transplantation, a 6% picrylethanol chloride solution was applied to the shaved abdomen. After that, the sample was administered once a day for 4 days. On the 4th day, Endo Shigen was administered again, and on the 9th day, a 1% chlorinated picryl olive oil solution was applied to the front and back of both ears for secondary sensitization.

After 2 hours, the degree of increase in ear thickness was measured, and the degree of delayed skin reaction was determined based on the rate of increase. Table 4 Taiichi 5 shows an immunostimulation test using Sarcoma 180 solid cancer.

The test method was to use Sarcoma180 in ICR mice (1
(Group of 5 animals) before subcutaneous transplantation, on the 6th day and on the 5th day, Example 2
The mixture of SF-1130-x2 substance and maltobentaose obtained in 1 was intraperitoneally administered, and on the 10th day after transplantation, the specimens were extracted and their size and weight were measured. Table -
As seen in Example 5, significant regression of the solid tumor was observed by pre-administration of the compound of the present invention. Table-5 On the other hand, Sa
rcoma180 was previously implanted into the mouse peritoneal cavity,
A subsequent therapeutic test using the SF-1130 1x2 substance was completely ineffective.

Therefore, it is clear that the SF-1130-x2 substance indirectly exhibits a curative effect by activating the immune resistance of the host. In recent years, it has been discovered that the immune resistance of patients with protozoan disease or cancer is significantly lower than normal, and so-called immunotherapy has begun to be used clinically to prevent this decline and increase the therapeutic effect. There is.

As immunostimulants for this purpose, BCG cells and various polysaccharides are well known, but all of them are polymers.
It has never been reported that oligosaccharides with a molecular weight below one level as shown in the present invention have an immunostimulatory effect, and this is truly unexpected. It is clear that such low-molecular-weight oligosaccharides are practically superior to polysaccharides in terms of absorption/exclusion, tuberculosis infectivity, frequency of allergic reactions, etc.
Furthermore, it is clear from the following test examples that the substance SF-1130-& exhibits a protective effect against bacterial infections and is effective as an immunostimulant.

That is, using JCL:ICR male mice with 8 mice per group, SF-1130-x2 substance was dissolved in a phosphate buffer solution of pH 7.2, and 50 9/K9 or 10 nails 9'k were administered per day.
A dose of 9 was administered intraperitoneally three times at 4 time intervals. Seven hours after the end of the final administration, the culture solution of Staphylococcus aureus Smith S-424 Vermilion was diluted with physiological saline.
A bacterial suspension supplemented with 5% mucin was inoculated intraperitoneally. Bacterial inoculation capacity is 0. I used it as an object/mouse. The number of surviving mice was observed for 5 days after inoculation, and the results shown in Table 6 below were obtained. SF-1130-x2 showed a reduction in bacterial inoculum L when administered with 50 9/k9 compared to the untreated control group
D5.

(which is a guideline for infection prevention effect) is approximately 11.4 times more effective.
An approximately 8.1-fold increase was observed when 9'k9 was administered for 10 days.
In addition, SF-1130-& the substance itself is as shown in Table-4.
Staphylococcus aureus has no antibacterial activity. Table 6 It is also known that immunostimulants are effective in treating autoimmune diseases such as arthritis (rheumatism).

This time, according to the test example shown below, SF-11 of the present invention
The 30-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, 50% of cocoons of bacteria, Mycobacterium butyricum, suspended in physiological saline were used.
．． A bacterial suspension containing 8 females/wing was subcutaneously injected into the right leg of SD rats (male, average weight 180 kg, 10 rats per group) to relieve inflammation.

SF-1130-1 day after bacterial injection for this bacterial infection.
×substance (SF-1130-×,) and SF-1130-×
2) in a 1:2 weight ratio with phosphate buffer (pH 7.
The solution dissolved in 2) was injected subcutaneously once a day for 10 consecutive days. The dose per dose was 1509/k9.

On the 14th day after inoculation of the bacteria, the signs of inflammation (intensity) that occurred in the legs, ears, nose, eyes, and tail of the rats were examined, evaluated on a 4-grade scale (score) from 0 to 3, and the average value was calculated. did. Furthermore, the volume of the left foot was measured to determine the anti-inflammatory efficacy of the SF-1130-x substance. For comparison, in place of the SF-1130-x substance, phenylbutazone was administered once daily to 20 females/day for 11 days.
It was administered orally at a dose of k9/day. Also, SF-1130
A control test was also conducted in exactly the same manner except that administration of -x and phenylbutazone was omitted. The results are shown in Table 7 below. ±-7 According to this test, SF-1130 1x2 substance was found to be effective in preventing experimental arthritis.

When administering SF-1130, intravenous infusion,
Local injection, intramuscular injection, thoracic or intraperitoneal injection is possible, daily or 2 to 3 times a week, 50 to 400 mo'k9, preferably 10
Administer around 09/k9.

Next, examples of the present invention will be shown.

Example 1 Strebutomyces myxogenes strain SF-1130 (Choken Bacteria No. 676) was mixed with 5.0% starch syrup and 2.5% soybean flour.
%, wheat pig sprouts 1.0%, and sodium chloride 0.25% in a liquid tower (pH 7.0), and cultured in an aerated rope frame for 64 hours at 2800 ml in a jar fermenter.

After the cultivation is completed, the culture solution is filtered through an acidic oven at pH 3, and the oven solution becomes 140%
I got it.

This reactor liquid was adsorbed onto a column packed with 20 strong acidic ion exchange resins (Dawex 50W x 2), washed thoroughly with water, and then eluted with 0.1N ammonia water (80% in total).
A fraction of the eluate having antibacterial activity against Ecoli K-1 was concentrated and dried to obtain 250 kg of brown powder.
The 150% flow-through solution was again adjusted to pH 3.0 with hydrochloric acid, and adsorbed onto a column filled with 20% Dowex 50W x 22 (H+) resin. After washing with water, 0.1N ammonia water,
A total of 80 particles were eluted, and the antibacterial active fraction was dried vertically to obtain 51 pieces of brown powder.

Dissolve 47 g of the powder initially obtained in 250 g of water and
After adjusting to H30, it was adsorbed on a column (5.5 x 26 cm) filled with Wako activated carbon 600', and after washing with water,
It was dissolved with 30% ethanol water and then with 35% methanol water, and among each fraction, the fraction 4, which had antibacterial activity against E. coli K-i waves, was concentrated and dried to form a yellowish brown powder. I got it.

This powder was dissolved again in 100% water, adjusted to pH 3 with hydrochloric acid, adsorbed on a column filled with DOWEX 50W x 2 (N ratio +) resin 600Z, thoroughly washed with water, and then 0.1N It was washed with aqueous ammonia, and the antibacterial active fraction was concentrated to dryness to obtain a yellowish brown powder.

Next, one night of this was dissolved in a buffer solution (0.1 M pyridine-formic acid solution, pH = 3.1) at 70 mm, and a column filled with pyridinium type Dowex 50W x 2 (200-400 mesh) at 500 mm was added. (3.5 x 60c)
and eluted with a buffer solution.

The eluate was fractionated into 10 fractions, and each fraction was subjected to high-pressure furnace paper electrophoresis.
Antibacterial activity fraction No. 1 shows a single spot (assuming the Rf of alanine is 1.00). Concentrate 63 to 79 to dryness to obtain SF-1130 1x, substance and SF-1130.
- White powder 380 and others were obtained as a mixture of 2 substances. Next, dissolve this powder in a small amount of water, add a mixed solvent (n-propanol/ethyl acetate/water = 6:1:3),
It was adsorbed onto a column (3x30c) packed with cellulose 200 and developed with a mixed solvent.

The eluate was divided into 7 fractions, and for each fraction,
Paper chromatography was performed using a mixed solvent of ethyl acetate, pyridine, and water (10:4:3), and R-raffinose was 0.57 (1.
Antibacterial activity fraction showing a single spot (as 00), fraction NO. 351-445 were concentrated to dryness to obtain SF-
150 powders of white powder of 1130-x2 material were obtained. At the same time, the pile fungus active fraction No. 1 showed a single spot of R roughinose 0.39. 503 to 550 were dried to dryness to obtain SF-1130-x, substance 70.

Furthermore, 9 parts of powder of SF-1130-x2 substance was dissolved in 6 parts of water, and a column (2 parts) packed with 15 parts of activated carbon was prepared.
After washing with water and eluting with 30% and 35% ethanol, the antibacterial active class 130 flower was concentrated to dryness to obtain white powder 120/9. Melting point: 195q○ (decomposed). SF-1130-x, substance powder 70-9 was treated in the same manner to obtain 55-9 as a white powder. Melting point 2
0300 (decomposition). 2nd Dawex 50 obtained
W x 2 (H+) resin ammonia melt 51 pieces were treated in the same way to obtain SF-1130-x, substance 210-9, S
9 of 520 F-1130-x2 materials were recovered.

Example 2 Streptomyces myxogenes strain SF-1130 (Choken Bacterial Serial No. 676) was inoculated into a liquid medium with the same composition as in Example 1 for 20 days, and incubated in a jar fermenter for 6 hours at 2 = 0. Aerated light azalea culture was performed.

After the cultivation was completed, the culture solution was drained in an acidic oven to obtain 15 pieces of oven solution.
This furnace liquid was poured into a column (6 x 3
6c) and washed thoroughly with water, then 50% of pH8.
E. coli K-12R was removed from the eluate using acetone.
The fraction with the fungal activity was concentrated to dryness to obtain 67.5 kg of yellowish brown powder. Of this, 45 ml of water was dissolved in 200 ml of water, and a column (5.6
×42 skin), after washing with water, 5, 10, 15, 20
, 25% ethanol water sequentially, and the eluate was diluted with 15% ethanol water.
I took it out one by one screaming. Antibacterial activity fraction No. 36
6-510 was concentrated to dryness to obtain white powder 5.7.
This white powder was dissolved in 15 ml of water, passed through a furnace, and 180 ml of ethanol was gradually added to the furnace solution to re-precipitate.
A mixture of SF-1130-1x2 material (10%) and maltobentaose (99%) was obtained in 4.6 hours.

[Brief explanation of the drawing]

Figures 1 and 2 show the infrared absorption spectrum (in KBr tablet) of the SF-1130 monomer substance of the present invention and the 100M
A curve diagram of a Hz hydrogen nuclear magnetic resonance spectrum (in heavy water) is shown. FIGS. 3 and 4 respectively show curve diagrams of SF-1130-x, material infrared absorption spectrum (in KBr tablet) and 100M hydrogen nuclear magnetic resonance absorption spectrum of water (in heavy water). Figure 2 Figure 3 Figure 4

Claims (1)

[Claims] 1. 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); specific optical rotation +155° (water), has no absorption maximum in the ultraviolet region, infrared absorption spectrum shown in Figure 1 of the attached drawings, hydrogen shown in Figure 2 Has a nuclear magnetic resonance absorption spectrum, is easily soluble in water and dimethyl sulfoxide, slightly soluble in alcohol, acetone, ethyl acetate,
It is insoluble in chloroform and benzene, and positive for silver nitrate, red tetrazolium, anthrone, ninhydrin, and Gray-Leapak reagents. Paper chromatography shows that it is insoluble in ethyl acetate, pyridine/water (10:4:
When using the developing solvent of 3), R-raffinose is 0.57,
n-butanol/pyridine/acetic acid/water (6:4:1:3
SF-1 is a white weakly basic powder with an R-raffinose of 0.33 when using a developing solvent of
130-x_2 substances. 2 SF-1130-x_ belonging to the genus Streptomyces
A new antibiotic SF-1130-x_2, which is characterized by culturing two substance-producing bacteria under aerobic conditions to accumulate SF-1130-x_2 substance in the culture solution and collecting it.
A method of manufacturing a substance. 3. An immunostimulant characterized by containing SF-1130-x_2 substance as an active ingredient.