JPS6129715B2 - - Google Patents
Info
- Publication number
- JPS6129715B2 JPS6129715B2 JP54092908A JP9290879A JPS6129715B2 JP S6129715 B2 JPS6129715 B2 JP S6129715B2 JP 54092908 A JP54092908 A JP 54092908A JP 9290879 A JP9290879 A JP 9290879A JP S6129715 B2 JPS6129715 B2 JP S6129715B2
- Authority
- JP
- Japan
- Prior art keywords
- reaction
- antibiotic
- genus
- producing
- culture
- 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.)
- Expired
Links
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 22
- 238000006243 chemical reaction Methods 0.000 claims description 11
- 241000894006 Bacteria Species 0.000 claims description 9
- 238000000862 absorption spectrum Methods 0.000 claims description 8
- 230000003115 biocidal effect Effects 0.000 claims description 6
- 238000004519 manufacturing process Methods 0.000 claims description 5
- 238000012258 culturing Methods 0.000 claims description 4
- 229910052739 hydrogen Inorganic materials 0.000 claims description 3
- 150000003839 salts Chemical class 0.000 claims description 3
- 229910021578 Iron(III) chloride Inorganic materials 0.000 claims description 2
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 claims description 2
- 230000002378 acidificating effect Effects 0.000 claims description 2
- 238000004042 decolorization Methods 0.000 claims description 2
- 238000000354 decomposition reaction Methods 0.000 claims description 2
- 238000000921 elemental analysis Methods 0.000 claims description 2
- RBTARNINKXHZNM-UHFFFAOYSA-K iron trichloride Chemical compound Cl[Fe](Cl)Cl RBTARNINKXHZNM-UHFFFAOYSA-K 0.000 claims description 2
- 238000002844 melting Methods 0.000 claims description 2
- 230000008018 melting Effects 0.000 claims description 2
- 230000007935 neutral effect Effects 0.000 claims description 2
- FEMOMIGRRWSMCU-UHFFFAOYSA-N ninhydrin Chemical compound C1=CC=C2C(=O)C(O)(O)C(=O)C2=C1 FEMOMIGRRWSMCU-UHFFFAOYSA-N 0.000 claims description 2
- 239000011591 potassium Substances 0.000 claims description 2
- 229910052700 potassium Inorganic materials 0.000 claims description 2
- 239000003242 anti bacterial agent Substances 0.000 claims 2
- 229940088710 antibiotic agent Drugs 0.000 claims 2
- 231100000252 nontoxic Toxicity 0.000 claims 1
- 230000003000 nontoxic effect Effects 0.000 claims 1
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 16
- UHOVQNZJYSORNB-UHFFFAOYSA-N Benzene Chemical compound C1=CC=CC=C1 UHOVQNZJYSORNB-UHFFFAOYSA-N 0.000 description 12
- 239000002609 medium Substances 0.000 description 11
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 6
- 241000233866 Fungi Species 0.000 description 6
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 5
- 239000002904 solvent Substances 0.000 description 5
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 4
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 3
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 3
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 3
- 230000001580 bacterial effect Effects 0.000 description 3
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 description 3
- 239000000741 silica gel Substances 0.000 description 3
- 229910002027 silica gel Inorganic materials 0.000 description 3
- 238000001228 spectrum Methods 0.000 description 3
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 229920001353 Dextrin Polymers 0.000 description 2
- 239000004375 Dextrin Substances 0.000 description 2
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 2
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose 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)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 2
- 229930006000 Sucrose Natural products 0.000 description 2
- 241000190111 Westerdykella dispersa Species 0.000 description 2
- 240000008042 Zea mays Species 0.000 description 2
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 description 2
- 235000002017 Zea mays subsp mays Nutrition 0.000 description 2
- 238000010521 absorption reaction Methods 0.000 description 2
- 230000000844 anti-bacterial effect Effects 0.000 description 2
- 229910052799 carbon Inorganic materials 0.000 description 2
- -1 chlorine salt Chemical class 0.000 description 2
- 235000005822 corn Nutrition 0.000 description 2
- 235000019425 dextrin Nutrition 0.000 description 2
- 235000013312 flour Nutrition 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 150000002823 nitrates Chemical class 0.000 description 2
- 229910052757 nitrogen Inorganic materials 0.000 description 2
- 238000000655 nuclear magnetic resonance spectrum Methods 0.000 description 2
- 235000015097 nutrients Nutrition 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- 241000894007 species Species 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 239000005720 sucrose Substances 0.000 description 2
- 238000004809 thin layer chromatography Methods 0.000 description 2
- OWEGMIWEEQEYGQ-UHFFFAOYSA-N 100676-05-9 Natural products OC1C(O)C(O)C(CO)OC1OCC1C(O)C(O)C(O)C(OC2C(OC(O)C(O)C2O)CO)O1 OWEGMIWEEQEYGQ-UHFFFAOYSA-N 0.000 description 1
- 229920001817 Agar Polymers 0.000 description 1
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 1
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- 241000588724 Escherichia coli Species 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- 108010068370 Glutens Proteins 0.000 description 1
- 244000068988 Glycine max Species 0.000 description 1
- 235000010469 Glycine max Nutrition 0.000 description 1
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 1
- FYYHWMGAXLPEAU-UHFFFAOYSA-N Magnesium Chemical compound [Mg] FYYHWMGAXLPEAU-UHFFFAOYSA-N 0.000 description 1
- GUBGYTABKSRVRQ-PICCSMPSSA-N Maltose Natural products O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@@H](CO)OC(O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-PICCSMPSSA-N 0.000 description 1
- PWHULOQIROXLJO-UHFFFAOYSA-N Manganese Chemical compound [Mn] PWHULOQIROXLJO-UHFFFAOYSA-N 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- 239000001888 Peptone Substances 0.000 description 1
- 108010080698 Peptones Proteins 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- PMZURENOXWZQFD-UHFFFAOYSA-L Sodium Sulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=O PMZURENOXWZQFD-UHFFFAOYSA-L 0.000 description 1
- 229920002472 Starch Polymers 0.000 description 1
- 241000209140 Triticum Species 0.000 description 1
- 235000021307 Triticum Nutrition 0.000 description 1
- 241000190113 Westerdykella Species 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 238000005377 adsorption chromatography Methods 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- PNEYBMLMFCGWSK-UHFFFAOYSA-N aluminium oxide Inorganic materials [O-2].[O-2].[O-2].[Al+3].[Al+3] PNEYBMLMFCGWSK-UHFFFAOYSA-N 0.000 description 1
- 150000003863 ammonium salts Chemical class 0.000 description 1
- 239000011260 aqueous acid Substances 0.000 description 1
- 239000012736 aqueous medium Substances 0.000 description 1
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 1
- GUBGYTABKSRVRQ-QUYVBRFLSA-N beta-maltose Chemical compound OC[C@H]1O[C@H](O[C@H]2[C@H](O)[C@@H](O)[C@H](O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@@H]1O GUBGYTABKSRVRQ-QUYVBRFLSA-N 0.000 description 1
- 239000011575 calcium Substances 0.000 description 1
- 229910052791 calcium Inorganic materials 0.000 description 1
- 229940041514 candida albicans extract Drugs 0.000 description 1
- 150000001722 carbon compounds Chemical class 0.000 description 1
- 239000005018 casein Substances 0.000 description 1
- BECPQYXYKAMYBN-UHFFFAOYSA-N casein, tech. Chemical compound NCCCCC(C(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(CC(C)C)N=C(O)C(CCC(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(C(C)O)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(COP(O)(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(N)CC1=CC=CC=C1 BECPQYXYKAMYBN-UHFFFAOYSA-N 0.000 description 1
- 235000021240 caseins Nutrition 0.000 description 1
- 230000034303 cell budding Effects 0.000 description 1
- 239000000460 chlorine Substances 0.000 description 1
- 229910052801 chlorine Inorganic materials 0.000 description 1
- 239000010941 cobalt Substances 0.000 description 1
- 229910017052 cobalt Inorganic materials 0.000 description 1
- GUTLYIVDDKVIGB-UHFFFAOYSA-N cobalt atom Chemical compound [Co] GUTLYIVDDKVIGB-UHFFFAOYSA-N 0.000 description 1
- 239000012141 concentrate Substances 0.000 description 1
- 235000012343 cottonseed oil Nutrition 0.000 description 1
- 238000004090 dissolution Methods 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 235000021312 gluten Nutrition 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 229910052742 iron Inorganic materials 0.000 description 1
- 239000011777 magnesium Substances 0.000 description 1
- 229910052749 magnesium Inorganic materials 0.000 description 1
- 239000011572 manganese Substances 0.000 description 1
- 229910052748 manganese Inorganic materials 0.000 description 1
- 238000001819 mass spectrum Methods 0.000 description 1
- 235000013372 meat Nutrition 0.000 description 1
- 238000000034 method Methods 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 235000013379 molasses Nutrition 0.000 description 1
- 229910017464 nitrogen compound Inorganic materials 0.000 description 1
- 150000002830 nitrogen compounds Chemical class 0.000 description 1
- 235000019319 peptone Nutrition 0.000 description 1
- 239000003208 petroleum Substances 0.000 description 1
- 235000021317 phosphate Nutrition 0.000 description 1
- 150000003013 phosphoric acid derivatives Chemical class 0.000 description 1
- 239000012286 potassium permanganate Substances 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 229910052708 sodium Inorganic materials 0.000 description 1
- GRONZTPUWOOUFQ-UHFFFAOYSA-M sodium;methanol;hydroxide Chemical compound [OH-].[Na+].OC GRONZTPUWOOUFQ-UHFFFAOYSA-M 0.000 description 1
- 238000001179 sorption measurement Methods 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
- 239000012138 yeast extract Substances 0.000 description 1
Landscapes
- Compounds Of Unknown Constitution (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Description
本発明は、ウエスターデイケラ
(Westerdykella)属に属する糸状菌M5070菌によ
る新規な抗生物質M5070およびその製法に関す
る。
本発明の新規な抗生物質M5070の物理化学的な
性質は次の通りである。
(1) 分子量 309(マススペクトルより)
(2) 融点 130℃(分解)
(3) 施光度 〔α〕24 D=−82°(C=0.5%メタノ
ール)
(4) 元素分析
C=65.91% H=8.56% N4.31%
(5) 分子式
本物質はC、H、N、OよりなりC17H27NO4
で示される。
(6) 紫外部吸収スペクトル
メタノール中で測定した紫外部吸収スペクト
ルは第1図(実線)に示す通りで、λmax265
mμ(E1%1cm=715)、λmax274mμ(E1%1cm
=
900)、λmax285mμ(E1%1cm=718)に特徴的
な吸収極大を有す。また0.1HCl−MeOH中お
よび0.1NNaOH−MeOH中の条件におけるその
スペクトルの変化は認められなかつた。(アル
カリ性下のスペクトルは第1図破線で示す通り
である)
(7) 赤外部吸収スペクトル
KBr法により測定した赤外部吸収スペクトル
は第2図に示す通りで、3350、2900、1740、
1650、1430、1370、1180、1100、1020、980cm
-1付近の各波長に吸収帯を有する。
(8) 核磁気共鳴スペクトル
第3図に示す通りである。(CDCl3中
100MHz、内部基準TMS)。
(9) 外 観
淡黄色〜黄白色(また、その塩素塩も同じ)
(10) 各種溶媒に対する溶解性
メタノール、エタノール、アセトン、酢酸エ
チル、クロロホルム、ベンゼン、ジエチルエー
テルに可溶、水、n−ヘキサン、石油エーテル
に不溶。
(11) 呈色反応
過マンガン酸カリウム脱色反応、ニンヒドリ
ン反応は陽性、塩化第二鉄反応、フエーリング
反応、モーリツシユ反応は陰性。
(12) 塩基性、中性、酸性の区別
弱塩基性
(13) 安定性
溶媒中60℃、15分間で、抗菌力の低下が認め
られた。またPH9以上ではすみやかに失活す
る。
(14) 薄層クロマトグラフイー〔東京化成社製ス
ポーツトフイルム(シリカゲル)使用〕
クロロホルム:メタノール=25:1
Rf=0.52
ベンゼン:メタノール=20:1
Rf=0.15
ベンゼン:酢酸エチル=1:1
Rf=0.05
また本発明の新規な抗生物質M5070の抗菌スペ
クトルは次の通りである。なお細菌については普
通寒天培地(Escherichia coli;ペプトン培地)、
真菌、酵母についてはサブロー培地を用い、ペー
パーデイスクによる阻止円の径にて示す(使用濃
度500γ/ml)
The present invention relates to a novel antibiotic M5070 produced by M5070, a filamentous fungus belonging to the genus Westerdykella, and a method for producing the same. The physicochemical properties of the novel antibiotic M5070 of the present invention are as follows. (1) Molecular weight 309 (from mass spectrum) (2) Melting point 130℃ (decomposition) (3) Light intensity [α] 24 D = -82° (C = 0.5% methanol) (4) Elemental analysis C = 65.91% H =8.56% N4.31% (5) Molecular formula This substance consists of C, H, N, and O. C 17 H 27 NO 4
It is indicated by. (6) Ultraviolet absorption spectrum The ultraviolet absorption spectrum measured in methanol is as shown in Figure 1 (solid line).
mμ (E 1 % 1 cm=715), λmax274mμ (E 1 % 1 cm
=
900), with a characteristic absorption maximum at λmax 285 mμ (E 1 % 1 cm = 718). Moreover, no change in the spectrum was observed under the conditions of 0.1HCl-MeOH and 0.1N NaOH-MeOH. (The spectrum under alkaline conditions is as shown by the broken line in Figure 1) (7) Infrared absorption spectrum The infrared absorption spectrum measured by the KBr method is as shown in Figure 2, 3350, 2900, 1740,
1650, 1430, 1370, 1180, 1100, 1020, 980cm
It has an absorption band at each wavelength around -1 . (8) Nuclear magnetic resonance spectrum As shown in Figure 3. (in CDCl 3
100MHz, internal reference TMS). (9) Appearance Pale yellow to yellowish white (same for its chlorine salt) (10) Solubility in various solvents Soluble in methanol, ethanol, acetone, ethyl acetate, chloroform, benzene, diethyl ether, water, n- Insoluble in hexane, petroleum ether. (11) Color reaction Positive for potassium permanganate decolorization reaction and ninhydrin reaction, negative for ferric chloride reaction, Fehring reaction, and Moritzhu reaction. (12) Distinction between basic, neutral, and acidic Weak basic (13) Stability A decrease in antibacterial activity was observed in a solvent at 60°C for 15 minutes. Moreover, it is quickly deactivated at pH 9 or higher. (14) Thin layer chromatography [using Sports Film (silica gel) manufactured by Tokyo Kasei Co., Ltd.] Chloroform: methanol = 25:1 Rf = 0.52 Benzene: methanol = 20:1 Rf = 0.15 Benzene: ethyl acetate = 1:1 Rf =0.05 Furthermore, the antibacterial spectrum of the novel antibiotic M5070 of the present invention is as follows. For bacteria, use ordinary agar medium (Escherichia coli; peptone medium),
For fungi and yeast, use Sabouraud medium and indicate the diameter of the inhibition circle on a paper disk (use concentration 500γ/ml)
以上の菌学的諾性質より本発明の糸状菌M5070
菌株は、閉じて口のない子のう殻を持ち、子のう
は32胞子、子のう胞子は出芽のための孔および裂
け目を持たないことから、ウエスターデイケム属
に属するものと認められる。
またウエスターデイケラ属は、1655年に、
Stolkによつて作られた属で、現在までに6種1
変種認められている。
ところで、その子のう殻、子のう、子のう胞子
の形、大きさ、その他の特色から、本菌の糸状菌
M5070菌株は、ウエスターデイケラ・デイスパー
サ(Westerdykella dispersa)の記載と良く一致
し、他の種とは一致せず、またウエスターデイケ
ラ・デイスパーサIFO8431、同NHL2530との比較
の結果も良く一致することから、本菌株はウエス
ターデイケラ・デイスパーサと同定され、ウエス
ターデイケラ・デイスパーサM5070と命名され
た。
〔参考文献;Trans Brit.mycol.Soc.、38、419
〜424(1955)、Can.J.Bot.、39.1633〜1666
(1961)、Ceska Mykologin、18.82〜84
(1964)、The Genera of Fungi Sporulating in
Pure Culture315pp(1974)〕
次いで、本発明のM5070を製造するに当つて、
上記のウエスターデイケラ・デイスパーサM5070
菌株はその一例であつて、ウエスターデイケラ属
に属するM5070生産菌はその変異株であつてもす
べて使用し得るもので、このような生産菌を通常
の微生物に培養に使用する培地成分を含む培地に
好気的に培養することによつて得られる。培地と
しては、固型培地または液体培地が用いられる
が、特に大量生産のためには液体培地、特に水性
培地が用いられる。
培地の栄養源としては、微生物の培養に通常用
いられるものが広く使用され得る。炭素源として
は同化可能な炭素化合物であればよく、例えばグ
ルコース、シユクロース、マルトース、スター
チ、デキストリン、モラセスなどが使用される。
窒素源としては利用可能な窒素化合物であればよ
く、例えばコーンスチープ・リカー、大豆粉、綿
実粉、小麦グルテン、ポリペプトン、肉エキス、
酵母エキス、カゼイン加水分解物、アンモニウム
塩、硝酸塩などが使用される。その他、リン酸
塩、硝酸塩、マグネシウム、カルシウム、カリウ
ム、ナトリウム、コバルト、鉄、マンガンなどの
塩類が必要に応じて使用される。
培養温度は菌が発育し、M5070を生産する範囲
内で適宜変更し得るが、特に好ましくは25〜30℃
である。培養時間は、条件によつて多少異なる
が、通常2〜5日程度であつて、M5070が最高力
価に達する時期を見計つて適当な時期に培養を終
了すればよい。
このようにして得られたM5070生産菌の培養物
中において、M5070はその菌体外に大部分産生さ
れている。
次いでこのM5070生産菌は培養物からM5070を
採取するのであるが、M5070が弱塩基性脂溶性物
質であることを利用して分離精製を行なうことが
簡便である。
M5070の分離精製手段の一例を示すと次の通り
である。まずM5070生産菌を前述の如く培養して
得られる培養物から菌体などの固形分を去して
培養液を、これを弱アルカリ性の条件下、クロ
ロホルム、酢酸エチル、ベンゼンなどの可溶性溶
媒にて抽出せしめ、これを濃縮し、さらにこれを
可溶性溶媒に溶解した後、シリカゲルやアルミナ
を充填した吸着クロマトグラフイーを行ない、そ
の活性区分を得、これを乾固して精製された
M5070を得ればよく、さらにこのM5070を可溶性
溶媒に溶解し、これに水を加えて酸類水溶液を加
えて水転せしめ、その水層を乾燥してM5070の酸
類との塩を得てもよい。
次に本発明の実施例を挙げて具体的に述べる
が、本発明はこれによつて何んら限定されるもの
ではない。
実施例 1
シユクロース1.5%、デキストリン1.5%、ポリ
ペプトン2%、コーン・スチープ・リカー2%、
KH2PO40.2%、MgSO4・7H2O0.1%、FeSO4・
7H2O0.01%、セトイト、(商品名ハイフロスーパ
ーセル)1%からなる培地100mlを500ml容三角フ
ラスコに分注し、120℃、15分間高圧減菌し、こ
れに、ウエスターデイケラ・デイスパーサM5070
菌株の斜面培養からの胞子を含む菌糸をかき取つ
て接種し、27℃、3日間振盪培養した。次いでこ
の培養を、同一組成からなる培地100ml含有500ml
容三角フラスコ60本に、1〜2mlづつ接種し、27
℃、4日間振盪培養した。培養後これを併合し、
培養物約5.5を得、さらにセライトを用いて
過して菌体を除去し、約5の培養液を得た
(この液のPHは7.8、M5070を300γ/ml含有)。
さらにこの培養液に1/2量のクロロホルムを
加え、PH9で抽出した。次いでそのクロロホルム
層を回収し、無水硫酸ナトリウムを用いて、脱水
後減圧濃縮し、乾燥し、さらにこれを少量のクロ
ロホルムに溶解せしめ、クロロホルムで充填した
シリカゲルカラム(直径2×30cm)に吸着せしめ
た。吸着後少量のクロロホルムで洗浄後クロロホ
ルム:メタノール=75:1の展開溶媒にて、展開
せしめ、1フラクシヨン12mlとして分画し、その
活性区分、フラクシヨンNo.37〜45を集め、濃縮
し、乾固して約836mgの淡黄色〜黄白色の精製さ
れたM5070を得た(回収率50%、またこの薄層ク
ロマトグラフイーにて単一スポツトを示した)。
実施例 2
400mgのM5070をベンゼン50mlに溶解せしめ、
これに同量の水を加えた後、これにPH3になるま
で0.5N塩酸を加え、その水層を回収し、同様の
操作を2回行ない、その層を集めて凍結乾燥して
約350mgのM5070の塩酸塩を得た。
From the above mycological properties, the filamentous fungus M5070 of the present invention
The strain is recognized as belonging to the Westerdychem genus because it has a closed, mouthless ascus, and the ascospores contain 32 spores, and the ascospores do not have holes or clefts for budding. In 1655, the genus Westerdycera was
The genus was created by Stolk, and to date there are 6 species and 1
Variants are recognized. By the way, the shape, size, and other characteristics of the asci, ascospores, and ascospores indicate that this fungus is a filamentous fungus.
The M5070 strain matches well with the description of Westerdykella dispersa, but does not match with other species, and the results of comparison with Westerdykella dispersa IFO8431 and NHL2530 also show good agreement. , this strain was identified as Westerdeichella dispersa and named Westerdeichella dispersa M5070. [References; Trans Brit.mycol.Soc., 38 , 419
~424 (1955), Can.J.Bot., 39 . 1633-1666
(1961), Ceska Mykologin, 18 . 82~84
(1964), The Genera of Fungi Sporulating in
Pure Culture 315pp (1974)] Next, in manufacturing M5070 of the present invention,
Westerdychella Dispersa M5070 above
Bacterial strains are one example, and all M5070-producing bacteria belonging to the genus Westerdeichella can be used, even if they are mutant strains. Obtained by culturing aerobically in a medium. As the medium, a solid medium or a liquid medium is used, and especially for mass production, a liquid medium, especially an aqueous medium, is used. As the nutrient source for the medium, a wide variety of nutrients commonly used for culturing microorganisms can be used. The carbon source may be any assimilable carbon compound, such as glucose, sucrose, maltose, starch, dextrin, and molasses.
The nitrogen source may be any available nitrogen compound, such as corn steep liquor, soybean flour, cottonseed flour, wheat gluten, polypeptone, meat extract,
Yeast extract, casein hydrolyzate, ammonium salts, nitrates, etc. are used. In addition, salts such as phosphates, nitrates, magnesium, calcium, potassium, sodium, cobalt, iron, and manganese are used as necessary. The culture temperature can be changed as appropriate within the range that allows the bacteria to grow and produce M5070, but is particularly preferably 25 to 30°C.
It is. The culture time varies somewhat depending on the conditions, but is usually about 2 to 5 days, and the culture may be terminated at an appropriate time by determining the time when M5070 reaches its maximum titer. In the culture of M5070-producing bacteria thus obtained, most of M5070 is produced outside the cells. Next, M5070 is collected from the culture of this M5070-producing bacterium, but it is convenient to separate and purify it by taking advantage of the fact that M5070 is a weakly basic fat-soluble substance. An example of separation and purification means for M5070 is as follows. First, M5070-producing bacteria are cultured as described above, and solid contents such as bacterial bodies are removed from the resulting culture to obtain a culture solution. After extraction, concentration, and dissolution in a soluble solvent, adsorption chromatography using silica gel or alumina was performed to obtain the active fraction, which was then dried and purified.
It is sufficient to obtain M5070, and it is also possible to dissolve this M5070 in a soluble solvent, add water to this, add an aqueous acid solution, perform water inversion, and dry the aqueous layer to obtain a salt of M5070 with an acid. . Next, the present invention will be specifically described with reference to examples, but the present invention is not limited thereto. Example 1 1.5% sucrose, 1.5% dextrin, 2% polypeptone, 2% corn steep liquor,
KH 2 PO 4 0.2%, MgSO 4・7H 2 O 0.1%, FeSO 4・
Dispense 100 ml of a medium consisting of 0.01% 7H 2 O and 1% Cetoite (trade name: Hyflo Supercell) into a 500 ml Erlenmeyer flask, sterilize it under high pressure at 120°C for 15 minutes, and add Westerdeichella dispersa to this. M5070
The mycelium containing spores from the slant culture of the strain was scraped and inoculated, and cultured with shaking at 27°C for 3 days. This culture was then transferred to 500 ml of medium containing 100 ml of the same composition.
Inoculate 1 to 2 ml each into 60 Erlenmeyer flasks, and inoculate 27
The cells were cultured with shaking at ℃ for 4 days. After culturing, merge this
A culture of about 5.5 was obtained, which was further filtered through Celite to remove bacterial cells, to obtain a culture solution of about 5 (PH of this solution was 7.8, containing 300 γ/ml of M5070). Furthermore, 1/2 amount of chloroform was added to this culture solution, and the mixture was extracted with PH9. The chloroform layer was then collected, dehydrated using anhydrous sodium sulfate, concentrated under reduced pressure, dried, and further dissolved in a small amount of chloroform and adsorbed on a silica gel column (diameter 2 x 30 cm) filled with chloroform. . After adsorption, wash with a small amount of chloroform, develop with a developing solvent of chloroform:methanol = 75:1, fractionate into 12 ml fraction, collect the active fraction, fractions No. 37 to 45, concentrate, and dry. Approximately 836 mg of pale yellow to yellowish white purified M5070 was obtained (recovery rate 50%, and thin layer chromatography showed a single spot). Example 2 400 mg of M5070 was dissolved in 50 ml of benzene,
After adding the same amount of water to this, add 0.5N hydrochloric acid to this until the pH becomes 3, collect the aqueous layer, repeat the same operation twice, collect the layer, freeze-dry it, and make about 350mg of it. The hydrochloride of M5070 was obtained.
第1図は本発明のM5070の紫外部吸収スペクト
ルを示し、第2図は本発明のM5070の赤外部吸収
スペクトルを示し、第3図は本発明のM5070の核
磁気共鳴スペクトルを示す。
FIG. 1 shows the ultraviolet absorption spectrum of M5070 of the present invention, FIG. 2 shows the infrared absorption spectrum of M5070 of the present invention, and FIG. 3 shows the nuclear magnetic resonance spectrum of M5070 of the present invention.
Claims (1)
生物質M5070およびその無毒性塩 ●分子量 309 ●融点 130℃(分解) ●施光度 〔α〕24 D=−82°(C=0.5%メタノー
ル) ●元素分析 C=65.91% H=8.56 N=4.31% ●分子式 C17H27NO4 ●紫外部吸収スペクトル 第1図に示す通り ●赤外部吸収スペクトル 第2図に示す通り ●外観 淡黄色〜黄白色 ●各種溶媒に対する溶解性 メタノール、エタノール、アセトン、酢酸エ
チル、クロロホルム、ベンゼン、ジエチルエー
テルに可溶、水、n−ヘキサン、石油エーテル
に不溶 ●呈色反応 過マンガン酸カリウム脱色反応、ニンヒドリ
ン反応は陽性、塩化第二鉄反応、フエーリング
反応、モーリツシユ反応は陰性 ●塩基性、中性、酸性の区別 弱塩基性 2 ウエスターデイケラ属に属する抗生物質
M5070生産菌を培地に培養し、その培養物より抗
生物質M5070を採取することを特徴とする新規な
抗生物質M5070の製法。 3 ウエスターデイケラ属に属する抗生物質
M5070生産菌が、ウエスターデイケラ・デイスパ
ーサM5070菌である特許請求の範囲第2項記載の
製法。[Claims] 1. A novel antibiotic M5070 and its non-toxic salt having the following physicochemical properties ●Molecular weight 309 ●Melting point 130℃ (decomposition) ●Degree of light application [α] 24 D = -82° (C=0.5% methanol) ●Elemental analysis C=65.91% H=8.56 N=4.31% ●Molecular formula C 17 H 27 NO 4 ●Ultraviolet absorption spectrum As shown in Figure 1 ●Infrared absorption spectrum As shown in Figure 2 Appearance: Pale yellow to yellowish white Potassium decolorization reaction, ninhydrin reaction are positive, ferric chloride reaction, Fehling reaction, Moritzhu reaction are negative ●Distinction between basic, neutral, and acidic Weak basic 2 Antibiotics belonging to the Westerdichella genus
A novel method for producing antibiotic M5070, which is characterized by culturing M5070-producing bacteria in a medium and collecting antibiotic M5070 from the culture. 3 Antibiotics belonging to the genus Westerdycera
3. The manufacturing method according to claim 2, wherein the M5070-producing bacterium is Westerdichella dispersa M5070 bacterium.
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP9290879A JPS5618592A (en) | 1979-07-20 | 1979-07-20 | Novel antibiotic, m5070 and its preparation |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP9290879A JPS5618592A (en) | 1979-07-20 | 1979-07-20 | Novel antibiotic, m5070 and its preparation |
Publications (2)
Publication Number | Publication Date |
---|---|
JPS5618592A JPS5618592A (en) | 1981-02-21 |
JPS6129715B2 true JPS6129715B2 (en) | 1986-07-08 |
Family
ID=14067575
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP9290879A Granted JPS5618592A (en) | 1979-07-20 | 1979-07-20 | Novel antibiotic, m5070 and its preparation |
Country Status (1)
Country | Link |
---|---|
JP (1) | JPS5618592A (en) |
Families Citing this family (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPS6128434A (en) * | 1984-07-19 | 1986-02-08 | Nippon Paint Co Ltd | Dispersion stabilizer and its use |
US5294725A (en) * | 1989-01-31 | 1994-03-15 | E. R. Squibb & Sons, Inc. | Scopularin |
US5270334A (en) * | 1989-01-31 | 1993-12-14 | E. R. Squibb & Sons, Inc. | 4-methoxy-5-methyl-pyran-3-ol natural products and derivatives thereof |
US5266593A (en) * | 1992-06-30 | 1993-11-30 | E. R. Squibb & Sons, Inc. | Substituted pyran derivatives having antiinfective activity |
CN110343620B (en) * | 2019-07-01 | 2020-08-14 | 福建农林大学 | Plant rhizosphere fungus F9 capable of adsorbing cadmium ions and application thereof |
-
1979
- 1979-07-20 JP JP9290879A patent/JPS5618592A/en active Granted
Also Published As
Publication number | Publication date |
---|---|
JPS5618592A (en) | 1981-02-21 |
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