JPH0153677B2 - - Google Patents

Description

[Detailed description of the invention]

The present invention relates to antibiotic AB-116. While searching for antibiotics produced by actinomycetes, the present inventors discovered that bacteria belonging to the genus Actinoplanes produce the antibiotic AB-116 and completed the present invention. The physicochemical properties and antibacterial spectrum of the antibiotic AB-116 of the present invention are as follows, and based on these data, the present substance is recognized as a new antibiotic. Physicochemical properties 1 Elemental analysis (experimental value %) C H N 44.08 5.32 19.18 2 Molecular weight of monoacetate: 381 (mass spectrometry) 3 Melting point: Although it does not show a clear melting point, it turns yellow at about 225℃ and becomes approx. Changes to brown at 258℃ 4 Specific rotation: [α] ²⁵ _D -14.0 (c = 1, in dimethyl sulfoxide) 5 Ultraviolet absorption spectrum: As shown in Figure 1, its characteristic absorption is shown in the table below. show.

[Table] 6 Infrared absorption spectrum: As shown in Figure 2, the main absorption positions are 3350, 1710 and 1630 cm ^-1 . 7 Solubility in solvents: Easily soluble; Soluble in dimethyl sulfoxide; Slightly soluble in pyridine, methanol, ethanol, water; Insoluble in n-butanol; Ethyl ether, n-hexane, chloroform, acetone, ethyl acetate8 Color reaction: Ninhydrin reaction; Positive 9 Distinction between basic, acidic and neutral: Basic 10 Color of substance: Colorless

【table】

[Table] Since there are no known antibiotics that match the above physicochemical properties and antibacterial spectrum, the antibiotic AB-116 of the present invention is recognized as a new substance. The antibiotic AB-116 of the present invention is a nucleic acid compound,
Its structure is estimated to be as follows. Examples of nucleic acid antibiotics include Agr.Biol.
chem., 38 (12) 2465-2469 (1974), but the elemental analysis values, melting point,
It can be clearly distinguished from AB-116 of the present invention in its infrared absorption spectrum and structure. The antibiotic AB-116 of the present invention is produced by culturing antibiotic AB-116-producing bacteria belonging to the genus Actinoplanes.
It is obtained by separating and purifying the target product from the culture. An example of an antibiotic AB-116-producing bacterium is Actinoplanes kanagawaensis strain 232-4, which the present inventors isolated and identified from the soil of Kamihara, Sagamihara City, Kanagawa Prefecture.
kanagawaensis232-4) and its mutant strains. This strain has been deposited at the Institute of Microbial Technology, Agency of Industrial Science and Technology as FERMP-6094 (deposit date: August 6, 1982). Next, the mycological properties of this strain are shown.

【table】

【table】

[Table] Shows color numbers.
Strain 232-4 is observed as primitive aerial mycelia on oatmeal agar, starch agar, sucrose/nitrate agar, glycerin/asparagine agar, etc.

[Table] Since this strain does not grow in glucose/peptone gelatin medium, the ability of this strain to liquefy gelatin cannot be determined.

[Table] V Cell wall composition The cell wall of strain 232-4 contains mesodiaminopimelic acid and lysine, but does not contain LL-diaminopimelic acid. Lechevaller's classification [Intr.J.Syst.Bact. 20 435
−443 (1970)], this bacterium is Cell wall type.
It is a type. Based on the above mycological properties, strain 232-4 forms subglobular sporangia on the basal hyphae, the spores are spherical and motile, and the cell wall is shaped, so it is believed that the actinoplanet of actinomycetes It is clear that it is a new species classified into a genus. A related species of this strain is Actinoplanes italicus A-5221 [Intr.J.Syst.Bact. 23 37 (1973)]
can be mentioned. However, strain 232-4 produces a weak pink soluble pigment on glycerol-asparagine agar and glucose-asparagine agar, whereas strain A-5221 produces a thierry-colored soluble pigment. In addition, the 232-4 strain forms abundant sporangia on most media, but the A-5221 strain only forms sporangia on starch agar and Mitsuku agar. From the above, the two can be clearly distinguished. The properties of microorganisms belonging to the genus Actinoplanes vary naturally or artificially, similar to those of other actinomycetes such as Streptomyces. 232
-4 strain is no exception. That is, these strains may mutate naturally or artificially, resulting in strains that differ from the above-mentioned characteristics. Antibiotic AB belonging to the genus Actinoplanes
Cultivation of the 116 producing bacteria can be carried out under aerobic conditions using an artificial or natural medium with appropriate nutritional sources. As a nutrient source for the culture medium, those commonly used for culturing actinomycetes are generally used. for example,
Carbon sources include glucose, rhamnose, arabinose, fructose, etc. Nitrogen sources include corn steep liquor, polypeptone, soybean flour, etc. In addition, salts such as common salt and calcium carbonate, antifoaming agents, etc. may be used as necessary. May be used. The target antibiotic AB-116 can be collected from the culture by appropriately combining conventional physicochemical means. Next, the present invention will be explained in more detail with reference to Examples. Example 1 5 ml of a medium (hereinafter referred to as A medium) consisting of 1.5% glucose, 1.5% soybean flour, 0.1% salt and 0.1% calcium carbonate (the remainder is water) was dispensed into test tubes and sterilized, followed by slant culture. One platinum loopful of the -4 strain was inoculated and cultured at 30°C for 5 days in a reciprocating shaking incubator. Further, 70 ml of A medium was dispensed into a sterilized rotary flask (500 ml volume), and 5 ml of the above-mentioned seed culture was inoculated, and cultured at 30°C on a rotary shaker for 3 days. Dispense 700 ml of medium A into a sterilized Erlenmeyer flask (3 volumes), inoculate with 70 ml of this inoculum, and similarly culture for 3 days. Next, the above seed culture 3.5 was transplanted into a sterilized stainless steel tank (200 volumes) by pouring 100 ml of a medium (PH 7.0) consisting of 1.5% glucose, 1.5% soybean flour, 0.3% salt, and 0.1% calcium carbonate into a sterilized stainless steel tank (200 volumes). ℃ under aeration and stirring (aeration 50/min, stirring 150 times/min, internal pressure 0.5Kg/cm ² )
Incubate for 120 hours. Celite (superior agent) for culture
Add diamond ions to the resulting solution.
After adding 6 volumes of Hp-20 and stirring at room temperature for 1 hour, the column was filled with Diaion Hp-20. column
Wash with 10% water and then elute with 10% acetone water. Distill the acetone under reduced pressure and use Amberlite IRC-50.
(H type) Adsorb into 500ml and wash with water. Elute with 0.1N hydrochloric acid and neutralize the eluate with 1N sodium hydroxide. This is passed through a Diaion Hp-20 (500 ml) column, and then eluted with 10% acetone water.
The eluate was concentrated under reduced pressure, passed through Sephadex G-10, and concentrated to dryness. The residue was dissolved in a small amount of 90% methanol, passed through Sephadex LH20, and the active fraction was collected and concentrated to dryness to obtain 500 mg of a single spot of AB-116 using thin layer chromatography.

[Brief explanation of drawings]

Figures 1 and 2 show the ultraviolet absorption spectrum and the infrared absorption spectrum measured by the KBr method of antibiotic AB-116, respectively.

Claims (1)

[Scope of Claims] 1 Antibiotic AB-116 produced by a bacterium belonging to the genus Actinoplanes and having the following properties: (1) Elemental analysis; Experimental values of elemental analysis are approximately as follows. C: 44.08 (%) H: 5.32 (%) N: 19.18 (%) (2) Molecular weight of AB-116 monoacetate; 381 (mass spectrometry) (3) Melting point: Shows a clear melting point However, it turns yellow at about 225℃ and turns brown at about 258℃. (4) Specific light absorption; [α] Around ²⁵ _D -14 (c=1, in dimethyl sulfoxide) (5) Ultraviolet absorption spectrum ;The maximum absorption wavelength of AB-116 aqueous solution is 234nm, 272nm and 298nm
(6) Infrared absorption spectrum; the main absorption position is
Around 3350, 1710 and 1630 cm ^-1 (7) Solubility in solvents: Easily soluble - Soluble in dimethyl sulfoxide - Slightly soluble in pyridine, methanol, ethanol, Slightly soluble in water - Insoluble in n-butanol - Ethyl ether, n-hexane, chloroform, Acetone, ethyl acetate (8) Color reaction; positive ninhydrin reaction (9) Distinction between basic, acidic, and neutral; basic (10) Color of substance; colorless.