JPH02229177A - New antibiotic MK1688 substance and its production method - Google Patents
New antibiotic MK1688 substance and its production methodInfo
- Publication number
- JPH02229177A JPH02229177A JP1049329A JP4932989A JPH02229177A JP H02229177 A JPH02229177 A JP H02229177A JP 1049329 A JP1049329 A JP 1049329A JP 4932989 A JP4932989 A JP 4932989A JP H02229177 A JPH02229177 A JP H02229177A
- Authority
- JP
- Japan
- Prior art keywords
- substance
- antibiotic
- culture
- present
- fusarium
- 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
- 239000000126 substance Substances 0.000 title claims description 46
- 230000003115 biocidal effect Effects 0.000 title claims description 19
- 238000004519 manufacturing process Methods 0.000 title claims description 7
- 241000894006 Bacteria Species 0.000 claims description 12
- UHPMCKVQTMMPCG-UHFFFAOYSA-N 5,8-dihydroxy-2-methoxy-6-methyl-7-(2-oxopropyl)naphthalene-1,4-dione Chemical compound CC1=C(CC(C)=O)C(O)=C2C(=O)C(OC)=CC(=O)C2=C1O UHPMCKVQTMMPCG-UHFFFAOYSA-N 0.000 claims description 11
- 241000223218 Fusarium Species 0.000 claims description 11
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 12
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 9
- 230000001580 bacterial effect Effects 0.000 description 9
- 238000000034 method Methods 0.000 description 7
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 6
- 230000000844 anti-bacterial effect Effects 0.000 description 5
- 238000000655 nuclear magnetic resonance spectrum Methods 0.000 description 5
- AKCRVYNORCOYQT-YFKPBYRVSA-N N-methyl-L-valine Chemical group CN[C@@H](C(C)C)C(O)=O AKCRVYNORCOYQT-YFKPBYRVSA-N 0.000 description 4
- 150000001875 compounds Chemical class 0.000 description 4
- 239000000470 constituent Substances 0.000 description 4
- IOLCXVTUBQKXJR-UHFFFAOYSA-M potassium bromide Chemical compound [K+].[Br-] IOLCXVTUBQKXJR-UHFFFAOYSA-M 0.000 description 4
- 241000233866 Fungi Species 0.000 description 3
- 244000068988 Glycine max Species 0.000 description 3
- 235000010469 Glycine max Nutrition 0.000 description 3
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 3
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 229920002472 Starch Polymers 0.000 description 3
- 238000000862 absorption spectrum Methods 0.000 description 3
- 229940024606 amino acid Drugs 0.000 description 3
- 150000001413 amino acids Chemical class 0.000 description 3
- 239000003242 anti bacterial agent Substances 0.000 description 3
- 238000004440 column chromatography Methods 0.000 description 3
- 239000013058 crude material Substances 0.000 description 3
- 238000010586 diagram Methods 0.000 description 3
- 239000000284 extract Substances 0.000 description 3
- 235000013312 flour Nutrition 0.000 description 3
- 239000007788 liquid Substances 0.000 description 3
- 238000001819 mass spectrum Methods 0.000 description 3
- 239000000741 silica gel Substances 0.000 description 3
- 229910002027 silica gel Inorganic materials 0.000 description 3
- 239000002904 solvent Substances 0.000 description 3
- 241000894007 species Species 0.000 description 3
- 239000008107 starch Substances 0.000 description 3
- 235000019698 starch Nutrition 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- RILPIWOPNGRASR-UHFFFAOYSA-N (2R,3S)-2-Hydroxy-3-methylpentanoic acid Natural products CCC(C)C(O)C(O)=O RILPIWOPNGRASR-UHFFFAOYSA-N 0.000 description 2
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 2
- 241000427940 Fusarium solani Species 0.000 description 2
- RILPIWOPNGRASR-WHFBIAKZSA-N L-Isoleucic acid Chemical compound CC[C@H](C)[C@H](O)C(O)=O RILPIWOPNGRASR-WHFBIAKZSA-N 0.000 description 2
- 241000699670 Mus sp. Species 0.000 description 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 2
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 2
- 241000209140 Triticum Species 0.000 description 2
- 235000021307 Triticum Nutrition 0.000 description 2
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 description 2
- 229940088710 antibiotic agent Drugs 0.000 description 2
- IQMUMKJNLGYVNS-UHFFFAOYSA-N butan-1-ol;methanol;hydrate Chemical compound O.OC.CCCCO IQMUMKJNLGYVNS-UHFFFAOYSA-N 0.000 description 2
- 238000001460 carbon-13 nuclear magnetic resonance spectrum Methods 0.000 description 2
- 238000004587 chromatography analysis Methods 0.000 description 2
- 235000012343 cottonseed oil Nutrition 0.000 description 2
- 238000012136 culture method Methods 0.000 description 2
- 239000012228 culture supernatant Substances 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- MIZMDSVSLSIMSC-OGLSAIDSSA-N enniatin Chemical compound CC(C)C1OC(=O)[C@H](C(C)C)N(C)C(=O)C(C(C)C)OC(=O)[C@H](C(C)C)N(C)C(=O)C(C(C)C)OC(=O)[C@H](C(C)C)N(C)C1=O MIZMDSVSLSIMSC-OGLSAIDSSA-N 0.000 description 2
- 229930191716 enniatin Natural products 0.000 description 2
- 238000000855 fermentation Methods 0.000 description 2
- 230000004151 fermentation Effects 0.000 description 2
- 235000012054 meals Nutrition 0.000 description 2
- 238000002844 melting Methods 0.000 description 2
- 230000008018 melting Effects 0.000 description 2
- 239000012528 membrane Substances 0.000 description 2
- 244000005700 microbiome Species 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 230000000877 morphologic effect Effects 0.000 description 2
- 230000007935 neutral effect Effects 0.000 description 2
- FEMOMIGRRWSMCU-UHFFFAOYSA-N ninhydrin Chemical compound C1=CC=C2C(=O)C(O)(O)C(=O)C2=C1 FEMOMIGRRWSMCU-UHFFFAOYSA-N 0.000 description 2
- 235000015097 nutrients Nutrition 0.000 description 2
- 239000003960 organic solvent Substances 0.000 description 2
- BASFCYQUMIYNBI-UHFFFAOYSA-N platinum Chemical compound [Pt] BASFCYQUMIYNBI-UHFFFAOYSA-N 0.000 description 2
- 239000000843 powder Substances 0.000 description 2
- OVARTBFNCCXQKS-UHFFFAOYSA-N propan-2-one;hydrate Chemical compound O.CC(C)=O OVARTBFNCCXQKS-UHFFFAOYSA-N 0.000 description 2
- 238000000746 purification Methods 0.000 description 2
- VWDWKYIASSYTQR-UHFFFAOYSA-N sodium nitrate Chemical compound [Na+].[O-][N+]([O-])=O VWDWKYIASSYTQR-UHFFFAOYSA-N 0.000 description 2
- 235000012424 soybean oil Nutrition 0.000 description 2
- 239000003549 soybean oil Substances 0.000 description 2
- 239000006188 syrup Substances 0.000 description 2
- 235000020357 syrup Nutrition 0.000 description 2
- 230000009261 transgenic effect Effects 0.000 description 2
- NWUYHJFMYQTDRP-UHFFFAOYSA-N 1,2-bis(ethenyl)benzene;1-ethenyl-2-ethylbenzene;styrene Chemical compound C=CC1=CC=CC=C1.CCC1=CC=CC=C1C=C.C=CC1=CC=CC=C1C=C NWUYHJFMYQTDRP-UHFFFAOYSA-N 0.000 description 1
- WFIYPADYPQQLNN-UHFFFAOYSA-N 2-[2-(4-bromopyrazol-1-yl)ethyl]isoindole-1,3-dione Chemical compound C1=C(Br)C=NN1CCN1C(=O)C2=CC=CC=C2C1=O WFIYPADYPQQLNN-UHFFFAOYSA-N 0.000 description 1
- 235000014469 Bacillus subtilis Nutrition 0.000 description 1
- DKPFZGUDAPQIHT-UHFFFAOYSA-N Butyl acetate Natural products CCCCOC(C)=O DKPFZGUDAPQIHT-UHFFFAOYSA-N 0.000 description 1
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 1
- ZAMOUSCENKQFHK-UHFFFAOYSA-N Chlorine atom Chemical compound [Cl] ZAMOUSCENKQFHK-UHFFFAOYSA-N 0.000 description 1
- 229920002261 Corn starch Polymers 0.000 description 1
- 229920001353 Dextrin Polymers 0.000 description 1
- 239000004375 Dextrin Substances 0.000 description 1
- 241000223221 Fusarium oxysporum 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
- 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
- 241001465754 Metazoa Species 0.000 description 1
- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical compound CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 description 1
- DFPAKSUCGFBDDF-UHFFFAOYSA-N Nicotinamide Chemical compound NC(=O)C1=CC=CN=C1 DFPAKSUCGFBDDF-UHFFFAOYSA-N 0.000 description 1
- 239000001888 Peptone Substances 0.000 description 1
- 108010080698 Peptones Proteins 0.000 description 1
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 1
- 229930006000 Sucrose Natural products 0.000 description 1
- 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 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
- 238000010521 absorption reaction Methods 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 230000002378 acidificating effect Effects 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 231100000215 acute (single dose) toxicity testing Toxicity 0.000 description 1
- 238000011047 acute toxicity test Methods 0.000 description 1
- 239000003463 adsorbent Substances 0.000 description 1
- 239000003905 agrochemical 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
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 1
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 description 1
- 229910052921 ammonium sulfate Inorganic materials 0.000 description 1
- 235000011130 ammonium sulphate Nutrition 0.000 description 1
- 239000010775 animal oil Substances 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
- 239000004202 carbamide Substances 0.000 description 1
- 235000013877 carbamide Nutrition 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 239000000460 chlorine Substances 0.000 description 1
- 229910052801 chlorine Inorganic materials 0.000 description 1
- WORJEOGGNQDSOE-UHFFFAOYSA-N chloroform;methanol Chemical compound OC.ClC(Cl)Cl WORJEOGGNQDSOE-UHFFFAOYSA-N 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
- 230000018842 conidium formation Effects 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 235000005822 corn Nutrition 0.000 description 1
- 239000008120 corn starch Substances 0.000 description 1
- 229940099112 cornstarch Drugs 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 230000034994 death Effects 0.000 description 1
- 231100000517 death Toxicity 0.000 description 1
- 235000019425 dextrin Nutrition 0.000 description 1
- 238000000502 dialysis Methods 0.000 description 1
- 238000009826 distribution Methods 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000002451 electron ionisation mass spectrometry Methods 0.000 description 1
- 238000000921 elemental analysis Methods 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 239000000499 gel Substances 0.000 description 1
- 238000002523 gelfiltration Methods 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 238000007912 intraperitoneal administration Methods 0.000 description 1
- 238000011835 investigation Methods 0.000 description 1
- 239000003456 ion exchange resin Substances 0.000 description 1
- 229920003303 ion-exchange polymer Polymers 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- 239000011777 magnesium Substances 0.000 description 1
- 229910052749 magnesium Inorganic materials 0.000 description 1
- 235000013372 meat Nutrition 0.000 description 1
- GBMDVOWEEQVZKZ-UHFFFAOYSA-N methanol;hydrate Chemical compound O.OC GBMDVOWEEQVZKZ-UHFFFAOYSA-N 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
- 230000000813 microbial effect Effects 0.000 description 1
- 235000013379 molasses Nutrition 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 239000003921 oil Substances 0.000 description 1
- 235000019198 oils Nutrition 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 235000019319 peptone Nutrition 0.000 description 1
- 230000004962 physiological condition Effects 0.000 description 1
- 229910052697 platinum Inorganic materials 0.000 description 1
- 229920001467 poly(styrenesulfonates) Polymers 0.000 description 1
- 239000011591 potassium Substances 0.000 description 1
- 229910052700 potassium Inorganic materials 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 238000012746 preparative thin layer chromatography Methods 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 238000010992 reflux Methods 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 238000011218 seed culture 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
- 235000010344 sodium nitrate Nutrition 0.000 description 1
- 239000004317 sodium nitrate Substances 0.000 description 1
- 238000000638 solvent extraction Methods 0.000 description 1
- 238000001179 sorption measurement Methods 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 235000013547 stew Nutrition 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- 239000013076 target substance Substances 0.000 description 1
- -1 tetrachloroform-methanol Chemical compound 0.000 description 1
- 238000004809 thin layer chromatography Methods 0.000 description 1
- 235000015112 vegetable and seed oil Nutrition 0.000 description 1
- 239000008158 vegetable oil Substances 0.000 description 1
- 239000000273 veterinary drug Substances 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
- 239000012138 yeast extract Substances 0.000 description 1
Landscapes
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Heterocyclic Carbon Compounds Containing A Hetero Ring Having Nitrogen And Oxygen As The Only Ring Hetero Atoms (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Preparation Of Compounds By Using 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】
(産業上の利用分野)
本発明は、エニアチン系の新規抗生物質MK1688物
質ならびにその製造法に関する。DETAILED DESCRIPTION OF THE INVENTION (Industrial Application Field) The present invention relates to a novel enniatin antibiotic MK1688 substance and a method for producing the same.
(従来の技術及び発明が解決しようとする問題点)従来
、微生物が生産する種々の抗生物質が知られており、医
薬品、動物薬、農薬等の分野で実用化されている。しか
しながら耐性菌の出現などの問題から、現在も新規な抗
生物質の出現が常に求められている。本発明は、新規な
抗生物質を提供するとともに、その製造法を確立するこ
とによって、この問題を解決しようとするものである。(Prior Art and Problems to be Solved by the Invention) Various antibiotics produced by microorganisms have been known and have been put to practical use in the fields of pharmaceuticals, veterinary drugs, agricultural chemicals, and the like. However, due to problems such as the emergence of resistant bacteria, there is a constant demand for the emergence of new antibiotics. The present invention aims to solve this problem by providing a new antibiotic and establishing a method for producing the same.
(問題点を解決するための手段)
本発明者らは、上述の期待にこたえるべく、抗菌活性を
有する物質の探索を続けたところ、フザリウム属に属す
るある菌株の培養物中に抗菌活性を有する物質が生産さ
れていることを見い出し、同物質を単離し、その物理化
学的性質を測定した結果、同物質はエニアチン系の化合
物であることがわかった。(Means for Solving the Problems) In order to meet the above expectations, the present inventors continued their search for substances with antibacterial activity, and found that a culture of a certain bacterial strain belonging to the genus Fusarium had antibacterial activity. As a result of discovering that a substance was produced, isolating the substance, and measuring its physicochemical properties, it was determined that the substance was an enniatin-based compound.
ニアチンA (PI,A,Plattnerら、ヘルヘ
チカ キミカ アクタ(Helv.Chim,Acta
)、第31巻、2192頁、1948年発行〕 ;偽エ
ニアチンA(pseudo−Ennfat in A
)(M.M.Shemyakinら、ジャーナル オブ
シュケイ キミイ(zh.Obshch,Khim.)
、第42巻、2320〜2334頁、1972年発行]
;シクロ(D2−ヒドロキ゛シー3−メチルペンタノ
イルーNメチル−D−バリルーD−2−ヒドロキシ−3
メチルペンタノイルーN−メチル−D−バリルD−2−
ヒドロキシ−3−メチルペンタノイルN−メチルーD−
パリル(M. M, S h e m yakin,
トエルブス ブロシイデインダスオブ ザ ヨー口
ピアン ペプチド シンポジウム(12th. Pro
c. Eur, Pept,Symp.)、346 〜
352頁、1972年発行〕などが知られている。Niatin A (PI, A, Plattner et al., Helv. Chim, Acta
), vol. 31, p. 2192, published in 1948] ; pseudo-Ennfat in A
) (MM Shemyakin et al., Journal Obshkei Kimii (zh. Obshch, Khim.)
, Vol. 42, pp. 2320-2334, published in 1972]
;cyclo(D2-hydroxy-3-methylpentanoyl-N-methyl-D-valyl-D-2-hydroxy-3
Methylpentanoyl N-methyl-D-valyl D-2-
Hydroxy-3-methylpentanoyl N-methyl-D-
Paril (M.M, Shemyakin,
Twelve Brosiderians of the Yokohama Peptide Symposium (12th. Pro
c. Eur, Pept, Symp. ), 346 ~
352 pages, published in 1972].
しかし、更に検討した結果、本発明の抗生物質MK16
88物質は構成アミノ酸がN−メチルL−バリンである
ことがわかり、上記の公知化合物とは構成アミノ酸が異
なることから、本発明の抗生物質は新規な化合物である
ことを確定するに至り、本発明を完成した。However, as a result of further investigation, the antibiotic MK16 of the present invention
It was found that the constituent amino acid of the 88 substance is N-methyl L-valine, and since the constituent amino acid is different from the above-mentioned known compounds, it has been determined that the antibiotic of the present invention is a new compound. Completed the invention.
即ち、本発明の要旨は、下記式(I)で表される新規抗
生物質MK1688物質、
及びフザリウム属に属する、抗生物質MK1688物質
生産菌を培養し、その培養物から抗生物質MK1688
物質を採取することを特徴とする上記式(1)で表され
る抗生物質MK1688物質の製造法に存する。That is, the gist of the present invention is to cultivate a novel antibiotic MK1688 substance represented by the following formula (I) and a bacteria producing the antibiotic MK1688 substance belonging to the genus Fusarium, and to obtain the antibiotic MK1688 from the culture.
The present invention relates to a method for producing the antibiotic MK1688 substance represented by the above formula (1), which comprises collecting the substance.
以下本発明を説明するに、本発明の新規抗生物質MK1
68B物質は、N−メチルーL−ハリンを構成アミノ酸
とする上記式( I’ )で表される新規化合物である
。To explain the present invention, the novel antibiotic MK1 of the present invention will be described below.
Substance 68B is a novel compound represented by the above formula (I') whose constituent amino acid is N-methyl-L-halin.
本発明の抗生物質MK168B物質は、フザリウム属に
属する該MK1688物質生産菌により生産される。The antibiotic MK168B substance of the present invention is produced by the MK1688 substance-producing bacterium belonging to the genus Fusarium.
かかる生産菌としては、例えば工業技術院微生物工業技
術研究所に、微工研菌寄第10564号(FERM
P−10564)として寄託されているフザリウム・オ
キシスポルム(Fusarium ox s o
rum)D338が挙げられる。その菌学的性質は下記
の通りである。As such producing bacteria, for example, the Institute of Microbial Technology, Agency of Industrial Science and Technology,
Fusarium oxysporum (P-10564) has been deposited as
rum) D338. Its mycological properties are as follows.
(1)形態学的特徴
コロニーは生育旺盛で、PDA培地上においてフェルト
状〜線毛状となり、その表面は白色、裏面は黄味白色〜
明るい黄橙色を呈する。(1) Morphological characteristics Colonies grow vigorously and become felt-like to fimbriae-like on PDA medium, with white on the surface and yellowish-white on the underside.
It has a bright yellow-orange color.
基底菌糸は隔壁を有し無色で、放射状に伸長、分枝し幅
8.2μmに至る。The basal hyphae have septa, are colorless, elongate and branch radially, and reach a width of 8.2 μm.
気生菌糸は豊富に形成される。Aerial hyphae are abundantly formed.
フイアライドは気生菌糸上に側生または分生子柄上に形
成され、長さは3.2〜43μmと短かく、先端に向か
って先が細くなっており、幅は基部で1.8〜2.7μ
mである。Phialides are formed on aerial hyphae or on conidiophores, and are as short as 3.2 to 43 μm in length, tapering toward the tip, and 1.8 to 2 mm wide at the base. .7μ
It is m.
分生子は大分生子と小分生子の2つの型を有し、分生子
形成様式はフイアロ型である。There are two types of conidia: macroconidia and microconidia, and the conidia formation mode is the phiaro type.
大分生子は多分技の分生子柄上に形成され、その形態は
、膜が薄く平滑な新月型で両端が尖っており、無色であ
る。Macroconidia are formed on polyconidiophores and are colorless, with a thin, smooth, crescent-shaped membrane, pointed at both ends.
また該大分生子は通常3〜5隔壁(まれに6〜7隔壁)
でその基部にはくちばし状のフソトセルを有する。The macroconidia usually have 3 to 5 septa (rarely 6 to 7 septa)
It has a beak-shaped fusotocell at its base.
上記大分生子の大きさは、3隔壁分生子で35〜60μ
mX4〜5.6μm、5隔壁分生子で51.6〜57μ
m X 4〜5.6μm、6〜7隔壁分生子で54〜7
3μm×4〜5.6μmである。The size of the above macroconidia is 35 to 60μ for 3-septate conidia.
mX4-5.6μm, 51.6-57μ with 5 septate conidia
m x 4-5.6 μm, 54-7 with 6-7 septate conidia
The size is 3 μm×4 to 5.6 μm.
小分生子は菌糸から側生じた分生子柄上に生し、その形
態は膜が薄く無色で平滑な円筒形〜楕円形である。Microconidia grow on conidiophores that arise laterally from hyphae, and are colorless, smooth, and cylindrical to oval in shape with a thin membrane.
また該小分生子は1〜2個の細胞から成り、その大きさ
は7.6〜27μm×2〜4μmである。Moreover, the microconidia consist of 1 to 2 cells and have a size of 7.6 to 27 μm×2 to 4 μm.
尚、完全世代は観察されなかった。Note that no complete generation was observed.
(2)生理的状質
■最適生育条件
■生育の範囲
pu : pH 2〜9 (LCA液体培地中で2週
間培養)
温度=10゜C〜30゜c (PDA培地上で2週間培
養)
?3)分類学的考察
■属レヘルの同定
本菌株(D338)は1)フイアロ型分生子形成様式を
持つ、2)大分生子は新月型で隔壁を有し、基部には明
瞭なフントセル(F(,■t Cell)を有する。(2) Physiological conditions ■ Optimum growth conditions ■ Growth range pu: pH 2 to 9 (cultured for 2 weeks in LCA liquid medium) Temperature = 10°C to 30°C (cultured for 2 weeks on PDA medium) ? 3) Taxonomic considerations ■Identification of the genus Leher This strain (D338) 1) has a phiaro-type conidiation mode, 2) the macroconidia are crescent-shaped and have septa, and the base has clear huntocells (F (,■t Cell).
上記菌学的性状を有する不完全糸状菌綱D{yphom
ycetes)の属について、H,L,Barnett
.とB.B.Hunterの“イラストレイテッド シ
エネラ オブ インパーフェクト フンギ(Illus
trated genera of Imper
fect Fungi)”記載の検索表に従って検索
した結果、本菌株(D 3 3.8 )はフザリウム属
に帰属することが判明した。Deuterofilamentous fungi class D{yphom having the above mycological properties
ycetes) by H. L. Barnett.
.. and B. B. Hunter's “Illustrated Sienera of Imperfect Funghi”
rated genera of Imper
As a result of searching according to the search table described in "Fect Fungi", it was found that this strain (D 3 3.8) belongs to the genus Fusarium.
■種レベルの同定
本菌株(D338)は1)大分生子と小分生子を持つ、
2)小分生子は単純なフイアライト型の分生子柄上に生
じ、その分生子柄は短い、3)小分生子は連鎖状には形
成されない、4)小分生子は円筒形〜長楕円形である。■ Species level identification This strain (D338) has 1) macroconidia and microconidia;
2) Microconidia occur on simple phialite-type conidiophores, and the conidiophores are short. 3) Microconidia are not formed in chains. 4) Microconidia are cylindrical to oblong. It is.
Booth (1971)の“ザ ジェヌスフザリウム
(The genus Fusarium)、1〜
237頁゛′の記載によれば、フザリウム属菌には47
種が知られている。“The genus Fusarium” by Booth (1971), 1-
According to the description on page 237', there are 47 Fusarium bacteria.
species are known.
本菌株(D338)の形態学的性状と上記文献記載の4
7菌種のものを対比したところ本菌株の性状は上記文献
に記載してあるフザリウム・オキシスボルムの性状とほ
とんど合致した。その中で近縁種のフザリウム・ソラニ
イ (F,solani)とは小分生子柄の長さの相違
によって識別された。従って本菌株(D338)はフザ
リウム・オキシスポルム(F.oxys orum)
と同定された。Morphological properties of this strain (D338) and 4 described in the above literature
When seven bacterial species were compared, the properties of this bacterial strain almost matched those of Fusarium oxysborum described in the above-mentioned literature. Among them, it was distinguished from the closely related species Fusarium solani (F. solani) by the difference in the length of the microconidiophore. Therefore, this strain (D338) is Fusarium oxysorum (F.oxys orum).
was identified.
一般に、フザリウム属(Fusarium属)菌は他の
菌類の場合にみられるようにその性状が変化しやすいが
、本発明においては、例えば、D338株の、またはこ
の株に由来する突然変異株(自然発生または誘発性)で
あっても、或いはその形質接合体または遺伝子組換え体
であっても、新規構成物質MK1688物質の生産能を
有するものはすべて本発明の方法に使用することができ
る。In general, the properties of Fusarium genus bacteria tend to change as seen in the case of other fungi, but in the present invention, for example, the D338 strain or a mutant strain derived from this strain (natural Any type of MK1688 that has the ability to produce the novel constituent MK1688 substance can be used in the method of the present invention, whether it is a transgenic or transgenic form (genetically or inducibly produced), or a transzygote or genetically recombinant thereof.
本発明においては、前記の菌を通常の微生物が利用しう
る栄養物を含有する培地で培養する。栄養源としてはグ
ルコース、水アメ、デキストリン、シュクロース、デン
プン、糖蜜、動・植物油等を使用できる。また窒素源と
して大豆粉、小麦胚芽、コーンスティーブ・リカー、綿
実粕、肉エキス、ペプトン、酵母エキス、硫酸アンモニ
ウム、硝酸ソーダ、尿素等を使用できる。その他必要に
応じて、ナトリウム、カリウム、カルシウム、マグネシ
ウム、コバルト、塩素、リン酸、硫酸及びその他のイオ
ンを生成することのできる無機塩類を添加することは有
効である。また菌の生育を助け抗生物質MK1688物
質の生産を促進するような有機及び無機物を適当に添加
することができる。In the present invention, the above bacteria are cultured in a medium containing nutrients that can be used by ordinary microorganisms. As a nutrient source, glucose, starch syrup, dextrin, sucrose, starch, molasses, animal/vegetable oil, etc. can be used. Also, soybean flour, wheat germ, corn stew liquor, cottonseed meal, meat extract, peptone, yeast extract, ammonium sulfate, sodium nitrate, urea, etc. can be used as the nitrogen source. In addition, it is effective to add inorganic salts capable of producing sodium, potassium, calcium, magnesium, cobalt, chlorine, phosphoric acid, sulfuric acid, and other ions as necessary. In addition, organic and inorganic substances that help the growth of bacteria and promote the production of the antibiotic MK1688 substance can be appropriately added.
培養法としては、好気的条件下での培養法、特q
に深部培養法が最も通している。培養に適当な温度は1
0〜30℃であるが多くの場合、20〜27℃付近で培
養する。抗生物質MK 1 6 8 8物質の生産は培
地や培養条件により異なるが、振とう培養、タンク培養
とも通常3〜10日の間でその蓄積が最高に達する。培
養物中のMK1688物質の蓄積量が最高になった時に
培養を停止し、培養液から目的物質を単離精製する。The most popular culture methods are those under aerobic conditions, especially deep culture methods. The appropriate temperature for culturing is 1.
The temperature is 0 to 30°C, but in most cases it is cultured at around 20 to 27°C. Production of the antibiotic MK1688 substance varies depending on the medium and culture conditions, but the accumulation usually reaches its maximum within 3 to 10 days in both shaking culture and tank culture. When the amount of MK1688 substance accumulated in the culture reaches the maximum, the culture is stopped, and the target substance is isolated and purified from the culture solution.
本発明において、MK1688物質の培養物からの採取
に当たっては、その性状を利用した通常の分離手段、例
えば、溶剤抽出法、イオン交換樹脂法、吸着または分配
力ラムクロマト法、ゲルろ過法、透析法、・沈澱法等を
単独でまたは適宜組み合わせて抽出精製することができ
る。例えば、MK168B物質は、培養菌体中からはア
セトン水またはメタノールー水で抽出される。また、培
養液中に蓄積されたMK1688物質は水と混ざらない
有機溶剤、例えば、ブタノール、酢酸エチル等で抽出す
ればMK1688物質は有機溶剤層に抽出される。MK
1688物質をさらに精製するには、シリカゲル(ワコ
ーゲルC−200、和光純薬工業社製等)、アルミナ等
の吸着剤やセファデックスLH−20 (ファルマシ
ア社製)等を用いるクロマトグラフィーを行うとよい。In the present invention, when collecting the MK1688 substance from a culture, conventional separation methods utilizing its properties are used, such as solvent extraction method, ion exchange resin method, adsorption or distribution force chromatography method, gel filtration method, and dialysis method. Extraction and purification can be carried out using , precipitation methods, etc. alone or in appropriate combinations. For example, the MK168B substance is extracted from cultured bacterial cells with acetone water or methanol-water. Furthermore, if the MK1688 substance accumulated in the culture solution is extracted with an organic solvent that does not mix with water, such as butanol or ethyl acetate, the MK1688 substance will be extracted into the organic solvent layer. M.K.
To further purify the 1688 substance, it is recommended to perform chromatography using an adsorbent such as silica gel (Wako Gel C-200, manufactured by Wako Pure Chemical Industries, Ltd.), alumina, or Sephadex LH-20 (manufactured by Pharmacia). .
かくして得られる前記式(r)で表される本発明の抗生
物質MK168B物質は、下記表1に示すように、ダラ
ム陽性菌に対して抗菌活性を有する。The antibiotic MK168B substance of the present invention represented by the formula (r) thus obtained has antibacterial activity against Durham-positive bacteria, as shown in Table 1 below.
15%、サングレイン0.25%、綿実粕0.5%、F
eSOa H 7HzO O. O O O 5%
、NiC1z ・68zO O.00005%、C
OCl2 ・6H20 0. 0 0 0 0 5%
およびCaCOz 0. 1%を含有する主醗酵培地(
p}l 6. 0 )を調製し、その8 0 m j+
を5 0 0mA三角フラスコ100本に分注し、12
1℃において20分間高圧滅菌する。この主醗酵培地に
前記種培養液を4 m Rずつ接種し、26゜Cにおい
て6日間、210回転にて振とう培養する。得られた培
養を遠心分離して、培養上清液4℃と培養菌体を得た。15%, sungrain 0.25%, cottonseed meal 0.5%, F
eSOa H 7HzO O. O O O 5%
, NiC1z ・68zO O. 00005%, C
OCl2 ・6H20 0. 0 0 0 0 5%
and CaCOz 0. The main fermentation medium containing 1% (
p}l 6. 0) and its 80 m j+
was dispensed into 100 500 mA Erlenmeyer flasks, and 12
Autoclave for 20 minutes at 1°C. This main fermentation medium is inoculated with 4 mR of the above-mentioned seed culture solution, and cultured with shaking at 210 rpm for 6 days at 26°C. The resulting culture was centrifuged to obtain a culture supernatant at 4°C and cultured bacterial cells.
■ 培養物の精製
上記■で得られた菌体約500g (湿重量)に、50
%アセトンー水(4β)を加え、30分間撹拌した後、
菌体を濾別し、減圧下アセトンを留去して菌体抽出液(
2.0β)を得た。この菌体抽出液と上記■で得た培養
上清(4E)を混合し、この混合液より酢酸エチル(6
l)で有効成分の抽出操作を行い、溶媒層を濃縮乾固し
た。得られた油状物質を、予めクロロホルムーメタノー
ル(20:1)で充填したシリカゲル力ラム(ワコーゲ
また、本発明の抗生物質MK1688物質は、jcl:
ICLマウス(体重25gの雄を3匹使用)への腹腔内
投与による急性毒性試験を行ったが、100mg/kg
の投与量で死亡例はなかった。■ Purification of culture Approximately 500 g (wet weight) of bacterial cells obtained in
After adding % acetone-water (4β) and stirring for 30 minutes,
The bacterial cells were filtered and the acetone was distilled off under reduced pressure to obtain the bacterial cell extract (
2.0β) was obtained. This bacterial cell extract was mixed with the culture supernatant (4E) obtained in step ① above, and ethyl acetate (6
The active ingredient was extracted in step 1), and the solvent layer was concentrated to dryness. The obtained oily substance was added to a silica gel column (wakoge) filled with chloroform-methanol (20:1) in advance.
An acute toxicity test was conducted by intraperitoneal administration to ICL mice (3 male mice weighing 25 g), but at 100 mg/kg.
There were no deaths at this dose.
以下に実施例を挙げて本発明を更に具体的に説明するが
、本発明の要旨を超えない限り、本発明は、以下の実施
例に限定されるものではない。The present invention will be described in more detail with reference to Examples below, but the present invention is not limited to the following Examples unless it exceeds the gist of the present invention.
実施例1
■ 培養
コーンスターチ3.0%、大豆粉3.5%、大豆油0.
2%、小麦胚芽1,5%、KJPOt0. 2%、Ca
CO30.5%を含有する種培地(pH 6. 0 )
を4. O m Qずつ2 0 0mAの三角フラスコ
10本に分注して、121℃、20分間高圧滅菌した。Example 1 ■ Cultured cornstarch 3.0%, soybean flour 3.5%, soybean oil 0.
2%, wheat germ 1.5%, KJPOt0. 2%, Ca
Seed medium containing 0.5% CO3 (pH 6.0)
4. The mixture was dispensed into 10 Erlenmeyer flasks at 200 mA each, and sterilized under high pressure at 121°C for 20 minutes.
次いで、フザリウム・オキシスボルムD338(微工研
菌寄第10564号〉を1白金耳ずつ植菌し、26℃で
3日間、210回転にて振とう培養した。Next, one platinum loopful of Fusarium oxysvorum D338 (Feikoken Bibori No. 10564) was inoculated and cultured at 26° C. for 3 days with shaking at 210 rpm.
別に、水アメ2.0%、大豆粉1.0%、大豆油0.ル
C−200、1 0 0 g)の上端に付し、ク四ロホ
ルムーメタノール(20:1)で展開して溶出液を15
gずつ分画した。活性画分(lkl25143)を濃縮
乾固すると7 9 6mgの油状物質つ
が得られた。この油状物質をセフプデソクスL Hjノ
20 (870mn)カラム(Φ3 5 mm x l
m,9Qcmの高さまでセファデソクスLH−20を
詰めた)の上端に付し、メタノールで展開して溶出液を
10gずつ分画した。活性画分(IIkl212B)を
濃縮乾固すると3 8 7mgのMK1688物質が得
られた。Separately, 2.0% starch syrup, 1.0% soybean flour, 0.0% soybean oil. C-200, 100 g) and developed with tetrachloroform-methanol (20:1), and the eluate was diluted with 15 g.
It was fractionated by g. The active fraction (lkl25143) was concentrated to dryness to yield 796 mg of oil. This oily substance was transferred to a Cefpadesox L Hj No 20 (870 mm) column (Φ3 5 mm x l
The eluate was applied to the upper end of a tube (packed with Sephadesox LH-20 to a height of 9 Q cm), developed with methanol, and the eluate was fractionated into 10 g portions. The active fraction (IIkl212B) was concentrated to dryness to obtain 387 mg of MK1688 substance.
尚、各工程での抗菌活性の測定は、被検菌としてハシル
ス サブテイリス(B.subt i l is)AT
CC 6633を使用して行った。The antibacterial activity in each step was measured using B. subtilis AT as the test bacterium.
Performed using CC 6633.
かくして得られたMK1688物質の理化学的性質は次
の通りであった。The physicochemical properties of the MK1688 substance thus obtained were as follows.
(1)色および形状:無色粉末
12)分子式: C 3b H b 3 N 30,(
3)元素分析値:C 63.02 H 9.5
8N6.08
(4)マススペクトル(El−MS):m/z681(
M1 )
(5》融点:吸湿性のため明確な融点を示さない。(1) Color and shape: colorless powder 12) Molecular formula: C 3b H b 3 N 30, (
3) Elemental analysis value: C 63.02 H 9.5
8N6.08 (4) Mass spectrum (El-MS): m/z681 (
M1) (5) Melting point: Does not show a clear melting point due to hygroscopicity.
(7)紫外部吸収スペクトル:末端吸収(8)赤外部吸
収スペクトル
(KBr cm−’):3430,2950,293
0 2860.1735.166’0,1460,1
415,1375,[295,1190.1125,1
100.1040,1010.87’O,.770
(9)’HNMRスペクトル(400MHz,CD3
0D)
δ(ppm):0.92 (3H,d,6.5Hz),
0.95 (3H.t,7.4Hz),0.99 <3
H,d,6.7Hz),1.0 7 (3H, d,
6.7Hz) .1.2 3 (I H, m)
1.4 7 (I H, m) ,
1.9 6 (L H, m) ,2.28
(IH. d septet , 1 0.
0 , 6. 7 H z )3.15 (3H,
s),
4.76 (IH, d, 10.OH2),5
.38 (LH, d, 6.5Hz) 。(7) Ultraviolet absorption spectrum: Terminal absorption (8) Infrared absorption spectrum (KBr cm-'): 3430, 2950, 293
0 2860.1735.166'0,1460,1
415,1375,[295,1190.1125,1
100.1040, 1010.87'O, . 770 (9)'HNMR spectrum (400MHz, CD3
0D) δ (ppm): 0.92 (3H, d, 6.5Hz),
0.95 (3H.t, 7.4Hz), 0.99 <3
H, d, 6.7Hz), 1.0 7 (3H, d,
6.7Hz). 1.2 3 (I H, m) 1.4 7 (I H, m) , 1.9 6 (L H, m) , 2.28
(IH. d septet, 10.
0, 6. 7 Hz)3.15 (3H,
s), 4.76 (IH, d, 10.OH2), 5
.. 38 (LH, d, 6.5Hz).
Q(II ’ 3C N M Rスペクトル(100
MHz,C D3 0 D)
δ (ppm) :12.5 Q. 15.9
q, 20.9 q. 21.3
q, 27.2 t, 30.1d, 33
.7 q. 3B.5 d, 64.5
d76.O d. 172.41 s,
172.44SO
Qll溶解性:メタノール、クロロホルム、酢酸エチル
、アセトンに溶け、水に溶けない。Q(II' 3C NMR spectrum (100
MHz, C D3 0 D) δ (ppm): 12.5 Q. 15.9
q, 20.9 q. 21.3
q, 27.2 t, 30.1d, 33
.. 7q. 3B. 5 d, 64.5
d76. O d. 172.41 s,
172.44SO Qll Solubility: Soluble in methanol, chloroform, ethyl acetate, acetone, insoluble in water.
αク 塩基性、酸性、中性の区別:中性物質上記理化学
的性質及び下記参考例lの結果から、得られたMK−1
688物質は前記式(I)で表わされる構造であること
が確認された。αku Distinction between basic, acidic, and neutral: Neutral substance MK-1 obtained from the above physical and chemical properties and the results of Reference Example 1 below
It was confirmed that substance 688 had a structure represented by the above formula (I).
l8
参考例I
MK1688物質(5 0mg)を6N HCI(3
ml)に溶解し、11’0℃で16時間加熱還流した。l8 Reference Example I MK1688 substance (50 mg) was dissolved in 6N HCI (3
ml) and heated under reflux at 11'0°C for 16 hours.
反応液は冷却後、濃縮乾固した。得られた油状物質は5
m itの水に溶解し、IN NaOHでpH4.
0に調製した後ダウエソクス50Wx2(H”″)(ダ
ウケミカル社製、5 m l )カラムに通過させた。After cooling, the reaction solution was concentrated to dryness. The oily substance obtained is 5
Dissolve in mit water and adjust to pH 4 with IN NaOH.
After adjusting to 0, it was passed through a Dow Sox 50Wx2 (H"") (manufactured by Dow Chemical Company, 5 ml) column.
通過液と水洗液(1 0ml)を混合し、濃縮乾固して
粗物質を得た。この粗物質は、水を展開溶媒とするセフ
ァデックスG−10(ファルマシア社製、4 4 0m
m!)のカラムクロマトグラフィーを行い、溶出液を5
gずつ分画した。The passing liquid and the washing liquid (10 ml) were mixed and concentrated to dryness to obtain a crude material. This crude material was prepared using Sephadex G-10 (manufactured by Pharmacia, 440 m
m! ) column chromatography, and the eluate was
It was fractionated by g.
1−ブタノールーメタノールー水(4 : 1 : 2
)を展開溶媒とするシリカゲル薄層クロマトグラフィー
上でRf値0.51に検出される物質を含む両分(Nl
30−35)を濃縮乾固すると2−ヒドロキシ−3−メ
チルペンタン酸(20.2mg)が得られた。1-butanol-methanol-water (4:1:2
) as a developing solvent, containing a substance detected at an Rf value of 0.51 on silica gel thin layer chromatography (Nl
30-35) was concentrated to dryness to obtain 2-hydroxy-3-methylpentanoic acid (20.2 mg).
ダウエックス50Wx2(H′″)カラムに吸着した成
分は0.5N HCIで溶出し、濃縮乾固して粗物質
を得た。この粗物質は、水を展開溶媒とするセファデソ
クスG − 1 0 (4 4 0ml!)のカラムク
ロマトグラフィーを行った後、分取用TLC(メルク社
製、Art.5744、展開溶媒:1−ブタノールーメ
タノールー水、4 : 1 : 2) )で精製してニ
ンヒドリン陽性の粗粉末を得た。この粗粉末は再び水を
展開溶媒とするセファデソクG 1 0 (’44
0mm2) (71カラムクロマトクラフィーを行い、
ニンヒドリン陽性画分を濃縮乾固してN−メチルーL−
バリン(11.9mg)を得た。The components adsorbed on the DOWEX 50Wx2 (H''') column were eluted with 0.5N HCI and concentrated to dryness to obtain a crude material. After column chromatography of 440 ml!), it was purified by preparative TLC (manufactured by Merck & Co., Art. 5744, developing solvent: 1-butanol-methanol-water, 4:1:2). A crude powder that was positive for ninhydrin was obtained.
0mm2) (71 column chromatography was performed,
The ninhydrin-positive fraction was concentrated to dryness to give N-methyl-L-
Valine (11.9 mg) was obtained.
これらの結果から、MK1688物質は、2一ヒドロキ
シ−3−メチルベンタン酸とN−メチルL−バリンを構
成成分とすることが分った。From these results, it was found that the MK1688 substance consists of 2-hydroxy-3-methylbentanoic acid and N-methyl L-valine.
〈2−ヒドロキシ−3−メチルペンタン酸〉マススペク
トル(El−MS):mlz 132(M4)
’H NMRスペクトル( 4 0 0 M H z
, D zO)δ (1)I)m) : 0.’8
6 (3H, d, ?.OH2’) ,0.
92 (3H,t,7;4Hz),1.3 1 (I
H.ddq.14.0.7. 0 H z )
(LH, ddq, 14.0.
7.0Hz),
(IH, dddq, 7.0,
7.0, 3.3Hz),
(IH, d, 3.3HZ) 。<2-Hydroxy-3-methylpentanoic acid> Mass spectrum (El-MS): mlz 132 (M4) 'H NMR spectrum (400 MHz
, D zO)δ (1)I)m): 0. '8
6 (3H, d, ?.OH2') ,0.
92 (3H, t, 7; 4Hz), 1.3 1 (I
H. ddq. 14.0.7. 0 Hz) (LH, ddq, 14.0. 7.0Hz), (IH, ddq, 7.0, 7.0, 3.3Hz), (IH, d, 3.3Hz).
(1 0 0MHz,Dz O)
q. 13.6 q.26.
d, 73.5 d, 178.7.4,
1.44
7.4,
1.86
7,0,
4.30
13C NMRスペクトル
δ(p pm) : 1 1.7
1t,38.6
9 S6
〈N−メチルーし−バリン〉
マススペクトル(E I −MS)
(M+)
比旋光度: 〔α).=+26.9
6N HCI)
:m/z 131
6(co.7
NMRスペクトル
(p pm) : 1.0 1
1.04
2.23
t,
(4 0 0 MH z , Dz O)(3H,
d, 7.0Hz),
(3H, d. 7.0Hz)
(IH. d septe
4. 5 , 7. 0 H z )2.7 1
(3H, s) ,3.47 (LH, d
, 4.5Hz) 。(100MHz, DzO) q. 13.6 q. 26. d, 73.5 d, 178.7.4, 1.44 7.4, 1.86 7,0, 4.30 13C NMR spectrum δ (p pm): 1 1.7 1t, 38.6 9 S6 <N-methyl-valine> Mass spectrum (EI-MS) (M+) Specific optical rotation: [α). = +26.9 6N HCI) : m/z 131 6 (co.7 NMR spectrum (p pm): 1.0 1 1.04 2.23 t, (4 0 0 MHz, DzO) (3H,
d, 7.0Hz), (3H, d. 7.0Hz) (IH.d septe 4.5, 7.0 Hz)2.7 1
(3H, s) ,3.47 (LH, d
, 4.5Hz).
I3C NMRスペクトル(1 0 0MHz,D2
0)δ (ppm) :18.O q, 1
8.5 q. 29.8 d, 33.3
q, 70.O d. 173.(発明
の効果)
本発明の新規抗生物質MK168B物質は、ダラム陽性
菌に対して抗菌活性を有するので、抗菌剤としての有用
性が期待できる。I3C NMR spectrum (100MHz, D2
0) δ (ppm): 18. Oq, 1
8.5 q. 29.8 d, 33.3
q, 70. O d. 173. (Effects of the Invention) The novel antibiotic substance MK168B of the present invention has antibacterial activity against Durham-positive bacteria, so it can be expected to be useful as an antibacterial agent.
第1図は、MK1688物質の臭化カリウム錠での赤外
部吸収スペクトルを示す図である。
第2図は、MK 1 6 8 8物質の重メタノール溶
液中での400MHz ’H NMRスペクトルを
示す図である。
第3図は、MK168B物質の重メタノール溶液中での
1 0 0MHz 13C NMRスペクトルを示
す図である。FIG. 1 is a diagram showing the infrared absorption spectrum of MK1688 substance in potassium bromide tablets. FIG. 2 is a diagram showing a 400 MHz 'H NMR spectrum of MK 1 6 8 8 substance in a heavy methanol solution. FIG. 3 is a diagram showing a 100 MHz 13C NMR spectrum of MK168B substance in a heavy methanol solution.
Claims (2)
88物質 ▲数式、化学式、表等があります▼ ( I )(1) Novel antibiotic MK16 represented by the following formula (I)
88 substances ▲ Contains mathematical formulas, chemical formulas, tables, etc. ▼ (I)
質生産菌を培養し、その培養物から抗生物質MK168
8物質を採取することを特徴とする請求項1記載の抗生
物質MK1688物質の製造法。(2) Cultivate bacteria that produce the antibiotic MK1688 belonging to the genus Fusarium, and use the culture to produce the antibiotic MK168.
The method for producing the antibiotic MK1688 substance according to claim 1, characterized in that eight substances are collected.
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP1049329A JP2815166B2 (en) | 1989-03-01 | 1989-03-01 | New antibiotic MK1688 and its production |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP1049329A JP2815166B2 (en) | 1989-03-01 | 1989-03-01 | New antibiotic MK1688 and its production |
Publications (2)
Publication Number | Publication Date |
---|---|
JPH02229177A true JPH02229177A (en) | 1990-09-11 |
JP2815166B2 JP2815166B2 (en) | 1998-10-27 |
Family
ID=12827948
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP1049329A Expired - Lifetime JP2815166B2 (en) | 1989-03-01 | 1989-03-01 | New antibiotic MK1688 and its production |
Country Status (1)
Country | Link |
---|---|
JP (1) | JP2815166B2 (en) |
-
1989
- 1989-03-01 JP JP1049329A patent/JP2815166B2/en not_active Expired - Lifetime
Also Published As
Publication number | Publication date |
---|---|
JP2815166B2 (en) | 1998-10-27 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
JP3111470B2 (en) | Novel polypeptide compound and method for producing the same | |
JPH0285296A (en) | Fr901228 substance and preparation thereof | |
JPH0320277A (en) | Antitumor substance BE-13793C | |
JPH02229177A (en) | New antibiotic MK1688 substance and its production method | |
JPS625990A (en) | Antibiotic substance and production thereof | |
JP3048513B2 (en) | 2,2'-bipyridine derivative, method for producing the same, and antitumor agent containing the derivative | |
JPH10120686A (en) | New polyene antibiotic substance 3874h1 to h6, their production and use thereof | |
US4902781A (en) | Novel tripetide derivatives | |
JP2868237B2 (en) | Novel bioactive substance OM-6519 and production method thereof | |
JP2856379B2 (en) | Protein phosphatase inhibitor and antitumor agent | |
JPH041179A (en) | Antitumor substance be-14106 | |
JPH02167092A (en) | Anti-tumor agent containing lisolipin x and production of lisolipin x | |
JPS62174099A (en) | Novel glycopeptide antibiotic substance pa-42867-a and pa-42867-b and production thereof | |
JPH1045789A (en) | New physiologically active substance na16887, its production and use thereof | |
JPH05194571A (en) | New antibiotic nk374186a, nk374186b, nk374186b3 and nk374186c3, their production and their use | |
JPH07277971A (en) | Antitumor agent, selective cytotoxic agent for human tumor cell, microbial strain producing heptelidic acid chlorohydrin and its production | |
JPS6348284A (en) | Novel antibiotic yp-02908l-a and production thereof | |
JPH04243894A (en) | New q-6402 compound and production thereof | |
JPH02221292A (en) | New substance 02-3, its use and production thereof | |
JPH0623192B2 (en) | Antibiotic TAN-868A and method for producing the same | |
JPH06135979A (en) | New substance nk374200, its production and use thereof | |
JPH0680607A (en) | Malbranicin | |
JPH01290673A (en) | Novel substance k3543r1, its use and production | |
JPH0892119A (en) | Antitumor agent | |
JPH0987185A (en) | New physiologically active substance nf03110, its production and use thereof |