JPS6316391B2 - - Google Patents
Info
- Publication number
- JPS6316391B2 JPS6316391B2 JP55128107A JP12810780A JPS6316391B2 JP S6316391 B2 JPS6316391 B2 JP S6316391B2 JP 55128107 A JP55128107 A JP 55128107A JP 12810780 A JP12810780 A JP 12810780A JP S6316391 B2 JPS6316391 B2 JP S6316391B2
- Authority
- JP
- Japan
- Prior art keywords
- acetone
- antibiotic
- culture
- salt
- hexane
- 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
- GQTQZTGDYKBXJP-UHFFFAOYSA-N Antibiotic TM 531B Natural products CC(CC(C)C(=O)C(=CC(C)C1OC2(CCC(C)(O2)C3CC(OC4CCC(O)C(C)O4)C(C)C5(OC(CC5C)C6OC(O)(CO)C(C)CC6C)O3)CC(O)C1C)C)C(=O)O GQTQZTGDYKBXJP-UHFFFAOYSA-N 0.000 claims description 9
- 150000003839 salts Chemical class 0.000 claims description 3
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 39
- UHOVQNZJYSORNB-UHFFFAOYSA-N Benzene Chemical compound C1=CC=CC=C1 UHOVQNZJYSORNB-UHFFFAOYSA-N 0.000 description 18
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 18
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 12
- 239000000284 extract Substances 0.000 description 7
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 6
- 238000000862 absorption spectrum Methods 0.000 description 6
- 239000013543 active substance Substances 0.000 description 6
- KDCIHNCMPUBDKT-UHFFFAOYSA-N hexane;propan-2-one Chemical compound CC(C)=O.CCCCCC KDCIHNCMPUBDKT-UHFFFAOYSA-N 0.000 description 6
- 239000012046 mixed solvent Substances 0.000 description 6
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 description 6
- 159000000000 sodium salts Chemical class 0.000 description 6
- 239000002904 solvent Substances 0.000 description 6
- 239000000126 substance Substances 0.000 description 6
- 239000006228 supernatant Substances 0.000 description 6
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 5
- 230000001580 bacterial effect Effects 0.000 description 5
- 239000003960 organic solvent Substances 0.000 description 5
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 4
- 241000894006 Bacteria Species 0.000 description 4
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 description 4
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 4
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 4
- 239000003242 anti bacterial agent Substances 0.000 description 4
- 230000000844 anti-bacterial effect Effects 0.000 description 4
- 239000002518 antifoaming agent Substances 0.000 description 4
- 230000003115 biocidal effect Effects 0.000 description 4
- 238000006243 chemical reaction Methods 0.000 description 4
- 239000000843 powder Substances 0.000 description 4
- 239000000741 silica gel Substances 0.000 description 4
- 229910002027 silica gel Inorganic materials 0.000 description 4
- 238000001460 carbon-13 nuclear magnetic resonance spectrum Methods 0.000 description 3
- 230000009422 growth inhibiting effect Effects 0.000 description 3
- 238000000425 proton nuclear magnetic resonance spectrum Methods 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 2
- OKKJLVBELUTLKV-MZCSYVLQSA-N Deuterated methanol Chemical compound [2H]OC([2H])([2H])[2H] OKKJLVBELUTLKV-MZCSYVLQSA-N 0.000 description 2
- 241000192125 Firmicutes Species 0.000 description 2
- 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 2
- 229910021380 Manganese Chloride Inorganic materials 0.000 description 2
- GLFNIEUTAYBVOC-UHFFFAOYSA-L Manganese chloride Chemical compound Cl[Mn]Cl GLFNIEUTAYBVOC-UHFFFAOYSA-L 0.000 description 2
- 235000019483 Peanut oil Nutrition 0.000 description 2
- PMZURENOXWZQFD-UHFFFAOYSA-L Sodium Sulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=O PMZURENOXWZQFD-UHFFFAOYSA-L 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 241000187747 Streptomyces Species 0.000 description 2
- 241000187391 Streptomyces hygroscopicus Species 0.000 description 2
- HEDRZPFGACZZDS-MICDWDOJSA-N Trichloro(2H)methane Chemical compound [2H]C(Cl)(Cl)Cl HEDRZPFGACZZDS-MICDWDOJSA-N 0.000 description 2
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 description 2
- 230000002378 acidificating effect Effects 0.000 description 2
- 150000003863 ammonium salts Chemical class 0.000 description 2
- 229940088710 antibiotic agent Drugs 0.000 description 2
- 229910000019 calcium carbonate Inorganic materials 0.000 description 2
- 229910052799 carbon Inorganic materials 0.000 description 2
- 239000012141 concentrate Substances 0.000 description 2
- 239000013078 crystal Substances 0.000 description 2
- 238000012258 culturing Methods 0.000 description 2
- 238000000921 elemental analysis Methods 0.000 description 2
- 238000010828 elution Methods 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 239000008103 glucose Substances 0.000 description 2
- 235000011187 glycerol Nutrition 0.000 description 2
- RUTXIHLAWFEWGM-UHFFFAOYSA-H iron(3+) sulfate Chemical compound [Fe+3].[Fe+3].[O-]S([O-])(=O)=O.[O-]S([O-])(=O)=O.[O-]S([O-])(=O)=O RUTXIHLAWFEWGM-UHFFFAOYSA-H 0.000 description 2
- 229910000360 iron(III) sulfate Inorganic materials 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 239000011565 manganese chloride Substances 0.000 description 2
- 235000002867 manganese chloride Nutrition 0.000 description 2
- 229940099607 manganese chloride Drugs 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 235000013372 meat Nutrition 0.000 description 2
- 238000002844 melting Methods 0.000 description 2
- 230000008018 melting Effects 0.000 description 2
- 230000000813 microbial effect Effects 0.000 description 2
- 229910052757 nitrogen Inorganic materials 0.000 description 2
- 239000000312 peanut oil Substances 0.000 description 2
- 229920001296 polysiloxane Polymers 0.000 description 2
- 229910052938 sodium sulfate Inorganic materials 0.000 description 2
- 235000011152 sodium sulphate Nutrition 0.000 description 2
- 238000012360 testing method Methods 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
- ZCYVEMRRCGMTRW-UHFFFAOYSA-N 7553-56-2 Chemical compound [I] ZCYVEMRRCGMTRW-UHFFFAOYSA-N 0.000 description 1
- 229920001817 Agar Polymers 0.000 description 1
- 229920001353 Dextrin Polymers 0.000 description 1
- 239000004375 Dextrin Substances 0.000 description 1
- 244000068988 Glycine max Species 0.000 description 1
- 235000010469 Glycine max Nutrition 0.000 description 1
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-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
- 239000001888 Peptone Substances 0.000 description 1
- 108010080698 Peptones Proteins 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- CDBYLPFSWZWCQE-UHFFFAOYSA-L Sodium Carbonate Chemical class [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 description 1
- 229920002472 Starch Polymers 0.000 description 1
- 240000008042 Zea mays Species 0.000 description 1
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 description 1
- 235000002017 Zea mays subsp mays Nutrition 0.000 description 1
- 238000005273 aeration Methods 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- 238000004458 analytical method Methods 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
- 159000000007 calcium salts Chemical class 0.000 description 1
- 239000004202 carbamide Substances 0.000 description 1
- 229920001429 chelating resin Polymers 0.000 description 1
- 239000004927 clay Substances 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 235000005822 corn Nutrition 0.000 description 1
- 238000012136 culture method Methods 0.000 description 1
- 239000000645 desinfectant Substances 0.000 description 1
- 235000019425 dextrin Nutrition 0.000 description 1
- 238000004821 distillation Methods 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 235000013312 flour Nutrition 0.000 description 1
- 239000000499 gel Substances 0.000 description 1
- 238000001802 infusion Methods 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 239000002054 inoculum Substances 0.000 description 1
- 229910017053 inorganic salt Inorganic materials 0.000 description 1
- 239000011630 iodine Substances 0.000 description 1
- 229910052740 iodine Inorganic materials 0.000 description 1
- 150000002505 iron Chemical class 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 238000000034 method Methods 0.000 description 1
- XELZGAJCZANUQH-UHFFFAOYSA-N methyl 1-acetylthieno[3,2-c]pyrazole-5-carboxylate Chemical compound CC(=O)N1N=CC2=C1C=C(C(=O)OC)S2 XELZGAJCZANUQH-UHFFFAOYSA-N 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 230000007935 neutral effect Effects 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 239000003921 oil Substances 0.000 description 1
- 235000019198 oils Nutrition 0.000 description 1
- 235000019319 peptone Nutrition 0.000 description 1
- XAEFZNCEHLXOMS-UHFFFAOYSA-M potassium benzoate Chemical compound [K+].[O-]C(=O)C1=CC=CC=C1 XAEFZNCEHLXOMS-UHFFFAOYSA-M 0.000 description 1
- 239000012286 potassium permanganate Substances 0.000 description 1
- 239000002243 precursor Substances 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 229920005989 resin Polymers 0.000 description 1
- 239000011347 resin Substances 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 238000001179 sorption measurement Methods 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- QAOWNCQODCNURD-UHFFFAOYSA-N sulfuric acid Substances OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 1
- 239000000273 veterinary drug Substances 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
- 229940088594 vitamin Drugs 0.000 description 1
- 229930003231 vitamin Natural products 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
Landscapes
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Heterocyclic Carbon Compounds Containing A Hetero Ring Having Oxygen Or Sulfur (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Description
本発明は新規な抗生物質に関し、更に詳しくは
ストレプトミセス属に属するTM−531株を好気
的条件下で培養することにより得られる抗生物質
TM−531Bに関する。
本発明者らは前記TM−531株の好気培養物中
に主としてグラム陽性菌に対して強い増殖抑制作
用を示す抗生物質が生産、蓄積されることを見出
し、培養物中より活性物質を抽出、精製して抗生
物質TM−531Bを得、本発明を完成した。
本発明において、抗生物質TM−531B生産菌
としてストレプトミセス属に属するストレプトミ
セス・ハイグロスコピカスTM−531を用いた。
この菌株は埼玉県大宮市の土壤から新たに分離し
たものであり、微生物の名称「放線菌TM−
531」、微生物寄託番号「微工研菌寄第4737号
（FERM−ＰNo.4737）」として工業技術院微生物
工業技術研究所に寄託されている。その菌学的性
状および在来の類似種との異同については特開昭
55−92387号明細書に詳しく記述されている。
本発明におけるTM−531Bの生産は、大略、
抗生物質を生産する場合に準じて行ない、各種栄
養物質を含む培地でTM−531B生産菌を好気的
条件下で培養することにより行なう。培地は液体
培地が好ましく、主として炭素源、窒素源、無機
塩よりなり、その他必要に応じて消泡剤、ビタミ
ン類、先駆物質を加えることができ、PHは７前後
に調整する。
炭素源としては、例えば、グルコース、マルト
ース、澱粉、デキストリン、グリセリン、落花生
油などを単独かまたは混合して用いる。窒素源と
しては、例えば、ペプトン、肉エキス、大豆粉、
オートミール、コーン・ステイープ・リカー、尿
素、アンモニウム塩などを単独かまたは混合して
用いる。無機塩としては、例えば、炭酸カルシウ
ム、塩化ナトリウム、硫酸第二鉄、塩化マンガン
などを単独かまたは混合して用いる。
消泡剤としてはシリコーン化合物などを用いる
ことができる。
培養方法としては振盪培養、通気深部培養など
の好気的培養が適しており、PH７前後で25〜40
℃、望ましくは28〜30℃で培養する。
培養により生成した抗生物質TM−531Bは、
例えば次の方法により単離することができる。
すなわち、培養終了後の培養物を遠心分離して
上澄液と菌体に分離する。
上澄液中の活性物質は酢酸エチル、ベンゼン、
クロロホルムなどの非水溶性有機溶媒で抽出する
か、吸着樹脂などに吸着させてアルコール、アセ
トンなどの有機溶媒で溶出する。必要に応じて、
濃縮後、適当な有機溶媒で更に抽出することもで
きる。
菌体中の活性物質は、アルコール、アセトンな
どの有機溶媒で抽出し、これを濃縮後、上澄液の
抽出溶媒と同じ溶媒に転溶する。
上澄液抽出液と菌体抽出液を合せ、減圧濃縮す
る。この濃縮物をベンゼンに溶解し、シリカゲ
ル・カラムに活性物質を吸着させ、ベンゼン、酢
酸エチルなどの有機溶媒で活性物質中のTM−
531およびデアネマイシンを溶出、除去する。
残りの活性物質をアセトンなどの有機溶媒で溶
出し、溶培溜去後、ヘキサン−アセトン（４：
１）混合溶媒に再溶解し、シリカゲル・カラムに
吸着させる。
このカラムをヘキサン−アセトン（４：１）混
合溶媒で洗浄後、活性物質をヘキサン−アセトン
（６：４）混合溶媒で溶出し、カラム容積の１〜
1.5ベツト量の区分（薄層クロマトグラフイーに
より確認）を集める。
これを常法により精製することにより、抗生物
質TM−531Bの結晶を得ることができる。精製
上または使用上、必要があればナトリウム塩、カ
リウム塩、カルシウム塩、鉄塩、アンモニウム塩
などに変換することもできる。
本発明の目的物質である抗生物質TM−531B
はその元素分析値、分子量、紫外線吸収スペクト
ル、赤外線吸収スペクトル、 1H−NMRスペク
トル、 13C−NMRスペクトルの解析よりその構
造式が次の如く決定された。また、その理化学的
性質および抗菌作用は次の通りである。
() 構造式
() 理化学的性質（ナトリウム塩として測定）
元素分析値 C46H75O14Naとして
理論値(%)：C63.16 H8.58 Na2.63
実測値(%)：C62.99 H8.71 Na2.51
分子量：874
融点：252.1〜253.3℃
比旋光度：〔ａ〕26 D＝＋37.8゜（Ｃ＝0.5、メタ
ノール）
紫外線吸収スペクトル（メタノール）
E1%1cn（232nｍ）＝160.0
赤外線吸収スペクトル
KBr錠で測定した結果を第１図に示す。
溶媒に対する溶解性：水に不溶。
メタノール、エタノール、アセトン、酢酸
エチル、ヘキサン、クロロホルム、ベンゼン
に可溶。
呈色反応：ヨード反応、過マンガン酸カリ
ウム反応およびバニリン−硫酸反応陽性
物質の色：無色
1H−NMRスペクトルおよび 13C−NMR
スペクトル：重メタノール添加の重クロロホ
ルム中、前者を100MHzで、後者を25.05MHz
で測定した結果をそれぞれ第２図および第３
図に示す。
塩基性、酸性、中性の区別：酸性
() 抗菌作用
抗生物質TM−531Bはグラム陽性菌に対し
てすぐれた増殖抑制作用を有するが、グラム陰
性菌、酵母に対しては増殖抑制作用を有しな
い。
以上のように、抗生物質TM−531Bは新規な
抗生物質であつて、医薬、動物薬、消毒殺菌剤と
して有用である。
次に抗生物質TM−531Bの抗菌作用を明らか
にする試験例と製造例を示す実施例を挙げて本発
明を具体的に説明する。
試験例
培地としてハートインフユージヨン・アガーを
用い、イノキユラム・サイズ106CFUで抗生物質
TM−531Bの各種細菌に対するMIC（最小発育阻
止濃度）を測定し、この抗菌作用を調べた。
その結果を次表に示す。
The present invention relates to a novel antibiotic, and more specifically to an antibiotic obtained by culturing the TM-531 strain belonging to the genus Streptomyces under aerobic conditions.
Regarding TM-531B. The present inventors discovered that an antibiotic that exhibits a strong growth-inhibitory effect mainly on Gram-positive bacteria is produced and accumulated in an aerobic culture of the TM-531 strain, and the active substance was extracted from the culture. , the antibiotic TM-531B was obtained and the present invention was completed. In the present invention, Streptomyces hygroscopicus TM-531, which belongs to the genus Streptomyces, was used as the antibiotic TM-531B-producing bacterium.
This strain was newly isolated from a clay pot in Omiya City, Saitama Prefecture, and the microorganism name is ``Actinobacterium TM-''.
531'' and has been deposited with the Institute of Microbial Technology, Agency of Industrial Science and Technology under the microbial deposit number ``FERM-P No. 4737''. Regarding its mycological properties and differences from similar native species, please refer to JP-A-Sho.
It is described in detail in the specification of No. 55-92387. The production of TM-531B in the present invention is roughly as follows:
This is carried out in the same manner as when producing antibiotics, by culturing TM-531B producing bacteria under aerobic conditions in a medium containing various nutrients. The medium is preferably a liquid medium, consisting mainly of a carbon source, a nitrogen source, and an inorganic salt, and antifoaming agents, vitamins, and precursors may be added as necessary, and the pH is adjusted to around 7. As the carbon source, for example, glucose, maltose, starch, dextrin, glycerin, peanut oil, etc. are used alone or in combination. Examples of nitrogen sources include peptone, meat extract, soybean flour,
Oatmeal, corn steep liquor, urea, ammonium salts, etc. may be used alone or in combination. Examples of inorganic salts include calcium carbonate, sodium chloride, ferric sulfate, manganese chloride, and the like, used alone or in combination. A silicone compound or the like can be used as an antifoaming agent. As a culture method, aerobic culture such as shaking culture and aerated deep culture is suitable.
Culture at 28-30°C, preferably 28-30°C. Antibiotic TM-531B produced by culture is
For example, it can be isolated by the following method. That is, the culture after completion of cultivation is centrifuged to separate the supernatant and the bacterial cells. The active substances in the supernatant are ethyl acetate, benzene,
Extract with a water-insoluble organic solvent such as chloroform, or adsorb onto an adsorption resin and elute with an organic solvent such as alcohol or acetone. as needed,
After concentration, it can be further extracted with a suitable organic solvent. The active substance in the bacterial cells is extracted with an organic solvent such as alcohol or acetone, concentrated, and then transferred to the same solvent as the extraction solvent of the supernatant. Combine the supernatant extract and the bacterial cell extract and concentrate under reduced pressure. This concentrate was dissolved in benzene, the active substance was adsorbed on a silica gel column, and the TM-
Elute and remove 531 and deanemycin. The remaining active substance was eluted with an organic solvent such as acetone, and after distillation of the solution medium, hexane-acetone (4:
1) Redissolve in a mixed solvent and adsorb onto a silica gel column. After washing this column with a hexane-acetone (4:1) mixed solvent, the active substance was eluted with a hexane-acetone (6:4) mixed solvent, and
Collect 1.5 bed volume fractions (confirmed by thin layer chromatography). By purifying this using a conventional method, crystals of antibiotic TM-531B can be obtained. If necessary for purification or use, it can be converted into sodium salt, potassium salt, calcium salt, iron salt, ammonium salt, etc. Antibiotic TM-531B, which is the object substance of the present invention
Its structural formula was determined as follows from the analysis of its elemental analysis values, molecular weight, ultraviolet absorption spectrum, infrared absorption spectrum, 1 H-NMR spectrum, and 13 C-NMR spectrum. In addition, its physicochemical properties and antibacterial effects are as follows. () Structural formula () Physical and chemical properties (measured as sodium salt) Elemental analysis value C 46 H 75 O 14 As Na Theoretical value (%): C63.16 H8.58 Na2.63 Actual value (%): C62.99 H8.71 Na2 .51 Molecular weight: 874 Melting point: 252.1-253.3°C Specific rotation: [a] 26 D = +37.8° (C = 0.5, methanol) Ultraviolet absorption spectrum (methanol) E 1 % 1cn (232nm) = 160.0 Infrared absorption spectrum Figure 1 shows the results measured using KB r tablets. Solubility in solvents: Insoluble in water. Soluble in methanol, ethanol, acetone, ethyl acetate, hexane, chloroform, and benzene. Color reaction: Iodine reaction, potassium permanganate reaction, and vanillin-sulfuric acid reaction positive Substance color: Colorless 1 H-NMR spectrum and 13 C-NMR
Spectrum: In heavy chloroform with heavy methanol added, the former at 100MHz, the latter at 25.05MHz
The measurement results are shown in Figures 2 and 3, respectively.
As shown in the figure. Distinction between basic, acidic, and neutral: acidic () Antibacterial action Antibiotic TM-531B has an excellent growth-inhibiting effect on Gram-positive bacteria, but it has a growth-inhibiting effect on Gram-negative bacteria and yeast. do not. As described above, antibiotic TM-531B is a new antibiotic and is useful as a medicine, veterinary drug, and disinfectant. Next, the present invention will be specifically explained with reference to examples showing test examples and manufacturing examples to clarify the antibacterial effect of antibiotic TM-531B. Test Example Heart Infusion Agar was used as the medium and antibiotics were added to Inoculum Size 10 6 CFU.
The MIC (minimum inhibitory concentration) of TM-531B against various bacteria was measured to examine its antibacterial activity. The results are shown in the table below.
【表】
実施例
グルコース２％、グリセリン２％、オートミー
ル２％、落花生油0.5％、肉エキス0.3％、炭酸カ
ルシウム0.5％、食塩0.3％、硫酸第二鉄0.04％、
塩化マンガン0.04％からなる無菌液体培地（PH
7.0）にストレプトミセス・ハイグロスコピカス
TM−531を接種し、30℃で72時間振盪培養した。
この培養液３を種菌として、250容発酵槽内
の前記と同一組成の培地200に植菌し、30℃で
120時間通気撹拌培養した。この際、状態に応じ
て必要があれば消泡剤としてKS−66（信越化学
製、シリコーン系消泡剤）などを添加することが
できる。こうして得た培溶液200を遠心分離し、
菌体と上澄液に分離した。
菌体をアセトン20で２回抽出し、上澄液はア
ンバーライトXAD−８（商品名、ローム・アン
ド・ハス社製）に吸着させ、洗浄後アセトンで溶
出した。
このアセトン抽出液とアセトン溶出液を合せ、
減圧濃縮した。得られた残渣より酢酸エチル２
で２回抽出し、この抽出液を合せ、硫酸ナトリウ
ムで脱水後、減圧溜去し、油状物約123ｇを得た。
この油状物をベンゼン0.5に溶解し、ベンゼ
ンで調製したシリカゲルカラム〔ワコーゲルＣ−
200（商品名、和光純薬製）；２〕に吸着させた。
ベンゼン５で洗浄し、酢酸エチル５で溶出さ
れた区分を除き、アセトン５で溶出した。
このアセトン区分を集め、溶媒を除去し、得ら
れた残渣をヘキサン−アセトン（４：１）の混合
溶媒0.1に溶解し、これと同じ溶媒で調製した
シリカゲルカラム（ワコーゲルＣ−200；0.5）
に吸着させた。続いてこれと同じ溶媒1.5を流
した後、ヘキサン−アセトン（６：４）の混合溶
媒で溶出し、0.5から0.75の間の溶出区分
〔薄層クロマトグラフイー（ヘキサン：アセトン
＝１：１）により確認〕を集め、濃縮乾涸して白
色粉末を得た。この粉末を酢酸エチル50mlに溶解
し、これを同量の希塩酸、水、飽和炭酸ナトリウ
ム水、水で順次各２回ずつ洗浄後、酢酸エチル層
を硫酸ナトリウムで脱水し、濃縮乾涸した。得ら
れた粉末を少量のアセトンに溶解し、セフアデツ
クスLH−20カラム（商品名、フアルマシア社
製）を用い、アセトンでゲル過を行なつた。得
られた活性区分を集め、濃縮乾涸して白色粉末を
得、これをヘキサン−アセトン（４：１）の混合
溶媒から結晶化し、無色のプリズム状結晶として
TM−531Bナトリウム塩173mgを得た。
融点 252.1〜253.3℃[Table] Examples Glucose 2%, Glycerin 2%, Oatmeal 2%, Peanut oil 0.5%, Meat extract 0.3%, Calcium carbonate 0.5%, Salt 0.3%, Ferric sulfate 0.04%,
Sterile liquid medium consisting of 0.04% manganese chloride (PH
7.0) Streptomyces hygroscopicus
TM-531 was inoculated and cultured with shaking at 30°C for 72 hours.
Using this culture solution 3 as a seed, inoculate medium 200 with the same composition as above in a 250-volume fermenter, and heat at 30℃.
Culture was carried out with aeration and stirring for 120 hours. At this time, if necessary depending on the situation, an antifoaming agent such as KS-66 (manufactured by Shin-Etsu Chemical, silicone antifoaming agent) can be added. Centrifuge 200ml of the culture solution obtained in this way,
The cells were separated into bacterial cells and supernatant. The bacterial cells were extracted twice with acetone 20, and the supernatant was adsorbed onto Amberlite XAD-8 (trade name, manufactured by Rohm & Hass), washed, and eluted with acetone. Combine this acetone extract and acetone eluate,
It was concentrated under reduced pressure. From the obtained residue, ethyl acetate 2
The extracts were combined, dehydrated with sodium sulfate, and then distilled under reduced pressure to obtain about 123 g of an oily substance. This oil was dissolved in 0.5% of benzene, and a silica gel column prepared with benzene [Wakogel C-
200 (trade name, manufactured by Wako Pure Chemical Industries, Ltd.); 2].
It was washed with 5 portions of benzene, the fraction eluted with 5 portions of ethyl acetate was removed, and the fraction was eluted with 5 portions of acetone. This acetone fraction was collected, the solvent was removed, the resulting residue was dissolved in 0.1 of a mixed solvent of hexane-acetone (4:1), and a silica gel column (Wako Gel C-200; 0.5) prepared with the same solvent was prepared.
was adsorbed to. Subsequently, after flowing 1.5 of the same solvent, elution was performed with a mixed solvent of hexane-acetone (6:4), and the elution range between 0.5 and 0.75 [thin layer chromatography (hexane:acetone = 1:1)] ] was collected and concentrated to dryness to obtain a white powder. This powder was dissolved in 50 ml of ethyl acetate and washed twice each with equal amounts of diluted hydrochloric acid, water, saturated sodium carbonate and water, and then the ethyl acetate layer was dehydrated over sodium sulfate and concentrated to dryness. The obtained powder was dissolved in a small amount of acetone and gel-filtered with acetone using a Sephadex LH-20 column (trade name, manufactured by Pharmacia). The obtained active fractions were collected and concentrated to dryness to obtain a white powder, which was crystallized from a mixed solvent of hexane-acetone (4:1) as colorless prismatic crystals.
173 mg of TM-531B sodium salt was obtained. Melting point 252.1-253.3℃
第１図はTM−531Bナトリウム塩のKBr錠によ
る赤外線吸収スペクトルを示し、第２図はTM−
531Bナトリウム塩の重メタノール添加重クロロ
ホルム溶液による 1H−NMRスペクトルを示し、
第３図は同 13C−NMRスペクトルを示す。
Figure 1 shows the infrared absorption spectrum of TM-531B sodium salt by KB r tablet, and Figure 2 shows the infrared absorption spectrum of TM-531B sodium salt.
1 H-NMR spectrum of 531B sodium salt in deuterated chloroform solution with added deuterated methanol is shown.
Figure 3 shows the same 13 C-NMR spectrum.
Claims (1)
Priority Applications (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP55128107A JPS5750992A (en) | 1980-09-16 | 1980-09-16 | Antibiotic tm-531b |
| EP81304209A EP0048585A1 (en) | 1980-09-16 | 1981-09-15 | Antibiotics TM-531 B and TM-531 C |
| US06/302,017 US4359583A (en) | 1980-09-16 | 1981-09-15 | Antibiotics TM-531 B and TM-531 C |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP55128107A JPS5750992A (en) | 1980-09-16 | 1980-09-16 | Antibiotic tm-531b |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPS5750992A JPS5750992A (en) | 1982-03-25 |
| JPS6316391B2 true JPS6316391B2 (en) | 1988-04-08 |
Family
ID=14976555
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP55128107A Granted JPS5750992A (en) | 1980-09-16 | 1980-09-16 | Antibiotic tm-531b |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPS5750992A (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH089501Y2 (en) * | 1993-04-28 | 1996-03-21 | 株式会社永代事業所 | Structure of joint part of casing pipe |
-
1980
- 1980-09-16 JP JP55128107A patent/JPS5750992A/en active Granted
Also Published As
| Publication number | Publication date |
|---|---|
| JPS5750992A (en) | 1982-03-25 |
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