JPH0139758B2 - - Google Patents
Info
- Publication number
- JPH0139758B2 JPH0139758B2 JP23328286A JP23328286A JPH0139758B2 JP H0139758 B2 JPH0139758 B2 JP H0139758B2 JP 23328286 A JP23328286 A JP 23328286A JP 23328286 A JP23328286 A JP 23328286A JP H0139758 B2 JPH0139758 B2 JP H0139758B2
- Authority
- JP
- Japan
- Prior art keywords
- uracil
- clostridium
- culture
- produced
- days
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired
Links
- ISAKRJDGNUQOIC-UHFFFAOYSA-N Uracil Chemical compound O=C1C=CNC(=O)N1 ISAKRJDGNUQOIC-UHFFFAOYSA-N 0.000 claims description 64
- 229940035893 uracil Drugs 0.000 claims description 32
- 241000193403 Clostridium Species 0.000 claims description 15
- 230000001580 bacterial effect Effects 0.000 claims description 11
- 238000004519 manufacturing process Methods 0.000 claims description 7
- 238000012258 culturing Methods 0.000 claims description 2
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 18
- 241000894006 Bacteria Species 0.000 description 15
- 239000002609 medium Substances 0.000 description 15
- 239000007789 gas Substances 0.000 description 12
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 11
- 244000005700 microbiome Species 0.000 description 11
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 9
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 9
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 9
- 229910052799 carbon Inorganic materials 0.000 description 9
- 239000001569 carbon dioxide Substances 0.000 description 9
- 229910002092 carbon dioxide Inorganic materials 0.000 description 9
- 238000000034 method Methods 0.000 description 9
- 230000003068 static effect Effects 0.000 description 8
- 239000007640 basal medium Substances 0.000 description 7
- 238000006243 chemical reaction Methods 0.000 description 7
- 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 6
- 210000004027 cell Anatomy 0.000 description 6
- 229910052757 nitrogen Inorganic materials 0.000 description 6
- 230000001954 sterilising effect Effects 0.000 description 6
- 241000193464 Clostridium sp. Species 0.000 description 5
- 229920001817 Agar Polymers 0.000 description 4
- 239000008272 agar Substances 0.000 description 4
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 4
- 229920005549 butyl rubber Polymers 0.000 description 4
- 239000001301 oxygen Substances 0.000 description 4
- 229910052760 oxygen Inorganic materials 0.000 description 4
- 241000894007 species Species 0.000 description 4
- LYCAIKOWRPUZTN-UHFFFAOYSA-N Ethylene glycol Chemical compound OCCO LYCAIKOWRPUZTN-UHFFFAOYSA-N 0.000 description 3
- 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 3
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 3
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- 235000011054 acetic acid Nutrition 0.000 description 3
- 238000000855 fermentation Methods 0.000 description 3
- 230000004151 fermentation Effects 0.000 description 3
- 239000008103 glucose Substances 0.000 description 3
- 239000007788 liquid Substances 0.000 description 3
- 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 2
- YBJHBAHKTGYVGT-ZKWXMUAHSA-N (+)-Biotin Chemical compound N1C(=O)N[C@@H]2[C@H](CCCCC(=O)O)SC[C@@H]21 YBJHBAHKTGYVGT-ZKWXMUAHSA-N 0.000 description 2
- XUCIJNAGGSZNQT-JHSLDZJXSA-N (R)-amygdalin 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[C@@H](C#N)C=2C=CC=CC=2)O1 XUCIJNAGGSZNQT-JHSLDZJXSA-N 0.000 description 2
- NLXLAEXVIDQMFP-UHFFFAOYSA-N Ammonia chloride Chemical compound [NH4+].[Cl-] NLXLAEXVIDQMFP-UHFFFAOYSA-N 0.000 description 2
- 241000304886 Bacilli Species 0.000 description 2
- 241000186570 Clostridium kluyveri Species 0.000 description 2
- SRBFZHDQGSBBOR-IOVATXLUSA-N D-xylopyranose Chemical compound O[C@@H]1COC(O)[C@H](O)[C@H]1O SRBFZHDQGSBBOR-IOVATXLUSA-N 0.000 description 2
- 229930091371 Fructose Natural products 0.000 description 2
- 239000005715 Fructose Substances 0.000 description 2
- 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 2
- SIKJAQJRHWYJAI-UHFFFAOYSA-N Indole Chemical compound C1=CC=C2NC=CC2=C1 SIKJAQJRHWYJAI-UHFFFAOYSA-N 0.000 description 2
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 2
- 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 2
- 241000193448 Ruminiclostridium thermocellum Species 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 229920002472 Starch Polymers 0.000 description 2
- 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 2
- 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 2
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 description 2
- 241001147786 [Clostridium] cellobioparum Species 0.000 description 2
- 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 2
- 229940089837 amygdalin Drugs 0.000 description 2
- YZLOSXFCSIDECK-UHFFFAOYSA-N amygdalin Natural products OCC1OC(OCC2OC(O)C(O)C(O)C2O)C(O)C(O)C1OC(C#N)c3ccccc3 YZLOSXFCSIDECK-UHFFFAOYSA-N 0.000 description 2
- PYMYPHUHKUWMLA-UHFFFAOYSA-N arabinose Natural products OCC(O)C(O)C(O)C=O PYMYPHUHKUWMLA-UHFFFAOYSA-N 0.000 description 2
- SRBFZHDQGSBBOR-UHFFFAOYSA-N beta-D-Pyranose-Lyxose Natural products OC1COC(O)C(O)C1O SRBFZHDQGSBBOR-UHFFFAOYSA-N 0.000 description 2
- 229940041514 candida albicans extract Drugs 0.000 description 2
- 238000012364 cultivation method Methods 0.000 description 2
- XBDQKXXYIPTUBI-UHFFFAOYSA-N dimethylselenoniopropionate Natural products CCC(O)=O XBDQKXXYIPTUBI-UHFFFAOYSA-N 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- YGHHWSRCTPQFFC-UHFFFAOYSA-N eucalyptosin A Natural products OC1C(O)C(O)C(CO)OC1OC1C(OC(C#N)C=2C=CC=CC=2)OC(CO)C(O)C1O YGHHWSRCTPQFFC-UHFFFAOYSA-N 0.000 description 2
- 229930182830 galactose Natural products 0.000 description 2
- 239000001963 growth medium Substances 0.000 description 2
- FUZZWVXGSFPDMH-UHFFFAOYSA-N hexanoic acid Chemical group CCCCCC(O)=O FUZZWVXGSFPDMH-UHFFFAOYSA-N 0.000 description 2
- 230000007062 hydrolysis Effects 0.000 description 2
- 238000006460 hydrolysis reaction Methods 0.000 description 2
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 description 2
- 230000000813 microbial effect Effects 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- BDERNNFJNOPAEC-UHFFFAOYSA-N propan-1-ol Chemical compound CCCO BDERNNFJNOPAEC-UHFFFAOYSA-N 0.000 description 2
- 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 2
- 239000002994 raw material Substances 0.000 description 2
- 230000001603 reducing effect Effects 0.000 description 2
- 238000007789 sealing Methods 0.000 description 2
- VWDWKYIASSYTQR-UHFFFAOYSA-N sodium nitrate Chemical compound [Na+].[O-][N+]([O-])=O VWDWKYIASSYTQR-UHFFFAOYSA-N 0.000 description 2
- 239000008107 starch Substances 0.000 description 2
- 235000019698 starch Nutrition 0.000 description 2
- KDYFGRWQOYBRFD-UHFFFAOYSA-N succinic acid Chemical compound OC(=O)CCC(O)=O KDYFGRWQOYBRFD-UHFFFAOYSA-N 0.000 description 2
- 235000000346 sugar Nutrition 0.000 description 2
- 229940088594 vitamin Drugs 0.000 description 2
- 229930003231 vitamin Natural products 0.000 description 2
- 235000013343 vitamin Nutrition 0.000 description 2
- 239000011782 vitamin Substances 0.000 description 2
- 239000012138 yeast extract Substances 0.000 description 2
- OWEGMIWEEQEYGQ-UHFFFAOYSA-N 100676-05-9 Natural products OC1C(O)C(O)C(CO)OC1OCC1C(O)C(O)C(O)C(OC2C(OC(O)C(O)C2O)CO)O1 OWEGMIWEEQEYGQ-UHFFFAOYSA-N 0.000 description 1
- OAKURXIZZOAYBC-UHFFFAOYSA-N 3-oxopropanoic acid Chemical compound OC(=O)CC=O OAKURXIZZOAYBC-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
- PLXMOAALOJOTIY-FPTXNFDTSA-N Aesculin Natural products OC[C@@H]1[C@@H](O)[C@H](O)[C@@H](O)[C@H](O)[C@H]1Oc2cc3C=CC(=O)Oc3cc2O PLXMOAALOJOTIY-FPTXNFDTSA-N 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
- 241000193830 Bacillus <bacterium> Species 0.000 description 1
- BVKZGUZCCUSVTD-UHFFFAOYSA-M Bicarbonate Chemical compound OC([O-])=O BVKZGUZCCUSVTD-UHFFFAOYSA-M 0.000 description 1
- 241000186146 Brevibacterium Species 0.000 description 1
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 description 1
- BVKZGUZCCUSVTD-UHFFFAOYSA-L Carbonate Chemical compound [O-]C([O-])=O BVKZGUZCCUSVTD-UHFFFAOYSA-L 0.000 description 1
- 102000016938 Catalase Human genes 0.000 description 1
- 108010053835 Catalase Proteins 0.000 description 1
- 241001112696 Clostridia Species 0.000 description 1
- GUBGYTABKSRVRQ-CUHNMECISA-N D-Cellobiose 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)OC(O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-CUHNMECISA-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
- SHZGCJCMOBCMKK-UHFFFAOYSA-N D-mannomethylose Natural products CC1OC(O)C(O)C(O)C1O SHZGCJCMOBCMKK-UHFFFAOYSA-N 0.000 description 1
- WQZGKKKJIJFFOK-QTVWNMPRSA-N D-mannopyranose Chemical compound OC[C@H]1OC(O)[C@@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-QTVWNMPRSA-N 0.000 description 1
- QWIZNVHXZXRPDR-UHFFFAOYSA-N D-melezitose Natural products O1C(CO)C(O)C(O)C(O)C1OC1C(O)C(CO)OC1(CO)OC1OC(CO)C(O)C(O)C1O QWIZNVHXZXRPDR-UHFFFAOYSA-N 0.000 description 1
- HMFHBZSHGGEWLO-SOOFDHNKSA-N D-ribofuranose Chemical compound OC[C@H]1OC(O)[C@H](O)[C@@H]1O HMFHBZSHGGEWLO-SOOFDHNKSA-N 0.000 description 1
- 239000004386 Erythritol Substances 0.000 description 1
- UNXHWFMMPAWVPI-UHFFFAOYSA-N Erythritol Natural products OCC(O)C(O)CO UNXHWFMMPAWVPI-UHFFFAOYSA-N 0.000 description 1
- 108010010803 Gelatin Proteins 0.000 description 1
- 229920002527 Glycogen Polymers 0.000 description 1
- 238000003794 Gram staining Methods 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
- SHZGCJCMOBCMKK-JFNONXLTSA-N L-rhamnopyranose Chemical compound C[C@@H]1OC(O)[C@H](O)[C@H](O)[C@H]1O SHZGCJCMOBCMKK-JFNONXLTSA-N 0.000 description 1
- PNNNRSAQSRJVSB-UHFFFAOYSA-N L-rhamnose Natural products CC(O)C(O)C(O)C(O)C=O PNNNRSAQSRJVSB-UHFFFAOYSA-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
- GUBGYTABKSRVRQ-PICCSMPSSA-N Maltose Natural products O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@@H](CO)OC(O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-PICCSMPSSA-N 0.000 description 1
- 229930195725 Mannitol Natural products 0.000 description 1
- 241000187708 Micromonospora Species 0.000 description 1
- PYMYPHUHKUWMLA-LMVFSUKVSA-N Ribose Natural products OC[C@@H](O)[C@@H](O)[C@@H](O)C=O PYMYPHUHKUWMLA-LMVFSUKVSA-N 0.000 description 1
- 229930006000 Sucrose Natural products 0.000 description 1
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 1
- QAOWNCQODCNURD-UHFFFAOYSA-L Sulfate Chemical compound [O-]S([O-])(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-L 0.000 description 1
- 108010046334 Urease Proteins 0.000 description 1
- 238000000862 absorption spectrum Methods 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- HMFHBZSHGGEWLO-UHFFFAOYSA-N alpha-D-Furanose-Ribose Natural products OCC1OC(O)C(O)C1O HMFHBZSHGGEWLO-UHFFFAOYSA-N 0.000 description 1
- WQZGKKKJIJFFOK-PHYPRBDBSA-N alpha-D-galactose Chemical compound OC[C@H]1O[C@H](O)[C@H](O)[C@@H](O)[C@H]1O WQZGKKKJIJFFOK-PHYPRBDBSA-N 0.000 description 1
- ZEMWIYASLJTEHQ-UHFFFAOYSA-J aluminum;sodium;disulfate;dodecahydrate Chemical compound O.O.O.O.O.O.O.O.O.O.O.O.[Na+].[Al+3].[O-]S([O-])(=O)=O.[O-]S([O-])(=O)=O ZEMWIYASLJTEHQ-UHFFFAOYSA-J 0.000 description 1
- 235000019270 ammonium chloride Nutrition 0.000 description 1
- 150000003863 ammonium salts Chemical class 0.000 description 1
- PYMYPHUHKUWMLA-WDCZJNDASA-N arabinose Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)C=O PYMYPHUHKUWMLA-WDCZJNDASA-N 0.000 description 1
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 1
- GUBGYTABKSRVRQ-QUYVBRFLSA-N beta-maltose Chemical compound OC[C@H]1O[C@H](O[C@H]2[C@H](O)[C@@H](O)[C@H](O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@@H]1O GUBGYTABKSRVRQ-QUYVBRFLSA-N 0.000 description 1
- 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
- 230000015572 biosynthetic process Effects 0.000 description 1
- 235000020958 biotin Nutrition 0.000 description 1
- 239000011616 biotin Substances 0.000 description 1
- 229960002685 biotin Drugs 0.000 description 1
- KGBXLFKZBHKPEV-UHFFFAOYSA-N boric acid Chemical compound OB(O)O KGBXLFKZBHKPEV-UHFFFAOYSA-N 0.000 description 1
- 239000004327 boric acid Substances 0.000 description 1
- 235000010338 boric acid Nutrition 0.000 description 1
- 238000009395 breeding Methods 0.000 description 1
- 230000001488 breeding effect Effects 0.000 description 1
- 239000001110 calcium chloride Substances 0.000 description 1
- 229910001628 calcium chloride Inorganic materials 0.000 description 1
- 229960002713 calcium chloride Drugs 0.000 description 1
- 239000004202 carbamide Substances 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 230000015271 coagulation Effects 0.000 description 1
- 238000005345 coagulation Methods 0.000 description 1
- GVPFVAHMJGGAJG-UHFFFAOYSA-L cobalt dichloride Chemical compound [Cl-].[Cl-].[Co+2] GVPFVAHMJGGAJG-UHFFFAOYSA-L 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 229910000365 copper sulfate Inorganic materials 0.000 description 1
- 229960000355 copper sulfate Drugs 0.000 description 1
- ARUVKPQLZAKDPS-UHFFFAOYSA-L copper(II) sulfate Chemical compound [Cu+2].[O-][S+2]([O-])([O-])[O-] ARUVKPQLZAKDPS-UHFFFAOYSA-L 0.000 description 1
- 229920001971 elastomer Polymers 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
- 229940009714 erythritol Drugs 0.000 description 1
- 235000019414 erythritol Nutrition 0.000 description 1
- 210000003495 flagella Anatomy 0.000 description 1
- 235000019253 formic acid Nutrition 0.000 description 1
- 239000008273 gelatin Substances 0.000 description 1
- 229920000159 gelatin Polymers 0.000 description 1
- 235000019322 gelatine Nutrition 0.000 description 1
- 235000011852 gelatine desserts Nutrition 0.000 description 1
- 229960001031 glucose Drugs 0.000 description 1
- 235000011187 glycerol Nutrition 0.000 description 1
- 229940096919 glycogen Drugs 0.000 description 1
- PZOUSPYUWWUPPK-UHFFFAOYSA-N indole Natural products CC1=CC=CC2=C1C=CN2 PZOUSPYUWWUPPK-UHFFFAOYSA-N 0.000 description 1
- RKJUIXBNRJVNHR-UHFFFAOYSA-N indolenine Natural products C1=CC=C2CC=NC2=C1 RKJUIXBNRJVNHR-UHFFFAOYSA-N 0.000 description 1
- 239000011261 inert gas Substances 0.000 description 1
- 150000002484 inorganic compounds Chemical class 0.000 description 1
- 229910010272 inorganic material Inorganic materials 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
- 229960000367 inositol Drugs 0.000 description 1
- 229910000358 iron sulfate Inorganic materials 0.000 description 1
- BAUYGSIQEAFULO-UHFFFAOYSA-L iron(2+) sulfate (anhydrous) Chemical compound [Fe+2].[O-]S([O-])(=O)=O BAUYGSIQEAFULO-UHFFFAOYSA-L 0.000 description 1
- 239000008101 lactose Substances 0.000 description 1
- 238000004811 liquid chromatography Methods 0.000 description 1
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 1
- 235000019341 magnesium sulphate Nutrition 0.000 description 1
- 229940099596 manganese sulfate Drugs 0.000 description 1
- 239000011702 manganese sulphate Substances 0.000 description 1
- 235000007079 manganese sulphate Nutrition 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
- 229940041290 mannose Drugs 0.000 description 1
- 238000004949 mass spectrometry Methods 0.000 description 1
- 239000013028 medium composition Substances 0.000 description 1
- QWIZNVHXZXRPDR-WSCXOGSTSA-N melezitose Chemical compound O([C@@]1(O[C@@H]([C@H]([C@@H]1O[C@@H]1[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O1)O)O)CO)CO)[C@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O QWIZNVHXZXRPDR-WSCXOGSTSA-N 0.000 description 1
- 238000005374 membrane filtration Methods 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
- 235000013336 milk Nutrition 0.000 description 1
- 239000008267 milk Substances 0.000 description 1
- 210000004080 milk Anatomy 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 235000019796 monopotassium phosphate Nutrition 0.000 description 1
- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 1
- 150000002823 nitrates Chemical class 0.000 description 1
- 229910017464 nitrogen compound Inorganic materials 0.000 description 1
- 150000002830 nitrogen compounds Chemical class 0.000 description 1
- QJGQUHMNIGDVPM-UHFFFAOYSA-N nitrogen group Chemical group [N] QJGQUHMNIGDVPM-UHFFFAOYSA-N 0.000 description 1
- 238000000655 nuclear magnetic resonance spectrum Methods 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
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 description 1
- 230000001766 physiological effect Effects 0.000 description 1
- 239000000049 pigment Substances 0.000 description 1
- 235000019260 propionic acid Nutrition 0.000 description 1
- IUVKMZGDUIUOCP-BTNSXGMBSA-N quinbolone Chemical compound O([C@H]1CC[C@H]2[C@H]3[C@@H]([C@]4(C=CC(=O)C=C4CC3)C)CC[C@@]21C)C1=CCCC1 IUVKMZGDUIUOCP-BTNSXGMBSA-N 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
- 229920002477 rna polymer Polymers 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
- 230000028327 secretion Effects 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 235000002639 sodium chloride Nutrition 0.000 description 1
- 239000011684 sodium molybdate Substances 0.000 description 1
- 235000015393 sodium molybdate Nutrition 0.000 description 1
- 239000004317 sodium nitrate Substances 0.000 description 1
- 235000010344 sodium nitrate Nutrition 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 239000001384 succinic acid Substances 0.000 description 1
- 235000011044 succinic acid Nutrition 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- 150000008163 sugars Chemical class 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 229940074410 trehalose Drugs 0.000 description 1
- 241001148471 unidentified anaerobic bacterium Species 0.000 description 1
- NWONKYPBYAMBJT-UHFFFAOYSA-L zinc sulfate Chemical compound [Zn+2].[O-]S([O-])(=O)=O NWONKYPBYAMBJT-UHFFFAOYSA-L 0.000 description 1
- 229910000368 zinc sulfate Inorganic materials 0.000 description 1
- 229960001763 zinc sulfate Drugs 0.000 description 1
Landscapes
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Description
(産業上の利用分野)
この発明は、クロストリジウム属に属する微生
物を用いてウラシルを製造する方法に関するもの
である、。ウラシルは医薬原料あるいは生化学試
薬として有用である。
(従来技術)
ウラシルは化学的にはホルミル酢酸と尿素を縮
合させて合成する。
従来、微生物の培養液中に、ウラシル系化合物
が他の核酸成分と共に分泌されることについて
は、例えばブレビバクテリウム・リクエフアシエ
ンス(Breuibacterium liquefaciens)(ザ・ジヤ
ーナル・オブ・バクテリオロジー(The Janal
of Bacteriolgy)84巻、1頁、1962年)、バチル
ス(Bacillus)属細菌(アグリカルチユラル・バ
イオロジカル・ケミストリー(Agricaltural
Biological Chemistry)、26巻、586頁、1962年)
またはその他の微生物(特公昭40−10957、52−
139786、53−38691)において知られている。微
生物がウラシルのみを生産する例としてブレビバ
クテリウム(Brevibacterium)属の菌(特公昭
57−30476)、ミクロモノスポラ
(Micromonospor)属の菌(特公昭57−18873)
がある。しかし、嫌気性菌が培養液中にウラシル
を蓄積することは知られていない。
(発明が解決しようとする問題点)
上記のような方法が知られているもののウラシ
ルの化学合成法は強烈な条件下の反応を必要とす
る欠点がある。また微生物的には解決すべき課題
はまだ多く、ウラシルの生産速度、蓄積濃度の高
い菌、あるいはウラシルのみを培養液中に蓄積
し、他のリボ核酸成分を含まない菌などを得るこ
とは重要である。そのためには公知の菌種にもと
づいた育種改良とならんで、ウラシルを製造する
新菌種の創製がきわめて重要な手段となる。
本発明はこのようなな目的のもとに検討を重ね
た結果、クロストリジウム(Clostridium)属に
属する微生物がウラシルを培地中に蓄積すること
を見出し、本発明に到達した。
(問題点を解決するための手段)
本発明はクロストリジウム属に属しウラシルを
生産する能力のある菌株を培養し、生成蓄積され
たウラシルを回収する事を特徴とするウラシルの
製造方法である。
本発明で用いられる微生物はクロストリジウム
属に属しウラシルを生産する能力を有する微生物
であり、この様な微生物は本発明実施例記載のも
のがはじめてである。実施例で用いられた微生物
は、偏性嫌気性有胞子桿菌である点でクロストリ
ジウム属に属する菌と考えられるが、グラム染色
性、炭素源の資化性など後で詳しく記す諸性質に
おいて公知の同属菌と相違しており、新菌種であ
ると考えられる。正式の種名はまだ付されていな
いので、本発明ではクロストリジウム・エスピー
No.S−1と表示する。次にクロストリジウム・エ
スピーNo.S−1(以下本菌と略記する)の創製法
および菌学的性質を示す。
(創製法)
本菌は下記の方法により分離した。すなわち第
1表に示す液体培地にソルボース1g/を加え
た培地5mlを試験管へ分注し滅菌後、嫌気グロー
ブボツクス中で給源を添加し、ブチルゴム栓で密
栓後、気相を嫌気ガスに置換し、30℃で静置培養
し、約10日毎に植え継ぎを行なつた。1回液体培
地で植え継いだのち、第1表の培地に寒天30g/
、ソルボース1g/を加えた寒天培地を用い
てロール・チユーブ法(メソツズ・イン・マイク
ロバイオロジー(Methods in Microbiolcgy)
3巻B、117頁(1969))を繰り返すことにより本
菌を得た。
(菌学的性質)
本発明の菌株の菌学的性質を示す、この菌学的
性質の検討には、「アンアエロブ・ラボラトリ
ー・マニユアル(Anaerobe Laboratory
Manual)第4版」(The V.I.P.Anaerobe
Laboratory Virginia Polytechnic Institute
and State University、Blacksburg(1972))お
よび「バージーズ・マニユアル・オブ・デターミ
ネイテイブ・バクテリオロジー(Bergey′s
Manual of Determinative Bacteriology)第8
版」「微生物の分類と同定」(長谷川武治著、学会
出版センター)に記載されている方法、培地組成
を用いた。
(顕微鏡的所見)
1 細胞の形および大きさ:単独もしくは3〜4
連の直桿菌。幅0.4〜0.8μm、長さ3〜8μm
2 鞭毛:周鞭毛を有する
3 胞子:あり、ターミナル
4 グラム染色:陰性
(培地組成)
第1表に例示する。
(Industrial Application Field) The present invention relates to a method for producing uracil using a microorganism belonging to the genus Clostridium. Uracil is useful as a pharmaceutical raw material or biochemical reagent. (Prior Art) Uracil is chemically synthesized by condensing formyl acetic acid and urea. Conventionally, the secretion of uracil-based compounds together with other nucleic acid components into the culture solution of microorganisms has been reported, for example, in Breuibacterium liquefaciens (The Journal of Bacteriology).
of Bacteriolgy (vol. 84, p. 1, 1962), Bacillus (Agricultural Biological Chemistry),
(Biological Chemistry), vol. 26, p. 586, 1962)
or other microorganisms (Japanese Patent Publication No. 40-10957, 52-139786, 53-38691). An example of a microorganism producing only uracil is a bacterium of the genus Brevibacterium (Tokukosho).
57-30476), bacteria of the genus Micromonospora (Special Publication No. 57-18873)
There is. However, it is not known that anaerobic bacteria accumulate uracil in the culture solution. (Problems to be Solved by the Invention) Although the above-mentioned methods are known, the chemical synthesis method for uracil has the drawback of requiring reaction under severe conditions. In addition, there are still many issues to be solved from a microbial perspective, and it is important to obtain bacteria that have high uracil production rates, high accumulation concentrations, or bacteria that accumulate only uracil in the culture solution and do not contain other ribonucleic acid components. It is. To this end, in addition to breeding and improvement based on known bacterial strains, the creation of new bacterial strains that produce uracil is an extremely important means. As a result of repeated studies based on these objectives, the present invention was achieved based on the discovery that microorganisms belonging to the genus Clostridium accumulate uracil in a culture medium. (Means for Solving the Problems) The present invention is a method for producing uracil, which is characterized by culturing a bacterial strain belonging to the genus Clostridium and having the ability to produce uracil, and collecting the produced and accumulated uracil. The microorganism used in the present invention belongs to the genus Clostridium and has the ability to produce uracil, and the microorganism described in the Examples of the present invention is the first such microorganism. The microorganisms used in the examples are considered to belong to the genus Clostridium in that they are obligate anaerobic spore-forming bacilli, but they are not well-known in terms of properties such as Gram stainability and ability to assimilate carbon sources, which will be described in detail later. It is different from bacteria of the same genus and is considered to be a new bacterial species. Since the official species name has not yet been assigned, this invention uses Clostridium sp.
Display No.S-1. Next, the production method and mycological properties of Clostridium sp. No. S-1 (hereinafter abbreviated as the present bacterium) will be described. (Creation method) This bacterium was isolated by the following method. That is, 5 ml of the liquid medium shown in Table 1 to which 1 g of sorbose was added was dispensed into test tubes, and after sterilization, a supply source was added in an anaerobic glove box, the tube was sealed with a butyl rubber stopper, and the gas phase was replaced with anaerobic gas. The cells were then cultured statically at 30°C, and subcultured approximately every 10 days. After sub-planting once in a liquid medium, add 30g of agar to the medium shown in Table 1.
, roll tube method using an agar medium supplemented with 1 g of sorbose (Methods in Microbiolcgy)
3B, p. 117 (1969)) was repeated to obtain this bacterium. (Mycological Properties) For examination of the mycological properties that indicate the mycological properties of the strain of the present invention, please refer to the “Anaerobe Laboratory Manual”.
Manual) 4th Edition” (The VIP Anaerobe
Laboratory Virginia Polytechnic Institute
and State University, Blacksburg (1972)) and Bergey's Manual of Determinative Bacteriology.
Manual of Determinative Bacteriology) No. 8
The method and culture medium composition described in "Classification and Identification of Microorganisms" (written by Takeharu Hasegawa, Gakkai Publishing Center) were used. (Microscopic findings) 1 Cell shape and size: Single or 3-4
Direct bacilli of Ren. Width: 0.4 to 0.8 μm, length: 3 to 8 μm 2 Flagella: having periflagella 3 Spores: present, terminal 4 Gram staining: negative (medium composition) Examples are shown in Table 1.
【表】【table】
【表】
(生育状態)
第1表の組成に30g/寒天、1g/ソルボ
ースを加えた寒天培地での生育は次の通りであ
る。
形 状:円形
周 縁:円滑
隆 起:わずかに盛上る
表 面:円滑
色 調:白
(生理的性質)
酸素に対する態度:偏性嫌気性
インドールの生産性:−
ゼラチンの液化:−
カタラーゼの生産性:−
デンプンの加水分解:−
エスクリンの加水分解:+
色素の生成:−
硫酸塩の還元性:−
ミルク:凝固、ペプトン化共に−
ウレアーゼ:−
ビタミンの要求性:なし
生育範囲:温度25〜40℃、最適温度35℃ PH
5.5〜9.0、最適PH7.0
(炭素源の質化性)
第1表の基本培地に下記炭素源(10g/)を
含む液体倍地5mlを直径18mmの試験管に加え、無
菌培地を作成し本菌を80μ植菌し、気相を窒素
(67%)と二酸化炭素(33%)を含む除菌ガスに
置換し、30℃で14日間静置培養した。生育は
600nmの濁度を分光計(スペクトロニツク20、
島津製作所製)で測定した。600nmの濁度が炭
素源を含まないコントロールとの差が0.1未満の
ものを「資化しない」、0.1以上0.2未満のものを
「わずかに資化する」0.2以上のものを「資化す
る」とした。
質化するもの:ラフイノース、ガラクトース、ラ
ムノース、グリコーゲン、トレハロース、マン
ノース、グルコース、アミグダリン、ソルボー
ス
資化しないもの:フルクトース、キシロース、リ
ボース、アラビノース、マルトース、シユクロ
ース、ラクトース、メリビオース、セロビオー
ス、メレジトース、マンニトール、デンプン、
アドニトール、エリスリトール、イノシトー
ル、メタノール、エタノール、n−プロパノー
ル、エチレングリコール、グリセロール、ギ
酸、酢酸、プロピオン酸、コハク酸
(糖などからの生産物)
第1表の基本培地に上記の試験で資化すること
が確かめられた糖を10g/添加し、気相を窒素
(67%)と二酸化炭素(33%)を含む除菌ガスに
置換し、本菌を植菌、30℃で静置培養した。すべ
ての炭素源において培地中には、主生産物として
酢酸、エタノールが生産された。
(従来の類似種との比較)
上記の菌学的性質から、このクロストリジウ
ム・エスピーNo.S−1は偏性嫌気性のグラム陰性
有胞子桿菌で、その主要醗酵代謝産物が酢酸、エ
タノールである菌株である。また、硫酸塩の還元
性もない。このようなことから、本菌はクロスト
リジウム(Clostridium)に属する菌株であると
考えられる。グラム陰性のクロストリジウムには
クロストリジウム・セロビオパルム
(Clostridium cellobioparum)、クロストリジウ
ム・サーモサツカロリテイカム(Costridium
thermosaccharolyticum)、クロストリジウム・
ブレビフアシエンス(Clostridium
brevifaciens)、クロストリジウム・クルイベリ
(Clostridium kluyveri)クロストリジウム・マ
ラコサム(Clostridium malacosomae)クロス
トリジウム・サーモセラム(Clostridium
thermocellum)がある。このうち、クロストリ
ジウム・サーモサツカロリテイカム、クロストリ
ジウム・サーモセラムは高温菌という点で本菌と
区別できる。クロストリジウム・ブレビフアシエ
ンス、クロストリジウム・マラコサムはアルカリ
菌という点で本菌と区別できる。そして、クロス
トリジウム・セロビオパルムは生産物が、酢酸の
みであり、エタノールを生産しないという点で、
クロストリジウム・クルイベリは生産物がカプロ
ン酸のみであるという点で区別できる。
以上のことから、本菌株はクロストリジウム属
に属する新菌種であると考えられるので、クロス
トリジウム・エスピーNo.S−1と命名した。さら
にこの菌株は工業技術院微生物工業技術研究所に
「微工研菌寄第8852号(FERM P−8852)」とし
て寄託した。
(培養方法)
培養方法は原則的には、一般の微生物の場合と
同様であるが、酸素の混入を防ぐことが必要であ
り、実験室的には、ゴム栓等で密栓した培養器中
で、静置あるいは振盪する方法が用いられる。や
や大きい規模では、通常用いられる醗酵槽がその
まま利用でき、装置内の酸素は、窒素などの不活
性気体などで置換することにより嫌気的な雰囲気
をつくることが可能である。醗酵槽の型式は特に
問わないが、普通に使用される撹拌混合槽のほ
か、一段あるいは多段の気泡塔型、ドラフトチユ
ーブ型の醗酵槽も利用できる。
培養に用いる炭素源は、資化する炭素源であれ
ばよく窒素源は塩化アンモニウムのごときアンモ
ニウム塩や硝酸ソーダのような硝酸塩のごとく、
通常の醗酵に用いうる各種の窒素化合物を用いる
ことができる。
その他必要に応じ、リン酸二水素カリ、硫酸マ
グネシウム、硫酸マンガン、塩化ナトリウム、硫
酸鉄、塩化コバルト、塩化カルシウム、硫酸亜
鉛、硫酸銅、明ばん、モリブデン酸ソーダ、硼酸
などの無機化合物、あるいはビオチンや酵母エキ
スなどのビタミン類を添加することは、通常行な
われる通りでである。
本菌株は休止菌体として使用し資化することが
できない二酸化炭素を原料としてウラシルを生産
する方法に使用することもできる。この場合には
培養で得られた菌体を遠心分離、膜濾過等通常用
いられる方法で培養液から分離し通常用いられる
培養液から炭素源を除いた培養液に懸濁させ、資
化する炭素源の代りに二酸化炭素ガスを供給し反
応させる。二酸化炭素ガガスの代りに、反応液中
に溶解二酸化炭素あるいは炭酸塩、炭酸水素塩と
して加えることもできる。培養とは異なり、反応
中酸素が存在してもウラシルを生産させることが
できる。
培地中に蓄積されたウラシルは、公知の技術に
より回収することができる。
以下具体例により本発明を説明する。
実施例 1
クロストリジウム・エスピーNo.S−1株を以下
のように培養した。第1表に示す培地に10g/
のソルボースを加え試験管へ5ml分注滅菌後、同
培地であらかじめ該菌株を前培養した培養液80μ
を嫌気グローブボツクス中で添加し、ブチルゴ
ム栓で密栓したのち気相を窒素(67%)と二酸化
炭素(33%)を含む除菌ガスに置換した。30℃で
10日間で静置培養を行なつたところ、培養液中に
ウラシル1.5mg/を得た。(ウラシルは培養液か
ら高速液クロで分散した後、質量分析、紫外線吸
収スペクトルおよびNMRスペクトルにより標品
と一致することを確認した)。
実施例 2
第1表の基本培地に10g/のアミグダリンを
加え実施例1と同様に培養を行なつた。静置培養
14日間で2.5mg/のウラシルを生産した。
実施例 3
第1表の基本培地に10g/のグルコースを加
え実施例1と同様に培養を行なつた。静成培養14
日間で1.9mg/ウラシルを生産した。
実施例 4
第1表の基本培地に10g/のフルクトースを
加え実施例1と同様に培養を行なつた。静置培養
14日間で2.2mg/のウラシルを生産した。
実施例 5
第1表の基本培地に10g/のグルコースを加
え実施例と同様に培養を行なつた。静置培養14日
間で1.1mg/ウラシルを生産した。
実施例 6
第1表の基本培地に10g/のガラクトースを
加え実施例1と同様に培養を行なつた。静置培養
14日間で1.3mg/のウラシルを生産した。
実施例 7
第1表の基本培地に10g/のトレハロースを
加え実施例1と同様に培養を行なつた。静置培養
14日間で1.7mg/のウラシルを生産した。
実施例 8
第1表の基本培地に10g/のラフイノースを
加え実施例1と同様に培養を行なつた。静置培養
14日間で1.5mg/のウラシルを生産した。
実施例 9
第1表に示す培地で10g/のソルボースを炭
素源として培養を行なつた菌体(乾燥重量6mg)
を遠心分離し、第1表に示す培地5mlに懸濁さ
せ、ブチルゴム栓で密栓した後、気相を窒素(67
%)と二酸化炭素(33%)を含む除菌ガスに置換
し、35℃で4日間反応させた。反応液中に30mg/
のウラシルを生成した。
実施例 10
実施例9と同様に菌体(乾燥重量1.6mg)を第
1表に示す培地5mlに懸濁させ、ブチルゴム栓で
密栓した後、気相を窒素(33%)、と二酸化炭素
(33%)空気(33%)を含む除菌ガスに置換し、
35℃で3日間反応させた。反応液中に5.2mg/
のウラシルを生成した。[Table] (Growth status) Growth on an agar medium containing the composition shown in Table 1 plus 30g/agar and 1g/sorbose is as follows. Shape: Circular Rim: Smooth Elevation: Slightly raised Surface: Smooth Color tone: White (physiological properties) Attitude towards oxygen: Obligately anaerobic Indole productivity: - Liquefaction of gelatin: - Catalase production Characteristics: - Starch hydrolysis: - Aesculin hydrolysis: + Pigment formation: - Sulfate reducing properties: - Milk: both coagulation and peptonization - Urease: - Vitamin requirement: None Growth range: Temperature 25~ 40℃, optimal temperature 35℃ PH
5.5 to 9.0, optimal PH7.0 (Characterization of carbon source) Add 5 ml of liquid medium containing the following carbon source (10 g/) to the basic medium shown in Table 1 to a test tube with a diameter of 18 mm to create a sterile medium. 80μ of this bacterium was inoculated, the gas phase was replaced with a sterilizing gas containing nitrogen (67%) and carbon dioxide (33%), and the culture was statically cultured at 30°C for 14 days. The growth is
Measure the turbidity at 600 nm using a spectrometer (Spectronic 20,
(manufactured by Shimadzu Corporation). If the turbidity at 600 nm differs from the control without carbon source by less than 0.1, it is "not assimilated," if it is 0.1 or more and less than 0.2, it is "slightly assimilated," and if it is 0.2 or more, it is "assimilated." And so. What is metabolized: raffinose, galactose, rhamnose, glycogen, trehalose, mannose, glucose, amygdalin, sorbose What is not metabolized: fructose, xylose, ribose, arabinose, maltose, sucrose, lactose, melibiose, cellobiose, melezitose, mannitol, starch ,
Adonitol, erythritol, inositol, methanol, ethanol, n-propanol, ethylene glycol, glycerol, formic acid, acetic acid, propionic acid, succinic acid (products from sugars, etc.) Assimilated into the basic medium shown in Table 1 in the above test 10 g of sugar was added, the gas phase was replaced with a sterilizing gas containing nitrogen (67%) and carbon dioxide (33%), and this bacteria was inoculated and cultured stationary at 30°C. Acetic acid and ethanol were produced as the main products in the medium using all carbon sources. (Comparison with conventional similar species) Based on the above mycological properties, this Clostridium sp. It is a bacterial strain. Moreover, it does not have the reducing property of sulfate. Based on these facts, this bacterium is considered to be a strain belonging to Clostridium. Gram-negative clostridia include Clostridium cellobioparum and Clostridium thermosatucaroliticum.
thermosaccharolyticum), Clostridium
Brevifuaciens (Clostridium)
brevifaciens), Clostridium kluyveri, Clostridium malacosomae, Clostridium thermocellum
thermocellum). Among these, Clostridium thermosatucaloriticum and Clostridium thermocellum can be distinguished from this bacterium in that they are thermophilic bacteria. Clostridium brevifuaciens and Clostridium malacosum can be distinguished from this bacterium in that they are alkaline bacteria. Clostridium cellobioparum only produces acetic acid and does not produce ethanol.
Clostridium kluyveri can be distinguished in that its only product is caproic acid. Based on the above, this strain is considered to be a new bacterial species belonging to the genus Clostridium, so it was named Clostridium sp. No. S-1. Furthermore, this strain was deposited with the Institute of Microbial Technology, Agency of Industrial Science and Technology as "FERM P-8852". (Cultivation method) The cultivation method is basically the same as that for general microorganisms, but it is necessary to prevent oxygen from entering, and in the laboratory, culture is carried out in an incubator tightly closed with a rubber stopper. , leaving it still or shaking it. On a slightly larger scale, a commonly used fermenter can be used as is, and an anaerobic atmosphere can be created by replacing the oxygen in the device with an inert gas such as nitrogen. The type of fermentation tank is not particularly limited, but in addition to commonly used stirring and mixing tanks, single-stage or multi-stage bubble column type, and draft tube type fermentation tanks can also be used. The carbon source used for culture may be any carbon source that can be assimilated, and the nitrogen source may be ammonium salts such as ammonium chloride or nitrates such as sodium nitrate.
Various nitrogen compounds that can be used in normal fermentation can be used. Other inorganic compounds such as potassium dihydrogen phosphate, magnesium sulfate, manganese sulfate, sodium chloride, iron sulfate, cobalt chloride, calcium chloride, zinc sulfate, copper sulfate, alum, sodium molybdate, boric acid, or biotin. Addition of vitamins such as yeast extract and yeast extract is generally carried out. This strain can also be used as a dormant bacterial cell in a method for producing uracil using carbon dioxide, which cannot be assimilated, as a raw material. In this case, the bacterial cells obtained by culture are separated from the culture solution by a commonly used method such as centrifugation or membrane filtration, and suspended in a culture solution in which the carbon source is removed from the normally used culture solution. Supply carbon dioxide gas instead of the source and cause the reaction to occur. Instead of carbon dioxide gas, dissolved carbon dioxide, carbonate, or hydrogen carbonate can also be added to the reaction solution. Unlike culture, uracil can be produced even in the presence of oxygen during the reaction. Uracil accumulated in the medium can be recovered by known techniques. The present invention will be explained below using specific examples. Example 1 Clostridium sp. No. S-1 strain was cultured as follows. 10g/
After adding 5 ml of sorbose to a test tube and sterilizing it, add 80μ of a culture solution in which the strain was pre-cultured in the same medium.
was added in an anaerobic glove box, and after sealing with a butyl rubber stopper, the gas phase was replaced with a sterilizing gas containing nitrogen (67%) and carbon dioxide (33%). at 30℃
When static culture was performed for 10 days, 1.5 mg of uracil was obtained in the culture solution. (Uracil was dispersed from the culture solution using high-speed liquid chromatography, and then confirmed to match the standard by mass spectrometry, ultraviolet absorption spectrum, and NMR spectrum.) Example 2 Culture was carried out in the same manner as in Example 1 by adding 10 g/amygdalin to the basal medium shown in Table 1. Static culture
2.5mg/uracil was produced in 14 days. Example 3 Culture was carried out in the same manner as in Example 1 by adding 10 g/glucose to the basal medium shown in Table 1. Static culture 14
1.9mg/uracil was produced per day. Example 4 Culture was carried out in the same manner as in Example 1 by adding 10 g/fructose to the basal medium shown in Table 1. Static culture
2.2 mg/uracil was produced in 14 days. Example 5 10 g/glucose was added to the basal medium shown in Table 1, and culture was carried out in the same manner as in Example. 1.1 mg/uracil was produced during 14 days of static culture. Example 6 Culture was carried out in the same manner as in Example 1 by adding 10 g/galactose to the basal medium shown in Table 1. Static culture
1.3 mg/uracil was produced in 14 days. Example 7 Culture was carried out in the same manner as in Example 1 by adding 10 g/trehalose to the basal medium shown in Table 1. Static culture
1.7 mg/uracil was produced in 14 days. Example 8 Culture was carried out in the same manner as in Example 1 by adding 10 g/raffinose to the basal medium shown in Table 1. Static culture
1.5mg/uracil was produced in 14 days. Example 9 Bacterial cells (dry weight 6 mg) were cultured in the medium shown in Table 1 using 10 g of sorbose as a carbon source.
was centrifuged, suspended in 5 ml of the medium shown in Table 1, and sealed with a butyl rubber stopper.
%) and carbon dioxide (33%), and reacted at 35°C for 4 days. 30mg/in the reaction solution
of uracil was produced. Example 10 In the same manner as in Example 9, bacterial cells (dry weight 1.6 mg) were suspended in 5 ml of the medium shown in Table 1, and after sealing with a butyl rubber stopper, the gas phase was replaced with nitrogen (33%) and carbon dioxide ( 33%) replaced with sterilizing gas containing air (33%),
The reaction was carried out at 35°C for 3 days. 5.2mg/in the reaction solution
of uracil was produced.
Claims (1)
る能力のある菌株を培養し、生成蓄積されたウラ
シルを回収する事を特徴とするウラシルの製造方
法。1. A method for producing uracil, which comprises culturing a bacterial strain belonging to the genus Clostridium and having the ability to produce uracil, and recovering the produced and accumulated uracil.
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP23328286A JPS6387992A (en) | 1986-10-02 | 1986-10-02 | Production of uracil |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP23328286A JPS6387992A (en) | 1986-10-02 | 1986-10-02 | Production of uracil |
Publications (2)
Publication Number | Publication Date |
---|---|
JPS6387992A JPS6387992A (en) | 1988-04-19 |
JPH0139758B2 true JPH0139758B2 (en) | 1989-08-23 |
Family
ID=16952650
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP23328286A Granted JPS6387992A (en) | 1986-10-02 | 1986-10-02 | Production of uracil |
Country Status (1)
Country | Link |
---|---|
JP (1) | JPS6387992A (en) |
-
1986
- 1986-10-02 JP JP23328286A patent/JPS6387992A/en active Granted
Also Published As
Publication number | Publication date |
---|---|
JPS6387992A (en) | 1988-04-19 |
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