JPS62135498A - 30-2 substance and preparation thereof - Google Patents
30-2 substance and preparation thereofInfo
- Publication number
- JPS62135498A JPS62135498A JP60275221A JP27522185A JPS62135498A JP S62135498 A JPS62135498 A JP S62135498A JP 60275221 A JP60275221 A JP 60275221A JP 27522185 A JP27522185 A JP 27522185A JP S62135498 A JPS62135498 A JP S62135498A
- Authority
- JP
- Japan
- Prior art keywords
- substance
- water
- bacillus
- producing
- ethanol
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Landscapes
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Peptides Or Proteins (AREA)
Abstract
Description
【発明の詳細な説明】
本発明は、新規な３０−２物質及びその製造法に関する
ものである。DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a novel 30-2 substance and a method for producing the same.
更に詳細には、本発明は、アミノ酸１２分子と未知酸１
分子からなる新規な抗真菌性３０−２物質及びその製造
法に関するものである。More specifically, the present invention provides 12 amino acid molecules and 1 unknown acid.
The present invention relates to a novel antifungal 30-2 substance consisting of molecules and a method for producing the same.
本発明者らは、バチルス属の生産する低分子抗菌物質を
求めて鋭意研究した結果、バチルス・エスピー３０−２
の培養物から新規な抗真菌性物質を単離することに成功
したのである。新たにｍ離された抗真菌性物質は３０−
２物質と命名された。As a result of intensive research in search of low-molecular antibacterial substances produced by the genus Bacillus, the present inventors discovered that Bacillus sp.
They succeeded in isolating a new antifungal substance from the culture. The newly released antifungal substance is 30-
Two substances were named.
３０−２物質の生産菌としては、３０−２物質を生産す
ればいかなる菌でもよいが、バチルス・エスピー３０−
２、ＦＥＲＭ Ｐ−８５２５を用いるのが好ましい。バ
チルス・エスピー３０−２は、ダラム染色−陽性、芽胞
染色−陽性、好気性−陽性によって特徴づけられる。As the 30-2 substance-producing bacteria, any bacteria may be used as long as it produces 30-2 substance, but Bacillus sp.
2. It is preferable to use FERM P-8525. Bacillus sp. 30-2 is characterized by Durham staining-positive, spore staining-positive, aerobic-positive.
３０−２物質生産菌の培養培地は、資化できる炭素源、
窒素源、ビタミン等を含有する栄養料等適宜含有したも
ので、３０−２物質を生産蓄積する培地であればいずれ
の培地でもよい。The culture medium of the 30-2 substance-producing bacteria contains an assimilable carbon source,
Any medium may be used as long as it contains appropriate nutrients such as a nitrogen source and vitamins and can produce and accumulate the 30-2 substance.
バチルス・エスピー３０−２の培養に適した培地は例え
ば次のものがあげられる。Examples of suitable media for culturing Bacillus sp. 30-2 include the following.
ポリペプトン ３０ｇ
酵母エキス ５ｇ
ＮａＣＱ ５ ｇ
脱イオン水 １ｆ１
（ＰＨ７，０）
培養は２５〜３０℃で、通気、攪拌、振どう等好気的に
４日間程度行なわれる。Polypeptone 30g Yeast extract 5g NaCQ 5g Deionized water 1f1 (PH7,0) Cultivation is carried out aerobically for about 4 days at 25-30°C with aeration, stirring, shaking, etc.
得られた培養液は濾過して、濾液を得、これに６Ｎ Ｈ
ＣＱを添加し、ｐｌ＋＝３．０に調整し、沈澱物を得る
。沈澱物は水に溶解し、ＮａＨＣＱｉを加え、Ｐ１１＝
７．０とし、次いでエタノールを８０％まで加える。The obtained culture solution was filtered to obtain a filtrate, and 6N H was added to this.
Add CQ and adjust pl+=3.0 to obtain a precipitate. The precipitate was dissolved in water, NaHCQi was added, P11=
7.0 and then add ethanol to 80%.
得られた上清液を真空乾燥し、乾燥物を蒸留水に溶解し
、再び６Ｎ ＨＣＩＩ！を加えてｐＨ＝３．０に調整し
、沈澱物を得る。得られた沈澱物を蒸留水に溶解し、Ｎ
ａＨＣＯａを加え、 ｐＨ＝７．０とし、蒸留水に対し
て透析する。透析内液を凍結乾燥することによって３０
−２物質粗精製物を黄色粉末として得る。The obtained supernatant liquid was vacuum dried, the dried product was dissolved in distilled water, and the mixture was again diluted with 6N HCII! was added to adjust the pH to 3.0 to obtain a precipitate. The obtained precipitate was dissolved in distilled water and N
Add aHCOa to pH=7.0 and dialyze against distilled water. By freeze-drying the dialysis fluid,
-2 Substance crudely purified product is obtained as a yellow powder.
得られた黄色粉末を水に溶解し、アセトニトリル系で高
速液体クロマトにかけ、これを単離し、凍結乾燥するこ
とによってほぼ純品の３０−２物質を白色無定形粉末と
して得る。The obtained yellow powder is dissolved in water, subjected to high performance liquid chromatography using an acetonitrile system, isolated, and freeze-dried to obtain substantially pure substance 30-2 as a white amorphous powder.
得られた３０−２物質の理化学的性質は次の通りである
。The physicochemical properties of the obtained substance 30-2 are as follows.
１、紫外線吸収スペクトルは第１図に示す通りである。1. The ultraviolet absorption spectrum is as shown in FIG.
２、溶解性：エタノール、メタノール、アセトン、水に
不溶で、希エタノール水、希メタノール水、 ＣｌＩＣ
Ｉ２．に可溶である。2. Solubility: Insoluble in ethanol, methanol, acetone, water, dilute ethanol water, dilute methanol water, ClIC
I2. It is soluble in
３、物質の色、性状：白色無定形粉末
４、呈色反応：ニンヒドリンテスト −５、本物質の
アミノ酸組成：本物質の加水分解６、 本物質は真菌に
対して強い抗菌性を示す。3. Color and properties of the substance: White amorphous powder 4. Color reaction: Ninhydrin test -5. Amino acid composition of this substance: Hydrolysis of this substance 6. This substance exhibits strong antibacterial properties against fungi.
サッカロミセス・セレビシエＩＡＭ ４１２５の生育を
８０μｔｒ、ＩＱで阻止する。Growth of Saccharomyces cerevisiae IAM 4125 is inhibited at 80 μtr, IQ.
７、精製法：ｐＨ＝３．０程度で沈澱させ、沈澱物をｐ
Ｈ＝７．０で溶解する工程をくりかえし、最後に高速液
体クロマトにかけることによってほぼ純品を得ることが
できる。高速液体クロマトにおける入２１０による吸光
度曲線は第２図に示す通りである。矢印が３０−２物質
を示している。7. Purification method: Precipitate at pH = about 3.0, and remove the precipitate from pH
By repeating the dissolution step at H=7.0 and finally applying it to high performance liquid chromatography, an almost pure product can be obtained. The absorbance curve obtained by input 210 in high performance liquid chromatography is as shown in FIG. The arrow indicates substance 30-2.
次に本発明の実施例を示す。Next, examples of the present invention will be shown.
実施例１゜
パチンｌ／Ｘ−エスピー３０−２． ＦＥｒｌＭ Ｐ−
８５２５を次の培地に接種して、２７℃で、４日間振ど
う培養した。Example 1゜Pachinl/X-SP 30-2. FErlM P-
8525 was inoculated into the following medium and cultured with shaking at 27°C for 4 days.
ポリペプトン ３０ｇ
酵母エキス ５ｇ
ＮａＣ２５ｇ
脱イオン水 ＩＱ
（ｐＨ７，０）
得られた培養液をｉ＋１過して、培養濾液とし、これを
６Ｎ ＨＣＱでｐ）Ｉ＝３．０として、しばらく放間す
ると沈澱物が得られるので、得られた沈澱物を蒸留水に
溶解し、これにＮａＨＣＯ，を加え、ＰＨ＝７．０とし
、更にエタノールを８０％まで加え、濾過する。得られ
た上清を真空乾燥し、乾固物を得る。この乾固物を蒸留
水に溶解し、 ６Ｎ ＨＯ２でｐ）Ｉ＝３．０とし、し
ばらく放置し、沈澱物を得る。この沈澱物を蒸留水に溶
解し、ＮａＨＣＯ３でｐＨ＝７．０とし、この溶液を蒸
留水に対して透析する。Polypeptone 30g Yeast extract 5g NaC 25g Deionized water IQ (pH 7,0) The obtained culture solution was filtered through i+1 to obtain a culture filtrate, which was mixed with 6N HCQ to set p)I=3.0, and when left for a while, a precipitate formed. The resulting precipitate is dissolved in distilled water, NaHCO is added thereto to adjust the pH to 7.0, ethanol is added up to 80%, and the mixture is filtered. The obtained supernatant is vacuum dried to obtain a dried product. This dried product was dissolved in distilled water, adjusted to p)I=3.0 with 6N HO2, and left to stand for a while to obtain a precipitate. The precipitate is dissolved in distilled water, the pH is brought to 7.0 with NaHCO3, and the solution is dialyzed against distilled water.
得られた透析液を凍結乾燥し、３０−２物質の粗精製物
を黄色粉末で得る。The obtained dialysate is freeze-dried to obtain a crude substance 30-2 as a yellow powder.
得られた黄色粉末を高速液体クロマトにかけ、第２図に
矢印で示す３０−２物質該当部分をとり、これを凍結乾
燥することにより、はぼ純品の３０−２物質を白色無定
形粉末として得た。The obtained yellow powder was subjected to high-performance liquid chromatography, and the portion corresponding to Substance 30-2 as shown by the arrow in Figure 2 was taken, and this was freeze-dried to obtain Substance 30-2, a Habo pure product, as a white amorphous powder. Obtained.
第１図は３０−２物質の紫外線吸収スペクトルを示す図
で、第２図は３０−２物質含有粗精製物の高速液体クロ
マトによる入２１０によ・る吸光度曲線を示す図である
。矢印は３０−２物質を示す。
代理人 弁理士 戸 １）親 男
第 ２ 図
Ｍ聞 （分）FIG. 1 is a diagram showing an ultraviolet absorption spectrum of substance 30-2, and FIG. 2 is a diagram showing an absorbance curve of a crude product containing substance 30-2 obtained by high-performance liquid chromatography. The arrow indicates substance 30-2. Agent Patent attorney 1) Parent Male 2nd figure M (minutes)
Claims (3)
不溶で、希エタノール水、希メタノール水、ＣＨＣｌ＿
３に可溶である。 ３、物質の色、性状：白色無定形粉末 ４、呈色反応：ニンヒドリンテスト− ５、本物質のアミノ酸組成：本物質の加水分解物のアミ
ノ酸分析によって次の結果を得た。 ▲数式、化学式、表等があります▼ ６、本物質は真菌に対して強い抗菌性を示す。 サッカロミセス・セレビシエＩＡＭ４１２５の生育を８
０μｇ／ｌで阻止する。(1) 30-2 substance having the following physical and chemical properties. 1. The ultraviolet absorption spectrum is as shown in FIG. 2. Solubility: Insoluble in ethanol, methanol, acetone, water, dilute ethanol water, dilute methanol water, CHCl_
It is soluble in 3. 3. Color and properties of the substance: White amorphous powder 4. Color reaction: Ninhydrin test - 5. Amino acid composition of this substance: The following results were obtained by amino acid analysis of the hydrolyzate of this substance. ▲There are mathematical formulas, chemical formulas, tables, etc.▼ 6. This substance exhibits strong antibacterial properties against fungi. Growth of Saccharomyces cerevisiae IAM4125 8
Inhibit at 0 μg/l.
、３０−２物質を採取することを特徴とする３０−２物
質の製造法。(2) A method for producing 30-2 substance, which comprises culturing 30-2 substance-producing bacteria belonging to the genus Bacillus and collecting 30-2 substance.
ス・エスピー３０−２である特許請求の範囲第２項記載
の製造法。(3) The production method according to claim 2, wherein the 30-2 substance-producing bacterium belonging to the genus Bacillus is Bacillus sp.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP60275221A JPS62135498A (en) | 1985-12-09 | 1985-12-09 | 30-2 substance and preparation thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP60275221A JPS62135498A (en) | 1985-12-09 | 1985-12-09 | 30-2 substance and preparation thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JPS62135498A true JPS62135498A (en) | 1987-06-18 |
Family
ID=17552390
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP60275221A Pending JPS62135498A (en) | 1985-12-09 | 1985-12-09 | 30-2 substance and preparation thereof |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPS62135498A (en) |
-
1985
- 1985-12-09 JP JP60275221A patent/JPS62135498A/en active Pending
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