JPH03251597A - Versaferrocide-based compound - Google Patents
Versaferrocide-based compoundInfo
- Publication number
- JPH03251597A JPH03251597A JP2046886A JP4688690A JPH03251597A JP H03251597 A JPH03251597 A JP H03251597A JP 2046886 A JP2046886 A JP 2046886A JP 4688690 A JP4688690 A JP 4688690A JP H03251597 A JPH03251597 A JP H03251597A
- Authority
- JP
- Japan
- Prior art keywords
- versaferroside
- substance
- methanol
- formula
- 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.)
- Pending
Links
- 150000001875 compounds Chemical class 0.000 title claims abstract description 13
- 239000000126 substance Substances 0.000 claims abstract description 28
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 claims abstract description 4
- 125000004435 hydrogen atom Chemical group [H]* 0.000 claims description 3
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 abstract description 21
- 241000244206 Nematoda Species 0.000 abstract description 13
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 abstract description 12
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 abstract description 6
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 abstract description 4
- 238000012136 culture method Methods 0.000 abstract description 4
- 241000233866 Fungi Species 0.000 abstract description 3
- 230000000704 physical effect Effects 0.000 abstract description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 abstract description 2
- 239000003795 chemical substances by application Substances 0.000 abstract 1
- 239000000463 material Substances 0.000 abstract 1
- 230000000007 visual effect Effects 0.000 abstract 1
- 239000002023 wood Substances 0.000 abstract 1
- 239000013543 active substance Substances 0.000 description 7
- 239000000203 mixture Substances 0.000 description 7
- 235000008331 Pinus X rigitaeda Nutrition 0.000 description 6
- 235000011613 Pinus brutia Nutrition 0.000 description 6
- 241000018646 Pinus brutia Species 0.000 description 6
- 239000002609 medium Substances 0.000 description 5
- 238000000655 nuclear magnetic resonance spectrum Methods 0.000 description 5
- HEDRZPFGACZZDS-MICDWDOJSA-N Trichloro(2H)methane Chemical compound [2H]C(Cl)(Cl)Cl HEDRZPFGACZZDS-MICDWDOJSA-N 0.000 description 4
- 230000000694 effects Effects 0.000 description 4
- 244000005700 microbiome Species 0.000 description 4
- 241000894006 Bacteria Species 0.000 description 3
- UHOVQNZJYSORNB-UHFFFAOYSA-N Benzene Chemical compound C1=CC=CC=C1 UHOVQNZJYSORNB-UHFFFAOYSA-N 0.000 description 3
- 240000001624 Espostoa lanata Species 0.000 description 3
- 235000009161 Espostoa lanata Nutrition 0.000 description 3
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 3
- 229920002472 Starch Polymers 0.000 description 3
- OMOVVBIIQSXZSZ-UHFFFAOYSA-N [6-(4-acetyloxy-5,9a-dimethyl-2,7-dioxo-4,5a,6,9-tetrahydro-3h-pyrano[3,4-b]oxepin-5-yl)-5-formyloxy-3-(furan-3-yl)-3a-methyl-7-methylidene-1a,2,3,4,5,6-hexahydroindeno[1,7a-b]oxiren-4-yl] 2-hydroxy-3-methylpentanoate Chemical compound CC12C(OC(=O)C(O)C(C)CC)C(OC=O)C(C3(C)C(CC(=O)OC4(C)COC(=O)CC43)OC(C)=O)C(=C)C32OC3CC1C=1C=COC=1 OMOVVBIIQSXZSZ-UHFFFAOYSA-N 0.000 description 3
- 238000000862 absorption spectrum Methods 0.000 description 3
- 239000001257 hydrogen Substances 0.000 description 3
- 229910052739 hydrogen Inorganic materials 0.000 description 3
- 238000000034 method Methods 0.000 description 3
- 239000001965 potato dextrose agar Substances 0.000 description 3
- 239000008107 starch Substances 0.000 description 3
- 235000019698 starch Nutrition 0.000 description 3
- FKHIFSZMMVMEQY-UHFFFAOYSA-N talc Chemical compound [Mg+2].[O-][Si]([O-])=O FKHIFSZMMVMEQY-UHFFFAOYSA-N 0.000 description 3
- 229920001817 Agar Polymers 0.000 description 2
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 2
- 244000068988 Glycine max Species 0.000 description 2
- 235000010469 Glycine max Nutrition 0.000 description 2
- 229910021586 Nickel(II) chloride Inorganic materials 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
- 238000009825 accumulation Methods 0.000 description 2
- PBCJIPOGFJYBJE-UHFFFAOYSA-N acetonitrile;hydrate Chemical compound O.CC#N PBCJIPOGFJYBJE-UHFFFAOYSA-N 0.000 description 2
- 239000008272 agar Substances 0.000 description 2
- TXHIDIHEXDFONW-UHFFFAOYSA-N benzene;propan-2-one Chemical compound CC(C)=O.C1=CC=CC=C1 TXHIDIHEXDFONW-UHFFFAOYSA-N 0.000 description 2
- 229910052799 carbon Inorganic materials 0.000 description 2
- 210000004027 cell Anatomy 0.000 description 2
- 238000004587 chromatography analysis Methods 0.000 description 2
- 238000012258 culturing Methods 0.000 description 2
- 230000002950 deficient Effects 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 229940079593 drug Drugs 0.000 description 2
- 239000000284 extract Substances 0.000 description 2
- 238000000855 fermentation Methods 0.000 description 2
- 230000004151 fermentation Effects 0.000 description 2
- 235000013312 flour Nutrition 0.000 description 2
- 238000004896 high resolution mass spectrometry Methods 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- QMMRZOWCJAIUJA-UHFFFAOYSA-L nickel dichloride Chemical compound Cl[Ni]Cl QMMRZOWCJAIUJA-UHFFFAOYSA-L 0.000 description 2
- 235000015097 nutrients Nutrition 0.000 description 2
- 239000006877 oatmeal agar Substances 0.000 description 2
- BASFCYQUMIYNBI-UHFFFAOYSA-N platinum Chemical compound [Pt] BASFCYQUMIYNBI-UHFFFAOYSA-N 0.000 description 2
- IOLCXVTUBQKXJR-UHFFFAOYSA-M potassium bromide Chemical compound [K+].[Br-] IOLCXVTUBQKXJR-UHFFFAOYSA-M 0.000 description 2
- 230000035755 proliferation Effects 0.000 description 2
- VWDWKYIASSYTQR-UHFFFAOYSA-N sodium nitrate Chemical compound [Na+].[O-][N+]([O-])=O VWDWKYIASSYTQR-UHFFFAOYSA-N 0.000 description 2
- 239000006188 syrup Substances 0.000 description 2
- 235000020357 syrup Nutrition 0.000 description 2
- WXCWPQMMDVSQOU-UHFFFAOYSA-N C1=CC=CC=C1.C(CCC)O.C(C)(=O)OCC Chemical compound C1=CC=CC=C1.C(CCC)O.C(C)(=O)OCC WXCWPQMMDVSQOU-UHFFFAOYSA-N 0.000 description 1
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 1
- ODINCKMPIJJUCX-UHFFFAOYSA-N Calcium oxide Chemical compound [Ca]=O ODINCKMPIJJUCX-UHFFFAOYSA-N 0.000 description 1
- ZAMOUSCENKQFHK-UHFFFAOYSA-N Chlorine atom Chemical compound [Cl] ZAMOUSCENKQFHK-UHFFFAOYSA-N 0.000 description 1
- 229920001353 Dextrin Polymers 0.000 description 1
- 239000004375 Dextrin Substances 0.000 description 1
- 241000196324 Embryophyta 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
- 240000008790 Musa x paradisiaca Species 0.000 description 1
- 235000018290 Musa x paradisiaca Nutrition 0.000 description 1
- GXCLVBGFBYZDAG-UHFFFAOYSA-N N-[2-(1H-indol-3-yl)ethyl]-N-methylprop-2-en-1-amine Chemical compound CN(CCC1=CNC2=C1C=CC=C2)CC=C GXCLVBGFBYZDAG-UHFFFAOYSA-N 0.000 description 1
- 240000007594 Oryza sativa Species 0.000 description 1
- 235000007164 Oryza sativa Nutrition 0.000 description 1
- 241001130173 Paralongidorus maximus Species 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
- PMZURENOXWZQFD-UHFFFAOYSA-L Sodium Sulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=O PMZURENOXWZQFD-UHFFFAOYSA-L 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
- 235000021307 Triticum Nutrition 0.000 description 1
- 244000098338 Triticum aestivum Species 0.000 description 1
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-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
- RTEXIPZMMDUXMR-UHFFFAOYSA-N benzene;ethyl acetate Chemical compound CCOC(C)=O.C1=CC=CC=C1 RTEXIPZMMDUXMR-UHFFFAOYSA-N 0.000 description 1
- MDHYEMXUFSJLGV-UHFFFAOYSA-N beta-phenethyl acetate Natural products CC(=O)OCCC1=CC=CC=C1 MDHYEMXUFSJLGV-UHFFFAOYSA-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
- 239000004202 carbamide Substances 0.000 description 1
- 235000013877 carbamide Nutrition 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
- GVPFVAHMJGGAJG-UHFFFAOYSA-L cobalt dichloride Chemical compound [Cl-].[Cl-].[Co+2] GVPFVAHMJGGAJG-UHFFFAOYSA-L 0.000 description 1
- 239000012230 colorless oil Substances 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 235000012343 cottonseed oil Nutrition 0.000 description 1
- 239000012228 culture supernatant Substances 0.000 description 1
- 235000019425 dextrin Nutrition 0.000 description 1
- 239000000706 filtrate Substances 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 239000012535 impurity Substances 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- MVZXTUSAYBWAAM-UHFFFAOYSA-N iron;sulfuric acid Chemical compound [Fe].OS(O)(=O)=O MVZXTUSAYBWAAM-UHFFFAOYSA-N 0.000 description 1
- 239000011777 magnesium Substances 0.000 description 1
- 229910052749 magnesium Inorganic materials 0.000 description 1
- 235000012054 meals Nutrition 0.000 description 1
- 235000013372 meat Nutrition 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
- 235000019319 peptone Nutrition 0.000 description 1
- 229910052697 platinum Inorganic materials 0.000 description 1
- 239000011591 potassium Substances 0.000 description 1
- 229910052700 potassium Inorganic materials 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 235000009566 rice Nutrition 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 239000000523 sample Substances 0.000 description 1
- 238000011218 seed culture Methods 0.000 description 1
- 238000010898 silica gel chromatography 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
- 239000002689 soil Substances 0.000 description 1
- 238000001179 sorption measurement Methods 0.000 description 1
- 235000012424 soybean oil Nutrition 0.000 description 1
- 239000003549 soybean oil Substances 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 239000010902 straw Substances 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 210000001179 synovial fluid Anatomy 0.000 description 1
- 239000013076 target substance Substances 0.000 description 1
- 235000015112 vegetable and seed oil Nutrition 0.000 description 1
- 239000008158 vegetable oil Substances 0.000 description 1
- 239000012138 yeast extract Substances 0.000 description 1
Landscapes
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
- Peptides Or Proteins (AREA)
Abstract
Description
【発明の詳細な説明】
(産業上の利用分野)
本発明は抗マツノザイセンチュウ活性を有するバーサフ
ェロサイド系化合物に関するものである。DETAILED DESCRIPTION OF THE INVENTION (Industrial Field of Application) The present invention relates to versaferroside compounds having antipine nematode activity.
(従来の技術及び発明が解決しようとする問題点)松枯
れ病は林業はもとより社会的な問題となっている。しか
しながら松枯れ病の原因であるマツノザイセンチュウを
駆除する効果的な薬剤は実用化には至っていない。(Problems to be Solved by the Prior Art and the Invention) Pine blight has become a problem not only in forestry but also in society. However, an effective drug to exterminate pine nematode, which causes pine blight, has not yet been put into practical use.
本発明者らは、こうした現状に鑑み、新規な抗マツノザ
イセンチュウ活性物質の探索を目的として、多数の微生
物を分離し、その産生ずる活性物質について単離、精製
を行った。In view of the current situation, the present inventors isolated a large number of microorganisms, and isolated and purified the active substances produced by the microorganisms, with the aim of searching for novel antipine nematode active substances.
その結果、特定の真菌類の培養物中に新規活性物質が産
生されることの知見を得て、その単離・精製を行い、し
かも本活性物質が良好な抗マツノザイセンチュウ活性を
有することを見いだし、本発明を完成するに至った。As a result, we found that a new active substance was produced in the culture of a specific fungus, isolated and purified it, and confirmed that this active substance had good antipine nematode activity. This discovery led to the completion of the present invention.
(問題点を解決するための手段)
即ち、本発明の要旨は、下記構造式で表されるバーサフ
ェロサイド系化合物に存する。(Means for Solving the Problems) That is, the gist of the present invention resides in a versaferroside compound represented by the following structural formula.
(式中、Rは水素原子またはメチル基を表わす。)本発
明のバーサフェロサイド系化合物は、前記構造式で表さ
れる。(In the formula, R represents a hydrogen atom or a methyl group.) The versaferroside compound of the present invention is represented by the above structural formula.
以下、本発明においては、上記式中、Rがメチル基を表
わすものをバーサフェロサイドA物質、またRが水素原
子を表わすものをバーサフェロサイドB物質という。Hereinafter, in the present invention, those in the above formula in which R represents a methyl group are referred to as versaferroside A substances, and those in which R represents a hydrogen atom are referred to as versaferroside B substances.
前記構造式で表されるバーサフェロサイド系化合物は、
後述のように、例えば無胞子不完全菌D1084 (微
工研菌寄第11289号(FERMp−11289))
を培養することによって得ることが出来る。The versaferroside compound represented by the above structural formula is
As will be described later, for example, the nonspore-deficient bacterium D1084 (FERMp-11289)
It can be obtained by culturing.
本発明に使用する微生物DI084株は天然の双子葉植
物体より新たに分離した真菌類(Eumycota)の
一種で、その菌学的性状は次の通りである。The microorganism strain DI084 used in the present invention is a type of fungus (Eumycota) newly isolated from a natural dicotyledonous plant, and its mycological properties are as follows.
D1084株の菌学的性状
ポテトデキストロース寒天(PDA)、オートミール寒
天(OA) 、麦芽寒天(MA)及び三浦培地(LCA
)上、27℃で生育は中程度、7日間の培養で、コロニ
ーの径は5〜6cm、白色綿毛状を呈する。顕微鏡下で
観察したところ、どの培地にも分生子等の特徴的な形態
は観察できなかった。さらに素寒天上に滅菌した稲藁や
バナナの皮を置いて植菌し、27℃で1ケ月間観察した
が、この場合も分生子などの形成は認められなかった。Mycological properties of strain D1084 Potato dextrose agar (PDA), oatmeal agar (OA), malt agar (MA), and Miura medium (LCA)
), the growth is moderate at 27°C, and after 7 days of culture, the colonies are 5-6 cm in diameter and white fluff-like. When observed under a microscope, no characteristic morphology such as conidia could be observed in any of the media. Furthermore, sterilized rice straw or banana peels were placed on the agar and inoculated, and the cells were observed at 27°C for one month, but no conidia were observed in this case either.
・生育温度(PDA上、1週間培養):15〜35°C
(最適温度27°C)
・生育pH(LCA上、1週間培養):2〜9、(最適
生育pH7〜8)
従って本菌株を無胞子不完全菌DI084株と呼称する
ことにした。・Growth temperature (cultured on PDA for 1 week): 15-35°C
(Optimum temperature 27°C) Growth pH (LCA, 1 week culture): 2-9, (Optimum growth pH 7-8) Therefore, this strain was designated as the apospore-deficient strain DI084.
本発明においては、前記の菌を通常の微生物が利用し得
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、水飴、デキストリン、シュクロース、澱粉、
糖蜜及び動・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、コーンステイープ・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
、硝酸ソーダ及び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、燐酸、硫酸及びその他のイオンを
生成する事の出来る無機塩類を添加することは有効であ
る。また菌の生育を助は活性物質バーサフェロサイド系
化合物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-mentioned bacteria are cultured in a medium containing nutrients that can be used by ordinary microorganisms. Nutrient sources include glucose, starch syrup, dextrin, sucrose, starch,
Molasses and animal/vegetable oils can be used. Also, soybean flour, wheat germ, cornstape liquor, cottonseed meal, meat extract, peptone, yeast extract, ammonium sulfate, sodium nitrate, urea, etc. can be used as nitrogen sources. 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 aid the growth of bacteria and promote the production of active substance versaferroside compounds can be appropriately added.
培養法としては、好気的条件下での培養法、特に深部培
養法が最も適している。培養に適当な温度は10〜30
°Cであるが多くの場合、20〜27℃付近で培養する
。バーサフェロサイド系化合物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。培養物中のバー
サフェロサイド系化合物の蓄積が最高になったときに培
養を停止し、培養液から目的物質を単離精製する。The most suitable culture method is a culture method under aerobic conditions, especially a deep culture method. The appropriate temperature for culturing is 10-30°C.
℃, but in most cases, it is cultured at around 20 to 27℃. The production of versaferroside compounds varies depending on the culture medium and culture conditions, but it is usually
Its accumulation reaches its maximum within 10 days. When the accumulation of versaferroside compounds in the culture reaches its maximum, the culture is stopped, and the target substance is isolated and purified from the culture solution.
本発明の前記構造式で表されるバーサフェロサイド系化
合物を培養物から単離、精製するには、不純物との溶解
度差を利用する手段、イオン結合力の差を利用する手段
、吸着親和力の差を利用する手段、分子量の差を利用す
る手段のいずれも、それぞれ単独、または適宜組み合わ
せて、あるいは反復して使用される。In order to isolate and purify the versaferroside compound represented by the above structural formula of the present invention from a culture, there are methods that utilize the difference in solubility with impurities, methods that utilize the difference in ionic binding strength, and methods that utilize the difference in adsorption affinity. The means that utilize the difference and the means that utilize the difference in molecular weight may be used alone, in appropriate combinations, or repeatedly.
すなわち、培養濾液を酢酸エチルで抽出し、減圧下で濃
縮する。That is, the culture filtrate is extracted with ethyl acetate and concentrated under reduced pressure.
濃縮液をシリカゲルカラムクロマトグラフィーフロリジ
ルクロマトグラフィー等を組み合わせて精製すると、純
粋なバーサフェロサイド系化合物が得られる。When the concentrated solution is purified using a combination of silica gel column chromatography, florisil chromatography, etc., a pure versaferroside compound can be obtained.
(実施例)
以下に実施例及び試験例を挙げてさらに具体的に本発明
を説明するが、本発明はその要旨を越えない限り以下の
実施例によって限定されるものではない。(Examples) The present invention will be described below in more detail with reference to Examples and Test Examples, but the present invention is not limited by the following Examples unless the gist thereof is exceeded.
実施例1
(1)培養
水飴4.0%、大豆粉2.0%、大豆油0.3%、ファ
マメディア1.0%、サングレイン0.5%、1化カル
シウム0.3%、硫酸鉄0.001%、塩化コバルト0
.0001%、塩化ニッケル0.0001%を含有する
培地(pH6,0)を40m1.ず”:)200m1.
三角フラスコ20本に分注し、121°Cにおいて20
分間高圧滅菌する。これにD1084系物質生産株、D
1084株を1白金耳ずつ植菌し、26°Cにおいて6
日間、210回転にて振とう培養する。Example 1 (1) Cultured starch syrup 4.0%, soybean flour 2.0%, soybean oil 0.3%, Famamedia 1.0%, sungrain 0.5%, calcium monoxide 0.3%, sulfuric acid Iron 0.001%, cobalt chloride 0
.. 0001% of nickel chloride and 0.0001% of nickel chloride (pH 6.0). zu”:)200m1.
Dispense into 20 Erlenmeyer flasks and incubate for 20 minutes at 121°C.
Sterilize under autoclave for minutes. In addition to this, D1084-based substance producing strain, D
1,084 strains were inoculated in one platinum loop, and incubated at 26°C for 6 days.
Culture with shaking at 210 rpm for 1 day.
別に、上記と同一組成の主発酵培地を調製し、その80
mI!、を500mf三角フラスコ100本に分注し、
121°Cにおいて20分間高圧滅菌する。この主発酵
培地に前記種培養液を4m7!ずつ接種し、26°Cに
おいて5日間、210回転にて振とう培養する。得られ
た培養物を遠心分離して、培養上清液を得た。Separately, prepare a main fermentation medium with the same composition as above, and
mI! , into 100 500mf Erlenmeyer flasks,
Autoclave at 121°C for 20 minutes. Add 4 m7 of the above seed culture solution to this main fermentation medium! The cells were inoculated and cultured at 26°C for 5 days with shaking at 210 rpm. The resulting culture was centrifuged to obtain a culture supernatant.
(2)培養物の精製
上記(1)で得られた培養上滑液7.32を等量の酢酸
エチルにて抽出した。(2) Purification of culture 7.32 g of the cultured synovial fluid obtained in (1) above was extracted with an equal volume of ethyl acetate.
抽出液を無水硫酸ナトリウムで脱水し、減圧上濃縮し、
3.3gの油状物質を得た。得られた油状物質をアビセ
ル(旭化成工業社製)に混合し、乾式で充填したワコー
ゲルC−200(和光純薬工業社製)100mj!の塔
の上にのせ、ベンゼン−酢酸エチル混液(1: 1)か
らベンゼン−酢酸エチル−ブタノール混液(1: 1
: 2)にて展開するクロマトグラフィーを行った。The extract was dehydrated with anhydrous sodium sulfate, concentrated under reduced pressure,
3.3 g of oil was obtained. The obtained oily substance was mixed with Avicel (manufactured by Asahi Kasei Industries, Ltd.) and dry-filled into Wakogel C-200 (manufactured by Wako Pure Chemical Industries, Ltd.) for 100mj! of benzene-ethyl acetate mixture (1:1) to benzene-ethyl acetate-butanol mixture (1:1).
: Chromatography developed in 2) was performed.
活性画分を集め、減圧上濃縮乾固し油状物質を得た。こ
の油状物質をアビセルに混合し、ベンゼンにて充填した
フロリジル(フロリジル社製)50mlの塔にのせ、ベ
ンゼン−アセトン混液にて展開した。The active fractions were collected and concentrated to dryness under reduced pressure to obtain an oily substance. This oily substance was mixed with Avicel, placed on a 50 ml column of Florisil (manufactured by Florisil) filled with benzene, and developed with a benzene-acetone mixture.
ベンゼン−アセトン混液(8:2)にて溶出する活性画
分を減圧上濃縮した。得られた油状物質はアセトニトリ
ル−水混液(8:2)にて平衡化したワコーゲルLC:
0DS−30K (和光純薬社製)にのせ、アセトニト
リル−水混液にて展開した。溶出した活性画分を集め、
減圧上濃縮すると、バーサフェロサイドA物[52■及
びバーサフェロサイドB物質153■をそれぞれ得た。The active fraction eluted with a benzene-acetone mixture (8:2) was concentrated under reduced pressure. The obtained oily substance was equilibrated with an acetonitrile-water mixture (8:2) on Wakogel LC:
It was placed on 0DS-30K (manufactured by Wako Pure Chemical Industries, Ltd.) and developed with an acetonitrile-water mixture. Collect the eluted active fraction,
Concentration under reduced pressure gave Versaferroside A [52] and Versaferroside B (153).
構造式を以下に示す。The structural formula is shown below.
また、得られたバーサフェロサイドA物質及びバーサフ
ェロサイドB物質の物性を以下に示す。Further, the physical properties of the obtained versaferroside A substance and versaferroside B substance are shown below.
■)外観:
無色油状物
2)分子量:
A物質:593 (HR−MS、593.3785)
B物質:607 (HR−MS、607.3942)
3)分子式:
%式%
)
:
)
)
5) 紫外部吸収スペクトル:
A物質:λwax (メタノール)nm(ε):21
0 (16,300)
B物質:λ@11+1 (メタノール)nm(C):
210 (18,200)
6)赤外部吸収スペクトル:
臭化カリウム錠剤中で測定したスペクトルは第1図(バ
ーサフェロサイドA物質)及び第2図(バーサフェロサ
イドB物質)に示す通りである。■) Appearance: Colorless oil 2) Molecular weight: Substance A: 593 (HR-MS, 593.3785)
Substance B: 607 (HR-MS, 607.3942)
3) Molecular formula: % formula % ) : ) ) 5) Ultraviolet absorption spectrum: Substance A: λwax (methanol) nm (ε): 21
0 (16,300) Substance B: λ@11+1 (methanol) nm (C):
210 (18,200) 6) Infrared absorption spectrum: The spectra measured in potassium bromide tablets are as shown in Figure 1 (versaferroside A substance) and Figure 2 (versaferroside B substance).
7)水素核磁気共鳴スペクトル:
重クロロホルム溶液中で測定した500MHz水素核磁
気共鳴スペクトルは第3図(バーサフェロサイドA物質
)及び第4図(バーサフェロサイドB物質)に示す通り
である。7) Hydrogen nuclear magnetic resonance spectrum: The 500 MHz hydrogen nuclear magnetic resonance spectrum measured in a deuterated chloroform solution is as shown in FIG. 3 (versaferroside A substance) and FIG. 4 (versaferroside B substance).
8)炭素核磁気共鳴スペクトル:
重クロロホルム溶液中で測定した125M)(z炭素核
磁気共鳴スペクトルは表1に示す通りである。8) Carbon nuclear magnetic resonance spectrum: The 125M)(z carbon nuclear magnetic resonance spectrum measured in deuterated chloroform solution is as shown in Table 1.
表 1
A物質 B物質
δ値 δ値
11.3 11.3
11.8 11.8
14.4 14.3
15.2 15.3
15.3 15.4
表 1
つづき
δ値
19.5
20.0
24.1
24.4
25.6
27.2
28.0
28.9
30.8
33.1
34.4
36.0
37.4
46.5
δ値
18.6
19.5
20.0
24.6
26.2
27.2
28.1
30.8
32.8
33.2
34.5
35.8
36.4
37.6
53.5
表1
つづき
δ値
53.6
55.4
57.9
60.7
76.1
168.8
169.7
171.0
171.0
173.5
173.7
δ値
53.9
55.5
57.9
60.4
76.0
168.4
169.7
171.0
172.0
173.6
173.7
9)溶解性:
メタノール、エタノール、酢酸エチル及びクロロホルム
に溶け、水に溶けない。Table 1 Substance A Substance B δ value δ value 11.3 11.3 11.8 11.8 14.4 14.3 15.2 15.3 15.3 15.4 Table 1 Continued δ value 19.5 20. 0 24.1 24.4 25.6 27.2 28.0 28.9 30.8 33.1 34.4 36.0 37.4 46.5 δ value 18.6 19.5 20.0 24. 6 26.2 27.2 28.1 30.8 32.8 33.2 34.5 35.8 36.4 37.6 53.5 Table 1 continued δ value 53.6 55.4 57.9 60. 7 76.1 168.8 169.7 171.0 171.0 173.5 173.7 δ value 53.9 55.5 57.9 60.4 76.0 168.4 169.7 171.0 172. 0 173.6 173.7 9) Solubility: Soluble in methanol, ethanol, ethyl acetate and chloroform, insoluble in water.
試験例1
(抗マツノザイセンチュウ試験)
前記の実施例で得られた試料物質バーサフェロサイドA
物質及びバーサフェロサイドB物質を各々100μg溶
解したメタノール溶液を、直径約5m11の球状に整形
した綿花に浸透させたのち乾燥した。該綿球に15,0
00頭/ m lに調製したマツノザイセンチュウ懸濁
液を100μ!浸透させ、予め表面にセンチュウの餌と
してのBotr工土±s cineraを生育させて
おいた直径4C1のシャーレ培地上に乗せた。5日間、
26°Cに保持したのち、マツノザイセンチュウによっ
て侵食されるシャーレ上のBotr tis ci
neraの観察を行った。Test Example 1 (Anti-pine nematode test) Sample material Versaferroside A obtained in the above example
A methanol solution in which 100 μg of each of the substance and Versaferroside B substance were dissolved was permeated into a cotton ball shaped into a sphere with a diameter of about 5 m11, and then dried. 15,0 on the cotton ball
100μ of a suspension of pine tree nematode prepared at 00 heads/ml. It was then placed on a petri dish medium with a diameter of 4C1 on which Botr construction soil ± s cinera as food for nematodes had been grown in advance. 5 days
Botr tis ci on a petri dish attacked by pine nematodes after being kept at 26°C
Nera was observed.
その結果、該活性物質を含む綿球をおいたシャーレ上で
はセンチュウによる侵食は認められなかった。As a result, no nematode attack was observed on the petri dish on which the cotton balls containing the active substance were placed.
また、それぞれの生存センチュウを集めて計数し、対照
区との比で増殖率を算出したところ、バーサフェロサイ
ドA物質に関しては3.3%、バーサフェロサイドB物
質に関しては1.2%であり、各物質ともセンチュウの
増殖を抑制していた。In addition, when each living nematode was collected and counted, and the proliferation rate was calculated in comparison with the control group, it was 3.3% for versaferroside A substance and 1.2% for versaferroside B substance. , each substance inhibited nematode proliferation.
(発明の効果)
本発明の新規抗マツノザイセンチュウ活性物質バーサフ
ェロサイド系化合物は、以上実施例で示した通り、該活
性を有しており、マツノザイセンチュウ駆除薬としての
有用性が期待される。(Effects of the Invention) The novel antipine nematode active substance versaferroside compound of the present invention has the activity as shown in the examples above, and is expected to be useful as a pine needle nematode exterminating drug. Ru.
第1図及び第2図はバーサフェロサイドA物賞及びB物
質の赤外部吸収スペクトルを示す図である。
第3図及び第4図はバーサフェロサイドA物質及びB物
質の水素核磁気共鳴スペクトルを示す図である。FIGS. 1 and 2 are diagrams showing infrared absorption spectra of Versaferroside A and B substances. FIGS. 3 and 4 are diagrams showing hydrogen nuclear magnetic resonance spectra of versaferroside A and B substances.
Claims (1)
物。 ▲数式、化学式、表等があります▼ (式中、Rは水素原子またはメチル基を表わす。)(1) Versaferroside compound represented by the following structural formula. ▲There are mathematical formulas, chemical formulas, tables, etc.▼ (In the formula, R represents a hydrogen atom or a methyl group.)
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP2046886A JPH03251597A (en) | 1990-02-27 | 1990-02-27 | Versaferrocide-based compound |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP2046886A JPH03251597A (en) | 1990-02-27 | 1990-02-27 | Versaferrocide-based compound |
Publications (1)
Publication Number | Publication Date |
---|---|
JPH03251597A true JPH03251597A (en) | 1991-11-11 |
Family
ID=12759846
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP2046886A Pending JPH03251597A (en) | 1990-02-27 | 1990-02-27 | Versaferrocide-based compound |
Country Status (1)
Country | Link |
---|---|
JP (1) | JPH03251597A (en) |
Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO1998054211A3 (en) * | 1997-05-29 | 1999-03-04 | Meiji Milk Prod Co Ltd | Cyclodepsipeptides and pharmaceutical compositions containing them |
CN110777149A (en) * | 2019-10-15 | 2020-02-11 | 山东农业大学 | Pine wood nematode tra-1 gene and application thereof in development interference |
-
1990
- 1990-02-27 JP JP2046886A patent/JPH03251597A/en active Pending
Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO1998054211A3 (en) * | 1997-05-29 | 1999-03-04 | Meiji Milk Prod Co Ltd | Cyclodepsipeptides and pharmaceutical compositions containing them |
CN110777149A (en) * | 2019-10-15 | 2020-02-11 | 山东农业大学 | Pine wood nematode tra-1 gene and application thereof in development interference |
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