JPS5832893A - Antibiotic ab-116 - Google Patents

Abstract

NEW MATERIAL:An antibiotic AB-116 having a presumed structure given as the formula and following physical and chemical properties: specific rotatory power: [alpha]D<25>=-14.0 (c=1 in dimethylsulfoxide); melting point: showing no definite melting point, yellowing at 225 deg.C, browning at 258 deg.C; molecular weight: 381 as monoacetate according to mass spectroscopy; elementary analyses in %: C 44.08, H 5.32, N 19.18; easily soluble in dimethylsulfoxide, soluble in pyridine, methanol, water, slightly soluble in n-butanol, insoluble in ethyl ether, n-hexane, chloroform or acetone; positive to ninhydrin reaction; colorless. USE:Antibiotic. PREPARATION:For exampel, a strain in Actinoplanes such as Actionoplanes kanagawaensis 232-4 strain (FERM P-6094) is aerobically cultured in an enriched medium.

Description

DETAILED DESCRIPTION OF THE INVENTION The present inventors, while searching for antibiotics produced by actinomycetes, 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'HN 44, 08 5.32 19.182, molecular weight: 381 (mass spectrometry) 3 Melting point: Does not show a clear melting point, but turns yellow at about 225°C , changes to brownish color at about 258℃ 4.Specific optical rotation: [α] -14.0 (c=1,
(in dimethyl sulfoxide) 5. Ultraviolet absorption spectrum: As shown in Figure 1, its characteristic absorption is shown in the following table.

(Left below) 6 Infrared absorption spectrum: As shown in Figure 2, the main absorption positions are 335.0, 1715 and 1630.
Cm 1.

7 Solubility in solvents: Easily soluble; Dimethyl sulfoxide soluble; Slightly soluble in pyridine, methanol, ethanol, water; Roubutanol insoluble; -r-f ether n-7 Xane, chloroform, acetone, ethyl acetate 3-8 Color reaction : Ninhydrin reaction; positive 9, basic/acidic/neutral distinction: basic 10, color of substance: colorless 17 Terrasima 〉100 Esierichia coli NIHJ JO-2100u u
P-510110011not K11
6
．． 25rrnP-51208ABPC-r5017
u P-51213NA-r
100u I7 K-1212.5 There is no known anti-inflammatory substance that matches the physicochemical properties and antibacterial spectrum of K-1212.5, so the antibiotic AB-116 of the present invention
is recognized as a new substance. Antibiotic AB-11 of the present invention
Although the structure of 6 is unknown, it is presumed to be a nucleic acid-based substance based on the above physical and chemical properties. Examples of nucleic acid antibiotics include Agr, Biol, chem, 38 (1
2) Those described in 2465-2469 (1974) can be mentioned, but they can be clearly distinguished from AB-116 of the present invention' in elemental analysis values, melting points, and infrared absorption spectra.

The antibiotic AB-116 of the present invention can be obtained by culturing antibiotic AB-116-producing bacteria belonging to the genus Actinoplanes and separating and purifying the target product from the culture. An example of an antibiotic AB-116-producing bacterium is Actinoplanes kinagawaensis strain 232-4 (Acti
noplanes kanagawaensis232
-4) and its mutant strains.

This strain has been deposited with the National Institute of Microbial Technology, Agency of Industrial Science and Technology as Fiber Engineering Research Institute No. 6094 (FERM p-6094) (deposited on August 6, 1982).

Next, the mycological properties of this strain are shown.

4- ■ Growth status on various media Strain 232-4 was grown on oatmeal agar, starch agar, sucrose/nitrate agar, and glycerin/
Primitive aerial hyphae can be seen on asparagine agar media, etc.

■ Physiological properties Since this strain does not grow in glucose-peptone gelatin medium, the ability of this strain to liquefy gelatin cannot be determined.

7- ■ Assimilation of carbon sources (on Pridham-Godelive agar) Carbon sources
Anabolic L-arabinose + D-xylose D-glucose + D-fructose + sucrose + inositol L-rhamnose + raffinose - D-mannitol + ■ Cell wall composition The cell wall of strain 232-4 contains mesodiaminopimelic acid and Contains lysine and does not contain LL-diaminopimelic acid.

Lechevaller's classification (Intr, J, 5ys
t, Bact, 20435-443 (1970)), this bacterium is Cell wall 8-type II.

Based on the above mycological properties, strain 232-4 forms hemocyte-shaped sporangia on the basal hyphae, the spores are spherical and motile, and the cell wall is shaped like a ■, indicating that the actinobacteria It is clear that this is a new species classified in the genus Planes.

Related species of this strain include Actinoplanes italicus A-5221 (Intr, J, 5yst, B
Act, 2337 (1973):].

However, strain 232-4 produces a weak pink soluble pigment on glycerol-asparagine agar and glucose-asparagine agar, whereas strain A-5221 produces a cherry-colored soluble pigment. In addition, the 232-4 strain forms abundant sporangia in most media, but the A-5 strain
Strain 221 only forms on starch agar and milk 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. The 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.

The bacteria producing the antibiotic AB-116 belonging to the genus Actinoplanes can be cultured under aerobic conditions using an artificial medium or a natural medium having an appropriate nutrient source. 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 stew liquor, polypeptone, soybean flour, etc., and salts such as common salt and calcium carbonate, and antifoaming agents as necessary. etc. may also be used.

The objective 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 Glucose 1. ．． A medium (
After dispensing 5me (hereinafter referred to as A medium) into test tubes and sterilizing them, one platinum loop of the 232-4 strain obtained from the slant culture was inoculated and cultured at 30°C for 5 days in a reciprocating shaking incubator. Further, 70 me of A medium was dispensed into a sterilized rotary flask (500 ml? capacity), and the above-mentioned seed culture 5 mi' was inoculated and cultured on a rotary shaker at 30'C for 3 days. Dispense 700 ml of A medium into a sterilized Erlenmeyer flask (capacity 3), inoculate 70 ml of this inoculum, and similarly culture for 3 days. Then glucose 15%.

Soy flour 15% 1 salt 03% and calcium carbonate 0.1
3.5 t of the seed culture was transferred to a sterilized stainless steel tank (200 ml capacity) by pouring 100 t of a medium (pH 7.0) consisting of min, internal pressure 0.5
ν'cm2) for 120 hours. Celite 11" (filter aid) is added to the culture and filtered. To the obtained P solution, 6 of Diamond Ion Hp-20 is added and stirred at room temperature for 1 hour = 11- Extrusion, Diamond Ion Hp-20 is added Fill the column.

The column was washed with 10 tons of water and then bathed with 10% acetone water. Acetone was distilled off under reduced pressure and Amberly) IRC
-50 (H type) Adsorb to 500+++e and wash with water. 0. Elute with IN hydrochloric acid and neutralize the eluate with IN sodium hydroxide. Add this to Diaion Hp--20 (50
0me) column and then elute with 10% acetone water. The eluate was concentrated under reduced pressure and transferred to Sephadex G-10.
and concentrate to dryness. The residue was dissolved in a small amount of 90% methanol, passed through Sephadex LH20, the active fraction was collected and concentrated to dryness, and a single spot of A was obtained using thin layer chromatography.
Obtain 500 ■ of B-116.

[Brief explanation of the drawing]

FIGS. 1 and 2 show the ultraviolet absorption spectrum and the infrared absorption spectrum measured by the KBr method of antibiotic AB-116, respectively. 12- Letter of amendment to postal procedures (spontaneous) July 2, 1980 1, Indication of the case 1982 Patent Application No. 129121 2, Name of the invention Antibiotic AB-116 3. Amendment Relationship with the case of the person who filed the patent application Address of the patent applicant: 3-25 Doshomachi, Higashi-ku, Osaka Name: 291
Dainippon Pharmaceutical Co., Ltd. Director: Tomio Fujiwara President 4, Agent address: 33-94 Enoki-cho, Suita-shi, Osaka Prefecture Dainippon Pharmaceutical Co., Ltd. Research Institute Name: Patent attorney ± 7652 Yushi Tsuboi: 5, Subject of amendment 11.6. Contents of amendment (1) The scope of claims is amended as shown in the attached sheet. (2) On page 2, line 6, 1-1 of the specification, “molecular weight:
” is corrected to be a molecule of 17 noacetate, Ll: Ni1. (3) Page 3 of the specification, lines 1 and 8 [r 17 in 1
15J is corrected to r1710.1. (11) In the last line of page 4 to line 3 of page 5 of the specification, “
Is the structure of the antibiotic AIIIG of the present invention unknown?
......-It is presumed to be a nucleic acid-based substance. ” has been amended as follows. 1. The antibiotic Al1-11G of the present invention is a nucleic acid compound, and its structure is estimated to be as follows. (5) In page 2, line 6 of the specification, the phrase ``in infrared absorption spectrum'' is corrected to 1 in r infrared absorption spectrum and structure. - The above claims are produced by a bacterium belonging to the genus Actinoplanes, and the specific optical rotation is [α]25-
14.0 (c = 1. in dimethyl sulfoxide), yellow at about 225°C, and antibiotic AB-116° at about 258°C.

Claims (1)

[Claims] It is produced by a bacterium belonging to the genus Actinoplanes, and its specific optical rotation is [α) sweet-
14,0 (c=1.'/2 in methyl sulfoxide), turns yellow at about 225°C, turns brown at about 258°C, and has a positive ninhydrin reaction, with a molecular weight of 381 (mass spectrometry). ) Antibiotic AB-116゜