JPS5867189A - Polyene anti-bacterial antibiotic and production thereof - Google Patents
Polyene anti-bacterial antibiotic and production thereofInfo
- Publication number
- JPS5867189A JPS5867189A JP57136457A JP13645782A JPS5867189A JP S5867189 A JPS5867189 A JP S5867189A JP 57136457 A JP57136457 A JP 57136457A JP 13645782 A JP13645782 A JP 13645782A JP S5867189 A JPS5867189 A JP S5867189A
- Authority
- JP
- Japan
- Prior art keywords
- antibiotic
- antifungal
- medium
- water
- producing
- 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.)
- Granted
Links
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- FBPFZTCFMRRESA-GUCUJZIJSA-N galactitol Chemical compound OC[C@H](O)[C@@H](O)[C@@H](O)[C@H](O)CO FBPFZTCFMRRESA-GUCUJZIJSA-N 0.000 description 1
- WHWDWIHXSPCOKZ-UHFFFAOYSA-N hexahydrofarnesyl acetone Natural products CC(C)CCCC(C)CCCC(C)CCCC(C)=O WHWDWIHXSPCOKZ-UHFFFAOYSA-N 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- CDAISMWEOUEBRE-GPIVLXJGSA-N inositol Chemical compound O[C@H]1[C@H](O)[C@@H](O)[C@H](O)[C@H](O)[C@@H]1O CDAISMWEOUEBRE-GPIVLXJGSA-N 0.000 description 1
- 238000001990 intravenous administration Methods 0.000 description 1
- 235000013372 meat Nutrition 0.000 description 1
- 230000010534 mechanism of action Effects 0.000 description 1
- 244000000010 microbial pathogen Species 0.000 description 1
- 230000007483 microbial process Effects 0.000 description 1
- 235000013336 milk Nutrition 0.000 description 1
- 239000008267 milk Substances 0.000 description 1
- 210000004080 milk Anatomy 0.000 description 1
- 239000002052 molecular layer Substances 0.000 description 1
- 239000003471 mutagenic agent Substances 0.000 description 1
- 231100000707 mutagenic chemical Toxicity 0.000 description 1
- 235000010298 natamycin Nutrition 0.000 description 1
- 239000004311 natamycin Substances 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 239000012074 organic phase Substances 0.000 description 1
- 238000004816 paper chromatography Methods 0.000 description 1
- 239000007793 ph indicator Substances 0.000 description 1
- 239000012071 phase Substances 0.000 description 1
- 238000002135 phase contrast microscopy Methods 0.000 description 1
- 239000011148 porous material Substances 0.000 description 1
- 210000002307 prostate Anatomy 0.000 description 1
- MUPFEKGTMRGPLJ-ZQSKZDJDSA-N raffinose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO[C@@H]2[C@@H]([C@@H](O)[C@@H](O)[C@@H](CO)O2)O)O1 MUPFEKGTMRGPLJ-ZQSKZDJDSA-N 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- CDAISMWEOUEBRE-UHFFFAOYSA-N scyllo-inosotol Natural products OC1C(O)C(O)C(O)C(O)C1O CDAISMWEOUEBRE-UHFFFAOYSA-N 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 229910052710 silicon Inorganic materials 0.000 description 1
- 239000010703 silicon Substances 0.000 description 1
- 229910052708 sodium Inorganic materials 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 239000002689 soil Substances 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 235000015096 spirit Nutrition 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- YWYZEGXAUVWDED-UHFFFAOYSA-N triammonium citrate Chemical compound [NH4+].[NH4+].[NH4+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O YWYZEGXAUVWDED-UHFFFAOYSA-N 0.000 description 1
- 210000004881 tumor cell Anatomy 0.000 description 1
- 239000008158 vegetable oil Substances 0.000 description 1
- 229930195724 β-lactose Natural products 0.000 description 1
Classifications
-
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/44—Preparation of O-glycosides, e.g. glucosides
- C12P19/60—Preparation of O-glycosides, e.g. glucosides having an oxygen of the saccharide radical directly bound to a non-saccharide heterocyclic ring or a condensed ring system containing a non-saccharide heterocyclic ring, e.g. coumermycin, novobiocin
- C12P19/62—Preparation of O-glycosides, e.g. glucosides having an oxygen of the saccharide radical directly bound to a non-saccharide heterocyclic ring or a condensed ring system containing a non-saccharide heterocyclic ring, e.g. coumermycin, novobiocin the hetero ring having eight or more ring members and only oxygen as ring hetero atoms, e.g. erythromycin, spiramycin, nystatin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/04—Antibacterial agents
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
- C12N1/205—Bacterial isolates
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- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/465—Streptomyces
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- Communicable Diseases (AREA)
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- Micro-Organisms Or Cultivation Processes Thereof (AREA)
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- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
(57)【要約】本公報は電子出願前の出願データであるた
め要約のデータは記録されません。(57) [Summary] This bulletin contains application data before electronic filing, so abstract data is not recorded.
Description
【発明の詳細な説明】
本発明はストレプトミセス属に属する新規做生物の酵素
プロセスによって従来知られていないある種のポリエン
抗真菌性抗生物質(antifungalantibi
otics )を製造する方法に関する◎また、本発明
は新規なテトラエン抗生物質に関する。DETAILED DESCRIPTION OF THE INVENTION The present invention is directed to the production of a polyene antifungal antibiotic, which has been previously unknown, by the enzymatic process of a novel organism belonging to the genus Streptomyces.
The present invention also relates to a novel tetraene antibiotic.
従来技術の記載
ポリエン抗生物質は極めて興味ある生物学的特性を与え
る弁膜二重結合を含有する構造を持った基本特性を有す
る物質である。Description of the Prior Art Polyene antibiotics are substances whose basic properties include a structure containing a valvular double bond which gives them very interesting biological properties.
これらの化合物としては、例えばアンホテリシンB(ア
メリカ特許第2,908,611および2゜908.6
12号明m書)、フィリッピン コンプレックス(ph
ilippine complex ) (英国特許第
788.486号明細書)、ナイスクチン(英国特許第
866,600号明細書)、テネセチンまたはナタマイ
シントして知られているピマリシン(英国特許第852
,888号明細書およびドイツ特許第1,024,20
6号明細書ン、カンジシジン(アメリカ特許第2,99
2,162号明細書)等を列挙する゛ことができる。こ
れらの物質はUVおよびIRスペクトルによりおよびク
ロマトグラフィー(ペーパー、薄層または高圧)により
互いに区別することができる。These compounds include, for example, amphotericin B (U.S. Pat. No. 2,908,611 and 2.908.6).
No. 12 Mei M), Philippine Complex (ph
ilippine complex) (UK Patent No. 788,486), Nyscutin (UK Patent No. 866,600), Pimaricin (also known as Tenesetine or Natamycin) (UK Patent No. 852)
, 888 and German Patent No. 1,024,20
Candicidin (U.S. Pat. No. 2,999)
2,162 specification), etc. These substances can be distinguished from each other by UV and IR spectra and by chromatography (paper, thin layer or high pressure).
ポリエン抗生物質はアクチノミセタレス(ActinO
−mycetales )目、特にストレプトミセス属
およびストレプトペルチシウム(Streptover
ticillinm)の微生物によって主として生ずる
。Polyene antibiotics are ActinO
-mycetales), especially the genus Streptomyces and Streptovertisium
ticillinm) microorganisms.
通常、これらの化合物の生合成は行われないが、しかし
1つの同じ培地において数種のポリエンの出現が観察さ
れる。Usually, biosynthesis of these compounds does not take place, but the appearance of several polyenes in one and the same medium is observed.
真核細胞(eukaryotic cells )にお
ける作用機構は膜ステロールにおいて作用する機構であ
り、これらのステロールにより与えられる選択的透過性
が変化する際に細胞の崩壊を生ずる。The mechanism of action in eukaryotic cells is one that operates on membrane sterols, resulting in cell collapse when the selective permeability conferred by these sterols is altered.
ポリエンは医薬の分野において広く用いられており、こ
れらポリエンの毒性は静脈内投与によるものであるけれ
ども、ポリエンは特別に毒性が存在しないから主として
局所的にまたは経口的に用−いられている。Polyenes are widely used in the pharmaceutical field, and although the toxicity of these polyenes is due to intravenous administration, since polyenes do not have any particular toxicity, they are mainly used topically or orally.
しかしながら、最近興味ある物質としてのポリエンが作
られるようになったことは、腫瘍細胞と正常細胞とにお
ける膜の透過性に及ぼす作用相互間にある差の存在する
ことが見出されるようになったことである。また、これ
らの化合物(カンジシジン)を経口投与する場合に血液
コレ不テロール レベルが減少する他の興味ある事実が
確められている。また、ある種のポリエンは前立線の肥
大に作用することが知られている。この事は、これらの
物質を生ずる微生物についての研究によって確められて
いる。However, the recent creation of polyenes as interesting substances has led to the discovery that there are some differences in their effects on membrane permeability in tumor cells and normal cells. It is. Another interesting fact has also been established that blood cholesterol levels are reduced when these compounds (candicidin) are administered orally. Additionally, certain polyenes are known to act on enlargement of the prostate gland. This has been confirmed by studies of the microorganisms that produce these substances.
発明の概要
本発明においては、特に抗真菌性活性を有する数種類の
ポリエン物質の生成について記載し、このうち顕著な物
質について出願人はAb 4 (+・0と名づけ、この
主な生物学的特性は菌類および酵母菌の生長を抑制する
特性を有していることであり、この特性によってこれら
のポリエン物質は動物および植物において病原性微生物
(上述するクラスに属する)により生ずる感染の処理に
用いることができる。SUMMARY OF THE INVENTION In the present invention, we describe the production of several polyene substances with particular antifungal activity, of which the applicant has named Ab 4 (+.0) and whose main biological properties have been described. have the property of inhibiting the growth of fungi and yeasts, and this property makes these polyene substances suitable for use in the treatment of infections caused by pathogenic microorganisms (belonging to the above-mentioned classes) in animals and plants. I can do it.
Ab 400をストレプトミセス属に属する好気性細菌
の培地で生成し、この生成物質をストレプトミセス属S
p、菌株5F−1[ムSA)と名づけだ。この微生物の
他の特性は他のポリエン物質を生成することであり、こ
のうち顕著なことはピマリシンと称するすでに知られた
テトラエンであることである。Ab 400 was produced in a culture medium of aerobic bacteria belonging to the genus Streptomyces, and the product was transformed into a culture of aerobic bacteria belonging to the genus Streptomyces.
p, and the strain was named 5F-1 [MuSA]. Another property of this microorganism is that it produces other polyene substances, the most prominent of which is the already known tetraene called pimaricin.
また、本発明においては上述する物質の抽出およびmg
、並びにAb 400の抗真菌性スペクトルについて説
明する。In addition, in the present invention, extraction of the above-mentioned substance and mg
, and the antifungal spectrum of Ab 400.
微生物の特性
使用した微生物はマドリード付近から採集したンチビオ
チブス ソシエタ アノニムに所属するコレクション
におよびANCIBII、フ88でスコツトランド、ア
バーデンのトレイ リサーチステーションのナショナル
コレクション オプインダストリアル バクテリア(
NOIB)に寄託した。Characteristics of the microorganisms The microorganisms used were collected from the vicinity of Madrid, in a collection belonging to the Antibiotics Societa Anonyme, and in the National Collection of Industrial Bacteria, Trey Research Station, Aberden, Scotland, in ANCIBII, F88.
NOIB).
シケ7. (5ykes )およびスキナー(5kin
rler )氏(1978年]、アカデミツク プレス
にょる「ザ アクチノミセ3r レス(The Act
inomycetales月襄示されている指針には、
微生物の毒性の確認および結論により一生物は渦巻状で
なく、また胞子のう状でもなく、鎖状に多数に胞子を含
有する胞子体を有する気菌糸本を有することから、微生
物がストレプトミセス属に属することを報告している。Shike 7. (5ykes) and Skinner (5kin
rler) (1978), Academic Press, ``The Act.
The guidelines shown in the inomycetales include:
As a result of the confirmation and conclusion of the toxicity of the microorganism, the organism is not spiral-shaped or sporangium-shaped, but has an aerial mycelium with chain-shaped sporophytes that contain many spores. It is reported that it belongs to
「インターナショナル ストレプトミセス プロジェク
トJ −I S P −(Int、 J、 5ist、
Baote−riology16 : ala 〜4
0,1966ンにシルリング(Shirling )
オよびゴトリエプ(Gottlieb)氏により記載さ
れた方法には種の特性について記載されている。位相差
顕微鏡下で、微生物5F−1(ASA )はl5P−a
およびl5P−4培地において1〜6個の幾分緻密なス
パイラルを有し、かつ10以上の0.8 X O,5μ
卵型胞子の鎖を有するらせん状胞子体?有することが観
察された0ISP−2およびl5P−Is培地において
、これらの胞子体は真のスパイラルを示さないが、しか
しときどき1つのスパイラルについて近接撮影した場合
に幾分!&密なホック(hooks )が観察された0
胞子は電子顕微錬下でいぼ状突起表面を示した。“International Streptomyces Project J-ISP-(Int, J, 5ist,
Baote-riology16: ala ~4
Shirling at 0,1966
The method described by O and Gottlieb describes the characteristics of the species. Under phase contrast microscopy, microorganism 5F-1 (ASA) is 15P-a
and 1 to 6 somewhat dense spirals in l5P-4 medium and 10 or more 0.8
Spiral sporophyte with chains of ovoid spores? In the 0ISP-2 and 15P-Is media which were observed to have these sporophytes do not show true spirals, but sometimes when one spiral is photographed up close! & Dense hooks observed 0
The spores showed a warty surface under electron microscopy.
異なる培地における培養の特性を表1に示す。The characteristics of the cultures in different media are shown in Table 1.
培養はすべて28℃で行った。色は2つの方法、すなわ
ち、E、Seguy氏によるEa、 P、 Leche
valiez(Paris ) (1986)
、 r (7ode Universal de
couleurs J (7) シー )による方法お
よび色についての正規の名前を用い、これらのサブジェ
クトネームを付ける方法によって示した。All cultures were performed at 28°C. Color can be determined in two ways: Ea by Mr. E. Seguy, P. Leche
valiez (Paris) (1986)
, r (7ode Universal de
couleurs J (7) C) and the regular names for colors, and the method of naming these subjects.
窒素(N)i(7)同化スベク) st (assim
ilat土Onspectrum )を表Hに示す。1
%のグルコースおよび1.5%の寒天(リュデマンに、
M、 (LuedemannG、M、 )氏、r In
t、 J、 5ist、 Baoteriol、 21
:240〜47(1971)J)を基培地として用い
た。Nitrogen (N) i (7) assimilation subek) st (assim
Ilat soil spectrum) are shown in Table H. 1
% glucose and 1.5% agar (Ludemann,
Mr. M. (LuedemannG, M.), r In
t, J, 5ist, Baoteriol, 21
:240-47 (1971) J) was used as the base medium.
炭素(C)源の同化スペクトルをシルリングE。Schilling E assimilation spectra of carbon (C) sources.
B、およびゴットリエプD。氏による方法(Int。B, and Gottlieb D. (Int.
J、 5ist、 Bacteriol、 16 :
31 B 〜340 (1966))によって得た。こ
の結果を表■に示す。J, 5ist, Bacteriol, 16:
31 B-340 (1966)). The results are shown in Table ■.
生理学的特性を表■に示す。Physiological properties are shown in Table ■.
要約すると、研死における菌株は次に示すような°系統
学的特性を有していた。In summary, the strains in Kenshin had the following phylogenetic characteristics.
表 ■
象の同化
対照−一−−〜−−−−−−−−−−−NH4No8−
−−−−−−−−−−−N−ZアミンA −−−−−−
−−−+酵母エキス −−−−−−−−−−++
÷+アスパラギン =−−−−−−−+
+++グルタ之ン酸 −−一−−−−−−
±秦28°Cで14日間
表 璽
炭素源 生長秦
対照−−−−−−−−−−−−−−−−−−−−D−グ
ルコース −−−−−−−−−−−−−++スクロース
ーーーーーーーーーー−−−−−一 ±イノ
シトール −−−−−−−−−−−−−−
±D−7ラクトース −−−−−−−−−−−++ラフ
ィノース −−−−−−−−−−−−−−
±L−アラビノース −−−−−−−−−−−D−キシ
四−スーーーーーーーーーーーーーD−マンニトール
−−−−−−−−−−−++L−ラムノース −一−−
−−−−−−−−D−ガラクトース −一−−−−−−
−−− ±D−アラビノース −−−−−
−−−−−−−α−メリビオース −−−−−−−−−
−−−グリセロール −一一−−−−−−−−−−皓
++β−ラクトース −−−−−−−−−−
−−−殿粉−−−−−−−−一−−−−−−−−−−−
+表 1 (続き)
D−リボース −−−−−−−−−−−−−−−−−七
ロビオース 脅−−”””=−−−−−−−−+十トレ
ハロース −””””””−−”−−−−−−++ソル
ボース −−−−−−−−−−−−−−−−−一−ソル
ビトールーーーーーー−−−一一一−−−−−−マンノ
ースー−−=−−”−””−”−”−−−−++ダルシ
トール −−−−−−−−−−−−−−−−−−メレジ
トース −−−−−−−−−−−−−−−−−一≧
脩
1英国特許第846,938号明細書(1960)2シ
ルリングE、B、およびブト91100氏「工nt。Table ■ Elephant assimilation control-1------------NH4No8-
-----------N-Z amine A -------
−−−+Yeast extract −−−−−−−−−−++
÷+asparagine =−−−−−−−+
+++Glutanoic acid −−−−−−−−
±Qin Table for 14 days at 28°C Table carbon source Growth Qin control −−−−−−−−−−−−−−−−−−−−−D-Glucose −−−−−−−−−−− −++Sucroseーーーーーーーーーーーーーーーーーーーーーー1 ±Inositol −−−−−−−−−−−−−−
±D-7 Lactose −−−−−−−−−−−++Raffinose −−−−−−−−−−−−−−
±L-arabinose ------------D-xy4-su-------D-mannitol
−−−−−−−−−−−++L−Rhamnose −1−−
−−−−−−−−D-galactose −1−−−−−−
--- ±D-arabinose ---
−−−−−−−α-Melibiose −−−−−−−−−
−−−Glycerol −1−−−−−−−−−−Hao
++β-lactose −−−−−−−−−−
−−−Starch −−−−−−−−−−−−−−−−
+Table 1 (Continued) D-Ribose −−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−10trehalose−−−−−−−−−−−−−−−−−−−−−−−−−− ”””−−”−−−−−−++Sorbose −−−−−−−−−−−−−−−−−1−Sorbitol−−−−−111−−−− −−Mannose −−=−−”−””−”−”−−−−++Dulcitol −−−−−−−−−−−−−−−−−−−Melezitose −−−−−−−−− ------------1≧ Shu 1 British Patent No. 846,938 (1960) 2 Schilling E, B, and Buto 91100 "Eng. nt.
J、 5ist、 Bacteriol、 Jl 8
: 69−189(1968)
8、シルリングE、B、およびテトラエンIL 氏r
Int。J, 5ist, Bacteriol, Jl 8
: 69-189 (1968) 8, Schilling E, B, and Tetraene IL Mr.
Int.
J* 5ist、 Bacterol、JAS!:
265−394(1972)弧この研凭所により得た結
果
a)スパイラル部分に属する。J* 5ist, Bacterol, JAS! :
265-394 (1972) Arc The results obtained by this laboratory a) belong to the spiral part.
b)いぼ状表面を持つ胞子を有する。b) It has spores with a warty surface.
C)メラノイド色素を生じない。C) Does not produce melanoid pigments.
d)胞子形成した気菌糸体の色は使用する培地によって
スミレ色糸か、または赤色系に属する08) pH指
示薬に作用しない可溶性赤(赤がかった褐色)色素を生
ずる。d) The color of the sporulated aerial mycelium is either violet or red depending on the medium used. 08) It produces a soluble red (reddish brown) pigment that does not act on pH indicators.
f)基質菌糸体の色は培地によって赤がかったクリ色か
らスミレ−クリ色に変化する。f) The color of the substrate mycelium changes from reddish chestnut to violet chestnut depending on the medium.
g)炭素源として次の材料を同化するニーグルコース
−グリセロールーイノシトール
−殿粉
一7ラクトース゛ −セロビオース−マンニ
トール −トレハロースーガラクトース
−マンノーススクロース(5uchr、ose
)およびう7/−スを同化するかどうかは明確でない
。g) Neglucose, which assimilates the following materials as a carbon source:
-Glycerol-inositol
- Starch - 7-lactose - Cellobiose - Mannitol - Trehalose-galactose
-mannose sucrose (5uchr, ose
) and 7/-s assimilated is not clear.
h)他の生理学的特性: L<ラチンの著しい加水分解を生ずる。h) Other physiological properties: L<results in significant hydrolysis of latin.
1 硝醗塩を亜硝酸塩に還元する。1. Reduce nitrate to nitrite.
& 牛乳カゼインを加水分解しない。& Does not hydrolyze milk casein.
44%Na0jまで耐える。Withstands up to 44% Na0j.
L )lVフイリツク(mesophyl土0〕であ
る。L) lV filtrate (mesophyl 0).
i)数種類のテトラエン抗真菌性抗生物質を生ずる。i) Generates several tetraene antifungal antibiotics.
「定l1Jllll菌学のペルゲイの便覧(Be3rl
lf9fB 1anualor D6termina
1=−ye BaCt8riOIOgy ) J
(1974年、第8版、ザ ウィリアムス アン
ド ゥィルキンス カンパニー)、および「ストレプト
ミ七スノ榛準菌株の協同記載(Cooperative
descriptionof Type 5trai
ns of StreptomyCes ) Jにシル
リングおよびゴトリエプ氏により発表された「インター
ナショナル ストレプトミセス プロゼクト」論文(r
Int、 J、 5ift、 Bacteriolコ
18:69〜189.1968:同文献18 + 2
!79〜892.1968:同文献19:891〜51
2.1969:および同文献22:265〜894.1
972)並びに1974年から現在まで発行されたこれ
らの文献におけるストレプトミセス種の系統学的特性を
観察して、5F−IAsA)に類似する微生物は見出す
ことができなかった。``Pergay's Handbook of Mycology (Be3rl)
lf9fB 1annualor D6termina
1=-ye BaCt8riOIOgy ) J
(1974, 8th edition, The Williams and Wilkins Company), and “Cooperative Description of Streptomyces spp.
Description of Type 5trai
ns of StreptomyCes) The paper ``International Streptomyces Project'' published by Schilling and Gotliep in J.
Int, J, 5ift, Bacteriol Co 18:69-189.1968: Ibid. 18 + 2
! 79-892.1968: Ibid. 19:891-51
2.1969: and the same document 22:265-894.1
972) and the phylogenetic characteristics of Streptomyces species in these documents published from 1974 to the present, no microorganisms similar to 5F-IAsA) could be found.
ト比較した。この研究結果を表Vに示し、ストレプトミ
セスsp、 SF−1(ASA )が他のピマリシン生
成物と相違することを硲めた。compared. The results of this study are shown in Table V and confirm that Streptomyces sp. SF-1 (ASA) is different from other pimaricin products.
ここに記載する微生物は野生菌株であるが、しかし自然
的に得られる突然変異体および突然変異誘発剤(mut
ageni、c agents ) kT−より誘発さ
れる突然変異体もまた共にまたは別にぎリエン抗生物質
のグループを生ずることができる。このために、後述す
るプロセスにおいて野生菌株およびその突然変異体子孫
(mutant descendants ) E用い
ることができる。The microorganisms described here are wild strains, but also naturally occurring mutants and mutagens.
mutants derived from kT-, together or separately, can give rise to a group of antibiotics. For this purpose, the wild-type strain and its mutant descendants can be used in the process described below.
発酵による生成
ここに記載するテトラエンは固体培地または液体培地に
おいて生成することができるが、液体培地は多量の化合
物を製造するのに極めて適当である。この液体培地では
抗生物質を表面培養および液内培養によって生成するこ
とができる。特に、液内i@養は多量の生成物を作るの
に最適である。Production by Fermentation The tetraenes described herein can be produced in solid or liquid media; liquid media are highly suitable for producing large amounts of the compound. In this liquid medium, antibiotics can be produced by surface culture and submerged culture. In particular, in-liquid i@culture is ideal for producing large amounts of product.
使用する培地は少なくとも次に示す材料を含む:a)同
化性炭素源、例えばグルコース、7ラクトース、マンニ
トール、グ゛、゛ ワール、殿粉、テキストリン、セロ
ビオース、トレハロース、マンノースおよびその混合物
。The medium used contains at least the following materials: a) Assimilable carbon sources such as glucose, 7-lactose, mannitol, guar, wal, starch, textrin, cellobiose, trehalose, mannose and mixtures thereof.
b)同化性炭素源、例えば植物性ケーキミール(大豆、
綿、粉砕ナツツ等)、コーン ステイ−7’ IJカー
、植物性または動物性ペプトン類、酵母、尿素、アンモ
ニア壇類等およびその混合物。b) Assimilable carbon sources, e.g. vegetable cake meal (soybean,
(cotton, crushed nuts, etc.), corn stay-7' IJ car, vegetable or animal peptones, yeast, urea, ammonia algae, etc., and mixtures thereof.
C) pH緩衝能を有する培地を生成する無機質IJ
i類または無機質元素を供給する無機質塩類。C) Inorganic IJ that produces a medium with pH buffering capacity
Inorganic salts that supply type i or inorganic elements.
d)抑泡剤として、または炭素源として用いる植物性ま
たは動物性油。シリコン。d) Vegetable or animal oils used as foam suppressants or as carbon sources. silicon.
本発明の方法に用いることのできるI)H醗酵範囲は6
.1〜8.4の範囲であり、最適な温度は25〜aO℃
の範囲である。The I)H fermentation range that can be used in the method of the present invention is 6
.. ranges from 1 to 8.4, and the optimal temperature is 25 to aO℃
is within the range of
規則的抗生物質醗酵プロセスを用いて醗酵により本発明
におけるテトラエンを作ることができる。The tetraenes of the present invention can be made by fermentation using a regular antibiotic fermentation process.
すなわち、培地を含有する傾斜管で菌株胞子体を凍結乾
燥胞子(1yOphilised 5pores )
”’C接種シ、最適温度で培養し、液内培養における生
長相をかかる管からの胞子および菌糸体で開始させる。That is, the strain sporophyte was lyophilized to 5 pores (1yOphilised 5pores) in a tilted tube containing the medium.
After inoculation and culturing at the optimum temperature, the growth phase in submerged culture is initiated with spores and mycelia from such tubes.
これらの段階は醗酵すべきプロス容量によって長くまた
は短くなる。大きい醗酵タンクを用いる場合には、段階
の数が5に達する。These stages can be longer or shorter depending on the process volume to be fermented. If large fermentation tanks are used, the number of stages reaches 5.
フラスコで醗酵する場合には、フラスコをサーモスタッ
ト制御チャンバーの適当な振動テーブル上で十分に珈盪
させるが;タンクで醗酵する場合には生成を最適レベル
に達成するために、タンクに通気、攪拌、pH制御、溶
解0.制御、温度制御および栄養素、アルカリおよび酸
添加する必要がある。すべてのプロセスは完全滅菌条件
下で行い異質微生物による汚染を避ける必要がある。培
地は容器において滅菌することができ、または容器が予
じめ滅菌されている場合にはすでに滅菌された容器に置
くことができる。When fermenting in flasks, the flasks are thoroughly shaken on a suitable shaking table in a thermostatically controlled chamber; when fermenting in tanks, the tanks are vented, stirred, and stirred to achieve optimal levels of production. pH control, dissolution 0. It is necessary to control, temperature control and add nutrients, alkalis and acids. All processes must be performed under completely sterile conditions to avoid contamination with foreign microorganisms. The medium can be sterilized in the container or placed in an already sterilized container if the container has been previously sterilized.
生成が一旦始まると、培地に十分な菌糸体が48時間よ
り生じ、この生成は醗酵の5日または6日まで徐々に増
加し、次いで増加が停止した。Once production started, sufficient mycelium was generated in the medium for 48 hours and the production gradually increased until day 5 or 6 of fermentation, then the increase stopped.
Ab 400に対する積率としてプロス中の抗生物質の
鎗を研究するために、ペーパーおよび高圧液体クロマト
グラフィーに単一活性物質を示し、かつサッカ!ミセス
セレビシェ−ATOO9767に対して0.5μg/
−の最小抑制濃度を有する哨製試肩を用いた。ピマリシ
ン標準として、オランダ、デルフトのコニンクリジケ
ニーデルランドシエ シスト アンド スピリッツ7ア
プリ ツ り (Koninklijke Nede
Rlandsche G15t −acSpiri
tusfabriek N、 V、 )で作った900
U/wI9の試料(バッチ9117A)を用いた。In order to study the antibiotic spear in the pros as a product factor for Ab 400, a single active substance was shown on paper and high pressure liquid chromatography, and Sacca! Mrs. Cereviche - 0.5μg/for ATOO9767
A test shoulder with a minimum inhibitory concentration of - was used. Koninklisike, Delft, Netherlands, as a pimaricin standard.
Koninklijke Nede Schist and Spirits 7 App Tour
Rlandsche G15t-acSpiri
900 made with tusfabriek N, V, )
A sample of U/wI9 (batch 9117A) was used.
すでに特許となっている微生物のプロスにAb400を
存在するごとについては記載されていないでの比較研究
を行い、この物質が従来の培地での醗酵において生じた
ものであるか否かを確めた。We conducted a comparative study on the presence of Ab400 in a patented microbial process, which was not described, to confirm whether this substance was produced during fermentation in a conventional culture medium. .
この結果を表■に示す。この表■から、ストレプトミセ
スsp、5F−1(ASA)が多量のAb 400およ
びピマリシンを生ずることからすでに特許された周知の
ピマリシン生成菌株と本発明の菌株とが明らかに相違す
ることがわかる。試験したすべての培地においては上記
2種の菌株のうち前者のAb 400が検出しつる程麿
に生成しないことがわかる。更に、かかる5F−1(A
SA)菌株の異なる特許は醗酵プロスにおいて強いリン
ゴ臭を生ずることである。The results are shown in Table ■. From Table 1, it can be seen that the strain of the present invention is clearly different from the well-known pimaricin-producing strain that has already been patented, since Streptomyces sp. 5F-1 (ASA) produces large amounts of Ab 400 and pimaricin. It can be seen that in all the media tested, Ab 400, the former of the two strains mentioned above, was not produced as quickly as it could be detected. Furthermore, such 5F-1 (A
SA) A different patent of the strain is that it produces a strong apple odor in the fermentation process.
表 ■
II SFMF−13培地は4%大豆粉、6%グルコ
ース、0.5%(NH,ン、aSO4、0,18%に、
HPO4,gH,O、1%caco、および0.5%大
豆油を含有し、pH6,2,を有する。Table II SFMF-13 medium contains 4% soybean flour, 6% glucose, 0.5% (NH, N, aSO4, 0.18%,
Contains HPO4,gH,O, 1% caco, and 0.5% soybean oil, and has a pH of 6.2.
MOH−1として知られている培地は英国特許@ssz
、ssa号明細書に記載され、2%グリセロール、0.
5%ファイトーン、0#5%ペプトン、0.8%酵母エ
キス、0.3%肉エキスおよび0.25@caco、を
含有し、かつpH7,6を有する。また、MFNa−1
として知られている培地はドイツ特許第1,024,2
06号明細書に記載され、5%大豆粉、1%グルコース
、0.5%(NH,5,、′o、、0.02%KH,P
04.1%0aOOB、0.5優大豆油および0.1%
コーン ステイブ リカーを含有し、かつpH6,8を
有する。The medium known as MOH-1 has a British patent @ssz
, SSA specification, 2% glycerol, 0.
Contains 5% phytone, 0#5% peptone, 0.8% yeast extract, 0.3% meat extract and 0.25@caco, and has a pH of 7.6. Also, MFNa-1
The medium known as
06 specification, 5% soybean flour, 1% glucose, 0.5% (NH, 5,, 'o, 0.02% KH, P
04.1% 0aOOB, 0.5% soybean oil and 0.1%
Contains corn stave liquor and has a pH of 6.8.
抽出および分離
一回の醗酵が終了し、p過したプロスの最小抑制濃度(
MIC)は、一般にサツカロミ七ス七しビシエーに対し
て172000であった。Extraction and separation After one fermentation, the minimum inhibitory concentration (
MIC) was generally 172,000 for Satsukalomi Shichisu Shichishi Vishie.
5〜6の範囲のpHの全プ四スをハイフロース−パー七
tv (H3/flo−8uparcall ) &:
通t、テr過シ、p過したプロスを水酸化ナトリウムで
pH8,0に調節し、次いでブタノールの如き水と非混
和性の脂肪族アルコールで抽出した。H3/flo-8uparcall all the liquids with a pH range of 5 to 6.
The filtered prosthesis was adjusted to pH 8.0 with sodium hydroxide and then extracted with a water-immiscible aliphatic alcohol such as butanol.
有機エキスを真空下で濃縮して容量を最初の容量のl/
20に減少し、これによって白色粉末が沈殿し、これは
「粗抗真菌性物質(raw antifungal月と
称されている。この沈殿をp過し、真空下常温で乾燥し
た。この得られた生成物は極めて薄い黄色の粉末であっ
て、このMIOは上述する酵母に対して約1/ 1,5
fl O,000(0,67mcg7’az )であ
った。この生成物の収量はp過したブロス1!当り0.
7gであった。The organic extract was concentrated under vacuum to reduce the volume to l/l of the initial volume.
20, thereby precipitating a white powder, which is referred to as "raw antifungal material". The precipitate was filtered and dried under vacuum at room temperature. The substance is an extremely pale yellow powder, and this MIO is about 1/1.5 that of the yeast mentioned above.
fl O,000 (0,67mcg7'az). The yield of this product is 1! Hit 0.
It was 7g.
かかる「粗抗真菌性物質」は比較的に水溶性で、ペーパ
ー り四マドグラフィーにより調へた場合に、サツカロ
ミセス セレビシェ−に対して活性である少なくとも2
種類の主抗生物質が存在することを確めた〇
「粗抗真菌性物質」を約4〜/−の割合で水に溶解し、
この溶液をpH10,2に調節し、+1”Cに放rlt
(24時間にわたり)した場合、粗抗真菌性物質は「抗
真菌性物質1 (antifungal 1) Jとし
て知られたl / 10,000,000 (0,1H
og/m)のMIOを有し、かつ上述するAb 400
に相当する白色物質を沈殿した。この抗真菌性物質lま
たはAb 400は最初の「粗抗真菌性物質」の6.6
優の収率で得た〇
メタノールにおける「抗真菌性物質l」の紫外スペクト
ルは、この物質がテトラエンであることを示し、また臭
化カリウムにおける赤外スペクトルは同一ではないけれ
どもテトラエン抗真菌性ピマリシンに類似することを示
した〇
ピマリシンはブタノール+エタノール+水(5:471
、V)クロマトグラフ糸において「抗真菌性物質1」よ
り低いRyを有する。Such "crude antifungal substances" are relatively water soluble and have at least 20% active against Saccharomyces cerevisiae when prepared by paper toweling.
It was confirmed that the main antibiotics of various types existed. 〇 "Crude antifungal substance" was dissolved in water at a ratio of about 4 to 1/-.
Adjust this solution to pH 10.2 and release to +1"C.
(over 24 hours), the crude antifungal substance was known as “antifungal 1 J”/10,000,000 (0,1H
Ab 400 with a MIO of
A white substance corresponding to . This antifungal substance l or Ab 400 is 6.6 of the initial “crude antifungal substance”.
The UV spectrum of "Antifungal" in methanol, obtained in excellent yield, shows that the material is a tetraene, and the infrared spectrum in potassium bromide, although not identical, shows that the tetraene antifungal pimaricin. Pimaricin was shown to be similar to butanol + ethanol + water (5:471
, V) has a lower Ry in the chromatographic thread than "Antifungal 1".
リクロソルプrp −18−カラム(Li0rosor
brp −18−column ) (5μン、0.0
6 M <えん酸アンモニウムの移動相、PH5,0+
アセトニトリル(8:1v/v)および800nmでの
横用による高圧液体クロマトグラフ(HPLC)、サツ
カロミセス セレビシェ−に対する抗生物質活性を有す
る74類の物質およびテトラエン紫外スペクトルを用い
て「粗抗真菌性物質」であることを確認した。Li0rosorp rp-18-column (Li0rosor
brp-18-column) (5 μn, 0.0
6 M <ammonium citrate mobile phase, PH5,0+
"crude antifungal" using acetonitrile (8:1 v/v) and high pressure liquid chromatography (HPLC) at 800 nm, class 74 substances with antibiotic activity against Satucharomyces cerevisiae and tetraene UV spectrum It was confirmed that
「抗真菌性物質1」の元素センテシマル組成(elem
entary centisimal composi
tion )はC54,5%、H7,1%、N2.9%
およびO(dir、)85.5%でC□H,40□、N
の最小実験式を与え、最小分子層は488.522であ
った。Elemental centesimal composition of "Antifungal Substance 1" (elem
entry centisimal composition
) is C54.5%, H7.1%, N2.9%
and O(dir,)85.5% C□H,40□,N
gave the minimum empirical formula of , and the minimum molecular layer was 488.522.
抗菌活性
Ab 400の抗菌活性について試験した。試験の結果
、細菌に対する活性および酵母に対するホ。Antibacterial Activity Ab 400 was tested for antibacterial activity. Test results showed activity against bacteria and yeast.
シテイプ活性はなかった:
カンジタ アルビカンス(trop
icalis)
次に、本発明を例について説明する。There was no siteip activity: Candida albicans (trop
icalis) Next, the invention will be explained by way of example.
例1
培地を次の組成から作った:
綿実粉・・・・・・・・・・・・・・・・・・・・
20 gコーン ステイープ リカー ・・・・−・
10 gデキストリン ・・・・・・・・・・・
・・、−10gグルコース・−・・・・・・・・・暑・
・・・N 50g(NH4)2SO4・・・・・
・・・・・・・・・・ 5gNaN0 ・・・・
・・・・・・・・・・・・・・・ 8.5gに、H
PO4・・・・・・・・・・・・・・・・・・ 1
gF8S0 .7HO・・・・・・・・・・・・・・
0.05 gm
MgSO4,7H,0・・・・・・・・・・・・・・
0.05 gMnSo4.H,O−・−−−−−−・
−・・−・0.05 gZnSo 、7HO・・・・
・・・・・・・◆・・ 0.1gm
蒸留水 ad ・・・・・・・・・・・・・・・・
1ooovpHはKOHにより6.2に調節した。Example 1 A medium was made from the following composition: Cottonseed flour.
20g Corn Steep Liquor ・・・・−・
10 g dextrin・・・・・・・・・・・・
・・・10g glucose・・・・・・・・・・・・・
...N 50g (NH4)2SO4...
・・・・・・・・・ 5gNaN0 ・・・・
・・・・・・・・・・・・・・・ 8.5g, H
PO4・・・・・・・・・・・・・・・・・・ 1
gF8S0. 7HO・・・・・・・・・・・・・・・
0.05 gm MgSO4,7H,0・・・・・・・・・・・・・・・
0.05 gMnSo4. H, O-・-------・
−・・−・0.05 gZnSo, 7HO・・・・
・・・・・・・◆・・ 0.1gm Distilled water ad ・・・・・・・・・・・・・・・・・・
1ooov pH was adjusted to 6.2 with KOH.
それぞれ200−の培地を各1,000−のエエレンマ
イヤーフラスコに入れた。Each 200-liter medium was placed in each 1,000-liter Erlenmeyer flask.
19の0aC01を各フラスコに添加した。19 of OaC01 was added to each flask.
各フラスコを121°Cで80分間にわたり滅菌した。Each flask was sterilized at 121°C for 80 minutes.
かようにして作った培地をストレプトミ七スsp、菌株
5F−1(ASA)胞子懸濁物で接種した。この懸濁物
は微生物を胞子形成する任意の固体培地において傾斜管
培養から作った。The medium thus prepared was inoculated with a spore suspension of Streptomys sp., strain 5F-1 (ASA). This suspension was made from slant tube cultures on any solid medium that sporulates microorganisms.
接11+径、この培地を5 C11行程および24 O
rpmの速度を有する軌道振動テーブル上で一定振動さ
せながらサーモスタットで8!8℃に制徊したチャンバ
ーで5日間にわたって培養した。かようにして作った接
種物(innoculum )を用いて次の組成からな
る醗酵培地を接種した:
大豆粉 °°°°°°°°0°””’ 40
gグルコース ・・・・・11■・・ 60
g(NH4ン、S04 ・・・・・・・・・・・
5 gK、HPO4・・・・・・・・・・
・・・・ 18 gCaOO8・・・・・・・・参
・・・・・・ 10 g蒸留水 ad 口・・・・
・・・−−−10001141)Hは、炭#1Mを添加
する前に、稀薄硫酸で6.2に調節した。11+ diameter, this medium was subjected to 5 C11 steps and 24 O
The cells were cultured for 5 days in a chamber controlled by a thermostat at 8-8° C. while being constantly vibrated on an orbital vibration table with a speed of rpm. The innoculum thus prepared was used to inoculate a fermentation medium consisting of the following composition: Soybean flour °°°°°°°°0°""' 40
g glucose...11■...60
g(NH4n, S04...
5 gK, HPO4・・・・・・・・・
...... 18 g CaOO8....... 10 g distilled water ad mouth...
...---10001141) H was adjusted to 6.2 with dilute sulfuric acid before adding charcoal #1M.
それぞれ200m1のナトリウムを1!フラスコに入れ
た。これらの各フラスコに1−の大豆油を添加した。各
フラスコを121°Cで80分簡にわたり滅菌した。200ml of sodium each! I put it in a flask. 1 - of soybean oil was added to each of these flasks. Each flask was sterilized briefly at 121°C for 80 minutes.
滅菌後、培地を5%の接種物で接種した。After sterilization, the medium was inoculated with a 5% inoculum.
この事はチャンバーにおいて接種物について記載したと
同じ特徴の振動テーブル上で28°Cで4日間にわたり
培養した。醗酵の末期におけるブロスの分析結果を次に
示す:
M5.5〜6.5;砂糖 2%;
アンモニウムN:100@9/l:全可溶性N:950
”9/ ’ ;および菌容積(mycelial v
olume)二 32゜
サツカロミセス セレビシェ−ATOO9767に対し
て、1/4000〜115000の最小抑制濃度を有し
;およびHPL(Eにおいて、標準として900U/■
のピマリシンを用いて300−500mcg/mのAb
400活性および1000〜12000mCg/−のピ
マリシン活性を示した。This was incubated for 4 days at 28°C on a vibrating table with the same characteristics as described for the inoculum in a chamber. The analysis of the broth at the end of the fermentation is as follows: M5.5-6.5; Sugar 2%; Ammonium N: 100 @ 9/l: Total soluble N: 950
"9/ '; and bacterial volume (mycelial v
olume) II 32゜Has a minimum inhibitory concentration of 1/4000 to 115000 against S. cerevisiae-ATOO9767; and 900 U/■ as standard in HPL (E)
300-500 mcg/m of Ab using pimaricin
400 activity and pimaricin activity of 1000-12000 mCg/-.
例2
培地を例1の接種物と同じ組成から作り、200−の割
合で1!エレンマイヤー フラスコに入れ、この培地を
】21°Cで80分間にわたり滅菌した。Example 2 A medium was prepared from the same composition as the inoculum of Example 1, with a ratio of 200-1! The medium was sterilized in Ellenmeyer flasks at 21°C for 80 minutes.
このようにして作った培地を特許の微生物の胞子の懸濁
物で接種した°。この培地を5(it行程および24
Orpmの揚動速度を有する軌動振動テーブル上で一定
振動させながらサーモスタット制御チャンバーにおいて
28℃で5日間にわたり培養した。The medium thus prepared was inoculated with a spore suspension of the proprietary microorganism. This medium was added for 5 (it steps and 24
Cultures were incubated for 5 days at 28° C. in a thermostatically controlled chamber with constant vibration on an orbital vibrating table with a lifting speed of Orpm.
かようにして作った接種物を用いて、0.5%の割合で
例1の醗酵培地と同じ組成を有する培地11を含有する
81g積の醗酵タンクで接種した。The inoculum thus prepared was used to inoculate an 81 g fermentation tank containing medium 11 having the same composition as the fermentation medium of Example 1 at a rate of 0.5%.
次の醗酵条件を用いた:温度28℃、通気0.8v/v
/m、撹拌40 Or、p、m、 、および醗酵時間9
0時間。この時、醗酵培地は500〜1100Ona
/−のピマリシン活性および800〜600mcg /
−のlb活性を有していた。同じタイプの標準を例1に
おけるように用いた。The following fermentation conditions were used: temperature 28°C, aeration 0.8v/v
/m, stirring 40 Or, p, m, and fermentation time 9
0 hours. At this time, the fermentation medium is 500-1100 Ona
Pimaricin activity of /- and 800-600 mcg /
- had lb activity. The same type of standard was used as in Example 1.
例8
pHIS、9T8540−の全プロスをハイフロース−
パーセル上で真空下で一過し、M I O−1/2,2
22を有する赤色がかった透明なp過ブ田ス2,150
−を得た。pHを25%水酸化ナトリウムで8.0にl
1節し、上記例と同様にして綿毛状の沈殿を戸別した。Example 8 pHIS, 9T8540- whole proso
Passed under vacuum on Purcel, MIO-1/2,2
Reddish transparent p-transparent with 22,150
I got -. Adjust pH to 8.0 with 25% sodium hydroxide.
The fluff-like precipitate was separated from house to house in the same manner as in the above example.
第1の抽出においては有機相を遠心分離して700−の
ブタナールを有するp過プロスを得た。水相は上述する
と同様に350−のブタナールで2回抽出した。全体の
透明な黄色I/)ブタナール抽出物を白色粉末の沈殿を
生ずる最初の容積の1720に真空下で濃縮し、濾過後
ブタナールで洗浄し、真空上常温で乾燥し、1.52g
(すなわち、f過プロス11当り0.719 )の「抗
真菌性物質」を得、この物質はMIO−1/1,450
゜000 (0,69mcg/m)を示した。In the first extraction, the organic phase was centrifuged to obtain p-perpros with 700-butanal. The aqueous phase was extracted twice with 350-butanal as described above. The total clear yellow I/) butanal extract was concentrated under vacuum to an initial volume of 1720 resulting in the precipitation of a white powder, washed with butanal after filtration and dried under vacuum at room temperature, 1.52 g
(i.e., 0.719 per f perpros 11), and this substance is MIO-1/1,450
°000 (0.69 mcg/m).
生成物を880−の水に溶解し、溶液のpHを水酸化ナ
トリウムで10.2に調節した。+1°Cで24時間後
、綿毛状沈殿を濾過し、水洗し、真空上常温で乾燥して
101■の抗真菌性物質1;6.7%の出発「粗抗真菌
性物質」を得たOこの抗真菌性物質lのMIOは1/1
0,000,000(o、1mcg /vd )であっ
た。The product was dissolved in 880-m water and the pH of the solution was adjusted to 10.2 with sodium hydroxide. After 24 hours at +1°C, the fluffy precipitate was filtered, washed with water and dried under vacuum at room temperature to yield 101 ■ of antifungal substance 1; 6.7% of the starting "crude antifungal substance". The MIO of this antifungal substance is 1/1
It was 0,000,000 (o, 1mcg/vd).
例4
pH5,6で8,800−の全ブロスをバイア0−スー
パーセル上で真空下で一過し、M M C−1/2!0
00を有する2、5QQmjの一過したブロスを得た。Example 4 A total broth of 8,800 at pH 5,6 was passed under vacuum over a Via 0-Supercell and MMC-1/2!0
A strained broth of 2,5QQmj with 0.00 was obtained.
このpHを水酸化ナトリウムの添加で8.0に調節し、
ブロスを同様にして再濾過した。このグロスを上記例に
記載するようにブタノールで抽出し、ブタノール抽出物
を真空下で最初の容積の1/20に濃縮した。沈殿した
「粗抗真菌性物質」を濾過し、ブタノールで洗浄し、真
空下において常温で乾燥してM I O−1/ 1,6
66,000 (0,60mcg//Lt)を有する生
成物を得た。The pH was adjusted to 8.0 by the addition of sodium hydroxide,
The broth was refiltered in the same way. This gloss was extracted with butanol as described in the example above and the butanol extract was concentrated under vacuum to 1/20 of its original volume. The precipitated "crude antifungal material" was filtered, washed with butanol and dried under vacuum at room temperature to give MIO-1/1,6
A product with a concentration of 66,000 (0,60 mcg//Lt) was obtained.
上記粗抗真菌性物質を500@jに溶解し、溶液のpH
を水酸化ナトリウムで10.2に調節し、溶液を+1℃
で24時間貯蔵した。乾燥衝、沈殿抗真菌性物質1は1
81m119(6,6%ンであり、そのMICは1 /
10,000,000 (0,1’導Og/m)であ
った。Dissolve the above crude antifungal substance in 500@j and adjust the pH of the solution.
was adjusted to 10.2 with sodium hydroxide and the solution was heated to +1°C.
It was stored for 24 hours. Dry, precipitate antifungal substance 1 is 1
81m119 (6.6%), its MIC is 1/
It was 10,000,000 (0,1' conductive Og/m).
ヱ1L
pH5,9で3+750WLtの全プロスをバイアtf
f−スーパーセル上で濾過し、MIO−2,500を有
するp過プロスを得た。このブロスを上記例に記載する
ようにしてpH8,0に調節し、渥過し、ブタナールで
抽出した。有機抽出物を濃縮した彼、MIO−1/2,
222,000 C0,45mcg/v4)を有する粗
抗真菌性物質1.9gを得た。この粗抗真菌性物質を4
75−の水に溶解した後(pH10,2)、1℃で24
時間後に1/10,000,000 (0,1mog
/ ml )を有する抗真菌性物質1 146mg(7
,7%)を得た。ヱ1L pH 5,9, 3+750WLt whole proso via tf
Filtration over f-Supercel yielded p-filtrate with MIO-2,500. The broth was adjusted to pH 8.0 as described in the example above, filtered and extracted with butanal. Concentrated organic extract, MIO-1/2,
1.9 g of crude antifungal substance having a concentration of 222,000 C0.45 mcg/v4) was obtained. This crude antifungal substance
After dissolving in water (pH 10,2) at 24°C at 1°C.
1/10,000,000 (0.1mog
/ ml) of antifungal substance 1 146 mg (7
, 7%).
特許出願人 アンチピオチコス。ソシェテ・アノ二ム第
1頁の続き
0発 明 者 ジョセ・マリア・フエルナンデズ・ソウ
サーフアロ
スペイン国ビラヌエバ・デ・う
・カニャダ(マドリッド)カス
ティ口・デ・マルピカ66Patent applicant Antipioticus. Sochete Anonymous Page 1 continued 0 Inventor Jose María Fernández Saucerjuaro Villanueva de U Cañada (Madrid), Spain 66
Claims (1)
と称する微生物により培地を醗酵させ、かかる抗生物質
を菌糸体力らおよび/または培地から抽出することを特
徴とするポリエン抗真菌性抗生物質の製造方法。 & 前記醗酵においては25〜80℃の温度で通気し、
培地には同化性炭素および窒素源、および無機塩を含有
する特許請求の範囲第1項記載のポリエン抗真菌性抗生
物質の製造方法。 & 炭素源としてグルコース、7ラクトース、マンニト
ール、グリセロール、’ll1mCテキストリン、セロ
ビオース、トレハロース、マンノースおよびその混合物
を用い、および窒素源として植物性ケーキミール、コー
ンステイープリカー、植物性または動物性ペプトン類、
酵母、尿素、アンモニウム塩またはその混合物を用いる
特許請求の範囲第2項記載のポリエン抗真菌性抗生物質
の製造方法。 表 生成発酵プロスを濾過し、pHを約8.0に調節し
、水分を抽出し、真空下で濃縮し、これにより抗生物質
錯体または粗抗真菌性物質を沈殿し、これからピマリシ
ンを分離する特許請求の範囲第1,2または8項記載の
ポリエン抗真菌性抗生物質の製造方法。 五 生成醗酵ブロスを濾過し、pHを約8.0に調節し
、水分を抽出し、真空下で濃縮し、これにより抗生物質
錯体または粗抗真菌性物質を沈殿し、この沈殿物を水に
約4s9/−の割合で溶解し、溶液のpHを約1O02
に調節し、次いで沈殿によりAb 400と名づけるテ
トラエン抗生物質を得る特許請求の範囲第1.2または
8項記載のポリエン抗真菌性抗生物質の製造方法。 & 生成醗酵プロスを濾過し、pHを約8.0に調節し
、水分を抽出し、真空下で濃縮し、これにより抗生物質
錯体または粗抗真菌性物質を沈殿し、この沈殿物を水に
約4s9/gLtの割合で溶解し、溶液のpHを約10
.2に調節し、次いで沈殿により得た0 54.5%、
H7,1%、N2.9%およびO35,5%の元素セン
テシマル組成およびサッカ四ミセス七しビシエーに対し
て1/10,000,000の最小抑制濃度を有するA
b 400と名づけるテトラエン抗真菌性抗生物質。[Claims] 1 Streftomyces sp, strain SF-1 (ASA)
1. A method for producing a polyene antifungal antibiotic, which comprises fermenting a medium using a microorganism known as a microorganism, and extracting the antibiotic from the mycelium and/or the medium. & In the fermentation, aeration is carried out at a temperature of 25 to 80°C,
2. The method for producing a polyene antifungal antibiotic according to claim 1, wherein the medium contains an assimilable carbon and nitrogen source and an inorganic salt. & Using glucose, lactose, mannitol, glycerol, 'll1mC textrin, cellobiose, trehalose, mannose and mixtures thereof as carbon sources and vegetable cake meal, cornstap liquor, vegetable or animal peptones as nitrogen sources ,
3. The method for producing a polyene antifungal antibiotic according to claim 2, using yeast, urea, ammonium salt, or a mixture thereof. Table Patent for filtering the resulting fermentation process, adjusting the pH to about 8.0, extracting the water and concentrating under vacuum, thereby precipitating the antibiotic complex or crude antifungal substance and separating pimaricin from it. A method for producing a polyene antifungal antibiotic according to claim 1, 2 or 8. 5. Filter the resulting fermentation broth, adjust the pH to about 8.0, extract the water and concentrate under vacuum, thereby precipitating the antibiotic complex or crude antifungal substance, and pouring this precipitate into water. Dissolve at a ratio of about 4s9/- and adjust the pH of the solution to about 1O02
A process for producing a polyene antifungal antibiotic according to claim 1.2 or 8, wherein the tetraene antibiotic designated as Ab 400 is obtained by precipitation. & Filter the resulting fermentation process, adjust the pH to about 8.0, extract the water and concentrate under vacuum, thereby precipitating the antibiotic complex or crude antifungal substance, and pouring this precipitate into water. Dissolve at a rate of approximately 4s9/gLt, and adjust the pH of the solution to approximately 10.
.. 2 and then obtained by precipitation 0 54.5%,
A with an elemental centesimal composition of H7.1%, N2.9% and O35.5% and a minimum inhibitory concentration of 1/10,000,000 against Sacca 4.
A tetraene antifungal antibiotic named b400.
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
ES504582A ES8301278A1 (en) | 1981-08-06 | 1981-08-06 | A process for the preparation of polyene antifungal antibiotics and new tetraene antifungal antibiotic |
ES504,582 | 1981-08-06 |
Publications (2)
Publication Number | Publication Date |
---|---|
JPS5867189A true JPS5867189A (en) | 1983-04-21 |
JPH0142274B2 JPH0142274B2 (en) | 1989-09-11 |
Family
ID=8482823
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP57136457A Granted JPS5867189A (en) | 1981-08-06 | 1982-08-06 | Polyene anti-bacterial antibiotic and production thereof |
Country Status (5)
Country | Link |
---|---|
JP (1) | JPS5867189A (en) |
DE (1) | DE3228306A1 (en) |
ES (1) | ES8301278A1 (en) |
FR (1) | FR2511034B1 (en) |
GB (1) | GB2106498B (en) |
Families Citing this family (9)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US5902579A (en) * | 1991-08-05 | 1999-05-11 | Bio-Technical Resources | Natamycin-containing streptomyces biomass and its use in animal feed |
US5231014A (en) * | 1991-08-05 | 1993-07-27 | Bio-Technical Resources | Fermentation process for producing natamycin |
US5686273A (en) * | 1991-08-05 | 1997-11-11 | Cultor Food Science, Inc. | Fermentation process for producing natamycin with additional carbon and nitrogen |
US5266347A (en) * | 1992-01-28 | 1993-11-30 | Ducoa L.P. | Antibiotic biomass animal feed compositions |
US5942611A (en) * | 1995-01-19 | 1999-08-24 | Cultor Ltd. | Process for natamycin recovery |
US6045815A (en) * | 1997-08-15 | 2000-04-04 | Board Of Regents, The University Of Texas System | Parenteral pimaricin as treatment of systemic infections |
US6844004B2 (en) | 1997-08-15 | 2005-01-18 | Board Of Regents, The University Of Texas System | Topical formulations of natamycin/pimaricin |
US20060275873A1 (en) * | 2003-05-22 | 2006-12-07 | Eliseo Recio Perez | Autoinducer compound to improve the productivity of natamycin streptomyces strains |
CN109913503B (en) * | 2018-12-27 | 2023-03-21 | 蚌埠学院 | Method for producing tetraene compound by utilizing streptomyces albidoflauvs Ah11601 |
-
1981
- 1981-08-06 ES ES504582A patent/ES8301278A1/en not_active Expired
-
1982
- 1982-07-27 GB GB08221659A patent/GB2106498B/en not_active Expired
- 1982-07-29 DE DE19823228306 patent/DE3228306A1/en not_active Ceased
- 1982-08-04 FR FR828213623A patent/FR2511034B1/en not_active Expired
- 1982-08-06 JP JP57136457A patent/JPS5867189A/en active Granted
Also Published As
Publication number | Publication date |
---|---|
ES504582A0 (en) | 1982-12-16 |
FR2511034A1 (en) | 1983-02-11 |
JPH0142274B2 (en) | 1989-09-11 |
DE3228306A1 (en) | 1983-11-10 |
FR2511034B1 (en) | 1985-07-26 |
ES8301278A1 (en) | 1982-12-16 |
GB2106498A (en) | 1983-04-13 |
GB2106498B (en) | 1985-01-30 |
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