CN86106265A - Method for producing mildew mycin - Google Patents
Method for producing mildew mycin Download PDFInfo
- Publication number
- CN86106265A CN86106265A CN86106265.5A CN86106265A CN86106265A CN 86106265 A CN86106265 A CN 86106265A CN 86106265 A CN86106265 A CN 86106265A CN 86106265 A CN86106265 A CN 86106265A
- Authority
- CN
- China
- Prior art keywords
- mildiomycin
- substratum
- streptoverticillium
- bacterial strain
- canavanine
- 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
- 238000004519 manufacturing process Methods 0.000 title claims description 13
- 241000187747 Streptomyces Species 0.000 claims abstract description 46
- FSBIGDSBMBYOPN-VKHMYHEASA-N L-canavanine Chemical compound OC(=O)[C@@H](N)CCONC(N)=N FSBIGDSBMBYOPN-VKHMYHEASA-N 0.000 claims abstract description 29
- RYVNIFSIEDRLSJ-UHFFFAOYSA-N 5-(hydroxymethyl)cytosine Chemical compound NC=1NC(=O)N=CC=1CO RYVNIFSIEDRLSJ-UHFFFAOYSA-N 0.000 claims abstract description 28
- FSBIGDSBMBYOPN-UHFFFAOYSA-N O-guanidino-DL-homoserine Natural products OC(=O)C(N)CCON=C(N)N FSBIGDSBMBYOPN-UHFFFAOYSA-N 0.000 claims abstract description 27
- LELJBJGDDGUFRP-UHFFFAOYSA-N serine hydroxamate Chemical compound OCC(N)C(=O)NO LELJBJGDDGUFRP-UHFFFAOYSA-N 0.000 claims abstract description 14
- 238000009825 accumulation Methods 0.000 claims abstract description 3
- KCIRYJNISRMYFI-UHFFFAOYSA-N mildiomycin Natural products NC(CO)C(=O)NC1C=CC(OC1C(O)(CC(O)CNC(=N)N)C(=O)O)N2CN=C(N)C(=C2)CO KCIRYJNISRMYFI-UHFFFAOYSA-N 0.000 claims description 76
- QKJJCZYFXJCKRX-HZHKWBLPSA-N 2-[(2s,3s,6r)-6-[4-amino-5-(hydroxymethyl)-2-oxopyrimidin-1-yl]-3-[[(2s)-2-amino-3-hydroxypropanoyl]amino]-3,6-dihydro-2h-pyran-2-yl]-5-(diaminomethylideneamino)-2,4-dihydroxypentanoic acid Chemical compound O1[C@H](C(O)(CC(O)CN=C(N)N)C(O)=O)[C@@H](NC(=O)[C@H](CO)N)C=C[C@@H]1N1C(=O)N=C(N)C(CO)=C1 QKJJCZYFXJCKRX-HZHKWBLPSA-N 0.000 claims description 74
- 230000001580 bacterial effect Effects 0.000 claims description 59
- 238000000034 method Methods 0.000 claims description 40
- 239000007788 liquid Substances 0.000 claims description 10
- 235000015097 nutrients Nutrition 0.000 claims description 10
- 238000012258 culturing Methods 0.000 abstract description 5
- 239000001963 growth medium Substances 0.000 abstract description 4
- 230000000895 acaricidal effect Effects 0.000 abstract description 3
- APNPVBXEWGCCLU-QNRZBPGKSA-N mycomycin Chemical compound OC(=O)C\C=C\C=C/C=C=CC#CC#C APNPVBXEWGCCLU-QNRZBPGKSA-N 0.000 abstract 2
- 239000000642 acaricide Substances 0.000 abstract 1
- 239000003242 anti bacterial agent Substances 0.000 abstract 1
- 230000000844 anti-bacterial effect Effects 0.000 abstract 1
- 239000003899 bactericide agent Substances 0.000 abstract 1
- 230000015572 biosynthetic process Effects 0.000 abstract 1
- 238000003898 horticulture Methods 0.000 abstract 1
- 238000003786 synthesis reaction Methods 0.000 abstract 1
- 229920001817 Agar Polymers 0.000 description 20
- 239000008272 agar Substances 0.000 description 20
- 239000000725 suspension Substances 0.000 description 16
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 12
- 239000000047 product Substances 0.000 description 12
- 241000894006 Bacteria Species 0.000 description 11
- 238000007747 plating Methods 0.000 description 11
- 239000000243 solution Substances 0.000 description 11
- 238000000855 fermentation Methods 0.000 description 10
- 230000004151 fermentation Effects 0.000 description 10
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 9
- 239000011259 mixed solution Substances 0.000 description 9
- 230000003068 static effect Effects 0.000 description 9
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 8
- 239000000463 material Substances 0.000 description 8
- 239000000203 mixture Substances 0.000 description 8
- 230000001954 sterilising effect Effects 0.000 description 8
- 238000004659 sterilization and disinfection Methods 0.000 description 8
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 8
- 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 7
- 239000007864 aqueous solution Substances 0.000 description 7
- -1 iron ion Chemical class 0.000 description 7
- 235000004400 serine Nutrition 0.000 description 7
- 239000000126 substance Substances 0.000 description 7
- MTCFGRXMJLQNBG-UHFFFAOYSA-N Serine Natural products OCC(N)C(O)=O MTCFGRXMJLQNBG-UHFFFAOYSA-N 0.000 description 6
- 229920002472 Starch Polymers 0.000 description 6
- 229930006000 Sucrose Natural products 0.000 description 6
- 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 6
- 240000008042 Zea mays Species 0.000 description 6
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 description 6
- 235000002017 Zea mays subsp mays Nutrition 0.000 description 6
- 239000002253 acid Substances 0.000 description 6
- 230000003115 biocidal effect Effects 0.000 description 6
- 229910052799 carbon Inorganic materials 0.000 description 6
- 235000005822 corn Nutrition 0.000 description 6
- 239000008103 glucose Substances 0.000 description 6
- 235000001727 glucose Nutrition 0.000 description 6
- 239000006916 nutrient agar Substances 0.000 description 6
- 235000019698 starch Nutrition 0.000 description 6
- 239000008107 starch Substances 0.000 description 6
- 239000005720 sucrose Substances 0.000 description 6
- 238000012360 testing method Methods 0.000 description 6
- PAWQVTBBRAZDMG-UHFFFAOYSA-N 2-(3-bromo-2-fluorophenyl)acetic acid Chemical compound OC(=O)CC1=CC=CC(Br)=C1F PAWQVTBBRAZDMG-UHFFFAOYSA-N 0.000 description 5
- 150000001875 compounds Chemical class 0.000 description 5
- 244000005700 microbiome Species 0.000 description 5
- 241001411320 Eriogonum inflatum Species 0.000 description 4
- JOYRKODLDBILNP-UHFFFAOYSA-N Ethyl urethane Chemical compound CCOC(N)=O JOYRKODLDBILNP-UHFFFAOYSA-N 0.000 description 4
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 4
- BAVYZALUXZFZLV-UHFFFAOYSA-N Methylamine Chemical compound NC BAVYZALUXZFZLV-UHFFFAOYSA-N 0.000 description 4
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 4
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 4
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 description 4
- 229910052921 ammonium sulfate Inorganic materials 0.000 description 4
- 235000011130 ammonium sulphate Nutrition 0.000 description 4
- 239000012531 culture fluid Substances 0.000 description 4
- 238000001914 filtration Methods 0.000 description 4
- 230000005764 inhibitory process Effects 0.000 description 4
- 235000012204 lemonade/lime carbonate Nutrition 0.000 description 4
- 239000012071 phase Substances 0.000 description 4
- 239000011591 potassium Substances 0.000 description 4
- 229910052700 potassium Inorganic materials 0.000 description 4
- 239000000843 powder Substances 0.000 description 4
- 238000011218 seed culture Methods 0.000 description 4
- 235000011121 sodium hydroxide Nutrition 0.000 description 4
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 3
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 3
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 3
- 229910002651 NO3 Inorganic materials 0.000 description 3
- NHNBFGGVMKEFGY-UHFFFAOYSA-N Nitrate Chemical compound [O-][N+]([O-])=O NHNBFGGVMKEFGY-UHFFFAOYSA-N 0.000 description 3
- 229910019142 PO4 Inorganic materials 0.000 description 3
- 238000010521 absorption reaction Methods 0.000 description 3
- 239000002585 base Substances 0.000 description 3
- 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 3
- 239000005018 casein Substances 0.000 description 3
- BECPQYXYKAMYBN-UHFFFAOYSA-N casein, tech. Chemical compound NCCCCC(C(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(CC(C)C)N=C(O)C(CCC(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(C(C)O)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(COP(O)(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(N)CC1=CC=CC=C1 BECPQYXYKAMYBN-UHFFFAOYSA-N 0.000 description 3
- 235000021240 caseins Nutrition 0.000 description 3
- 239000012141 concentrate Substances 0.000 description 3
- 235000012343 cottonseed oil Nutrition 0.000 description 3
- AUZONCFQVSMFAP-UHFFFAOYSA-N disulfiram Chemical compound CCN(CC)C(=S)SSC(=S)N(CC)CC AUZONCFQVSMFAP-UHFFFAOYSA-N 0.000 description 3
- 239000000706 filtrate Substances 0.000 description 3
- KWIUHFFTVRNATP-UHFFFAOYSA-N glycine betaine Chemical compound C[N+](C)(C)CC([O-])=O KWIUHFFTVRNATP-UHFFFAOYSA-N 0.000 description 3
- XEEYBQQBJWHFJM-UHFFFAOYSA-N iron Substances [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 3
- 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 3
- 229910000360 iron(III) sulfate Inorganic materials 0.000 description 3
- 235000012054 meals Nutrition 0.000 description 3
- CVGSWVDBRMDWPX-UHFFFAOYSA-N methanetriamine;hydrochloride Chemical compound Cl.NC(N)N CVGSWVDBRMDWPX-UHFFFAOYSA-N 0.000 description 3
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 description 3
- 231100000350 mutagenesis Toxicity 0.000 description 3
- 235000019198 oils Nutrition 0.000 description 3
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 3
- 239000010452 phosphate Substances 0.000 description 3
- 238000001556 precipitation Methods 0.000 description 3
- 238000002360 preparation method Methods 0.000 description 3
- MEIRRNXMZYDVDW-MQQKCMAXSA-N (2E,4E)-2,4-hexadien-1-ol Chemical compound C\C=C\C=C\CO MEIRRNXMZYDVDW-MQQKCMAXSA-N 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 2
- 239000001763 2-hydroxyethyl(trimethyl)azanium Substances 0.000 description 2
- 235000019743 Choline chloride Nutrition 0.000 description 2
- DYDCUQKUCUHJBH-UWTATZPHSA-N D-Cycloserine Chemical compound N[C@@H]1CONC1=O DYDCUQKUCUHJBH-UWTATZPHSA-N 0.000 description 2
- ROSDSFDQCJNGOL-UHFFFAOYSA-N Dimethylamine Chemical compound CNC ROSDSFDQCJNGOL-UHFFFAOYSA-N 0.000 description 2
- 241000196324 Embryophyta Species 0.000 description 2
- 241000221785 Erysiphales Species 0.000 description 2
- BDAGIHXWWSANSR-UHFFFAOYSA-M Formate Chemical compound [O-]C=O BDAGIHXWWSANSR-UHFFFAOYSA-M 0.000 description 2
- 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 2
- XGEGHDBEHXKFPX-UHFFFAOYSA-N N-methyl urea Chemical compound CNC(N)=O XGEGHDBEHXKFPX-UHFFFAOYSA-N 0.000 description 2
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 description 2
- 150000007513 acids Chemical class 0.000 description 2
- 239000003513 alkali Substances 0.000 description 2
- 235000011114 ammonium hydroxide Nutrition 0.000 description 2
- 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 2
- 210000004027 cell Anatomy 0.000 description 2
- OEYIOHPDSNJKLS-UHFFFAOYSA-N choline Chemical compound C[N+](C)(C)CCO OEYIOHPDSNJKLS-UHFFFAOYSA-N 0.000 description 2
- 229960001231 choline Drugs 0.000 description 2
- SGMZJAMFUVOLNK-UHFFFAOYSA-M choline chloride Chemical compound [Cl-].C[N+](C)(C)CCO SGMZJAMFUVOLNK-UHFFFAOYSA-M 0.000 description 2
- 229960003178 choline chloride Drugs 0.000 description 2
- 238000004587 chromatography analysis Methods 0.000 description 2
- 239000011248 coating agent Substances 0.000 description 2
- 238000000576 coating method Methods 0.000 description 2
- 229910000365 copper sulfate Inorganic materials 0.000 description 2
- ARUVKPQLZAKDPS-UHFFFAOYSA-L copper(II) sulfate Chemical compound [Cu+2].[O-][S+2]([O-])([O-])[O-] ARUVKPQLZAKDPS-UHFFFAOYSA-L 0.000 description 2
- 238000010790 dilution Methods 0.000 description 2
- 239000012895 dilution Substances 0.000 description 2
- PXEDJBXQKAGXNJ-QTNFYWBSSA-L disodium L-glutamate Chemical compound [Na+].[Na+].[O-]C(=O)[C@@H](N)CCC([O-])=O PXEDJBXQKAGXNJ-QTNFYWBSSA-L 0.000 description 2
- 239000006185 dispersion Substances 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 150000004675 formic acid derivatives Chemical class 0.000 description 2
- 235000011187 glycerol Nutrition 0.000 description 2
- 238000000227 grinding Methods 0.000 description 2
- 239000004615 ingredient Substances 0.000 description 2
- 229910017053 inorganic salt Inorganic materials 0.000 description 2
- 229910052742 iron Inorganic materials 0.000 description 2
- 239000002609 medium Substances 0.000 description 2
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 description 2
- 235000013336 milk Nutrition 0.000 description 2
- 239000008267 milk Substances 0.000 description 2
- 210000004080 milk Anatomy 0.000 description 2
- 238000002156 mixing Methods 0.000 description 2
- 229940057059 monascus purpureus Drugs 0.000 description 2
- 235000013923 monosodium glutamate Nutrition 0.000 description 2
- 238000002703 mutagenesis Methods 0.000 description 2
- 230000035772 mutation Effects 0.000 description 2
- 125000004433 nitrogen atom Chemical group N* 0.000 description 2
- QJGQUHMNIGDVPM-UHFFFAOYSA-N nitrogen group Chemical group [N] QJGQUHMNIGDVPM-UHFFFAOYSA-N 0.000 description 2
- 230000003287 optical effect Effects 0.000 description 2
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 description 2
- 229920000053 polysorbate 80 Polymers 0.000 description 2
- 238000003825 pressing Methods 0.000 description 2
- BDERNNFJNOPAEC-UHFFFAOYSA-N propan-1-ol Chemical compound CCCO BDERNNFJNOPAEC-UHFFFAOYSA-N 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 150000003839 salts Chemical class 0.000 description 2
- 229940073490 sodium glutamate Drugs 0.000 description 2
- QEMXHQIAXOOASZ-UHFFFAOYSA-N tetramethylammonium Chemical compound C[N+](C)(C)C QEMXHQIAXOOASZ-UHFFFAOYSA-N 0.000 description 2
- GETQZCLCWQTVFV-UHFFFAOYSA-N trimethylamine Chemical compound CN(C)C GETQZCLCWQTVFV-UHFFFAOYSA-N 0.000 description 2
- 239000003643 water by type Substances 0.000 description 2
- HDTRYLNUVZCQOY-UHFFFAOYSA-N α-D-glucopyranosyl-α-D-glucopyranoside Natural products OC1C(O)C(O)C(CO)OC1OC1C(O)C(O)C(O)C(CO)O1 HDTRYLNUVZCQOY-UHFFFAOYSA-N 0.000 description 1
- DPEYHNFHDIXMNV-UHFFFAOYSA-N (9-amino-3-bicyclo[3.3.1]nonanyl)-(4-benzyl-5-methyl-1,4-diazepan-1-yl)methanone dihydrochloride Chemical compound Cl.Cl.CC1CCN(CCN1Cc1ccccc1)C(=O)C1CC2CCCC(C1)C2N DPEYHNFHDIXMNV-UHFFFAOYSA-N 0.000 description 1
- HMVYERAUBSAVAX-UHFFFAOYSA-N 1-nitro-1-nitrosoguanidine Chemical compound NC(=N)N(N=O)[N+]([O-])=O HMVYERAUBSAVAX-UHFFFAOYSA-N 0.000 description 1
- PGZLPGBQBBTYDR-UHFFFAOYSA-N 2-hydrazinyl-1h-pteridin-4-one Chemical class C1=CN=C2C(=O)NC(NN)=NC2=N1 PGZLPGBQBBTYDR-UHFFFAOYSA-N 0.000 description 1
- OSWFIVFLDKOXQC-UHFFFAOYSA-N 4-(3-methoxyphenyl)aniline Chemical compound COC1=CC=CC(C=2C=CC(N)=CC=2)=C1 OSWFIVFLDKOXQC-UHFFFAOYSA-N 0.000 description 1
- 241000186361 Actinobacteria <class> Species 0.000 description 1
- GUBGYTABKSRVRQ-XLOQQCSPSA-N Alpha-Lactose Chemical compound O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H](CO)O[C@H](O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-XLOQQCSPSA-N 0.000 description 1
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 1
- RYGMFSIKBFXOCR-UHFFFAOYSA-N Copper Chemical compound [Cu] RYGMFSIKBFXOCR-UHFFFAOYSA-N 0.000 description 1
- DYDCUQKUCUHJBH-UHFFFAOYSA-N D-Cycloserine Natural products NC1CONC1=O DYDCUQKUCUHJBH-UHFFFAOYSA-N 0.000 description 1
- RFSUNEUAIZKAJO-VRPWFDPXSA-N D-Fructose Natural products OC[C@H]1OC(O)(CO)[C@@H](O)[C@@H]1O RFSUNEUAIZKAJO-VRPWFDPXSA-N 0.000 description 1
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 description 1
- 229920001353 Dextrin Polymers 0.000 description 1
- 239000004375 Dextrin Substances 0.000 description 1
- UNXHWFMMPAWVPI-UHFFFAOYSA-N Erythritol Natural products OCC(O)C(O)CO UNXHWFMMPAWVPI-UHFFFAOYSA-N 0.000 description 1
- RFSUNEUAIZKAJO-ARQDHWQXSA-N Fructose Chemical compound OC[C@H]1O[C@](O)(CO)[C@@H](O)[C@@H]1O RFSUNEUAIZKAJO-ARQDHWQXSA-N 0.000 description 1
- 239000001828 Gelatine Substances 0.000 description 1
- 244000068988 Glycine max Species 0.000 description 1
- 235000010469 Glycine max Nutrition 0.000 description 1
- SQUHHTBVTRBESD-UHFFFAOYSA-N Hexa-Ac-myo-Inositol Natural products CC(=O)OC1C(OC(C)=O)C(OC(C)=O)C(OC(C)=O)C(OC(C)=O)C1OC(C)=O SQUHHTBVTRBESD-UHFFFAOYSA-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
- 229920001202 Inulin Polymers 0.000 description 1
- LKDRXBCSQODPBY-AMVSKUEXSA-N L-(-)-Sorbose Chemical compound OCC1(O)OC[C@H](O)[C@@H](O)[C@@H]1O LKDRXBCSQODPBY-AMVSKUEXSA-N 0.000 description 1
- SRBFZHDQGSBBOR-HWQSCIPKSA-N L-arabinopyranose Chemical compound O[C@H]1COC(O)[C@H](O)[C@H]1O SRBFZHDQGSBBOR-HWQSCIPKSA-N 0.000 description 1
- DCXYFEDJOCDNAF-REOHCLBHSA-N L-asparagine Chemical compound OC(=O)[C@@H](N)CC(N)=O DCXYFEDJOCDNAF-REOHCLBHSA-N 0.000 description 1
- OUYCCCASQSFEME-QMMMGPOBSA-N L-tyrosine Chemical compound OC(=O)[C@@H](N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-QMMMGPOBSA-N 0.000 description 1
- GUBGYTABKSRVRQ-QKKXKWKRSA-N Lactose Natural products OC[C@H]1O[C@@H](O[C@H]2[C@H](O)[C@@H](O)C(O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@H]1O GUBGYTABKSRVRQ-QKKXKWKRSA-N 0.000 description 1
- FYYHWMGAXLPEAU-UHFFFAOYSA-N Magnesium Chemical compound [Mg] FYYHWMGAXLPEAU-UHFFFAOYSA-N 0.000 description 1
- 229930195725 Mannitol Natural products 0.000 description 1
- 241000511739 Melampyrum Species 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical class CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 description 1
- UEEJHVSXFDXPFK-UHFFFAOYSA-N N-dimethylaminoethanol Chemical compound CN(C)CCO UEEJHVSXFDXPFK-UHFFFAOYSA-N 0.000 description 1
- VZUNGTLZRAYYDE-UHFFFAOYSA-N N-methyl-N'-nitro-N-nitrosoguanidine Chemical compound O=NN(C)C(=N)N[N+]([O-])=O VZUNGTLZRAYYDE-UHFFFAOYSA-N 0.000 description 1
- 239000001888 Peptone Substances 0.000 description 1
- 108010080698 Peptones Proteins 0.000 description 1
- 244000046052 Phaseolus vulgaris Species 0.000 description 1
- 235000010627 Phaseolus vulgaris Nutrition 0.000 description 1
- WCUXLLCKKVVCTQ-UHFFFAOYSA-M Potassium chloride Chemical compound [Cl-].[K+] WCUXLLCKKVVCTQ-UHFFFAOYSA-M 0.000 description 1
- MUPFEKGTMRGPLJ-RMMQSMQOSA-N Raffinose Natural products O(C[C@H]1[C@@H](O)[C@H](O)[C@@H](O)[C@@H](O[C@@]2(CO)[C@H](O)[C@@H](O)[C@@H](CO)O2)O1)[C@@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@@H](CO)O1 MUPFEKGTMRGPLJ-RMMQSMQOSA-N 0.000 description 1
- JVWLUVNSQYXYBE-UHFFFAOYSA-N Ribitol Natural products OCC(C)C(O)C(O)CO JVWLUVNSQYXYBE-UHFFFAOYSA-N 0.000 description 1
- CZMRCDWAGMRECN-UHFFFAOYSA-N Rohrzucker Natural products OCC1OC(CO)(OC2OC(CO)C(O)C(O)C2O)C(O)C1O CZMRCDWAGMRECN-UHFFFAOYSA-N 0.000 description 1
- 241001558929 Sclerotium <basidiomycota> Species 0.000 description 1
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 1
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 1
- 244000062793 Sorghum vulgare Species 0.000 description 1
- 241000334075 Streptomyces rimofaciens Species 0.000 description 1
- HDTRYLNUVZCQOY-WSWWMNSNSA-N Trehalose Natural products O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-WSWWMNSNSA-N 0.000 description 1
- MUPFEKGTMRGPLJ-UHFFFAOYSA-N UNPD196149 Natural products OC1C(O)C(CO)OC1(CO)OC1C(O)C(O)C(O)C(COC2C(C(O)C(O)C(CO)O2)O)O1 MUPFEKGTMRGPLJ-UHFFFAOYSA-N 0.000 description 1
- HCHKCACWOHOZIP-UHFFFAOYSA-N Zinc Chemical compound [Zn] HCHKCACWOHOZIP-UHFFFAOYSA-N 0.000 description 1
- 239000011149 active material Substances 0.000 description 1
- 239000003463 adsorbent Substances 0.000 description 1
- HDTRYLNUVZCQOY-LIZSDCNHSA-N alpha,alpha-trehalose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-LIZSDCNHSA-N 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
- 150000001408 amides Chemical class 0.000 description 1
- 229940024606 amino acid Drugs 0.000 description 1
- 235000001014 amino acid Nutrition 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 150000003863 ammonium salts Chemical class 0.000 description 1
- SRBFZHDQGSBBOR-UHFFFAOYSA-N beta-D-Pyranose-Lyxose Natural products OC1COC(O)C(O)C1O SRBFZHDQGSBBOR-UHFFFAOYSA-N 0.000 description 1
- DLRVVLDZNNYCBX-ZZFZYMBESA-N beta-melibiose Chemical compound O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@@H]1OC[C@@H]1[C@@H](O)[C@H](O)[C@@H](O)[C@H](O)O1 DLRVVLDZNNYCBX-ZZFZYMBESA-N 0.000 description 1
- 230000003570 biosynthesizing effect Effects 0.000 description 1
- 230000001851 biosynthetic effect Effects 0.000 description 1
- 239000011575 calcium Substances 0.000 description 1
- 229910052791 calcium Inorganic materials 0.000 description 1
- 239000004202 carbamide Substances 0.000 description 1
- 239000000919 ceramic Substances 0.000 description 1
- 229920001429 chelating resin Polymers 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 238000005345 coagulation Methods 0.000 description 1
- 230000015271 coagulation Effects 0.000 description 1
- 229910017052 cobalt Inorganic materials 0.000 description 1
- 239000010941 cobalt Substances 0.000 description 1
- GUTLYIVDDKVIGB-UHFFFAOYSA-N cobalt atom Chemical compound [Co] GUTLYIVDDKVIGB-UHFFFAOYSA-N 0.000 description 1
- 230000002508 compound effect Effects 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 229910052802 copper Inorganic materials 0.000 description 1
- 239000010949 copper Substances 0.000 description 1
- 239000007799 cork Substances 0.000 description 1
- 238000005336 cracking Methods 0.000 description 1
- 238000013016 damping Methods 0.000 description 1
- 235000019425 dextrin Nutrition 0.000 description 1
- 238000004090 dissolution Methods 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 238000006911 enzymatic reaction Methods 0.000 description 1
- UNXHWFMMPAWVPI-ZXZARUISSA-N erythritol Chemical compound OC[C@H](O)[C@H](O)CO UNXHWFMMPAWVPI-ZXZARUISSA-N 0.000 description 1
- XHCADAYNFIFUHF-TVKJYDDYSA-N esculin Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1OC(C(=C1)O)=CC2=C1OC(=O)C=C2 XHCADAYNFIFUHF-TVKJYDDYSA-N 0.000 description 1
- 230000002349 favourable effect Effects 0.000 description 1
- 210000003495 flagella Anatomy 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 235000019253 formic acid Nutrition 0.000 description 1
- 235000011389 fruit/vegetable juice Nutrition 0.000 description 1
- FBPFZTCFMRRESA-GUCUJZIJSA-N galactitol Chemical compound OC[C@H](O)[C@@H](O)[C@@H](O)[C@H](O)CO FBPFZTCFMRRESA-GUCUJZIJSA-N 0.000 description 1
- 239000000499 gel Substances 0.000 description 1
- 229920000159 gelatin Polymers 0.000 description 1
- 235000019322 gelatine Nutrition 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 229960005150 glycerol Drugs 0.000 description 1
- 235000013882 gravy Nutrition 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 239000003112 inhibitor Substances 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 229910001959 inorganic nitrate Inorganic materials 0.000 description 1
- 229960000367 inositol Drugs 0.000 description 1
- CDAISMWEOUEBRE-GPIVLXJGSA-N inositol Chemical compound O[C@H]1[C@H](O)[C@@H](O)[C@H](O)[C@H](O)[C@@H]1O CDAISMWEOUEBRE-GPIVLXJGSA-N 0.000 description 1
- JYJIGFIDKWBXDU-MNNPPOADSA-N inulin Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)OC[C@]1(OC[C@]2(OC[C@]3(OC[C@]4(OC[C@]5(OC[C@]6(OC[C@]7(OC[C@]8(OC[C@]9(OC[C@]%10(OC[C@]%11(OC[C@]%12(OC[C@]%13(OC[C@]%14(OC[C@]%15(OC[C@]%16(OC[C@]%17(OC[C@]%18(OC[C@]%19(OC[C@]%20(OC[C@]%21(OC[C@]%22(OC[C@]%23(OC[C@]%24(OC[C@]%25(OC[C@]%26(OC[C@]%27(OC[C@]%28(OC[C@]%29(OC[C@]%30(OC[C@]%31(OC[C@]%32(OC[C@]%33(OC[C@]%34(OC[C@]%35(OC[C@]%36(O[C@@H]%37[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O%37)O)[C@H]([C@H](O)[C@@H](CO)O%36)O)[C@H]([C@H](O)[C@@H](CO)O%35)O)[C@H]([C@H](O)[C@@H](CO)O%34)O)[C@H]([C@H](O)[C@@H](CO)O%33)O)[C@H]([C@H](O)[C@@H](CO)O%32)O)[C@H]([C@H](O)[C@@H](CO)O%31)O)[C@H]([C@H](O)[C@@H](CO)O%30)O)[C@H]([C@H](O)[C@@H](CO)O%29)O)[C@H]([C@H](O)[C@@H](CO)O%28)O)[C@H]([C@H](O)[C@@H](CO)O%27)O)[C@H]([C@H](O)[C@@H](CO)O%26)O)[C@H]([C@H](O)[C@@H](CO)O%25)O)[C@H]([C@H](O)[C@@H](CO)O%24)O)[C@H]([C@H](O)[C@@H](CO)O%23)O)[C@H]([C@H](O)[C@@H](CO)O%22)O)[C@H]([C@H](O)[C@@H](CO)O%21)O)[C@H]([C@H](O)[C@@H](CO)O%20)O)[C@H]([C@H](O)[C@@H](CO)O%19)O)[C@H]([C@H](O)[C@@H](CO)O%18)O)[C@H]([C@H](O)[C@@H](CO)O%17)O)[C@H]([C@H](O)[C@@H](CO)O%16)O)[C@H]([C@H](O)[C@@H](CO)O%15)O)[C@H]([C@H](O)[C@@H](CO)O%14)O)[C@H]([C@H](O)[C@@H](CO)O%13)O)[C@H]([C@H](O)[C@@H](CO)O%12)O)[C@H]([C@H](O)[C@@H](CO)O%11)O)[C@H]([C@H](O)[C@@H](CO)O%10)O)[C@H]([C@H](O)[C@@H](CO)O9)O)[C@H]([C@H](O)[C@@H](CO)O8)O)[C@H]([C@H](O)[C@@H](CO)O7)O)[C@H]([C@H](O)[C@@H](CO)O6)O)[C@H]([C@H](O)[C@@H](CO)O5)O)[C@H]([C@H](O)[C@@H](CO)O4)O)[C@H]([C@H](O)[C@@H](CO)O3)O)[C@H]([C@H](O)[C@@H](CO)O2)O)[C@@H](O)[C@H](O)[C@@H](CO)O1 JYJIGFIDKWBXDU-MNNPPOADSA-N 0.000 description 1
- 229940029339 inulin Drugs 0.000 description 1
- 239000008101 lactose Substances 0.000 description 1
- 239000007791 liquid phase Substances 0.000 description 1
- 239000011777 magnesium Substances 0.000 description 1
- 229910052749 magnesium Inorganic materials 0.000 description 1
- WPBNNNQJVZRUHP-UHFFFAOYSA-L manganese(2+);methyl n-[[2-(methoxycarbonylcarbamothioylamino)phenyl]carbamothioyl]carbamate;n-[2-(sulfidocarbothioylamino)ethyl]carbamodithioate Chemical compound [Mn+2].[S-]C(=S)NCCNC([S-])=S.COC(=O)NC(=S)NC1=CC=CC=C1NC(=S)NC(=O)OC WPBNNNQJVZRUHP-UHFFFAOYSA-L 0.000 description 1
- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 description 1
- 239000000594 mannitol Substances 0.000 description 1
- 235000010355 mannitol Nutrition 0.000 description 1
- 235000013372 meat Nutrition 0.000 description 1
- HEBKCHPVOIAQTA-UHFFFAOYSA-N meso ribitol Natural products OCC(O)C(O)C(O)CO HEBKCHPVOIAQTA-UHFFFAOYSA-N 0.000 description 1
- 239000002207 metabolite Substances 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 235000019713 millet Nutrition 0.000 description 1
- 235000013379 molasses Nutrition 0.000 description 1
- 239000002808 molecular sieve Substances 0.000 description 1
- 239000000178 monomer Substances 0.000 description 1
- 230000000877 morphologic effect Effects 0.000 description 1
- ZIUHHBKFKCYYJD-UHFFFAOYSA-N n,n'-methylenebisacrylamide Chemical compound C=CC(=O)NCNC(=O)C=C ZIUHHBKFKCYYJD-UHFFFAOYSA-N 0.000 description 1
- YPHQUSNPXDGUHL-UHFFFAOYSA-N n-methylprop-2-enamide Chemical compound CNC(=O)C=C YPHQUSNPXDGUHL-UHFFFAOYSA-N 0.000 description 1
- 150000007523 nucleic acids Chemical class 0.000 description 1
- 102000039446 nucleic acids Human genes 0.000 description 1
- 108020004707 nucleic acids Proteins 0.000 description 1
- 150000007524 organic acids Chemical class 0.000 description 1
- 239000003960 organic solvent Substances 0.000 description 1
- 235000019319 peptone Nutrition 0.000 description 1
- 230000001766 physiological effect Effects 0.000 description 1
- 239000000049 pigment Substances 0.000 description 1
- 239000002243 precursor Substances 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- OVARTBFNCCXQKS-UHFFFAOYSA-N propan-2-one;hydrate Chemical class O.CC(C)=O OVARTBFNCCXQKS-UHFFFAOYSA-N 0.000 description 1
- 235000018102 proteins Nutrition 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 238000012207 quantitative assay Methods 0.000 description 1
- 150000003242 quaternary ammonium salts Chemical class 0.000 description 1
- MUPFEKGTMRGPLJ-ZQSKZDJDSA-N raffinose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO[C@@H]2[C@@H]([C@@H](O)[C@@H](O)[C@@H](CO)O2)O)O1 MUPFEKGTMRGPLJ-ZQSKZDJDSA-N 0.000 description 1
- 229920005989 resin Polymers 0.000 description 1
- 239000011347 resin Substances 0.000 description 1
- HEBKCHPVOIAQTA-ZXFHETKHSA-N ribitol Chemical compound OC[C@H](O)[C@H](O)[C@H](O)CO HEBKCHPVOIAQTA-ZXFHETKHSA-N 0.000 description 1
- CQRYARSYNCAZFO-UHFFFAOYSA-N salicyl alcohol Chemical compound OCC1=CC=CC=C1O CQRYARSYNCAZFO-UHFFFAOYSA-N 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- CDAISMWEOUEBRE-UHFFFAOYSA-N scyllo-inosotol Natural products OC1C(O)C(O)C(O)C(O)C1O CDAISMWEOUEBRE-UHFFFAOYSA-N 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 150000003355 serines Chemical class 0.000 description 1
- 229940099992 seromycin Drugs 0.000 description 1
- 239000000741 silica gel Substances 0.000 description 1
- 229910002027 silica gel Inorganic materials 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 229910052708 sodium Inorganic materials 0.000 description 1
- 239000001632 sodium acetate Substances 0.000 description 1
- 235000017281 sodium acetate Nutrition 0.000 description 1
- 229960004249 sodium acetate Drugs 0.000 description 1
- URGAHOPLAPQHLN-UHFFFAOYSA-N sodium aluminosilicate Chemical compound [Na+].[Al+3].[O-][Si]([O-])=O.[O-][Si]([O-])=O URGAHOPLAPQHLN-UHFFFAOYSA-N 0.000 description 1
- 239000001509 sodium citrate Substances 0.000 description 1
- 229940074404 sodium succinate Drugs 0.000 description 1
- ZDQYSKICYIVCPN-UHFFFAOYSA-L sodium succinate (anhydrous) Chemical compound [Na+].[Na+].[O-]C(=O)CCC([O-])=O ZDQYSKICYIVCPN-UHFFFAOYSA-L 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 239000002594 sorbent Substances 0.000 description 1
- 238000001179 sorption measurement Methods 0.000 description 1
- 229940032147 starch Drugs 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 235000000346 sugar Nutrition 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- HRXKRNGNAMMEHJ-UHFFFAOYSA-K trisodium citrate Chemical compound [Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O HRXKRNGNAMMEHJ-UHFFFAOYSA-K 0.000 description 1
- 229940038773 trisodium citrate Drugs 0.000 description 1
- OUYCCCASQSFEME-UHFFFAOYSA-N tyrosine Natural products OC(=O)C(N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-UHFFFAOYSA-N 0.000 description 1
- 238000009423 ventilation Methods 0.000 description 1
- 229940088594 vitamin Drugs 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
- 229930003231 vitamin Natural products 0.000 description 1
- 150000003722 vitamin derivatives Chemical class 0.000 description 1
- 229960003487 xylose Drugs 0.000 description 1
- 239000011701 zinc Substances 0.000 description 1
- 229910052725 zinc Inorganic materials 0.000 description 1
Landscapes
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The mildew, which is used as an antibacterial agent, bactericide or acaricide in agriculture and horticulture, can be produced industrially conveniently by culturing a mildew-producing strain belonging to the genus Streptoverticillium and resistant to serine hydroxamate or canavanine in a culture medium. The culture medium preferably contains at least 3 millimoles of N-methyl compounds and/or 5-hydroxymethylcytosine to promote synthesis of the strain and accumulation of the mycomycin, and the mycomycin is recovered from the culture.
Description
The present invention is relevant with the fermentation process of producing mildiomycin.
Mildiomycin is one to have the compound of structural formula (I); Belong to streptoverticillium by cultivation
Belong to the bacterial strain that produces mildiomycin, it is found and called after " antibiotic B-98891 " first as antibiotic.We concentrate on attention on its activity, and particularly it is as a kind of prevention of various plants and treatment Powdery Mildew medicine or as acaricidal effect.Therefore, wish to improve the output of mildiomycin, be included in and contain on 3 mmoles (mmole/liter) the N-methyl compound substratum at least or contain the fermentation process of cultivating the bacterial strain that produces mildiomycin on the substratum of 5-hydroxymethyl cytosine.(U.S.P.4007267; U.S.P.4334022; U.S.P.4296107; British Patent No. 1507193; " antibiotic magazine " 31 volumes, 6 phase 511-518 pages or leaves, 519-524 page or leaf (1978); " sterilant science magazine " 43 phases of volume, 349-353 page or leaf (1979); " tetrahedron chemistry " 37 volumes, 1317-1327 page or leaf (1981); Or the like).
Yet, the fermentation process of production mildiomycin as far as is known, the ability that the product mildiomycin bacterial strain that is adopted is produced mildiomycin is limited, thereby also limit by improving the productive rate that the material that contains in the substratum improves mildiomycin, so these methods can not satisfy the needs of large-scale industrialization ground production mildiomycin.
The inventor of this invention found that through diligent in one's studies to improving bacterial strain product mildiomycin ability; When handle produces the bacterial strain of mildiomycin to having structural formula
The Serine hydroxamate of (II) or to having structural formula
When the canavanine of (III) had resistance, even if use ordinary culture medium, the ability that bacterial strain is produced mildiomycin also obtained beyond thought raising, thereby makes the production efficiency of mildiomycin become higher.According to this discovery, finished this invention.
More precisely, the invention relates to:
(1) a kind of method of producing mildiomycin, it is characterized in that belonging to streptoverticillium by on substratum, cultivating, produce the bacterial strain of mildiomycin, because Serine hydroxamate or canavanine in the anti-substratum of this bacterial strain, thereby cause the synthetic and accumulation mildiomycin of said bacterial strain, and from last culture liquid, reclaim mildiomycin.
(2) method of the described production mildiomycin of above-mentioned applying for a patent (1) is characterized in that containing at least in the substratum N-methyl compound of 3 mmoles.
(3) apply for a patent the method for (1) or (2) described production mildiomycin, it is characterized in that containing in the substratum 5-hydroxymethyl cytosine.
(4) schizogenesis streptoverticillium (Streptoverticillium rimofaciens) anti-Serine hydroxamate of energy or canavanine also can produce mildiomycin.
Prepare in the method for mildiomycin in the present invention, application be to belong to streptoverticillium, the bacterial strain (here or claim later on " bacterial strain of the present invention ") of the product mildiomycin of anti-Serine hydroxamate or canavanine characteristic is arranged.For instance, bacterial strain of the present invention has following characteristic:
(1) morphological characteristic:
On various conventional substratum, bacterial strain of the present invention forms aerial hyphae, is colyliform tool two step cone branches.The synthetic agar substratum as, though seldom also can find fibrillae of spores ring and hook-shaped fibrillae of spores on the glucose, l-asparagine agar.Spore is shaped as avette to the garden tubular, and size is 0.6~0.8 * 0.7~1.3 microns.General spore produces and is chain, contains 3 to 16, the smooth or warty of each spore surface.On conventional substratum, do not observe sporocyst, flagellum, sclerotium or the like.
(2) cultural characters:
The cultural characters of bacterial strain of the present invention is as shown in table 1.If no special instructions, description is in the result after cultivating in 21 days under 28 ℃.The color designation that shows with bracket () in the table is according to " chromatogram handbook, the 4th edition, Container Corporation of America.
(3) physiological property:
(1) growth temperature range
In 15~38 ℃ of scopes, grow, grow better and the temperature that produces aerial hyphae is 28~36 ℃.
(2) gelatine liquefication (cultivating 28 days for 24 ℃)
Do not liquefy.
(3) starch hydrolysis: the positive
(4) nitrate reduction: negative in bacterium nitrate nutrient solution (ISP-No.8) and Cha Shi nutrient solution (Czapek ' s solution).
(5) skimming milk coagulation reaction: the positive
Peptonization: the positive
(6) melanic generation:
Positive (peptone, yeast juice, iron ion nutrient agar)
Negative (tyrosine nutrient agar)
(7) utilization of carbon source (on the PridhamGottlieb nutrient agar)
ⅰ) utilize goodish carbon source:
Inositol, D-semi-lactosi, D-glucose, maltose, D-seminose, starch, glycerine, sodium-acetate, sodium succinate, Trisodium Citrate.
ⅱ) the carbon source of general using:
D-fructose, trehalose.
The carbon source of ⅲ) not utilizing:
Tetrahydroxybutane, ribitol, D-sorbyl alcohol, D-N.F,USP MANNITOL, melampyrum, D-wood sugar, L-arabinose, L-sorbose, rhamnosyl, sucrose, lactose, melibiose, raffinose, saligenin, Vitamin C2, inulin.
(8) generate at the pigment on the protein substratum: a little less than
(9) cracking on meat soup-nutrient agar: do not have
(10) to the resistance of Serine hydroxamate or canavanine:
The minimum concentration that suppresses mycelial growth; Anti-Serine hydroxamate is 40 mcg/ml at least, and anti-canavanine is 100 mcg/ml at least.
(4) other characteristic:
According to " fermentation engineering magazine " 63 volumes, 1 phase, 17-21 page or leaf (1985) is gone up the Enzymology method of describing, and the ratio vigor that produces 5-hydroxymethyl cytosine is 0.18 nanomole/minute/milligram albumen or bigger.
This bacterial strain with above-mentioned characteristic is the schizogenesis streptoverticillium, for example schizogenesis streptoverticillium C
R 3The anti-Serine hydroxamate of-182(), schizogenesis streptoverticillium CV
RAnti-Serine hydroxamate of-48(and canavanine) and schizogenesis streptoverticillium CV
RThe anti-canavanine of-16().Above-mentioned schizogenesis streptoverticillium C
R 3-182 in preservation Osaka on the 28th fermentation research institute June in 1985 (IFO), preserving number is IFO-14448, simultaneously on July 8th, 1985 it is deposited in the fermentation research institute (FRI) of industrial science and Technical Board, preserving number is FERM P-8334, schizogenesis streptoverticillium CV
ROn June 28th ,-48 1985 was deposited in IFO, and preserving number is IFO-14449, and on July 8th, 1985 was deposited in FRI, and preserving number is FERM P-8333, schizogenesis streptoverticillium CV
R-16 are deposited in IFO on July 15th, 1986, and preserving number is IFO-14527, and on July 25th, 1986 was deposited in FRI, and preserving number is FERMP-8874.
In the strain culturing of producing this method of mildiomycin, can utilize the absorbable carbon source of common microorganism, the substratum that digestible nitrogenous source and inorganic salt or the like are formed.During as needs, can add trace nutrient in the substratum, growth stimulant, precursor and other are on a small quantity with regard to active material.Common assimilable carbon source comprises, as starch, dextrin, glucose, maltose, sucrose, molasses, corn steep liquor, millet gel, and organic acid such as acetic acid, succsinic acid or the like or fat and oil or polyhydroxy-alcohol such as glycerine, these materials can be separately with or combined utilization.Digestible nitrogenous source comprises inorganic nitrate such as various ammonium salt, nitrate and urea and organic crude substance such as yeast leach liquor, casein, and gravy, cottonseed meal, corn steep liquor, bean powder or the like, they both can be singly with also uniting use.Substratum also includes metal-salts such as inorganic salt, iron, magnesium, manganese, cobalt, copper, sodium, potassium, calcium, zinc when needs.The microorganism of usefulness if necessary then should contain an amount of particular nutrient in the substratum, as amino acid, VITAMIN, nucleic acid base or the like material in addition.Be not particularly limited as long as can reach these concentration that add thing of purpose of the present invention, can from the general scope of using of conventional fermenting process, suitably select suitable concn.
In order to obtain the higher output of mildiomycin, the N-methyl compound is combined in the substratum.This N-methyl compound can be included in have in its molecule be no less than 1 can be by 1 to 4 methyl substituted nitrogen-atoms.Better be to have 1 by one or more methyl substituted those compounds in molecule, (wherein nitrogen-atoms is by 3 methyl substituted, promptly particularly to have the trimethylammonium amino group
Quaternary ammonium salt best.In addition, in substratum, have a group and can change into N-OH
3The compound of group, as, N
1N-methylene bis acrylamide also can be used as said N-methyl compound.The common molecular weight of N-methyl compound is preferably in the 90-130 scope in the 50-1000 scope.The N-methyl compound can be water-soluble or water-insoluble, but the application of easily water-soluble N-methyl compound is comparatively favourable.The example of this N-methyl compound has; N-methyl acid acid amides, N-methylamino compound, N-methylamine, N-ammonium methyl compound, N
1N-methyl bisacrylamide or the like.N-methyl acid amides has, for example: N
1The N-N,N-DIMETHYLACETAMIDE, N-methyl-acrylamide or the like, N-methylamino compound has, for example, N-methyl urea, 1,1,3,3-4-methyl urea, 2-dimethylaminoethanol or the like; The N-methylamine for example has, Trimethylamine, dimethyl amine or the like; N-ammonium methyl compound has, for example, Yelkin TTS, choline, trimethyl-glycine, tetramethyl-ammonium, or the like, above material can both advantageously use.Use N-ammonium methyl compound, particularly choline, trimethyl-glycine or tetramethyl-ammonium can obtain satisfied result.These N-methyl compounds both can use separately also can two kinds, several mix use.In above-mentioned scope, in order to make the N-methyl compound that contains in the substratum more than 3 mmoles, the present invention also comprises a kind of method, that is: in substratum, add a large amount of materials that contains the N-methyl compound, as: the beet sugar that contains trimethyl-glycine is close, contains the soya-bean milk of Yelkin TTS or contains egg of Yelkin TTS or the like.The concentration of N-methyl compound selects the superior to select in the growth scope that does not suppress microorganism used therefor in the substratum, and is adjustable to greater than 3 mmoles.Suitable concentration is that 4 mmoles are to 200 mmoles, optimal concentration is 7 to 50 mmoles, surpassing under the greater concn of 200 mmoles, adding N-methyl compound effect that is obtained and the effect that is lower than N-methyl compound acquisition under 200 millimolar concentrations relatively, tending to not obvious.Contain the selection of the crude substance consumption of N-methyl compound, make its N-methyl compound concentration reach above-mentioned requirements, but this concentration need use the natural goods quality more used than the routine cultivation big, microbial growth is subjected to the influence of composition beyond the N-methyl compound in the crude substance on the contrary thus, thereby stops the generation of mildiomycin.At this moment, preferably N-methyl compound and crude substance are united use.Consider easy to operate and validity, add the time of N-methyl compound in the substratum.Preferably before microbe inoculation is in the substratum, but the N-methyl compound also can add by the appropriate time in culturing process.
In addition, in the method for the invention,, can add 5-hydroxymethyl cytosine in the substratum or a kind ofly can in substratum, can generate the 5-hydroxymethyl cytosine material in order to obtain more mildiomycin.5-hydroxymethyl cytosine or in substratum, can generate the concentration of 5-hydroxymethyl cytosine material, what represent with 5-hydroxymethyl cytosine is 0.01~1.0%(weight/volume in right amount), the suitableeest is 0.03~0.5%(weight/volume).The time that adds these materials in the substratum is preferably in before effective cultivation begins, but also can add the suitable period in culturing process.
Though cultivate on the surface is feasible, the deep layer of ventilation is cultivated more suitable usually.When the deep layer of ventilating was cultivated, the PH of substratum suited in little acid in the scope of little alkali, and culture temperature preferably remains on 15-40 ℃, especially at 24-34 ℃.These culture condition certainly by the special bacterial strain of applied microorganism, control and select by external conditions and internal factor, to obtain satisfied result.In any case, generally beginning to cultivate the nutrient solution that the back can obtain to contain a large amount of mildiomycins in 4~14 days.In order from nutrient solution, to reclaim mildiomycin, preferably use the ordinary method of separate microorganism metabolite from nutrient solution.For example, because mildiomycin is a kind of water-soluble alkali, its major part is present in the liquid portion of nutrient solution, by filtering or centrifugal can removing cell from nutrient solution earlier.Then the liquid portion that obtains is like this added suitable sorbent material such as gac, polymeric adsorbent, Zeo-karb, activated alumina or silica gel, perhaps use molecular sieve, active part is adsorbed in the above.Then, with water miscible aqueous solutions of organic solvent such as acetone, methyl alcohol, ethanol, propyl alcohol, butanols or the like, and the aqueous solution acid, that ealkaline buffer makes the aqueous solution and inorganic or organic salt removes the wash-out active part as solvent.These separating steps also can suitably cooperate and carry out, and active part can concentrate and pulverize to obtain the mildiomycin of monomer or salt form.
Produce in the present invention and employedly in the method for mildiomycin to belong to the bacterial strain that the anti-Serine hydroxamate of streptoverticillium or canavanine produce mildiomycin, can be by the product mildiomycin bacterial strain inducing that belongs to streptoverticillium being mutated into anti-Serine hydroxamate or the canavanine bacterial strain obtains.Carry out the bacterial strain that streptoverticillium produces mildiomycin that belongs to of induced mutation, commonly used is that schizogenesis streptoverticillium E5-24-5(is called " parent strain " here or later on).This parent strain has been deposited in IFO on June 12nd, 1981, and preserving number is IFO-14125, and also has been deposited in FRI on June 24th, 1981, and preserving number is FERM P-6052.In order to cause that the bacterial strain sudden change produces the bacterial strain of anti-Serine hydroxamate or canavanine, the mutation method of using for example, application belongs to spore or the mycelia that streptoverticillium produces the mildiomycin bacterial strain and carries out uviolizing, or uses and belong to bacterial strain that streptoverticillium produces mildiomycin and have structural formula and be
The N-methyl N of (IV) '-method of nitro-N-nitrosoguanidine contact, and containing its structural formula belonging to the strain growth that streptoverticillium produces mildiomycin
Method on the substratum of the D-4-amino-3-isoxazolidinone of (V) (popular name " seromycin ") perhaps suitably is used in combination these methods.
For example, when taking to produce the spore of mildiomycin bacterial strain or mycelia when suddenling change with the method for uviolizing with belonging to streptoverticillium, the spore of parent strain or mycelia are with 10~100 watts, and preferably 30~60 watts ultraviolet lamp carries out uviolizing.Normally 130~350 millimicrons of ultraviolet wavelengths are preferably 190~290 millimicrons.Common 30 seconds to 5 minutes of irradiation time is preferably 60 seconds to 90 seconds.Shine again after should being suspended in the spore of parent strain and mycelia in the sterilized water.For example, the spore of parent strain is suspended in the sterilized water, puts 30 watts of ultraviolet lamps below 30 centimeters, stir 90 minutes (wavelength 2537 of irradiation down
) with mutagenesis.
When adopting a bacterial strain that belongs to streptoverticillium product mildiomycin to suddenly change, preferably the spore of bacterial strain or individual cells are contacted with compound (IV) with the method that N-methyl-N '-nitro-N-nitrosoguanidine (IV) contacts.Contact available any mode and carry out, normally with mixing mutually.Temperature range when contacting is preferably 10 ℃ to 40 ℃ from 0 ℃ to 50 ℃, and the required time scope is often from 10 seconds to 3 hours, preferably 10 minutes to 2 hours.Produce the spore or unicellular being dispersed in the suitable dispersion substratum of mildiomycin bacterial strain, this substratum by, 1 to the 9%(weight/volume) sucrose, 1 is to the 6%(weight/volume) Sodium Glutamate, 0.001~0.1%(weight/volume) tween-80 is (by Kao Atlas Co., Ltd. produce) and sterilized water composition, substratum is in advance 110~130 ℃ of high pressure steam sterilizations 10~30 minutes.Compound (IV) may be dissolved in back use in damping fluid such as the Trisaminomethane-hydrochloride buffer (pH7.5), and its concentration is 1.0 to 5000 mcg/ml, is preferably 100 to 200 mcg/ml.In addition, allow in employing and to belong to streptoverticillium and produce the mildiomycin strain when being grown in method on the substratum that contains ring-type Serine (V) and suddenling change, be on the substratum of 10~100 mcg/ml producing the mildiomycin strain culturing containing ring-type Serine (V) amount preferably, the suitableeest is 20~60 mcg/ml.Other culture condition can suitably be selected from the aforesaid culture condition relevant with cultivating anti-Serine hydroxamate or canavanine product mildiomycin bacterial strain.
From the mutant that the method with above-mentioned mutagenesis obtains, select anti-Serine hydroxamate or canavanine to produce the bacterial strain of mildiomycin.Also can select according to normally used screening method in actinomycetes field.For example handle the growth bacterial strain coating that obtains or culture transferring to containing Serine hydroxamate (40 to 200 mcg/ml spore suspension, spore mixed solution or through sudden change, be preferably 60 to 120 mcg/ml) or canavanine (100 to 300 mcg/ml, be preferably 110 to 240 mcg/ml) amount be enough to stop on the substratum of parent strain growth, then, separate the bacterial strain that is grown on the substratum.Perhaps, suspension, mixed solution or the growth bacterial strain of handling acquisition through suddenling change are coated on and contain the biosynthetic inhibitor of 5-hydroxymethyl cytosine-amino butterfly cry of certain animals (2 to 50 mcg/ml, be preferably 5 to 20 mcg/ml) substratum on, allow strain growth then, then downcut an agar, measure antibiotic vigor (the mensuration bacterium: red yeast) of this piece agar with ordinary method, screen those and show antibiotic energetic bacterial strain, thereby determine to filter out the good product mildiomycin bacterial strain of Serine hydroxamate resistance.Produce anti-Serine hydroxamate of mildiomycin bacterial strain or canavanine salt method by repeating above-mentioned acquisition, the performance that anti-Serine hydroxamate or canavanine produce the mildiomycin bacterial strain is further enhanced.To the Serine hydroxamate or to the resistant mutation of canavanine, produce the resistance that the mildiomycin bacterial strain also can obtain anti-Serine hydroxamate and canavanine simultaneously through above-mentioned product mildiomycin bacterial strain.These bacterial strains are also included within the type that the invention belongs to anti-Serine hydroxamate of streptoverticillium or canavanine bacterial strain.
By the method for the invention described above, the mildiomycin that makes is used as the effective ingredient that prevents and treat the Powdery Mildew preparation on the plant safely, or as sterilization or acaricidal effective ingredient.(U.S.P.4007267, British Patent No. 1507193)
Belong to streptoverticillium among the present invention, the product mildiomycin bacterial strain of anti-Serine hydroxamate or canavanine except having Serine hydroxamate or the high resistance of canavanine, has high vigor in the 5-hydroxymethyl cytosine biosynthesizing.
Test examples 1
By 2% sucrose, 0.5% Zulkovsky starch, 0.12% ammonium nitrate, the Serine hydroxamate and the canavanine of the various different concns of adding in the nutrient agar (pH7.0) that 0.25% potassium primary phosphate, 0.005% sal epsom, 0.0025% casamino acids, 0.0025% yeast leach liquor and 2.0% agar powder (% refers to weight/volume) are formed.The substratum that is mixed with like this, is poured in the culture dish after 15 minutes through 120 ℃ of high pressure steam sterilizations, 20 milliliters in every ware.After the cooling, the spore of the following test strain of coating on every ware substratum.Culture dish is placed on 28 ℃ and cultivated 10 days down, has determined that the promptly minimum inhibition growth concentration of MIC is as shown in table 2.
Test strain:
(1) schizogenesis streptoverticillium E5-24-5
(2) schizogenesis streptoverticillium C
R 3-182
(3) schizogenesis streptoverticillium CV
R-48
(4) schizogenesis streptoverticillium CV
R-16
Table 2
The minimum growth concentration (mcg/ml) that suppresses of bacterial strain
Serine hydroxamate canavanine
(1) 40 100
(2) 70 120
(3) 100 200
(4) 40 400
Test examples 2
Preparation is by 2.5% glucose, 2.0% corn steep liquor, 1.0% cottonseed meal (trade(brand)name: Proflo, Trader Oil Mill Co.), the substratum formed of 0.3% ammonium sulfate, 0.05% sal epsom and 0.3% lime carbonate (% refers to weight/volume), to wherein dripping the 20%(weight/volume) the caustic soda aqueous solution is 7.0 to regulate pH.The substratum that makes is like this poured in 200 milliliters of triangular flasks (using the urethanum bottle stopper), and 25 milliliters every bottle, then 120 ℃ of high pressure steam sterilizations 15 minutes.Inoculate schizogenesis streptoverticillium E5-24-5 on these substratum respectively, C
R 3-182, CV
R-48 and CV
RThe spore of-16 bacterial classifications, shaking by swirling is cultivated 20 hours to obtain seed culture fluid under 28 ℃ of 200rpm.Prepare the substratum of forming by 11% glucose, 3.0% sorbyl alcohol, 3.0% casein, 0.5% corn steep liquor, 20%Preflo, 0.5% ammonium sulfate, 0.5% ammonium nitrate, 0.1% choline chloride 60,0.005% sal epsom, 0.005% ferric sulfate and 0.003% copper sulfate (% refers to weight/volume) then, to wherein dripping the 20%(weight/volume) the caustic soda aqueous solution is 7.0 to regulate pH.Adding lime carbonate in the substratum makes its concentration reach the 1%(weight/volume) then thorough mixing.The substratum that makes is like this poured in 200 milliliters of triangular flasks (using the urethanum bottle stopper), and 25 milliliters every bottle, thereupon 120 ℃ of high pressure steam sterilizations 20 minutes.Add 2 milliliters of above-mentioned seed culture fluids in every flask culture base (25 milliliters), changeed wave and culture 3~5 days with 200 at 28 ℃.Then, prepare cell-free extract, carry out the mensuration of enzyme reaction and 5-hydroxymethyl cytosine, obtain the vigor of result's product 5-hydroxymethyl cytosine as shown in table 3 according to 17 pages of described steps of (1985) " fermentation industry monthly magazine " 63 volumes.
Table 3
Bacterial strain produces the vigor of 5-hydroxymethyl cytosine
*
(nanomole/minute/milligram albumen)
E5-24-5 0.17
C
R 3-182 0.24
CV
R-48 0.27
CV
R-16 0.19
*3 days, 4 days, the mean value after the cultivation in 5 days.
Example 1
(1) at schizogenesis streptoverticillium E5-24-5(FERM P-6052, IFO-14125) spore and aerial hyphae are collected in the front on the slant medium, and are placed on the dispersion substratum of being made up of 3% sucrose, 2% Sodium Glutamate and 0.01% tween-80 (through 120 ℃ of high pressure steam sterilizations 15 minutes).Mixed solution fully grinds with ceramic homogenizer, and it is spore and aerial hyphae suspension of debris liquid (25 milliliters) that lapping liquid obtains filtrate with No. 101 filter paper of Toyo (by Toyo Filter Paper Co., Ltd. produces) filtration.
(2) in the suspension that makes by (1), add isopyknic be dissolved in Trisaminomethane-hydrochloride buffer (0.1M, pH7.5) the N-methyl N in '-nitro-N-nitrosoguanidine, make its concentration reach 2 mg/ml.Mixed solution shook 90 minutes at 28 ℃.The mixed solution that obtains like this is coated on contains on the Agar Plating (pH7.0) that concentration is 30 mcg/ml ring-type Serines, 28 ℃ of static cultivations 14 days, other compositions of this plate culture medium were: 2% sucrose, 0.5% Zulkovsky starch, 0.2% ammonium nitrate, 0.1% potassium primary phosphate, 0.05% Repone K, 0.05% sal epsom, 0.0015% ferric sulfate and 2.0% agar powder (% refers to weight/volume).In the bacterium colony that grows, isolate the bacterium colony that 100 forms normally have aerial hyphae.These bacterium colonies are used aseptic Glass rod grinding respectively, and each is ground adds 10 ml sterile waters in the bacterium colony, get filtrate with No. 101 filter paper filterings of Toyo then.Filtrate being coated on do not contained on the Agar Plating of ring-type Serine 28 ℃ of static cultivations 10 days.The bacterium colony that obtains is transferred to (the substratum composition is identical with above-mentioned Agar Plating composition) on the agar slant respectively.28 ℃ of static cultivations 10 days.Collect aerial hyphae and spore from the shaving of slant culture primary surface, then by the suspension described in above-mentioned (1), grinding, with steps such as filter paper filterings, make 100 suspension samples.
(3) get 100 suspension samples (viable bacteria amount: about 1 * 10 that (2) step prepared
5) separate application is on the whole surface of the Agar Plating that respectively contains 10 mcg/ml aminopterins of 100 reference numerals, 28 ℃ are descended static cultivation 10 days.Utilize cork drill to get a garden column piece (6 millimeters of diameters, about 4 millimeters in height), and transfer on the substratum of its antibiotic vigor of mensuration (the test bacterium: red yeast, the nutrient agar substratum, pH7.0).The substratum of handling is like this spent the night 30 ℃ of placements.Wherein there are 5 suspension samples to show bigger (diameter: 14 millimeters or bigger) inhibition zone (suppressing the growth circle).
It is 1 * 10 that 100 suspension samples that (4) (2) steps obtained make each sample contain viable count with the sterilized water dilution respectively
5/ milliliter.The suspension that each sample is got after the dilution of 20 microlitres is coated on the corresponding encoded Agar Plating that respectively contains 40 mg/ml Serine hydroxamates, and 28 ℃ are descended static cultivation 14 days.In 100 samples, observe the growth on 7 substratum, wherein there are 5 samples all to show the as above big like that inhibition zone in (3) step.In 5 samples, there is a mutant strain to form as above described maximum inhibition zone of (3) step, is designated as schizogenesis streptoverticillium C after the separation
R 1-18.
Example 2
With the C that obtains in the example 1
R 1-18 bacterial strains replace schizogenesis streptoverticillium E5-24-5, and the same procedure that goes on foot by (1) and (2) of example 1 prepares 150 suspension samples of this bacterial strain.According in the example 1 (3) and (4) one step process, from these samples, isolate schizogenesis streptoverticillium C
R 2-125 (as long as use contains the Agar Plating that concentration is the Serine hydroxamate of 50 mg/ml in (4) step)
Example 3
The C that makes with example 2
R 2-125 bacterial strains replace schizogenesis streptoverticillium E5-24-5, prepare 200 suspension samples of this bacterial strain by (1) and (2) in the example 1 step same procedure.From these samples, isolate schizogenesis streptoverticillium C by (3) in the example 1 and (4) one step process
R 3-182(FERM P-8334, IFO-14448) (as long as use contains the Agar Plating that concentration is 6.0 mg/ml Serine hydroxamates in (4) step).
Example 4
C with example 3 acquisitions
R 3-182 bacterial strains replace schizogenesis streptoverticillium E5-24-5, and (1) and (2) the step same procedure of pressing in the example 1 prepares 300 suspension samples from this bacterial strain.From these samples, isolate schizogenesis streptoverticillium C by (3) in the example 1 and (4) one step process
R 4-257 (as long as use contains the Agar Plating that concentration is 80 mcg/ml Serine hydroxamates in (4) step).
Example 5
C with example 4 acquisitions
R 4-257 bacterial strains replace schizogenesis streptoverticillium E5-24-5, and (1) and (2) the step same procedure of pressing in the example 1 prepares 100 suspension samples from this bacterial strain.Each suspension sample is coated on contains on the Agar Plating that concentration is 190 mg/ml canavanine.100 samples were 28 ℃ of following static cultivations 14 days.The bacterium colony that occurs in sample only moved receive by 2.0% base sugar, on the substratum that 0.5% Zulkovsky starch, 0.12% ammonium nitrate, 0.25% potassium primary phosphate, 0.005% sal epsom, 0.0025% casamino acids, 0.0025% yeast leach liquor and 2.0% agar (% refers to weight/volume) are formed (pH7.0), the mutant strain CV that static cultivation acquisition in 10 days has anti-Serine hydroxamate and canavanine characteristic under 28 ℃
R-48(FERM P-8333 IFO-14449).
Example 6
At composition by 2.5% glucose, 2% corn steep liquor, 1.0% cottonseed meal (trade(brand)name: Proflo is produced by Trader Oil Mill Co.).Dropping 20%(weight/volume in the substratum that 0.3% ammonium sulfate, 0.05% sal epsom and 0.3% lime carbonate (% refers to weight/volume) are formed) the caustic soda aqueous solution makes pH reach 7.0.Add 25 milliliters of substratum in 200 milliliters of triangular flasks (using the urethanum bottle stopper), 120 ℃ of following high pressure steam sterilizations 15 minutes.Schizogenesis streptoverticillium E5-24-5, C
R 3-182 and CV
RThe spore of-48 bacterial strains is inoculated on the substratum respectively, and at 28 ℃, 200 commentaries on classics rotating speed were down cultivated 20 hours down, obtained each required seed culture fluid.Then, preparation contains the 11%(weight/volume) substratum of glucose, 3.0% sorbyl alcohol, 3.0% casein food grade, 0.5% corn steep liquor, 2.0%Proflo, 0.5% ammonium sulfate, 0.5% ammonium nitrate, 0.1% choline chloride 60,0.005% manganous sulfate, 0.005% ferric sulfate and 0.003% copper sulfate (% refers to weight/volume), drip the 20%(weight/volume therein) the caustic soda aqueous solution so that pH reaches 7.0.Adding lime carbonate in this substratum makes its concentration reach the 1%(weight/volume), mixed solution is fully shaken, then in each 200 milliliters of triangular flask (using the urethanum bottle stopper), add 25 milliliters of substratum, made fermention medium in 20 minutes in 120 ℃ of high pressure steam sterilizations.Insert 2 milliliters of above-mentioned seed culture fluids in every bottle of substratum that makes like this.Under 28 ℃ of 200 commentaries on classics, shook fermentation culture 10 days.After having cultivated, go up the efficient liquid-phase chromatography method quantitative assay mildiomycin of describing according to " fermentation engineering magazine " 62 volumes 537 pages (1984).The mildiomycin production test of carrying out as stated above, gained test-results such as table 4 are listed.
Table 4
Bacterial strain produces the quantity (mcg/ml) of mildiomycin
E5-24-5 3887
C
R 3-182 5755
CV
R-48 8000
Example 7
Under the condition identical, schizogenesis streptoverticillium CV with example 6
R-48 ferment.Collect fermented liquid, the supernatant liquor of 1 liter of fermented liquid, then by lifting away from sub-exchange resin Amberlite IRC-50(H by 2.5 through centrifugal acquisition
+Type) (Rohm ﹠amp; Haas Co.) post of dressing up.Post washes with water, uses 2.5 liters of 0.5%(weight/volume again) ammoniacal liquor carries out wash-out.Active part (by the method for quantitatively determining of example 6) allows it flow in the post of being dressed up by 250 milliliters of chromatographies gac (by Takeda Chemical Industries, Ltd. produces), makes it absorption.With 1.25 liters of acetone-waters (3: 7) mixed solution wash-out.The active part of collection and concentrated wash-out.Dropwise 5 00 ml methanol is impelled thick mildiomycin precipitation in concentrated solution.Filter and collect powdery mildiomycin raw product.With the such powder of water dissolution 6.5 grams, and it is flow through by 125 milliliters of Amberlite CG-50(H
+Type) (by Rohm ﹠amp; Haas Co. production) post of dressing up.Post cleans with 50 ml waters, then with 1.25 liters of 0.5%(weight/volume) the ammoniacal liquor wash-out.Collect active part and go up the adsorption column of dressing up by 250 milliliters of activated carbon.Carry out branch's wash-out with 1.25 liters of acetone-0.1 equivalent concentration formic acid (2: 8), concentrate active part.Add methyl alcohol and make the mildiomycin precipitation in concentrated solution, use the centrifuging collecting precipitation, drying under reduced pressure obtains 5.5 gram mildiomycin formate.In the physico-chemical property of this formate, fusing point, specific optical rotation ((α)
24 DConcentration 1.0 in the water, concentration 1.0 in 0.1 equivalent HCl) and 271 millimicrons and 280 millimicrons locate ultraviolet absorption value and meet fully that the 523rd page of " antibiotic magazine " 1978 31 6 phase of volume are reported.
Example 8
In the suspension that uses schizogenesis streptoverticillium E5-24-5 to make by (1) step of example 1, add isopyknic (0.1 mole of Trisaminomethane-hydrochloride buffer that is dissolved in, pH7.5) the N-methyl-N ' in-nitro-N-nitroso guanidine solution makes its concentration reach 2 mg/ml.Shook 90 minutes under 28 ℃ of the mixed solutions.The above-mentioned mixed solution that obtains like this is coated on the Agar Plating that contains 300 mcg/ml L-canavanine 28 ℃ of static cultivations 14 days.The gained bacterium colony moves respectively receives (the substratum composition is identical with above-mentioned Agar Plating) on the slant agar substratum.Static cultivation obtained 14 mutant strains with anti-canavanine characteristic in 10 days under 28 ℃.
Under the condition identical with example 6, the mutant strain of these anti-canavanine and parent strain (E5-24-5) ferment, relatively the productive rate of their mildiomycins.In them, be separated to a mutant strain of performance maximum output, and volume is schizogenesis streptoverticillium CV
R-16(FERM P-8874, IFO-14527).Parent strain and mutant strain CV
R-16 to produce the amount of mildiomycin as stated above as shown in table 5.
Table 5
Bacterial strain produces the quantity (mcg/ml) of mildiomycin
E5-24-5 3900
CV
R-16 4680
Clearly find out mutant strain CV from last table
R-16 production mildiomycin abilities are more much higher than parent strain.
Example 9
Under the condition identical, schizogenesis streptoverticillium CV with example 6
R-16 ferment.Collect fermented liquid (1 liter), press example 7 identical step process fermented liquids and also separate and the purifying mildiomycin.Obtain 3.1 gram mildiomycin formate by these steps.In the physico-chemical property of this formate, the meeting fully of the ultraviolet absorption value at fusing point, specific optical rotation and 271 millimicrons and 280 millimicrons places and bibliographical information (document was said in example 7).
The present invention produces the method for mildiomycin, comprise and belong to streptoverticillium has the product mildiomycin of resistance to serine hydroxamate or canavanine the cultivation of bacterial strain, dynamical production mildiomycin, therefore be one a large amount of produce mildiomycins at industrial superior fermentation process.
Errata
Errata
Claims (4)
1, produces the method for mildiomycin, be characterised in that and cultivate the bacterial strain belong to streptoverticillium and the Serine hydroxamate in the nutrient solution or canavanine are had resistance, impel the synthetic and accumulation mildiomycin of said bacterial strain, and from the culture liquid of gained, reclaim mildiomycin.
2, the method for the production mildiomycin described in claim 1, it is characterised in that the N-methyl compound that will contain at least 3 mmoles in the substratum.
3, as the method for claim (1) or the described production mildiomycin of claim (2), be characterised in that in the substratum and will contain 5-hydroxymethyl cytosine.
4, there is the schizogenesis streptoverticillium of resistance can produce mildiomycin to Serine hydroxamate or canavanine.
Applications Claiming Priority (4)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP183343/85 | 1985-08-20 | ||
JP18334385 | 1985-08-20 | ||
JP178402/86 | 1986-07-29 | ||
JP61178402A JPH0673465B2 (en) | 1985-08-20 | 1986-07-29 | Mildiomycin production method |
Publications (2)
Publication Number | Publication Date |
---|---|
CN86106265A true CN86106265A (en) | 1987-08-12 |
CN1038770C CN1038770C (en) | 1998-06-17 |
Family
ID=26498581
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN 86106265 Expired - Lifetime CN1038770C (en) | 1985-08-20 | 1986-08-20 | Method of producing mildiomycin |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN1038770C (en) |
-
1986
- 1986-08-20 CN CN 86106265 patent/CN1038770C/en not_active Expired - Lifetime
Also Published As
Publication number | Publication date |
---|---|
CN1038770C (en) | 1998-06-17 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN1052512C (en) | Fermentation | |
CN1091472A (en) | The Stereoselective reduction of ketone | |
CN101037658A (en) | Bacillus subtilis ZJB-063 and its application | |
CN1246156A (en) | Novel bacterial strains and use thereof in fermentation processes for 2-keto-L-gulonic acid production | |
CN1076965A (en) | New fungal bacterial strain and the application in production of antibiotics thereof | |
CN1415758A (en) | Technique for producing raw material of vancomycin hydrochloride | |
CN1044005C (en) | Process for production of amide compounds and microorganisms for use therein | |
CN1072262C (en) | Process for production of amide compounds using microorganism | |
CN116218690A (en) | Curvularia robusta producing cloth Lei Feide bacteria A and fermentation method thereof | |
CN1048282C (en) | Process for producing 2-keto-L-gulonic acid | |
CN1027084C (en) | Process for preparing L-sorbose | |
CN1106018A (en) | Method of producing vitamin B12 | |
CN1038770C (en) | Method of producing mildiomycin | |
CN1038669A (en) | The preparation method of L-L-Ala | |
CN1162538C (en) | Method for producing L-phenylalanine | |
CN1260004A (en) | Strain producing remarkable amount of epsilon-poly-L-Lysine and process for producing same | |
JPH06141888A (en) | Production of d-mandelic acid | |
JPH04234991A (en) | New microorganism | |
KR940004000B1 (en) | Production of mildiomycin | |
CN109312298B (en) | Thiamine miehei bacillus strain and application thereof | |
CN1022765C (en) | Preparation method of cell variety mycomycin of new antibiotic | |
CN1050407A (en) | Method with producing glucose isomerase with acidophilic streptomycete | |
CN87104545A (en) | Preparation method of difluorobenzamide | |
SU1703689A1 (en) | Strain of fungus penicillium canescens producent of fructodofuranosidase | |
CN1179793A (en) | Method for producing organic acids and amino acids |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
C06 | Publication | ||
PB01 | Publication | ||
C10 | Entry into substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
C14 | Grant of patent or utility model | ||
C17 | Cessation of patent right |