JPS6122952B2 - - Google Patents
Info
- Publication number
- JPS6122952B2 JPS6122952B2 JP59054611A JP5461184A JPS6122952B2 JP S6122952 B2 JPS6122952 B2 JP S6122952B2 JP 59054611 A JP59054611 A JP 59054611A JP 5461184 A JP5461184 A JP 5461184A JP S6122952 B2 JPS6122952 B2 JP S6122952B2
- Authority
- JP
- Japan
- Prior art keywords
- glycerin
- nad
- dihydroxyacetone
- glycerol dehydrogenase
- receptors
- 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
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 claims abstract description 128
- 102000004190 Enzymes Human genes 0.000 claims abstract description 61
- 108090000790 Enzymes Proteins 0.000 claims abstract description 61
- 238000000034 method Methods 0.000 claims abstract description 31
- 101000892220 Geobacillus thermodenitrificans (strain NG80-2) Long-chain-alcohol dehydrogenase 1 Proteins 0.000 claims abstract description 29
- 150000003626 triacylglycerols Chemical class 0.000 claims abstract description 16
- 244000005700 microbiome Species 0.000 claims abstract description 11
- 241000589236 Gluconobacter Species 0.000 claims abstract description 8
- 241000589220 Acetobacter Species 0.000 claims abstract description 7
- 230000008569 process Effects 0.000 claims abstract description 6
- 241000589516 Pseudomonas Species 0.000 claims abstract description 4
- 241000228212 Aspergillus Species 0.000 claims abstract description 3
- 241000588698 Erwinia Species 0.000 claims abstract description 3
- 241000588748 Klebsiella Species 0.000 claims abstract description 3
- 241000228143 Penicillium Species 0.000 claims abstract description 3
- 241000235527 Rhizopus Species 0.000 claims abstract description 3
- 241000607720 Serratia Species 0.000 claims abstract 2
- 235000011187 glycerol Nutrition 0.000 claims description 57
- RXKJFZQQPQGTFL-UHFFFAOYSA-N dihydroxyacetone Chemical compound OCC(=O)CO RXKJFZQQPQGTFL-UHFFFAOYSA-N 0.000 claims description 40
- 230000000694 effects Effects 0.000 claims description 27
- 229940120503 dihydroxyacetone Drugs 0.000 claims description 20
- RXGJTUSBYWCRBK-UHFFFAOYSA-M 5-methylphenazinium methyl sulfate Chemical compound COS([O-])(=O)=O.C1=CC=C2[N+](C)=C(C=CC=C3)C3=NC2=C1 RXGJTUSBYWCRBK-UHFFFAOYSA-M 0.000 claims description 17
- 238000006243 chemical reaction Methods 0.000 claims description 17
- CCBICDLNWJRFPO-UHFFFAOYSA-N 2,6-dichloroindophenol Chemical compound C1=CC(O)=CC=C1N=C1C=C(Cl)C(=O)C(Cl)=C1 CCBICDLNWJRFPO-UHFFFAOYSA-N 0.000 claims description 13
- 238000002835 absorbance Methods 0.000 claims description 9
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 claims description 6
- 230000007423 decrease Effects 0.000 claims description 6
- 125000003831 tetrazolyl group Chemical group 0.000 claims description 6
- 230000015572 biosynthetic process Effects 0.000 claims description 5
- 239000001301 oxygen Substances 0.000 claims description 5
- 229910052760 oxygen Inorganic materials 0.000 claims description 5
- 150000003904 phospholipids Chemical class 0.000 claims description 4
- 238000002360 preparation method Methods 0.000 claims description 4
- 235000002837 Acetobacter xylinum Nutrition 0.000 claims description 3
- 235000021588 free fatty acids Nutrition 0.000 claims description 3
- VMGAPWLDMVPYIA-HIDZBRGKSA-N n'-amino-n-iminomethanimidamide Chemical compound N\N=C\N=N VMGAPWLDMVPYIA-HIDZBRGKSA-N 0.000 claims description 3
- SOWBFZRMHSNYGE-UHFFFAOYSA-N oxamic acid Chemical compound NC(=O)C(O)=O SOWBFZRMHSNYGE-UHFFFAOYSA-N 0.000 claims description 3
- BOLDJAUMGUJJKM-LSDHHAIUSA-N renifolin D Natural products CC(=C)[C@@H]1Cc2c(O)c(O)ccc2[C@H]1CC(=O)c3ccc(O)cc3O BOLDJAUMGUJJKM-LSDHHAIUSA-N 0.000 claims description 3
- 241000287231 Serinus Species 0.000 claims description 2
- 238000012258 culturing Methods 0.000 claims description 2
- 230000009467 reduction Effects 0.000 claims description 2
- 239000003153 chemical reaction reagent Substances 0.000 claims 4
- 239000002824 redox indicator Substances 0.000 claims 3
- 241001136169 Komagataeibacter xylinus Species 0.000 claims 1
- 238000004519 manufacturing process Methods 0.000 claims 1
- 235000015097 nutrients Nutrition 0.000 abstract description 2
- 239000000243 solution Substances 0.000 description 15
- 239000000370 acceptor Substances 0.000 description 10
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 9
- XEEYBQQBJWHFJM-UHFFFAOYSA-N iron Substances [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 8
- 239000012528 membrane Substances 0.000 description 8
- 101710088194 Dehydrogenase Proteins 0.000 description 7
- 230000008859 change Effects 0.000 description 7
- 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 6
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 6
- 230000001580 bacterial effect Effects 0.000 description 6
- 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 6
- 239000000872 buffer Substances 0.000 description 6
- 239000001257 hydrogen Substances 0.000 description 6
- 229910052739 hydrogen Inorganic materials 0.000 description 6
- 235000000177 Indigofera tinctoria Nutrition 0.000 description 5
- 239000000975 dye Substances 0.000 description 5
- 229940097275 indigo Drugs 0.000 description 5
- COHYTHOBJLSHDF-UHFFFAOYSA-N indigo powder Natural products N1C2=CC=CC=C2C(=O)C1=C1C(=O)C2=CC=CC=C2N1 COHYTHOBJLSHDF-UHFFFAOYSA-N 0.000 description 5
- 239000002609 medium Substances 0.000 description 5
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 description 4
- 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 4
- 239000000463 material Substances 0.000 description 4
- 239000000758 substrate Substances 0.000 description 4
- 230000007704 transition Effects 0.000 description 4
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 3
- 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 3
- 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 3
- RGHNJXZEOKUKBD-SQOUGZDYSA-M D-gluconate Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@@H](O)C([O-])=O RGHNJXZEOKUKBD-SQOUGZDYSA-M 0.000 description 3
- 102100034289 Deoxynucleoside triphosphate triphosphohydrolase SAMHD1 Human genes 0.000 description 3
- 241000589232 Gluconobacter oxydans Species 0.000 description 3
- 108700040097 Glycerol dehydrogenases Proteins 0.000 description 3
- 101000641031 Homo sapiens Deoxynucleoside triphosphate triphosphohydrolase SAMHD1 Proteins 0.000 description 3
- 108090001060 Lipase Proteins 0.000 description 3
- 102000004882 Lipase Human genes 0.000 description 3
- 239000004367 Lipase Substances 0.000 description 3
- 238000011088 calibration curve Methods 0.000 description 3
- 210000004027 cell Anatomy 0.000 description 3
- 239000005515 coenzyme Substances 0.000 description 3
- 238000007796 conventional method Methods 0.000 description 3
- 238000001514 detection method Methods 0.000 description 3
- 239000003599 detergent Substances 0.000 description 3
- 239000012154 double-distilled water Substances 0.000 description 3
- 230000002255 enzymatic effect Effects 0.000 description 3
- 238000002523 gelfiltration Methods 0.000 description 3
- 125000004435 hydrogen atom Chemical group [H]* 0.000 description 3
- 150000002500 ions Chemical class 0.000 description 3
- 229910052742 iron Inorganic materials 0.000 description 3
- 235000019421 lipase Nutrition 0.000 description 3
- 238000000746 purification Methods 0.000 description 3
- 238000011002 quantification Methods 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- UFTFJSFQGQCHQW-UHFFFAOYSA-N triformin Chemical compound O=COCC(OC=O)COC=O UFTFJSFQGQCHQW-UHFFFAOYSA-N 0.000 description 3
- WJFKNYWRSNBZNX-UHFFFAOYSA-N 10H-phenothiazine Chemical compound C1=CC=C2NC3=CC=CC=C3SC2=C1 WJFKNYWRSNBZNX-UHFFFAOYSA-N 0.000 description 2
- TZMSYXZUNZXBOL-UHFFFAOYSA-N 10H-phenoxazine Chemical compound C1=CC=C2NC3=CC=CC=C3OC2=C1 TZMSYXZUNZXBOL-UHFFFAOYSA-N 0.000 description 2
- BFSVOASYOCHEOV-UHFFFAOYSA-N 2-diethylaminoethanol Chemical compound CCN(CC)CCO BFSVOASYOCHEOV-UHFFFAOYSA-N 0.000 description 2
- 244000235858 Acetobacter xylinum Species 0.000 description 2
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- FBPFZTCFMRRESA-FSIIMWSLSA-N D-Glucitol Natural products OC[C@H](O)[C@H](O)[C@@H](O)[C@H](O)CO FBPFZTCFMRRESA-FSIIMWSLSA-N 0.000 description 2
- FBPFZTCFMRRESA-JGWLITMVSA-N D-glucitol Chemical compound OC[C@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-JGWLITMVSA-N 0.000 description 2
- RGHNJXZEOKUKBD-SQOUGZDYSA-N D-gluconic acid Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@@H](O)C(O)=O RGHNJXZEOKUKBD-SQOUGZDYSA-N 0.000 description 2
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 2
- 108090000371 Esterases Proteins 0.000 description 2
- 108010013563 Lipoprotein Lipase Proteins 0.000 description 2
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 description 2
- XJLXINKUBYWONI-DQQFMEOOSA-N [[(2r,3r,4r,5r)-5-(6-aminopurin-9-yl)-3-hydroxy-4-phosphonooxyoxolan-2-yl]methoxy-hydroxyphosphoryl] [(2s,3r,4s,5s)-5-(3-carbamoylpyridin-1-ium-1-yl)-3,4-dihydroxyoxolan-2-yl]methyl phosphate Chemical compound NC(=O)C1=CC=C[N+]([C@@H]2[C@H]([C@@H](O)[C@H](COP([O-])(=O)OP(O)(=O)OC[C@@H]3[C@H]([C@@H](OP(O)(O)=O)[C@@H](O3)N3C4=NC=NC(N)=C4N=C3)O)O2)O)=C1 XJLXINKUBYWONI-DQQFMEOOSA-N 0.000 description 2
- 238000010521 absorption reaction Methods 0.000 description 2
- 230000009471 action Effects 0.000 description 2
- 210000004369 blood Anatomy 0.000 description 2
- 239000008280 blood Substances 0.000 description 2
- 210000000170 cell membrane Anatomy 0.000 description 2
- 239000001913 cellulose Substances 0.000 description 2
- 229920002678 cellulose Polymers 0.000 description 2
- 238000004440 column chromatography Methods 0.000 description 2
- 239000013078 crystal Substances 0.000 description 2
- 230000001419 dependent effect Effects 0.000 description 2
- 238000006911 enzymatic reaction Methods 0.000 description 2
- 210000003527 eukaryotic cell Anatomy 0.000 description 2
- 238000000855 fermentation Methods 0.000 description 2
- 230000004151 fermentation Effects 0.000 description 2
- 238000001502 gel electrophoresis Methods 0.000 description 2
- 239000008103 glucose Substances 0.000 description 2
- 239000001963 growth medium Substances 0.000 description 2
- 229910052588 hydroxylapatite Inorganic materials 0.000 description 2
- JORABGDXCIBAFL-UHFFFAOYSA-M iodonitrotetrazolium chloride Chemical compound [Cl-].C1=CC([N+](=O)[O-])=CC=C1N1[N+](C=2C=CC(I)=CC=2)=NC(C=2C=CC=CC=2)=N1 JORABGDXCIBAFL-UHFFFAOYSA-M 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 235000010355 mannitol Nutrition 0.000 description 2
- 238000005259 measurement Methods 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- XYJRXVWERLGGKC-UHFFFAOYSA-D pentacalcium;hydroxide;triphosphate Chemical compound [OH-].[Ca+2].[Ca+2].[Ca+2].[Ca+2].[Ca+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O XYJRXVWERLGGKC-UHFFFAOYSA-D 0.000 description 2
- 229950000688 phenothiazine Drugs 0.000 description 2
- 230000000704 physical effect Effects 0.000 description 2
- 235000018102 proteins Nutrition 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 150000003839 salts Chemical class 0.000 description 2
- 210000002966 serum Anatomy 0.000 description 2
- 229960002920 sorbitol Drugs 0.000 description 2
- 239000012086 standard solution Substances 0.000 description 2
- 150000005846 sugar alcohols Polymers 0.000 description 2
- AWDBHOZBRXWRKS-UHFFFAOYSA-N tetrapotassium;iron(6+);hexacyanide Chemical compound [K+].[K+].[K+].[K+].[Fe+6].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-] AWDBHOZBRXWRKS-UHFFFAOYSA-N 0.000 description 2
- PUWAVSCMFIGOJQ-DFWYDOINSA-N (2s)-2-aminopentanedioic acid;propane-1,2,3-triol Chemical compound OCC(O)CO.OC(=O)[C@@H](N)CCC(O)=O PUWAVSCMFIGOJQ-DFWYDOINSA-N 0.000 description 1
- 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
- HNLXNOZHXNSSPN-UHFFFAOYSA-N 2-[2-[2-[2-[2-[2-[2-[4-(2,4,4-trimethylpentan-2-yl)phenoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethanol Chemical compound CC(C)(C)CC(C)(C)C1=CC=C(OCCOCCOCCOCCOCCOCCOCCO)C=C1 HNLXNOZHXNSSPN-UHFFFAOYSA-N 0.000 description 1
- QFVHZQCOUORWEI-UHFFFAOYSA-N 4-[(4-anilino-5-sulfonaphthalen-1-yl)diazenyl]-5-hydroxynaphthalene-2,7-disulfonic acid Chemical compound C=12C(O)=CC(S(O)(=O)=O)=CC2=CC(S(O)(=O)=O)=CC=1N=NC(C1=CC=CC(=C11)S(O)(=O)=O)=CC=C1NC1=CC=CC=C1 QFVHZQCOUORWEI-UHFFFAOYSA-N 0.000 description 1
- UOZOCOQLYQNHII-UHFFFAOYSA-N 6-bromo-2-(6-bromo-3-hydroxy-1H-indol-2-yl)indol-3-one Chemical compound [O-]c1c([nH]c2cc(Br)ccc12)C1=[NH+]c2cc(Br)ccc2C1=O UOZOCOQLYQNHII-UHFFFAOYSA-N 0.000 description 1
- 108010021809 Alcohol dehydrogenase Proteins 0.000 description 1
- 102000007698 Alcohol dehydrogenase Human genes 0.000 description 1
- 102000005369 Aldehyde Dehydrogenase Human genes 0.000 description 1
- 108020002663 Aldehyde Dehydrogenase Proteins 0.000 description 1
- 102000016893 Amine Oxidase (Copper-Containing) Human genes 0.000 description 1
- 108010028700 Amine Oxidase (Copper-Containing) Proteins 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 1
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- 108010058733 Choline dehydrogenase Proteins 0.000 description 1
- 102100032363 Choline dehydrogenase, mitochondrial Human genes 0.000 description 1
- RGHNJXZEOKUKBD-UHFFFAOYSA-N D-gluconic acid Natural products OCC(O)C(O)C(O)C(O)C(O)=O RGHNJXZEOKUKBD-UHFFFAOYSA-N 0.000 description 1
- 101000925662 Enterobacteria phage PRD1 Endolysin Proteins 0.000 description 1
- 229930091371 Fructose Natural products 0.000 description 1
- 239000005715 Fructose Substances 0.000 description 1
- 241001518248 Gluconobacter cerinus Species 0.000 description 1
- 102100022119 Lipoprotein lipase Human genes 0.000 description 1
- 102000043296 Lipoprotein lipases Human genes 0.000 description 1
- JLVVSXFLKOJNIY-UHFFFAOYSA-N Magnesium ion Chemical compound [Mg+2] JLVVSXFLKOJNIY-UHFFFAOYSA-N 0.000 description 1
- GUBGYTABKSRVRQ-PICCSMPSSA-N Maltose Natural products O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@@H](CO)OC(O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-PICCSMPSSA-N 0.000 description 1
- 229930195725 Mannitol Natural products 0.000 description 1
- FBWADIKARMIWNM-UHFFFAOYSA-N N-3,5-dichloro-4-hydroxyphenyl-1,4-benzoquinone imine Chemical compound C1=C(Cl)C(O)=C(Cl)C=C1N=C1C=CC(=O)C=C1 FBWADIKARMIWNM-UHFFFAOYSA-N 0.000 description 1
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 1
- 229920005654 Sephadex Polymers 0.000 description 1
- 239000012507 Sephadex⢠Substances 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
- 229930006000 Sucrose Natural products 0.000 description 1
- 240000008042 Zea mays Species 0.000 description 1
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 description 1
- 235000002017 Zea mays subsp mays Nutrition 0.000 description 1
- DGEZNRSVGBDHLK-UHFFFAOYSA-N [1,10]phenanthroline Chemical compound C1=CN=C2C3=NC=CC=C3C=CC2=C1 DGEZNRSVGBDHLK-UHFFFAOYSA-N 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 230000002776 aggregation Effects 0.000 description 1
- 238000004220 aggregation Methods 0.000 description 1
- 239000003905 agrochemical Substances 0.000 description 1
- 108010048916 alcohol dehydrogenase (acceptor) Proteins 0.000 description 1
- 150000001298 alcohols Chemical class 0.000 description 1
- 150000001299 aldehydes Chemical class 0.000 description 1
- 150000001412 amines Chemical class 0.000 description 1
- 150000003863 ammonium salts Chemical class 0.000 description 1
- 238000013103 analytical ultracentrifugation Methods 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
- 230000008033 biological extinction Effects 0.000 description 1
- 210000001124 body fluid Anatomy 0.000 description 1
- 239000010839 body fluid Substances 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 229910052791 calcium Inorganic materials 0.000 description 1
- 239000011575 calcium Substances 0.000 description 1
- 229940041514 candida albicans extract Drugs 0.000 description 1
- 239000004202 carbamide Substances 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 150000001722 carbon compounds Chemical class 0.000 description 1
- 125000002057 carboxymethyl group Chemical group [H]OC(=O)C([H])([H])[*] 0.000 description 1
- 239000005018 casein Substances 0.000 description 1
- 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 1
- 235000021240 caseins Nutrition 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 238000010668 complexation reaction Methods 0.000 description 1
- 235000005822 corn Nutrition 0.000 description 1
- 230000001086 cytosolic effect Effects 0.000 description 1
- 238000004042 decolorization Methods 0.000 description 1
- SYELZBGXAIXKHU-UHFFFAOYSA-N dodecyldimethylamine N-oxide Chemical compound CCCCCCCCCCCC[N+](C)(C)[O-] SYELZBGXAIXKHU-UHFFFAOYSA-N 0.000 description 1
- 238000000921 elemental analysis Methods 0.000 description 1
- 230000007071 enzymatic hydrolysis Effects 0.000 description 1
- 238000006047 enzymatic hydrolysis reaction Methods 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- 239000003925 fat Substances 0.000 description 1
- 229940050410 gluconate Drugs 0.000 description 1
- 239000000174 gluconic acid Substances 0.000 description 1
- 235000012208 gluconic acid Nutrition 0.000 description 1
- 238000000703 high-speed centrifugation Methods 0.000 description 1
- 230000002209 hydrophobic effect Effects 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 150000002484 inorganic compounds Chemical class 0.000 description 1
- 229910010272 inorganic material Inorganic materials 0.000 description 1
- DCYOBGZUOMKFPA-UHFFFAOYSA-N iron(2+);iron(3+);octadecacyanide Chemical compound [Fe+2].[Fe+2].[Fe+2].[Fe+3].[Fe+3].[Fe+3].[Fe+3].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-] DCYOBGZUOMKFPA-UHFFFAOYSA-N 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- QTWZICCBKBYHDM-UHFFFAOYSA-N leucomethylene blue Chemical compound C1=C(N(C)C)C=C2SC3=CC(N(C)C)=CC=C3NC2=C1 QTWZICCBKBYHDM-UHFFFAOYSA-N 0.000 description 1
- 238000004811 liquid chromatography Methods 0.000 description 1
- 239000007791 liquid phase Substances 0.000 description 1
- 229910001425 magnesium ion Inorganic materials 0.000 description 1
- 159000000003 magnesium salts Chemical class 0.000 description 1
- FDZZZRQASAIRJF-UHFFFAOYSA-M malachite green Chemical compound [Cl-].C1=CC(N(C)C)=CC=C1C(C=1C=CC=CC=1)=C1C=CC(=[N+](C)C)C=C1 FDZZZRQASAIRJF-UHFFFAOYSA-M 0.000 description 1
- 229940107698 malachite green Drugs 0.000 description 1
- 239000000594 mannitol Substances 0.000 description 1
- 235000013372 meat Nutrition 0.000 description 1
- 238000003808 methanol extraction Methods 0.000 description 1
- CXKWCBBOMKCUKX-UHFFFAOYSA-M methylene blue Chemical group [Cl-].C1=CC(N(C)C)=CC2=[S+]C3=CC(N(C)C)=CC=C3N=C21 CXKWCBBOMKCUKX-UHFFFAOYSA-M 0.000 description 1
- 229960000907 methylthioninium chloride Drugs 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 230000007935 neutral effect Effects 0.000 description 1
- 229930027945 nicotinamide-adenine dinucleotide Natural products 0.000 description 1
- BOPGDPNILDQYTO-NNYOXOHSSA-N nicotinamide-adenine dinucleotide Chemical compound C1=CCC(C(=O)N)=CN1[C@H]1[C@H](O)[C@H](O)[C@@H](COP(O)(=O)OP(O)(=O)OC[C@@H]2[C@H]([C@@H](O)[C@@H](O2)N2C3=NC=NC(N)=C3N=C2)O)O1 BOPGDPNILDQYTO-NNYOXOHSSA-N 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 229910017464 nitrogen compound Inorganic materials 0.000 description 1
- 150000002830 nitrogen compounds Chemical class 0.000 description 1
- 150000007524 organic acids Chemical class 0.000 description 1
- 235000005985 organic acids Nutrition 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 230000001590 oxidative effect Effects 0.000 description 1
- 230000036284 oxygen consumption Effects 0.000 description 1
- 150000003013 phosphoric acid derivatives Chemical class 0.000 description 1
- 210000002381 plasma Anatomy 0.000 description 1
- 238000002264 polyacrylamide gel electrophoresis Methods 0.000 description 1
- 229940093429 polyethylene glycol 6000 Drugs 0.000 description 1
- 229910052700 potassium Inorganic materials 0.000 description 1
- 239000011591 potassium Substances 0.000 description 1
- 239000002244 precipitate Substances 0.000 description 1
- 238000011085 pressure filtration Methods 0.000 description 1
- 238000004445 quantitative analysis Methods 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 238000007127 saponification reaction Methods 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 159000000000 sodium salts Chemical class 0.000 description 1
- 230000007928 solubilization Effects 0.000 description 1
- 238000005063 solubilization Methods 0.000 description 1
- 239000000600 sorbitol Substances 0.000 description 1
- 238000004611 spectroscopical analysis Methods 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 239000004094 surface-active agent Substances 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 239000001003 triarylmethane dye Substances 0.000 description 1
- ZVAPIIDBWWULJN-UHFFFAOYSA-N tyrian purple Natural products N1C2=CC(Br)=CC=C2C(=O)C1=C1C(=O)C2=CC=C(Br)C=C2N1 ZVAPIIDBWWULJN-UHFFFAOYSA-N 0.000 description 1
- 238000002604 ultrasonography Methods 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
- 238000010626 work up procedure Methods 0.000 description 1
- 239000012138 yeast extract Substances 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/0004—Oxidoreductases (1.)
- C12N9/0006—Oxidoreductases (1.) acting on CH-OH groups as donors (1.1)
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/26—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving oxidoreductase
- C12Q1/32—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving oxidoreductase involving dehydrogenase
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/61—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving triglycerides
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
- Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10S435/00—Chemistry: molecular biology and microbiology
- Y10S435/8215—Microorganisms
- Y10S435/822—Microorganisms using bacteria or actinomycetales
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
- Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10S435/00—Chemistry: molecular biology and microbiology
- Y10S435/8215—Microorganisms
- Y10S435/822—Microorganisms using bacteria or actinomycetales
- Y10S435/823—Acetobacter
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Genetics & Genomics (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- General Health & Medical Sciences (AREA)
- Microbiology (AREA)
- Molecular Biology (AREA)
- Biotechnology (AREA)
- General Engineering & Computer Science (AREA)
- Biochemistry (AREA)
- Analytical Chemistry (AREA)
- Physics & Mathematics (AREA)
- Immunology (AREA)
- Biophysics (AREA)
- Medicinal Chemistry (AREA)
- Biomedical Technology (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Enzymes And Modification Thereof (AREA)
Description
ãçºæã®è©³çŽ°ãªèª¬æã
æ¬çºæã¯æ°èŠã®NADïŒïŒ°ïŒâç¬ç«æ§ã°ãªã»ãªã³
ããããã²ããŒãŒã«é¢ãããã°ãªã»ãªã³ãããã
ã²ããŒãŒã¯ãžããããã·ã¢ã»ãã³ã®åœ¢æäžã«ã°ãª
ã»ãªã³ã®åå¿ãæ¥è§Šããã
ããããã²ããŒãŒã«é¢ãããã°ãªã»ãªã³ãããã
ã²ããŒãŒã¯ãžããããã·ã¢ã»ãã³ã®åœ¢æäžã«ã°ãª
ã»ãªã³ã®åå¿ãæ¥è§Šããã
åŸæ¥å
¬ç¥ã®ã°ãªã»ãªã³ããããã²ããŒãŒã¯è£é
µ
çŽ ãšããŠé žååã®NADãŸãã¯NADPãå¿ èŠãšã
ããã°ãªã»ãªã³ïŒâNADïŒïŒ°ïŒâãªãã·ãã¬ãã¯
ã¿ãŒãŒïŒE.C.1.1.1.6ãE.C.1.1.1.72ãŸãã¯E.
C.1.1.1.156ïŒã§ãããåå¿ã¯æ¬¡ã®åå¿åŒã«ãã€ãŠ
è¡šããããšãã§ããïŒ åŒ(1)ã¯å¹³è¡¡åå¿ã§ãããããªãã¡NADïŒïŒ°ïŒâ
äŸåæ§ã°ãªã»ãªã³ããããã²ããŒãŒã¯åæ§ã«ããŠ
ãžããããã·ã¢ã»ãã³ããã°ãªã»ãªã³ã®åœ¢æãæ¥
觊ããããããã€ãŠã°ãªã»ãªã³ãããã¯ã°ãªã»ãª
ã³ããããã²ããŒãŒæŽ»æ§ãå®éããããã«åŒ(1)ã
å©çšããå Žåã«ã¯éå°éã®NADïŒïŒ°ïŒã®æ·»å ã«
ãã平衡ããžããããã·ã¢ã»ãã³ã®åŽãžã·ããã
ããå¿ èŠããããããã¯ãã®å®éæ¹æ³ãçžå¯Ÿçã«
æéãšè²»çšã®ããããã®ãšãããå°ããªã°ãªã»ãª
ã³éãããã¯ã°ãªã»ãªã³ããããã²ããŒãŒæŽ»æ§ã¯
ãã®æ¹æ³ã§ã¯ç²Ÿç¢ºã«ã¯æž¬å®ããããšãã§ããªãã
æŽã«æ°ŽçŽ ãç°¡åã«ã¯NADïŒïŒ°ïŒïŒšãã奜é©ãªæ
瀺è¬âã¬ããã¯ã¹ç³»ã«ç§»ããªãã®ãæ¬ ç¹ã§ããã
ãã®ããã«æ°ŽçŽ ãNADHãããã«ãã¶ã³ã®åœ¢æäž
ã«ããã©ãŸãªãŠã å¡©ã«ç§»ãããã«ããïŒã€ã®é µ
çŽ ãäŸãã°ãžã¢ãã©ãŒãŒãå¿ èŠã§ããã
çŽ ãšããŠé žååã®NADãŸãã¯NADPãå¿ èŠãšã
ããã°ãªã»ãªã³ïŒâNADïŒïŒ°ïŒâãªãã·ãã¬ãã¯
ã¿ãŒãŒïŒE.C.1.1.1.6ãE.C.1.1.1.72ãŸãã¯E.
C.1.1.1.156ïŒã§ãããåå¿ã¯æ¬¡ã®åå¿åŒã«ãã€ãŠ
è¡šããããšãã§ããïŒ åŒ(1)ã¯å¹³è¡¡åå¿ã§ãããããªãã¡NADïŒïŒ°ïŒâ
äŸåæ§ã°ãªã»ãªã³ããããã²ããŒãŒã¯åæ§ã«ããŠ
ãžããããã·ã¢ã»ãã³ããã°ãªã»ãªã³ã®åœ¢æãæ¥
觊ããããããã€ãŠã°ãªã»ãªã³ãããã¯ã°ãªã»ãª
ã³ããããã²ããŒãŒæŽ»æ§ãå®éããããã«åŒ(1)ã
å©çšããå Žåã«ã¯éå°éã®NADïŒïŒ°ïŒã®æ·»å ã«
ãã平衡ããžããããã·ã¢ã»ãã³ã®åŽãžã·ããã
ããå¿ èŠããããããã¯ãã®å®éæ¹æ³ãçžå¯Ÿçã«
æéãšè²»çšã®ããããã®ãšãããå°ããªã°ãªã»ãª
ã³éãããã¯ã°ãªã»ãªã³ããããã²ããŒãŒæŽ»æ§ã¯
ãã®æ¹æ³ã§ã¯ç²Ÿç¢ºã«ã¯æž¬å®ããããšãã§ããªãã
æŽã«æ°ŽçŽ ãç°¡åã«ã¯NADïŒïŒ°ïŒïŒšãã奜é©ãªæ
瀺è¬âã¬ããã¯ã¹ç³»ã«ç§»ããªãã®ãæ¬ ç¹ã§ããã
ãã®ããã«æ°ŽçŽ ãNADHãããã«ãã¶ã³ã®åœ¢æäž
ã«ããã©ãŸãªãŠã å¡©ã«ç§»ãããã«ããïŒã€ã®é µ
çŽ ãäŸãã°ãžã¢ãã©ãŒãŒãå¿ èŠã§ããã
æ¬çºæã®èª²é¡ã¯ãåèšã®æ¬ ç¹ãæãããããã
ã€ãŠã°ãªã»ãªã³ãç°¡åãã€å®äŸ¡ã«å®éãããã€å°
éã®ã°ãªã»ãªã³ãã§ãããããå®éçã«æ€åºãã
ããšãã§ãããæ°èŠã®ã°ãªã»ãªã³ããããã²ããŒ
ãŒãèŠåºãããšã§ãã€ãã
ã€ãŠã°ãªã»ãªã³ãç°¡åãã€å®äŸ¡ã«å®éãããã€å°
éã®ã°ãªã»ãªã³ãã§ãããããå®éçã«æ€åºãã
ããšãã§ãããæ°èŠã®ã°ãªã»ãªã³ããããã²ããŒ
ãŒãèŠåºãããšã§ãã€ãã
ãã®èª²é¡ã¯æ¬çºæã«ããNADïŒïŒ°ïŒâç¬ç«æ§ã°
ãªã»ãªã³ããããã²ããŒãŒã®æäŸã«ãã解決ãã
ããæ°èŠã®é µçŽ ã¯æ¬¡ã®åå¿ãæ¥è§Šããæ°èŠã®ã°ãª
ã»ãªã³ããããã²ããŒãŒã§ããïŒ ãã®éïŒã€ã®ãããã³ããã³é»åã¯è£é µçŽ ãšã
ãŠã®NADãããã¯NADPãäžç·ã«äœ¿çšããã«å¥œ
é©ãªæ°ŽçŽ å容äœããã³é»åå容äœã«ç§»ããããå
å¿(2)ã¯å®å šã«ãžããããã·ã¢ã»ãã³åœ¢æã®æ¹åã«
é²è¡ããæ°èŠé µçŽ ã¯ã°ãªã»ãªã³ãããã³ããªã°ãª
ã»ãªããã°ãªã»ãªã³ãéé¢èèªé žã«ããåã®åŸç°¡
åãã€ç²Ÿç¢ºã«å®éããã®ãå¯èœã«ããã
ãªã»ãªã³ããããã²ããŒãŒã®æäŸã«ãã解決ãã
ããæ°èŠã®é µçŽ ã¯æ¬¡ã®åå¿ãæ¥è§Šããæ°èŠã®ã°ãª
ã»ãªã³ããããã²ããŒãŒã§ããïŒ ãã®éïŒã€ã®ãããã³ããã³é»åã¯è£é µçŽ ãšã
ãŠã®NADãããã¯NADPãäžç·ã«äœ¿çšããã«å¥œ
é©ãªæ°ŽçŽ å容äœããã³é»åå容äœã«ç§»ããããå
å¿(2)ã¯å®å šã«ãžããããã·ã¢ã»ãã³åœ¢æã®æ¹åã«
é²è¡ããæ°èŠé µçŽ ã¯ã°ãªã»ãªã³ãããã³ããªã°ãª
ã»ãªããã°ãªã»ãªã³ãéé¢èèªé žã«ããåã®åŸç°¡
åãã€ç²Ÿç¢ºã«å®éããã®ãå¯èœã«ããã
ãã®ããã«ããŠããããŠå°éã®ã°ãªã»ãªã³ãã
ããã€ãŠãŸãããªã°ãªã»ãªããå®éããããšãå¯
èœã§ãããæ¬çºæã«ããé µçŽ ãçšããŠã®ã°ãªã»ãª
ã³ãããã¯ããªã°ãªã»ãªãã®åŸ®éå®éã¯çµç¹æ¹æ³
ã«ããã®ã¿ãªãããåååŠçæ¹æ³ã«ãã€ãŠãè¡ãª
ãããšãã§ããã
ããã€ãŠãŸãããªã°ãªã»ãªããå®éããããšãå¯
èœã§ãããæ¬çºæã«ããé µçŽ ãçšããŠã®ã°ãªã»ãª
ã³ãããã¯ããªã°ãªã»ãªãã®åŸ®éå®éã¯çµç¹æ¹æ³
ã«ããã®ã¿ãªãããåååŠçæ¹æ³ã«ãã€ãŠãè¡ãª
ãããšãã§ããã
æ¬çºæã«ããNADïŒïŒ°ïŒâç¬ç«æ§ã°ãªã»ãªã³ã
ãããã²ããŒãŒã¯æ¬¡ã®æ§è³ªã«ãã€ãŠè©³çŽ°ã«ç¹æ§ã¥
ããããšãã§ããïŒ ïŒ äœçš è©²é µçŽ ã¯ã°ãªã»ãªã³ãè±æ°ŽçŽ ããŠãžãããã
ã·ã¢ã»ãã³ã«ãããã®éåé¢ãããïŒã€ã®ãã
ãã³ããã³ïŒã€ã®é»åã¯è£é µçŽ ãšããŠã®NAD
ïŒïŒ°ïŒãäžç·ã«äœ¿çšããã«å¥œé©ãªå容äœã«ç§»ã
ããã
ãããã²ããŒãŒã¯æ¬¡ã®æ§è³ªã«ãã€ãŠè©³çŽ°ã«ç¹æ§ã¥
ããããšãã§ããïŒ ïŒ äœçš è©²é µçŽ ã¯ã°ãªã»ãªã³ãè±æ°ŽçŽ ããŠãžãããã
ã·ã¢ã»ãã³ã«ãããã®éåé¢ãããïŒã€ã®ãã
ãã³ããã³ïŒã€ã®é»åã¯è£é µçŽ ãšããŠã®NAD
ïŒïŒ°ïŒãäžç·ã«äœ¿çšããã«å¥œé©ãªå容äœã«ç§»ã
ããã
ïŒ åºè³ªç¹ç°æ§
該é
µçŽ ã¯ããããŠç¹ç°çã«ã°ãªã»ãªã³ãšåå¿
ãããç¹ã«æ§é çã«è¿äŒŒã®ãåæ§ã«ããŠéåžžäœ
液äžã«åºçŸããç©è³ªã«å¯ŸããŠã¯æŽ»æ§ãèªããã
ãªãããŸãã¯å ããªæŽ»æ§ããèªããããªãã次
è¡šã«çš®ã ã®åºè³ªã«å¯Ÿãã掻æ§ãæãããã®éã°
ãªã»ãªã³ã«å¯Ÿãã掻æ§ã100ãšããã
ãããç¹ã«æ§é çã«è¿äŒŒã®ãåæ§ã«ããŠéåžžäœ
液äžã«åºçŸããç©è³ªã«å¯ŸããŠã¯æŽ»æ§ãèªããã
ãªãããŸãã¯å ããªæŽ»æ§ããèªããããªãã次
è¡šã«çš®ã ã®åºè³ªã«å¯Ÿãã掻æ§ãæãããã®éã°
ãªã»ãªã³ã«å¯Ÿãã掻æ§ã100ãšããã
è¡š
çš®ã
ã®åºè³ªã«å¯Ÿããæ¬çºæã«ããã°ãªã»ãªã³
ããããã²ããŒãŒã®çžå¯Ÿç掻æ§ïŒã°ãªã»ãªã³ïŒ
100ïŒ ã°ãªã»ãªã³ 100 âãœã«ãããŒã« 45 âãã³ãããŒã« 26 âã°ã«ã³ãŒã¹ ïŒ ïŒ€âãã©ã¯ããŒã¹ ïŒ ãšã¿ããŒã« ïŒ ïŒ æé©ãªPHå€ããã³å®å®ãªPHç¯å² æ¬çºæã«ããé µçŽ ã®æé©ãªPHå€ã¯PH7.0ã8.5
ã§ãããæé«ã®æŽ»æ§ã¯ãããPH7.5ã§åŸãã
ããé µçŽ ã¯PH6.0ã10.0ã®ç¯å²ã§å®å®ã§ããã
ããããã²ããŒãŒã®çžå¯Ÿç掻æ§ïŒã°ãªã»ãªã³ïŒ
100ïŒ ã°ãªã»ãªã³ 100 âãœã«ãããŒã« 45 âãã³ãããŒã« 26 âã°ã«ã³ãŒã¹ ïŒ ïŒ€âãã©ã¯ããŒã¹ ïŒ ãšã¿ããŒã« ïŒ ïŒ æé©ãªPHå€ããã³å®å®ãªPHç¯å² æ¬çºæã«ããé µçŽ ã®æé©ãªPHå€ã¯PH7.0ã8.5
ã§ãããæé«ã®æŽ»æ§ã¯ãããPH7.5ã§åŸãã
ããé µçŽ ã¯PH6.0ã10.0ã®ç¯å²ã§å®å®ã§ããã
ïŒ é
µçŽ äœçšã®æé©æž©åºŠ
æ¬çºæã«ããé
µçŽ ã¯æž©åºŠ25ã37âã§æé«ã®æŽ»
æ§ãæã€ã
æ§ãæã€ã
ïŒ æå¶ããã³æŽ»æ§å
ãªããµã¡ãŒãã¯é
µçŽ ã«å¯ŸããŠè»œãæå¶äœçšã
æã€ããªã³è質ã¯æ¬çºæã«ããã°ãªã»ãªã³ãã
ããã²ããŒãŒã®æŽ»æ§ãçŽ20ïŒ å¢å ãããã
æã€ããªã³è質ã¯æ¬çºæã«ããã°ãªã»ãªã³ãã
ããã²ããŒãŒã®æŽ»æ§ãçŽ20ïŒ å¢å ãããã
ïŒ ç©æ§
èçµåé
µçŽ ã¯ããã®é
µçŽ ã®çæ°Žæ§éšåãèã®
ãã¹ããªãããããã€ãŠçŽ°èèäžã«å®å šã«äœçœ®
ããŠããã®ã§ãããã¯äžè¬ã«çé¢æŽ»æ§å€ã®å©ã
ãªãã«å¯æº¶ååã³ç²Ÿè£œããããšãå°é£ã§ããã
æ¬çºæã®ã°ãªã»ãªã³ããããã²ããŒãŒã¯å žåç
ãªèçµåé µçŽ ã§ããããããäžåºŠå¯æº¶ååã³ç²Ÿ
補ããããšãé µçŽ æº¶æ¶²äžã«å¥œé©ãªçé¢æŽ»æ§å€ã®
ååšãªãã§ã¯ããã®æŽ»æ§ãä¿æããããšã¯å°é£
ã§ããããµã€ããŸã«ïŒcytosolïŒé µçŽ ã®å Žåã«
ã¯ãã²ã«é»æ°æ³³åãã²ã«ãéåã³åæçšè¶ é å¿
åé¢ãå å«ããåŸæ¥æ³ã«ãããååéåã³é µçŽ
ãµããŠãããæ§é ãåŸãããããããããã®ã
ããªåŸæ¥æ³ã¯èçµåã°ãªã»ãªã³ããããã²ããŒ
ãŒã®åŒ·çãªçæ°Žæ§ã®ããã«ãããã«é©çšããã
ãšãã§ããªããããªã¢ã¯ãªã«ã¢ããã²ã«é»æ°æ³³
ååã³SDSã²ã«é»æ°æ³³åäžã§è©²é µçŽ ã¯ã²ã«ã«ã©
ã ã®é éšã§åéãããåæçšè¶ é å¿åé¢è£ 眮ã®
é å¿åé¢ç®¡äžã§è©²é µçŽ ã¯åéããæ©æ¢°ãæ倧å
転æ°ïŒ59780r.p.m.ïŒã«éããåã«ãã§ã«æ²æ®¿
ããããã®ãããªå·šå€§ãªåéã¯ã²ã«ãéã«ãã
ååéã®æ±ºå®ã«ãããŠãèŠããããåŸã€ãŠãæ¬
çºæã®é µçŽ ã®ååéãçµæ¶æ§é åã³å çŽ åæã®
ããŒã¿ãŒãæããããšã¯äžå¯èœã§ããããâã¡ãœ
ããã»ã€ã³ã»ãšã³ãã¢ãã®ãŒïŒMethods in
EnzymologyïŒâ第89å·»ãã¢ã«ãããã¯ã»ãã¬
ã¹ïŒAcademic Prâ²essïŒã1982幎ïŒâé µçŽ ïŒ©âã
ã¢ã°ãªã«ã«ããŠã©ã«ã»ã±ãã«ã«ã»ãœãµã€ãšããŒ
ïŒAgricultural Chemical SocietyïŒãæ¥æ¬ã
1985幎ïŒâã¹ãã©ã¯ãã€ãŒã»ã¢ã³ãã»ãã¢ã³ã¯
ã·ãšã³ãºã»ãªãã»ã»ã«ã©ãŒã»ã¡ã³ãã©ã³
ïŒStructure and Functions of Cellular
MembraneïŒâã¢ã«ãããã¯ã»ãã¬ã¹ã»ã»ã³ã¿
ãŒïŒAcademic Press CenterïŒãæ¥æ¬ã1982幎
åç §ãã
ãã¹ããªãããããã€ãŠçŽ°èèäžã«å®å šã«äœçœ®
ããŠããã®ã§ãããã¯äžè¬ã«çé¢æŽ»æ§å€ã®å©ã
ãªãã«å¯æº¶ååã³ç²Ÿè£œããããšãå°é£ã§ããã
æ¬çºæã®ã°ãªã»ãªã³ããããã²ããŒãŒã¯å žåç
ãªèçµåé µçŽ ã§ããããããäžåºŠå¯æº¶ååã³ç²Ÿ
補ããããšãé µçŽ æº¶æ¶²äžã«å¥œé©ãªçé¢æŽ»æ§å€ã®
ååšãªãã§ã¯ããã®æŽ»æ§ãä¿æããããšã¯å°é£
ã§ããããµã€ããŸã«ïŒcytosolïŒé µçŽ ã®å Žåã«
ã¯ãã²ã«é»æ°æ³³åãã²ã«ãéåã³åæçšè¶ é å¿
åé¢ãå å«ããåŸæ¥æ³ã«ãããååéåã³é µçŽ
ãµããŠãããæ§é ãåŸãããããããããã®ã
ããªåŸæ¥æ³ã¯èçµåã°ãªã»ãªã³ããããã²ããŒ
ãŒã®åŒ·çãªçæ°Žæ§ã®ããã«ãããã«é©çšããã
ãšãã§ããªããããªã¢ã¯ãªã«ã¢ããã²ã«é»æ°æ³³
ååã³SDSã²ã«é»æ°æ³³åäžã§è©²é µçŽ ã¯ã²ã«ã«ã©
ã ã®é éšã§åéãããåæçšè¶ é å¿åé¢è£ 眮ã®
é å¿åé¢ç®¡äžã§è©²é µçŽ ã¯åéããæ©æ¢°ãæ倧å
転æ°ïŒ59780r.p.m.ïŒã«éããåã«ãã§ã«æ²æ®¿
ããããã®ãããªå·šå€§ãªåéã¯ã²ã«ãéã«ãã
ååéã®æ±ºå®ã«ãããŠãèŠããããåŸã€ãŠãæ¬
çºæã®é µçŽ ã®ååéãçµæ¶æ§é åã³å çŽ åæã®
ããŒã¿ãŒãæããããšã¯äžå¯èœã§ããããâã¡ãœ
ããã»ã€ã³ã»ãšã³ãã¢ãã®ãŒïŒMethods in
EnzymologyïŒâ第89å·»ãã¢ã«ãããã¯ã»ãã¬
ã¹ïŒAcademic Prâ²essïŒã1982幎ïŒâé µçŽ ïŒ©âã
ã¢ã°ãªã«ã«ããŠã©ã«ã»ã±ãã«ã«ã»ãœãµã€ãšããŒ
ïŒAgricultural Chemical SocietyïŒãæ¥æ¬ã
1985幎ïŒâã¹ãã©ã¯ãã€ãŒã»ã¢ã³ãã»ãã¢ã³ã¯
ã·ãšã³ãºã»ãªãã»ã»ã«ã©ãŒã»ã¡ã³ãã©ã³
ïŒStructure and Functions of Cellular
MembraneïŒâã¢ã«ãããã¯ã»ãã¬ã¹ã»ã»ã³ã¿
ãŒïŒAcademic Press CenterïŒãæ¥æ¬ã1982幎
åç §ãã
åŸã€ãŠãåèšã®ç©æ§ã«ä»£ããç¹åŸŽã瀺ãïŒ
(a) 該é
µçŽ ã¯ããã«ãããªã³ããã³ïŒPQQïŒ
ãè£æ¬ ååå£ãšããŠæãããPQQã¯æåã«
1979幎ã«ã¡ãããããŒãã¹
ïŒmethylotrophsïŒã®ã¡ã¿ããŒã«ããããã²
ããŒãŒäžã«èŠãåºãããããã®åŸPQQã¯é ¢
é žèããã®ã¢ã«ã³ãŒã«ããããã²ããŒãŒ
ïŒEC1.1.99.8ïŒåã³ã¢ã«ãããããããã²ã
ãŒãŒïŒEC1.2.99.3ïŒãé ¢é žèåã³ä»ã®é žåæ§
èããã®ïŒ€âã°ã«ã³ãŒã¹ããããã²ããŒãŒ
ïŒEC1.1.99.17ïŒãããœã€ãã¢ãã¹ããã®ã¢ã
ã³ããããã²ããŒãŒïŒEC1.4.99.3ïŒãçæ žçŽ°
èããã®é å«æã¢ãã³ãªãã·ããŒãŒ
ïŒEC1.4.3.6ïŒåã³ã³ãªã³ããããã²ããŒãŒ
ïŒEC1.1.99.1ïŒã®ãããªä»£è¬ã«éèŠãªããã€
ãã®çš®é¡ã®ããããã²ããŒãŒã®è£æ¬ ååå£ã§
ããããšãå ±åãããŠãããæ¬çºæã®é µçŽ ã
è£æ¬ ååå£ãšããŠPQQãå«æããŠãããšã
ãäºå®ã¯ãä»ã®NADïŒïŒ°ïŒâäŸåã°ãªã»ãªã³
ããããã²ããŒãŒãšã®æ¯èŒã«ãããŠãæãç¹
城ã®ããæ§è³ªã§ãããPQQã¯ã¡ã¿ããŒã«æœ
åºïŒ90%V/VïŒã«ããé µçŽ èçœããé€å»ãã
ããšãã§ããé µçŽ èçœãžã®PQQã®éå ±æçµ
åã瀺ããŠãããPQQæº¶è§£é µçŽ ïŒã¢ãã»ã°
ãªã»ãªã³ããããã²ããŒãŒïŒã¯å€æ¥PQQã
æ·»å ããããšã«ãã掻æ§åããããšãã§ã
ãã
ãè£æ¬ ååå£ãšããŠæãããPQQã¯æåã«
1979幎ã«ã¡ãããããŒãã¹
ïŒmethylotrophsïŒã®ã¡ã¿ããŒã«ããããã²
ããŒãŒäžã«èŠãåºãããããã®åŸPQQã¯é ¢
é žèããã®ã¢ã«ã³ãŒã«ããããã²ããŒãŒ
ïŒEC1.1.99.8ïŒåã³ã¢ã«ãããããããã²ã
ãŒãŒïŒEC1.2.99.3ïŒãé ¢é žèåã³ä»ã®é žåæ§
èããã®ïŒ€âã°ã«ã³ãŒã¹ããããã²ããŒãŒ
ïŒEC1.1.99.17ïŒãããœã€ãã¢ãã¹ããã®ã¢ã
ã³ããããã²ããŒãŒïŒEC1.4.99.3ïŒãçæ žçŽ°
èããã®é å«æã¢ãã³ãªãã·ããŒãŒ
ïŒEC1.4.3.6ïŒåã³ã³ãªã³ããããã²ããŒãŒ
ïŒEC1.1.99.1ïŒã®ãããªä»£è¬ã«éèŠãªããã€
ãã®çš®é¡ã®ããããã²ããŒãŒã®è£æ¬ ååå£ã§
ããããšãå ±åãããŠãããæ¬çºæã®é µçŽ ã
è£æ¬ ååå£ãšããŠPQQãå«æããŠãããšã
ãäºå®ã¯ãä»ã®NADïŒïŒ°ïŒâäŸåã°ãªã»ãªã³
ããããã²ããŒãŒãšã®æ¯èŒã«ãããŠãæãç¹
城ã®ããæ§è³ªã§ãããPQQã¯ã¡ã¿ããŒã«æœ
åºïŒ90%V/VïŒã«ããé µçŽ èçœããé€å»ãã
ããšãã§ããé µçŽ èçœãžã®PQQã®éå ±æçµ
åã瀺ããŠãããPQQæº¶è§£é µçŽ ïŒã¢ãã»ã°
ãªã»ãªã³ããããã²ããŒãŒïŒã¯å€æ¥PQQã
æ·»å ããããšã«ãã掻æ§åããããšãã§ã
ãã
(b) 粟補ããæ¬çºæã®é
µçŽ ãEDTAã§åŠçãã
ãšãé µçŽ æŽ»æ§ã®å®å šãªåªå€±ãã¿ããããå€æ¥
ã®PQQåã³ãã°ãã·ãŠã ã€ãªã³ããã®åŸæ·»
å ãããšãé µçŽ æŽ»æ§ã¯EDTAåŠçã®åãšã»ãŒ
åãã¬ãã«ãŸã§äžæããã
ãšãé µçŽ æŽ»æ§ã®å®å šãªåªå€±ãã¿ããããå€æ¥
ã®PQQåã³ãã°ãã·ãŠã ã€ãªã³ããã®åŸæ·»
å ãããšãé µçŽ æŽ»æ§ã¯EDTAåŠçã®åãšã»ãŒ
åãã¬ãã«ãŸã§äžæããã
(c) ãžã¡ãã«ããã·ã«ã¢ãã³ãªãã·ãã§ã®å¯æº¶
ååã³ããªãšãã¬ã³ã°ãªã³ãŒã«6000ã§ã®åé¢
ã«ãã100å粟補ãåŸããããããããåèš
ã®åŒ·ãçæ°Žæ§ã«ãã粟補ããé µçŽ ã®ç²Ÿè£œåºŠã
蚌æããåºæºããªããããããªãããè©Šéšã
è¡ãªã€ãŠãä»ã®é µçŽ 掻æ§ã¯ãã¯ãå šã芳å¯ã
ãããè©²é µçŽ ãèãã粟補ãããŠããããšã¯
æããã§ãããæ¬çºæã®é µçŽ ã¯å€ãã®çš®é¡ã®
å€äŸ¡ã¢ã«ã³ãŒã«ãé žåãããããšã¿ããŒã«ã
ã¢ã«ããããâã°ã«ã³ãŒã¹ãâãã«ã¯ã
ãŒã¹ãâã°ã«ã³ããŒãåã¯ïŒâã±ãââ
ã°ã«ã³ããŒããé žåããªãããããã€ãŠãæ¬
çºæã®é µçŽ ã¯å°ãªããšããããã®åºè³ªã®é žå
ã觊åªããé µçŽ ã«ããæ±æãããŠããªããæŽ
ã«ãâã°ã«ã³ãŒã¹ãâãã«ã¯ããŒã¹åã¯
âã°ã«ã³ããŒãã«å¯Ÿããé µçŽ æŽ»æ§ã粟補ã®
éã«æ¶å€±ããã
ååã³ããªãšãã¬ã³ã°ãªã³ãŒã«6000ã§ã®åé¢
ã«ãã100å粟補ãåŸããããããããåèš
ã®åŒ·ãçæ°Žæ§ã«ãã粟補ããé µçŽ ã®ç²Ÿè£œåºŠã
蚌æããåºæºããªããããããªãããè©Šéšã
è¡ãªã€ãŠãä»ã®é µçŽ 掻æ§ã¯ãã¯ãå šã芳å¯ã
ãããè©²é µçŽ ãèãã粟補ãããŠããããšã¯
æããã§ãããæ¬çºæã®é µçŽ ã¯å€ãã®çš®é¡ã®
å€äŸ¡ã¢ã«ã³ãŒã«ãé žåãããããšã¿ããŒã«ã
ã¢ã«ããããâã°ã«ã³ãŒã¹ãâãã«ã¯ã
ãŒã¹ãâã°ã«ã³ããŒãåã¯ïŒâã±ãââ
ã°ã«ã³ããŒããé žåããªãããããã€ãŠãæ¬
çºæã®é µçŽ ã¯å°ãªããšããããã®åºè³ªã®é žå
ã觊åªããé µçŽ ã«ããæ±æãããŠããªããæŽ
ã«ãâã°ã«ã³ãŒã¹ãâãã«ã¯ããŒã¹åã¯
âã°ã«ã³ããŒãã«å¯Ÿããé µçŽ æŽ»æ§ã粟補ã®
éã«æ¶å€±ããã
æ¬çºæã«ããã°ãªã»ãªã³ããããã²ããŒãŒã¯å€
æ°ã®åŸ®çç©ãäŸãã°ããœã€ãã¢ãã¹ããŒã©ãã¢ã
ã¯ã¬ãã·ãšã©ããšã«ãŠã€ãã¢ãã¢ã¹ãã«ã®ã«ã¹ã
ããã·ãªãŠã ããªãŸãã¹ãã¢ã»ããã¯ã¿ãŒããã³
ã°ã«ã³ããã¯ã¿ãŒå±ããç£çããããšãã§ããã
æ¬çºæã«ããé µçŽ ã¯ã¢ã»ããã¯ã¿ãŒããã³ã°ã«ã³
ããã¯ã¿ãŒå±ã®åŸ®çç©ãäŸãã°ã¢ã»ããã¯ã¿ãŒã»
ãã·ãªãã¹ïŒAcetobacter xylinusïŒDSM2623
ïŒIFOâ3288ïŒãã°ã«ã³ããã¯ã¿ãŒã»ã»ãªãã¹
ïŒGluconobacter cerinusïŒDSM2622ïŒIFOâ
3268ïŒããã³ã°ã«ã³ããã¯ã¿ãŒã»ã€ã³ãã¹ããªãŠ
ã¹ïŒGluconobacter industriusïŒDSM2621ïŒIFO
â3260ïŒããç¹ã«è¯å¥œãªåçã§ç£çããããåèš
ã®åŸ®çç©æ ªã¯ãã€ããšã»ã¶ã ã«ã³ã°ã»ããŠã¢ã»ã
ã¯ããªã«ã¬ããºã¡ã³ïŒDeutsche Sammlung fušr
MikroorganismenïŒã«åèšã®DSMçªå·ã§ããŸã
ã¯çºé µç 究æïŒIustitut fušr Fermentationã
OsakaïŒã«ã«ãã³å ã«æããIFOçªå·ã§å¯èšãã
ãŠããã
æ°ã®åŸ®çç©ãäŸãã°ããœã€ãã¢ãã¹ããŒã©ãã¢ã
ã¯ã¬ãã·ãšã©ããšã«ãŠã€ãã¢ãã¢ã¹ãã«ã®ã«ã¹ã
ããã·ãªãŠã ããªãŸãã¹ãã¢ã»ããã¯ã¿ãŒããã³
ã°ã«ã³ããã¯ã¿ãŒå±ããç£çããããšãã§ããã
æ¬çºæã«ããé µçŽ ã¯ã¢ã»ããã¯ã¿ãŒããã³ã°ã«ã³
ããã¯ã¿ãŒå±ã®åŸ®çç©ãäŸãã°ã¢ã»ããã¯ã¿ãŒã»
ãã·ãªãã¹ïŒAcetobacter xylinusïŒDSM2623
ïŒIFOâ3288ïŒãã°ã«ã³ããã¯ã¿ãŒã»ã»ãªãã¹
ïŒGluconobacter cerinusïŒDSM2622ïŒIFOâ
3268ïŒããã³ã°ã«ã³ããã¯ã¿ãŒã»ã€ã³ãã¹ããªãŠ
ã¹ïŒGluconobacter industriusïŒDSM2621ïŒIFO
â3260ïŒããç¹ã«è¯å¥œãªåçã§ç£çããããåèš
ã®åŸ®çç©æ ªã¯ãã€ããšã»ã¶ã ã«ã³ã°ã»ããŠã¢ã»ã
ã¯ããªã«ã¬ããºã¡ã³ïŒDeutsche Sammlung fušr
MikroorganismenïŒã«åèšã®DSMçªå·ã§ããŸã
ã¯çºé µç 究æïŒIustitut fušr Fermentationã
OsakaïŒã«ã«ãã³å ã«æããIFOçªå·ã§å¯èšãã
ãŠããã
æ¬çºæã«ããé
µçŽ ãç£çããããã«ã奜é©ãªåŸ®
çç©ãå ¬ç¥æ¹æ³ã«ããå¹å°ã§å¹é€ãããå¹å°ãšã
ãŠã¯çŽ°èå¹é€ã®æ é€æºãšããŠäžè¬ã«ç¥ãããŠãã
åžžçšã®å¹å°ãå©çšããããšãã§ããã
çç©ãå ¬ç¥æ¹æ³ã«ããå¹å°ã§å¹é€ãããå¹å°ãšã
ãŠã¯çŽ°èå¹é€ã®æ é€æºãšããŠäžè¬ã«ç¥ãããŠãã
åžžçšã®å¹å°ãå©çšããããšãã§ããã
ççŽ æºãšããŠç¹ã«çš®ã
ã®ä»£è¬å¯èœãªççŽ åå
ç©ãäŸãã°ã°ã«ã³ãŒã¹ããã©ã¯ããŒã¹ããµã¯ããŒ
ã¹ããã«ããŒã¹ãâååæ°ïŒã®ãµãã«ãªããã¡
ã©ã¹ïŒMelasseïŒããã³ææ©é žïŒäŸãã°ã°ã«ã³ã³
é žããã³é ¢é žïŒãã¢ã«ã³ãŒã«ïŒäŸãã°ã°ãªã»ãªã³
ããã³ãšã¿ããŒã«ïŒããã³ç³ã¢ã«ã³ãŒã«ïŒäŸãã°
ãã³ãããããã³ãœã«ãããŒã«ïŒãå©çšããããš
ãã§ããã
ç©ãäŸãã°ã°ã«ã³ãŒã¹ããã©ã¯ããŒã¹ããµã¯ããŒ
ã¹ããã«ããŒã¹ãâååæ°ïŒã®ãµãã«ãªããã¡
ã©ã¹ïŒMelasseïŒããã³ææ©é žïŒäŸãã°ã°ã«ã³ã³
é žããã³é ¢é žïŒãã¢ã«ã³ãŒã«ïŒäŸãã°ã°ãªã»ãªã³
ããã³ãšã¿ããŒã«ïŒããã³ç³ã¢ã«ã³ãŒã«ïŒäŸãã°
ãã³ãããããã³ãœã«ãããŒã«ïŒãå©çšããããš
ãã§ããã
çªçŽ æºãšããŠã¯çš®ã
ã®çªçŽ ååç©ãäŸãã°é
µæ¯
ãšã¯ã¹ãã©ã¯ããåžã«ãŒã€ã³ãã³ãŒã³æµžæŒ¬æ¶²ãè
ãšã¯ã¹ãã©ã¯ãïŒå€©ç¶åæïŒãå°¿çŽ ããã³ã¢ã³ã¢
ããŠã å¡©ã奜é©ã§ããã
ãšã¯ã¹ãã©ã¯ããåžã«ãŒã€ã³ãã³ãŒã³æµžæŒ¬æ¶²ãè
ãšã¯ã¹ãã©ã¯ãïŒå€©ç¶åæïŒãå°¿çŽ ããã³ã¢ã³ã¢
ããŠã å¡©ã奜é©ã§ããã
奜é©ãªç¡æ©ååç©ãšããŠå¹å°ã«ãããªãŠã å¡©ã
ã«ãªãŠã å¡©ãã«ã«ã·ãŠã å¡©ããã°ãã·ãŠã 塩䞊ã³
ã«çš®ã ã®ãªã³é žå¡©ãæ·»å ããããšãã§ããã
ã«ãªãŠã å¡©ãã«ã«ã·ãŠã å¡©ããã°ãã·ãŠã 塩䞊ã³
ã«çš®ã ã®ãªã³é žå¡©ãæ·»å ããããšãã§ããã
æ¬çºæã«ããé
µçŽ ã補é ããããã®åŸ®çç©ã®å¹
é€ã¯æå©ã«æ¶²çžäžã§å®æœãããããããå·¥æ¥çèŠ
æš¡ã§ã¯æ°Žäžå¹é€ãç¹ã«åªããŠããã
é€ã¯æå©ã«æ¶²çžäžã§å®æœãããããããå·¥æ¥çèŠ
æš¡ã§ã¯æ°Žäžå¹é€ãç¹ã«åªããŠããã
æ¬çºæã«ããé
µçŽ ãå¹é€ããããã®çŽ°èå¹å°ã¯
æå©ã«æž©åºŠ28ã32âã§ä¿æããããããããã®æž©
床ã¯ã埮çç©ãæé·ãããã€ææã®é µçŽ ãç£çã
åŸãéãå€ããŠãããã
æå©ã«æž©åºŠ28ã32âã§ä¿æããããããããã®æž©
床ã¯ã埮çç©ãæé·ãããã€ææã®é µçŽ ãç£çã
åŸãéãå€ããŠãããã
å¹é€æéã¯å¹é€æ¡ä»¶ã«å·Šå³ããããéåžžå¹é€ã
çŽïŒãïŒæ¥åŸã«çµãããããå Žåã«æ倧ã®é µçŽ å
çãåŸãããã
çŽïŒãïŒæ¥åŸã«çµãããããå Žåã«æ倧ã®é µçŽ å
çãåŸãããã
å¹é€æº¶æ¶²ã®PHå€ã¯ç¹ã«éèŠã§ã¯ãªããPHå€ã¯æ
å©ã«PHïŒãïŒã«èª¿ç¯ãããã
å©ã«PHïŒãïŒã«èª¿ç¯ãããã
æå€ã®é
µçŽ ãæãã现èç©è³ªã®åŸåŠçã¯åžžæ³ã«
ããè¡ãªã€ãŠããã现èç©è³ªã¯äœå§æ¿ŸéãŸãã¯é
å¿åé¢ã«ãã液ç¶åªäœããåé¢ããããšãã§ã
ãã埮çç©ã®çŽ°èã®ç Žå£ã¯åæ§ã«å ¬ç¥æ¹æ³ãå©çš
ãããäŸãã°éãã现èç©è³ªã奜é©ãªç·©è¡æ¶²ã«æž
æ¿ããããã€è¶ é³æ³¢ããã¬ã³ããã¬ã¹ãŸãã¯ãã€
ãïŒDynoïŒâãã«ã«ãã€ãŠç Žå£ããããšãã§ã
ããæ¬çºæã«ããé µçŽ ã¯èãã©ã¯ã·ãšã³äžã«ååš
ãããã®ãã©ã¯ã·ãšã³ã¯é«éé å¿åé¢
ïŒ27000rpmã§90åïŒã«ããåé¢ããããšãã§ã
ããå Žåã«ãã奜é©ãªç·©è¡æ¶²ç³»äžã®å¥œé©ãªæž æµå€
ã®æ·»å ã«ããé µçŽ ãèãã©ã¯ã·ãšã³ãã溶åºãã
ãããšãã§ããã
ããè¡ãªã€ãŠããã现èç©è³ªã¯äœå§æ¿ŸéãŸãã¯é
å¿åé¢ã«ãã液ç¶åªäœããåé¢ããããšãã§ã
ãã埮çç©ã®çŽ°èã®ç Žå£ã¯åæ§ã«å ¬ç¥æ¹æ³ãå©çš
ãããäŸãã°éãã现èç©è³ªã奜é©ãªç·©è¡æ¶²ã«æž
æ¿ããããã€è¶ é³æ³¢ããã¬ã³ããã¬ã¹ãŸãã¯ãã€
ãïŒDynoïŒâãã«ã«ãã€ãŠç Žå£ããããšãã§ã
ããæ¬çºæã«ããé µçŽ ã¯èãã©ã¯ã·ãšã³äžã«ååš
ãããã®ãã©ã¯ã·ãšã³ã¯é«éé å¿åé¢
ïŒ27000rpmã§90åïŒã«ããåé¢ããããšãã§ã
ããå Žåã«ãã奜é©ãªç·©è¡æ¶²ç³»äžã®å¥œé©ãªæž æµå€
ã®æ·»å ã«ããé µçŽ ãèãã©ã¯ã·ãšã³ãã溶åºãã
ãããšãã§ããã
æ¬çºæã«ããã°ãªã»ãªã³ããããã²ããŒãŒã¯é
µ
çŽ ã®åé¢ã®ããã®åžžæ³ã«ããåŸããã溶液ããåŸ
ããããæ¬çºæã«ããé µçŽ ã¯äŸãã°æå©ã«ã€ãªã³
亀æäœãäŸãã°ã«ã«ããã·ã¡ãã«âãŸãã¯DEAE
âã»ã«ããŒã¹ãçšããŠã«ã©ã ã¯ãããã°ã©ãã€ãŒ
ã«ãããæ°Žé žåãªã³ç°ç³ãçšããŠã«ã©ã ã¯ããã
ã°ã©ãã€ãŒã«ãããŸãã¯ã»ãã¢ããã¯ã¹ãçšããŠ
ã²ã«æ¿Ÿéã«ããçŽç²åœ¢ã§åé¢ããããšãã§ããã
çŽ ã®åé¢ã®ããã®åžžæ³ã«ããåŸããã溶液ããåŸ
ããããæ¬çºæã«ããé µçŽ ã¯äŸãã°æå©ã«ã€ãªã³
亀æäœãäŸãã°ã«ã«ããã·ã¡ãã«âãŸãã¯DEAE
âã»ã«ããŒã¹ãçšããŠã«ã©ã ã¯ãããã°ã©ãã€ãŒ
ã«ãããæ°Žé žåãªã³ç°ç³ãçšããŠã«ã©ã ã¯ããã
ã°ã©ãã€ãŒã«ãããŸãã¯ã»ãã¢ããã¯ã¹ãçšããŠ
ã²ã«æ¿Ÿéã«ããçŽç²åœ¢ã§åé¢ããããšãã§ããã
æ¬çºæã«ããé
µçŽ ã®æŽ»æ§ã¯åºè³ªãšããŠã°ãªã»ãª
ã³ãããšããžã³ã¡ãã¹ã«ããšãŒãããã³ïŒã»ïŒâ
ãžã¯ãã«ããšããŒã«ã€ã³ãããšããŒã«ãçšããŠ25
âãPH7.5ã§åœéåäœïŒU/mgïŒã§è¡šããã
ã³ãããšããžã³ã¡ãã¹ã«ããšãŒãããã³ïŒã»ïŒâ
ãžã¯ãã«ããšããŒã«ã€ã³ãããšããŒã«ãçšããŠ25
âãPH7.5ã§åœéåäœïŒU/mgïŒã§è¡šããã
æ¬çºæã«ããNADïŒïŒ°ïŒâç¬ç«æ§ã°ãªã»ãªã³ã
ãããã²ããŒãŒã¯æå©ã«åŒ(2)ã«ããã°ãªã»ãªã³ã®
枬å®ã«äœ¿çšããããšãã§ããããã®åå¿åŒã«ãã
ã°ã°ãªã»ãªã³ïŒååããæ°ŽçŽ ååïŒåãé€å»ã
ãããžããããã·ã¢ã»ãã³ïŒååã圢æãããïŒ
åã®ãããã³ããã³ïŒåã®é»åã¯å¥œé©ãªæ°ŽçŽ å容
äœïŒïŒ¡ïŒã«ç§»è¡ãããã次ã®æ¡å€§åå¿åŒãåŸãã
ãïŒ åå¿(3)ã«ããã°é žååå容äœïŒïŒ¡ïŒã®æžå°ãŸã
ã¯éå åå容äœïŒïŒ¡ïŒã®å¢å ãŸãã¯åœ¢æããããž
ããããã·ã¢ã»ãã³ã®éã枬å®ããããšã«ããã°
ãªã»ãªã³ãã°ãªã»ãªã³âããããã²ããŒãŒæŽ»æ§ã
å®éããããšãã§ããã
ãããã²ããŒãŒã¯æå©ã«åŒ(2)ã«ããã°ãªã»ãªã³ã®
枬å®ã«äœ¿çšããããšãã§ããããã®åå¿åŒã«ãã
ã°ã°ãªã»ãªã³ïŒååããæ°ŽçŽ ååïŒåãé€å»ã
ãããžããããã·ã¢ã»ãã³ïŒååã圢æãããïŒ
åã®ãããã³ããã³ïŒåã®é»åã¯å¥œé©ãªæ°ŽçŽ å容
äœïŒïŒ¡ïŒã«ç§»è¡ãããã次ã®æ¡å€§åå¿åŒãåŸãã
ãïŒ åå¿(3)ã«ããã°é žååå容äœïŒïŒ¡ïŒã®æžå°ãŸã
ã¯éå åå容äœïŒïŒ¡ïŒã®å¢å ãŸãã¯åœ¢æããããž
ããããã·ã¢ã»ãã³ã®éã枬å®ããããšã«ããã°
ãªã»ãªã³ãã°ãªã»ãªã³âããããã²ããŒãŒæŽ»æ§ã
å®éããããšãã§ããã
ãããã€ãŠæ¬çºæã®ç®çã¯æŽã«ã°ãªã»ãªã³ã®å®
éæ³ã§ããã該æ¹æ³ã¯ãã°ãªã»ãªã³ãNAD
ïŒïŒ°ïŒâç¬ç«æ§ã°ãªã»ãªã³ããããã²ããŒãŒã®ååš
ã§ãžããããã·ã¢ã»ãã³ã«å€ãããã®éïŒã€ã®ã
ããã³ããã³ïŒã€ã®é»åã¯çŽæ¥å¥œé©ãªå容äœã«ç§»
ããããã€é žååå容äœã®æžå°ãããã¯éå åå
容äœã®å¢å ãããã¯ãžããããã·ã¢ã»ãã³ã®å¢å
ãå®éããããšããæãã
éæ³ã§ããã該æ¹æ³ã¯ãã°ãªã»ãªã³ãNAD
ïŒïŒ°ïŒâç¬ç«æ§ã°ãªã»ãªã³ããããã²ããŒãŒã®ååš
ã§ãžããããã·ã¢ã»ãã³ã«å€ãããã®éïŒã€ã®ã
ããã³ããã³ïŒã€ã®é»åã¯çŽæ¥å¥œé©ãªå容äœã«ç§»
ããããã€é žååå容äœã®æžå°ãããã¯éå åå
容äœã®å¢å ãããã¯ãžããããã·ã¢ã»ãã³ã®å¢å
ãå®éããããšããæãã
å容äœïŒïŒ¡ïŒãšããŠã¯é»åããã³ãããã³ãåž
åãåŸãå šãŠã®ç©è³ªã奜é©ã§ãããå容äœãé µçŽ
åå¿ã®æ瀺è¬ãšããŠäœ¿çšããå Žåã«ã¯è©²å容äœã¯
é žååããéå åã«ç§»è¡ããéã«æž¬å®å¯èœãªå€å
ã瀺ããªããã°ãªããªããé žååããéå åã«ç§»
è¡ããéã«å¯èŠã®è²å€åãçæ³çã«ã¯ç¡è²ããç
è²ãžããŸãã¯ãã®éã®å€åã瀺ãå容äœãåªããŠ
ãããå容äœãã¬ããã¯ã¹éçšã§æž¬å®ãåŸãå€å
ã瀺ããªãå Žåã«ã¯ã第ïŒã®å容äœâã¬ããã¯ã¹
ç³»ã«æž¬å®å¯èœãªã¬ããã¯ã¹éçšãæãã第ïŒã®ã¬
ããã¯ã¹ç³»ãçµåããããšãã§ããã
åãåŸãå šãŠã®ç©è³ªã奜é©ã§ãããå容äœãé µçŽ
åå¿ã®æ瀺è¬ãšããŠäœ¿çšããå Žåã«ã¯è©²å容äœã¯
é žååããéå åã«ç§»è¡ããéã«æž¬å®å¯èœãªå€å
ã瀺ããªããã°ãªããªããé žååããéå åã«ç§»
è¡ããéã«å¯èŠã®è²å€åãçæ³çã«ã¯ç¡è²ããç
è²ãžããŸãã¯ãã®éã®å€åã瀺ãå容äœãåªããŠ
ãããå容äœãã¬ããã¯ã¹éçšã§æž¬å®ãåŸãå€å
ã瀺ããªãå Žåã«ã¯ã第ïŒã®å容äœâã¬ããã¯ã¹
ç³»ã«æž¬å®å¯èœãªã¬ããã¯ã¹éçšãæãã第ïŒã®ã¬
ããã¯ã¹ç³»ãçµåããããšãã§ããã
æ¬çºæã«ãã䜿çšãããæå³ã«ãããå容äœãš
ããŠæ°Žãããã¯ä»ã®éå åé žçŽ åœ¢ã«å€ããé žçŽ ã
䜿çšããããšãã§ãããé žçŽ æ¶è²»éã¯åžžæ³ã§ãäŸ
ãã°é žçŽ é»æ¥µãŸãã¯ã¯ãŒã«ãã«ã°æ€åºèšãçšããŠ
枬å®ããããšãã§ãããé žçŽ ã®æ¶è²»éã¯è©Šæäžã«
ååšããã°ãªã»ãªã³éã«æ¯äŸããã
ããŠæ°Žãããã¯ä»ã®éå åé žçŽ åœ¢ã«å€ããé žçŽ ã
䜿çšããããšãã§ãããé žçŽ æ¶è²»éã¯åžžæ³ã§ãäŸ
ãã°é žçŽ é»æ¥µãŸãã¯ã¯ãŒã«ãã«ã°æ€åºèšãçšããŠ
枬å®ããããšãã§ãããé žçŽ ã®æ¶è²»éã¯è©Šæäžã«
ååšããã°ãªã»ãªã³éã«æ¯äŸããã
è©Šæäžã«ååšããã°ãªã»ãªã³ã®éã¯å
¬ç¥æ¹æ³ã§
圢æããããžããããã·ã¢ã»ãã³ã®æž¬å®ã«ãã枬
å®ããããšãã§ããããã®å Žåãåæ§ã«äžé£ã®å ¬
ç¥æ¹æ³ãå©çšããããšãã§ãããäŸãã°ãµã³ããŒ
ã¹ïŒE.B.SandersïŒããã³ã·ãŠãŒãã«ãïŒJ.
SchubertïŒå ±èãâã¢ããªãã€ã«ã«ã»ã±ãã¹ããª
ïŒAnal.Chem.ïŒâã第43å·»ïŒ1971幎ïŒã59é 以äžãã
圢æããããžããããã·ã¢ã»ãã³ã®æž¬å®ã«ãã枬
å®ããããšãã§ããããã®å Žåãåæ§ã«äžé£ã®å ¬
ç¥æ¹æ³ãå©çšããããšãã§ãããäŸãã°ãµã³ããŒ
ã¹ïŒE.B.SandersïŒããã³ã·ãŠãŒãã«ãïŒJ.
SchubertïŒå ±èãâã¢ããªãã€ã«ã«ã»ã±ãã¹ããª
ïŒAnal.Chem.ïŒâã第43å·»ïŒ1971幎ïŒã59é 以äžãã
ç¹ã«é
žååããéå
åã«ç§»è¡ããéã«è²å€åã
ããå容äœã®äœ¿çšãæå©ã§ããããããè²å€åã
åå æ³ãçšããŠç°¡åãã€ä¿¡é ŒãåŸãæ¹æ³ã§åå æ³
ã«ãã枬å®ããããšãã§ããã
ããå容äœã®äœ¿çšãæå©ã§ããããããè²å€åã
åå æ³ãçšããŠç°¡åãã€ä¿¡é ŒãåŸãæ¹æ³ã§åå æ³
ã«ãã枬å®ããããšãã§ããã
以äžã«ãã®ã¬ããã¯ã¹éçšãçŽæ¥æž¬å®å¯èœã§ã
ãããããïŒã€ã®ã¬ããã¯ã¹ç³»ãšçµåããŠæž¬å®å¯
èœã«ããããšã®ã§ãããæ¬çºæã«ãã䜿çšå¯èœãª
å容äœç³»ã®äŸãè¥å¹²æããïŒ ïŒ Fe2+ïŒFe3+âç³» ããã«ã¯ãããµã·ã¢ãéïŒïŒé žã«ãªãŠã ã
䜿çšããããšãã§ããããã¯é»åã®åžååŸç¡è²
ã®ãããµã·ã¢ãéïŒïŒé žã«ãªãŠã ã«å€ããã
ããããéïŒïŒã€ãªã³ã«ãã€ãŠæ·±éè²ã«çè²
ããé¯äœã®ãã«ãªã³ãã«ãŒã圢æãããã
ãããããïŒã€ã®ã¬ããã¯ã¹ç³»ãšçµåããŠæž¬å®å¯
èœã«ããããšã®ã§ãããæ¬çºæã«ãã䜿çšå¯èœãª
å容äœç³»ã®äŸãè¥å¹²æããïŒ ïŒ Fe2+ïŒFe3+âç³» ããã«ã¯ãããµã·ã¢ãéïŒïŒé žã«ãªãŠã ã
䜿çšããããšãã§ããããã¯é»åã®åžååŸç¡è²
ã®ãããµã·ã¢ãéïŒïŒé žã«ãªãŠã ã«å€ããã
ããããéïŒïŒã€ãªã³ã«ãã€ãŠæ·±éè²ã«çè²
ããé¯äœã®ãã«ãªã³ãã«ãŒã圢æãããã
çŽæ¥éïŒïŒå¡©ã䜿çšããŠããããéå
æã«ç
ããéïŒïŒã€ãªã³ã¯å¥œé©ãªææãäŸãã°ãã¹
âãžã¡ãã«ã°ãªãªãã·ã ãŸãã¯ãŸãïŒã»10âã
ãšãã³ãããªã³ãšã®é¯åœ¢æã«ããæ€åºããããš
ãã§ããã
ããéïŒïŒã€ãªã³ã¯å¥œé©ãªææãäŸãã°ãã¹
âãžã¡ãã«ã°ãªãªãã·ã ãŸãã¯ãŸãïŒã»10âã
ãšãã³ãããªã³ãšã®é¯åœ¢æã«ããæ€åºããããš
ãã§ããã
ïŒ ãžâãŸãã¯ããªã¢ãªãŒã«ã¡ã¿ã³ææ
ãã®ã¬ããã¯ã¹ç³»ã«ã€ããŠã¯äžè¬çãªåå¿åŒ
ã以äžã®ããã«ããŠè¡šããããšãã§ããïŒ ãã®ææé¡ã®ä»£è¡šãšããŠæ¬¡ã®ãã®ãæããã
ãïŒãã©ã«ã€ãã°ãªãŒã³ãã¯ãªã¹ã¿ã«ãã€ãªã¬
ãããã¯ãã·ãŒãã«ãŒãããã¯ãããã®ãã€ã³
å¡©åºã
ã以äžã®ããã«ããŠè¡šããããšãã§ããïŒ ãã®ææé¡ã®ä»£è¡šãšããŠæ¬¡ã®ãã®ãæããã
ãïŒãã©ã«ã€ãã°ãªãŒã³ãã¯ãªã¹ã¿ã«ãã€ãªã¬
ãããã¯ãã·ãŒãã«ãŒãããã¯ãããã®ãã€ã³
å¡©åºã
ïŒ ããšããžã³ã¡ãã¹ã«ããšãŒãïŒPMSïŒ
äžè¬åŒïŒ
PMSã®é
žååããéå
åãžã®ç§»è¡ã¯æž¬å®å°
é£ã§ããããã®ã¬ããã¯ã¹ç³»ãä»ã®ã¬ããã¯ã¹
ç³»ãšçµåããã®ãæå©ã§ãããããã©ãŸãªãŠã
å¡©ãšã®çµåãæå©ã§ãããšèšŒæãããããã®äž
è¬çãªåå¿åŒã¯ä»¥äžã®ããã«ããŠè¡šããããšã
ã§ããïŒ (8) PMSïŒéå åïŒïŒããã©ãŸãªãŠã å¡©â ïŒé žååãç¡è²ïŒ PMSïŒé žååïŒïŒãã«ãã¶ã³ ïŒéå åãçè²ïŒ äŸãã°ããã©ãŸãªãŠã å¡©ãšããŠæ¬¡ã®ãã®ã䜿
çšããããšãã§ããïŒ INTïŒïŒâïŒïœâãšãŒãããšãã«ïŒâïŒâïŒïœâ
ãããããšãã«ïŒâïŒâããšãã«âããã©ãŸ
ãªãŠã âã¯ããªã MTTïŒïŒâïŒ4â²ã»5â²âãžã¡ãã«ãã¢ãŸãªã«âïŒ
âïŒâïŒã»ïŒâãžããšãã«âããã©ãŸãªãŠã
âãããã NBTïŒïŒã»2â²âãžïŒïœâãããããšãã«ïŒâ
ïŒã»5â²âãžããšãã«âïŒã»3â²âïŒïŒã»3â²âãžã¡
ããã·âïŒã»4â²âãžããšãã¬ã³ïŒâãžããã©
ãŸãªãŠã ã¯ããªã PMSã¯æå©ã«ã¯ïŒã»ïŒâãžã¯ãã«ããšããŒ
ã«ã€ã³ãããšããŒã«ïŒDCIPïŒãšçµåããããš
ãã§ããããã®å Žå次ã®ã¬ããã¯ã¹åŒã該åœã
ãïŒ (9) PMSïŒéå åïŒïŒDCIPïŒé žååïŒâ PMSïŒé žååïŒïŒDCIPïŒéå åïŒ ïŒ ã€ã³ãžãŽãããã¯ã€ã³ãžãŽèªå°äœ ã€ã³ãžãŽããã³ãã®èªå°äœã¯åæ§ã«ããŠæ°ŽçŽ
å容äœãšããŠäœçšããã次ã®äžè¬åŒã該åœã
ãïŒ æ¬çºæã«ãã䜿çšå¯èœãªã€ã³ãžãŽèªå°äœã¯äŸ
ãã°ïŒã»6â²âãžããã ã€ã³ãžãŽããã³ïŒã»5â²ã»
ïŒã»7â²âããã©ããã ã€ã³ãžãŽã§ããã
é£ã§ããããã®ã¬ããã¯ã¹ç³»ãä»ã®ã¬ããã¯ã¹
ç³»ãšçµåããã®ãæå©ã§ãããããã©ãŸãªãŠã
å¡©ãšã®çµåãæå©ã§ãããšèšŒæãããããã®äž
è¬çãªåå¿åŒã¯ä»¥äžã®ããã«ããŠè¡šããããšã
ã§ããïŒ (8) PMSïŒéå åïŒïŒããã©ãŸãªãŠã å¡©â ïŒé žååãç¡è²ïŒ PMSïŒé žååïŒïŒãã«ãã¶ã³ ïŒéå åãçè²ïŒ äŸãã°ããã©ãŸãªãŠã å¡©ãšããŠæ¬¡ã®ãã®ã䜿
çšããããšãã§ããïŒ INTïŒïŒâïŒïœâãšãŒãããšãã«ïŒâïŒâïŒïœâ
ãããããšãã«ïŒâïŒâããšãã«âããã©ãŸ
ãªãŠã âã¯ããªã MTTïŒïŒâïŒ4â²ã»5â²âãžã¡ãã«ãã¢ãŸãªã«âïŒ
âïŒâïŒã»ïŒâãžããšãã«âããã©ãŸãªãŠã
âãããã NBTïŒïŒã»2â²âãžïŒïœâãããããšãã«ïŒâ
ïŒã»5â²âãžããšãã«âïŒã»3â²âïŒïŒã»3â²âãžã¡
ããã·âïŒã»4â²âãžããšãã¬ã³ïŒâãžããã©
ãŸãªãŠã ã¯ããªã PMSã¯æå©ã«ã¯ïŒã»ïŒâãžã¯ãã«ããšããŒ
ã«ã€ã³ãããšããŒã«ïŒDCIPïŒãšçµåããããš
ãã§ããããã®å Žå次ã®ã¬ããã¯ã¹åŒã該åœã
ãïŒ (9) PMSïŒéå åïŒïŒDCIPïŒé žååïŒâ PMSïŒé žååïŒïŒDCIPïŒéå åïŒ ïŒ ã€ã³ãžãŽãããã¯ã€ã³ãžãŽèªå°äœ ã€ã³ãžãŽããã³ãã®èªå°äœã¯åæ§ã«ããŠæ°ŽçŽ
å容äœãšããŠäœçšããã次ã®äžè¬åŒã該åœã
ãïŒ æ¬çºæã«ãã䜿çšå¯èœãªã€ã³ãžãŽèªå°äœã¯äŸ
ãã°ïŒã»6â²âãžããã ã€ã³ãžãŽããã³ïŒã»5â²ã»
ïŒã»7â²âããã©ããã ã€ã³ãžãŽã§ããã
ïŒ ããšãããµãžã³âãããã¯ããšããã¢ãžã³æ
æ ããšãããµãžã³ãããã¯ããšããã¢ãžã³ææ
ãæ°ŽçŽ ååãåžåãããã€ãã®éçè²ã®é žåå
ããç¡è²ã®éå åã«å€ããã
æ ããšãããµãžã³ãããã¯ããšããã¢ãžã³ææ
ãæ°ŽçŽ ååãåžåãããã€ãã®éçè²ã®é žåå
ããç¡è²ã®éå åã«å€ããã
ãã®ææã«ã€ããŠã¯äŸãã°ïŒåååœãæ°ŽçŽ å
åïŒåãåžåããŠãã€ã³ã¡ãã¬ã³ãã«ãŒã«å€ã
ãã¡ãã¬ã³ãã«ãŒãæããããã
åïŒåãåžåããŠãã€ã³ã¡ãã¬ã³ãã«ãŒã«å€ã
ãã¡ãã¬ã³ãã«ãŒãæããããã
åèšã®ã°ãªã»ãªã³å®éæ¹æ³ã¯ã°ãªã»ãªã³ã«å€
æãããç©è³ªã®å®é䞊ã³ã«ãããåå¿ãæ¥è§Šã
ãé µçŽ ã®å®éã«ãæå©ã«äœ¿çšãåŸãã
æãããç©è³ªã®å®é䞊ã³ã«ãããåå¿ãæ¥è§Šã
ãé µçŽ ã®å®éã«ãæå©ã«äœ¿çšãåŸãã
åèšã®çšéã®äŸãšããŠè¡æ¶²ãè¡æŒ¿ãŸãã¯è¡æž
äžã®ããªã°ãªã»ãªãã®å®éã詳説ããããã®ã
ãã«ããªã°ãªã»ãªãïŒäžæ§èèªïŒã奜é©ãªãªã
ãŒãŒãçšããŠå ¬ç¥æ¹æ³ã§å æ°Žå解ãããã€éé¢
ã®ã°ãªã»ãªã³ãæ¬çºæã«ããã°ãªã»ãªã³ããã
ãã²ããŒãŒããã³å¥œé©ãªé»åå容äœããã³æ°ŽçŽ
å容äœãçšããŠåèšã®ããã«ããŠå€æãããã
åå¿åŒã¯ä»¥äžã®ããã«ããŠè¡šããããšãã§ã
ãïŒ ããªã°ãªã»ãªãã®é µçŽ å æ°Žå解ã¯å ¬ç¥æ¹æ³ã«
ããå®æœããããããªã°ãªã»ãªããããåãã
ããã®é µçŽ ãšããŠã¯æå©ã«åžè²©ã®ãªãã¿ã³ãã¯
質ãªããŒãŒïŒE.C.3.1.1 34ïŒãŸãã¯ãªããŒãŒ
ïŒE.C.3.1.1.3ïŒãšãšã¹ãã©ãŒãŒïŒE.C.3.1.1.1ïŒ
ã®æ··åç©ãå©çšããããšãã§ããã
äžã®ããªã°ãªã»ãªãã®å®éã詳説ããããã®ã
ãã«ããªã°ãªã»ãªãïŒäžæ§èèªïŒã奜é©ãªãªã
ãŒãŒãçšããŠå ¬ç¥æ¹æ³ã§å æ°Žå解ãããã€éé¢
ã®ã°ãªã»ãªã³ãæ¬çºæã«ããã°ãªã»ãªã³ããã
ãã²ããŒãŒããã³å¥œé©ãªé»åå容äœããã³æ°ŽçŽ
å容äœãçšããŠåèšã®ããã«ããŠå€æãããã
åå¿åŒã¯ä»¥äžã®ããã«ããŠè¡šããããšãã§ã
ãïŒ ããªã°ãªã»ãªãã®é µçŽ å æ°Žå解ã¯å ¬ç¥æ¹æ³ã«
ããå®æœããããããªã°ãªã»ãªããããåãã
ããã®é µçŽ ãšããŠã¯æå©ã«åžè²©ã®ãªãã¿ã³ãã¯
質ãªããŒãŒïŒE.C.3.1.1 34ïŒãŸãã¯ãªããŒãŒ
ïŒE.C.3.1.1.3ïŒãšãšã¹ãã©ãŒãŒïŒE.C.3.1.1.1ïŒ
ã®æ··åç©ãå©çšããããšãã§ããã
è¡æ¶²äžã®ã°ã«ã³ãŒã¹ã¯æ¬çºæã«ããNAD
ïŒïŒ°ïŒâç¬ç«æ§ã°ãªã»ãªã³ããããã²ããŒãŒãçš
ãããã°ãªã»ãªã³ãšããªã°ãªã»ãªãã®å®éã«åœ±
é¿ãäžããªãã
ïŒïŒ°ïŒâç¬ç«æ§ã°ãªã»ãªã³ããããã²ããŒãŒãçš
ãããã°ãªã»ãªã³ãšããªã°ãªã»ãªãã®å®éã«åœ±
é¿ãäžããªãã
以äžå®æœäŸã«ã€ãæ¬çºæã詳述ããïŒ
äŸ ïŒ
NADïŒïŒ°ïŒâç¬ç«æ§ã°ãªã»ãªã³ããããã²ããŒ
ãŒïŒã°ã«ã³ããã¯ã¿ãŒã»ã€ã³ãã¹ããªãŠã¹
DSM2621ïŒIFOâ3260ïŒããïŒ ã°ã«ã³ããã¯ã¿ãŒã»ã€ã³ãã¹ããªãŠã¹
DSM2621ãã°ãªã»ãªã³âã°ã«ã¿ãã³é žâåªäœäž
ã§å ¬ç¥æ¹æ³ã«ããå¹é€ããã现èç©è³ªãéããã
ã€ãã¬ã³ãã»ãã¬ã¹ã§å§å1000Kg/cm2ã§ç Žå£ã
ããç Žå£ãããªãã€ã现èãé å¿åé¢ïŒ5000ïœïŒ
ã«ããé€å»ããã现èãã¢ãžããŒããè¶ é å¿åé¢
æ©ïŒ68000ïœïŒã§90åé å¿åé¢ããã现èèãã©
ã¯ã·ãšã³ãæéããããããã0.05Mâããªã¹â
HClâç·©è¡æ¶²ïŒPH8.0ïŒãçšããŠã¿ã³ãã¯è³ª10mg/
mlã®å«éã«èª¿ç¯ãããæçµæ¿åºŠ0.5ïŒ ã«ãããã
ã«åžžçšã®ã奜é©ãªæž æµå€ãäŸãã°ããªãã³
ïŒTritonïŒÃ100ãæ·»å ãããåŸãããæ··åç©ãïŒ
âã§60åæ¹æããŠé µçŽ ã溶液ã«ãããåŒç¶ã60å
éé å¿åé¢ããïŒ68000ïœïŒãäžæŸã¿æ¶²ããDEAE
âã»ã«ããŒãŒã¹ïŒPH7.5ïŒããã³ããããã·ã«ã¢
ãã¿ã€ãïŒPH7.5ïŒãçšããŠã¯ãããã°ã©ãã€ãŒ
ããããšã«ããã¿ã³ãã¯è³ªïŒmgåœã50Uã®æŽ»æ§ã
æãããäžæ§ãªé µçŽ 調å€ãåŸãããã
ãŒïŒã°ã«ã³ããã¯ã¿ãŒã»ã€ã³ãã¹ããªãŠã¹
DSM2621ïŒIFOâ3260ïŒããïŒ ã°ã«ã³ããã¯ã¿ãŒã»ã€ã³ãã¹ããªãŠã¹
DSM2621ãã°ãªã»ãªã³âã°ã«ã¿ãã³é žâåªäœäž
ã§å ¬ç¥æ¹æ³ã«ããå¹é€ããã现èç©è³ªãéããã
ã€ãã¬ã³ãã»ãã¬ã¹ã§å§å1000Kg/cm2ã§ç Žå£ã
ããç Žå£ãããªãã€ã现èãé å¿åé¢ïŒ5000ïœïŒ
ã«ããé€å»ããã现èãã¢ãžããŒããè¶ é å¿åé¢
æ©ïŒ68000ïœïŒã§90åé å¿åé¢ããã现èèãã©
ã¯ã·ãšã³ãæéããããããã0.05Mâããªã¹â
HClâç·©è¡æ¶²ïŒPH8.0ïŒãçšããŠã¿ã³ãã¯è³ª10mg/
mlã®å«éã«èª¿ç¯ãããæçµæ¿åºŠ0.5ïŒ ã«ãããã
ã«åžžçšã®ã奜é©ãªæž æµå€ãäŸãã°ããªãã³
ïŒTritonïŒÃ100ãæ·»å ãããåŸãããæ··åç©ãïŒ
âã§60åæ¹æããŠé µçŽ ã溶液ã«ãããåŒç¶ã60å
éé å¿åé¢ããïŒ68000ïœïŒãäžæŸã¿æ¶²ããDEAE
âã»ã«ããŒãŒã¹ïŒPH7.5ïŒããã³ããããã·ã«ã¢
ãã¿ã€ãïŒPH7.5ïŒãçšããŠã¯ãããã°ã©ãã€ãŒ
ããããšã«ããã¿ã³ãã¯è³ªïŒmgåœã50Uã®æŽ»æ§ã
æãããäžæ§ãªé µçŽ 調å€ãåŸãããã
äŸ ïŒ
äŸïŒã«æãã埮çç©ã°ã«ã³ããã¯ã¿ãŒã»ã€ã³ã
ã¹ããªãŠã¹DSM2621ã®ä»£ããã«ã¢ã»ããã¯ã¿
ãŒã»ãã·ãªãã¹DSM2623ãããã¯ã°ã«ã³ããã¯
ã¿ãŒã»ã»ãªãã¹DSM2622ãäŸïŒã«èšèŒã®æ¹æ³ã§
å¹é€ãããã€åŸåŠçããã42U/mgãããã¯46U/
mgã®æŽ»æ§ãæã€é µçŽ 調å€ãåŸãããã
ã¹ããªãŠã¹DSM2621ã®ä»£ããã«ã¢ã»ããã¯ã¿
ãŒã»ãã·ãªãã¹DSM2623ãããã¯ã°ã«ã³ããã¯
ã¿ãŒã»ã»ãªãã¹DSM2622ãäŸïŒã«èšèŒã®æ¹æ³ã§
å¹é€ãããã€åŸåŠçããã42U/mgãããã¯46U/
mgã®æŽ»æ§ãæã€é µçŽ 調å€ãåŸãããã
äŸ ïŒ
æ¬çºæã«ããã°ãªã»ãªã³ããããã²ããŒãŒã®é
µ
çŽ æŽ»æ§ã®å®é ããŠãããã«ããšããžã³ã¡ãã¹ã«ããšãŒã0.6
ÎŒã¢ã«ãïŒã»ïŒâãžã¯ãã«ããšããŒã«ã€ã³ãããš
ããŒã«0.6ÎŒã¢ã«ããã³ããªã¹âHClâç·©è¡æ¶²
ïŒPH7.5ïŒ100ÎŒã¢ã«ãè£ å ¥ããããã®æŽ»æ§ã0.01
ã0.1åäœã«ãããå®éãã¹ãé µçŽ èª¿å€ãæ·»å ã
ããïŒåºŠèžæºããæ°Žãæ·»å ããŠæçµå®¹é2.9mlã
補é ããã25âã§ïŒåã®ææž©ä¿æåŸã°ãªã»ãªã³
100ÎŒã¢ã«ãæ·»å ããŠé µçŽ åå¿ãéå§ãããã
ïŒã»ïŒâãžã¯ãã«ããšããŒã«ã€ã³ãããšããŒã«ã®
è±è²ã®é床ã600nïœã§ã®åžåã枬å®ããããšã«
ããè¿œæ±ãããé µçŽ æŽ»æ§ã®èšç®ããPH7.5ã«ãã
ãïŒã»ïŒâãžã¯ãã«ããšããŒã«ã€ã³ãããšããŒã«
ã®ïŒãã«ã¢ã«ã®åžå ä¿æ°ãå€100ãšããŠè¡ãªãã
åžå å€åâ³E600ïŒ2.33åãè¡ãªãé µçŽ éãïŒåäœ
ãšããã
çŽ æŽ»æ§ã®å®é ããŠãããã«ããšããžã³ã¡ãã¹ã«ããšãŒã0.6
ÎŒã¢ã«ãïŒã»ïŒâãžã¯ãã«ããšããŒã«ã€ã³ãããš
ããŒã«0.6ÎŒã¢ã«ããã³ããªã¹âHClâç·©è¡æ¶²
ïŒPH7.5ïŒ100ÎŒã¢ã«ãè£ å ¥ããããã®æŽ»æ§ã0.01
ã0.1åäœã«ãããå®éãã¹ãé µçŽ èª¿å€ãæ·»å ã
ããïŒåºŠèžæºããæ°Žãæ·»å ããŠæçµå®¹é2.9mlã
補é ããã25âã§ïŒåã®ææž©ä¿æåŸã°ãªã»ãªã³
100ÎŒã¢ã«ãæ·»å ããŠé µçŽ åå¿ãéå§ãããã
ïŒã»ïŒâãžã¯ãã«ããšããŒã«ã€ã³ãããšããŒã«ã®
è±è²ã®é床ã600nïœã§ã®åžåã枬å®ããããšã«
ããè¿œæ±ãããé µçŽ æŽ»æ§ã®èšç®ããPH7.5ã«ãã
ãïŒã»ïŒâãžã¯ãã«ããšããŒã«ã€ã³ãããšããŒã«
ã®ïŒãã«ã¢ã«ã®åžå ä¿æ°ãå€100ãšããŠè¡ãªãã
åžå å€åâ³E600ïŒ2.33åãè¡ãªãé µçŽ éãïŒåäœ
ãšããã
äŸ ïŒ
ããšããžã³ã¡ãã¹ã«ããšãŒãããã³ïŒã»ïŒâãž
ã¯ãã«ããšããŒã«ã€ã³ãããšããŒã«ãçšããŠã°
ãªã»ãªã³ã®å®é ïŒ èŒæ£æ²ç·ã®äœæ ã°ãªã»ãªã³æº¶æ¶²ïŒ100ÎŒã¢ã«ïŒïŒããïŒåºŠ
èžæºæ°ŽãçšããŠæº¶æ¶²0.1mläžã«ã°ãªã»ãªã³
0.01ã0.02ã0.04ã0.06ã0.08ã0.10ã0.50ÎŒã¢
ã«ãå«æããïŒçš®ã®æšæºæº¶æ¶²ã補é ãããåã°
ãªã»ãªã³æšæºæº¶æ¶²ã«æŽ»æ§10Uãæããé µçŽ æº¶æ¶²
0.1mlãæ·»å ãããïŒåºŠèžæºæ°Žã®æ·»å ã«ããã
ããã2.8mlã®æçµå®¹éã«èª¿ç¯ããã25âã§ïŒ
åææž©ä¿æããã
ã¯ãã«ããšããŒã«ã€ã³ãããšããŒã«ãçšããŠã°
ãªã»ãªã³ã®å®é ïŒ èŒæ£æ²ç·ã®äœæ ã°ãªã»ãªã³æº¶æ¶²ïŒ100ÎŒã¢ã«ïŒïŒããïŒåºŠ
èžæºæ°ŽãçšããŠæº¶æ¶²0.1mläžã«ã°ãªã»ãªã³
0.01ã0.02ã0.04ã0.06ã0.08ã0.10ã0.50ÎŒã¢
ã«ãå«æããïŒçš®ã®æšæºæº¶æ¶²ã補é ãããåã°
ãªã»ãªã³æšæºæº¶æ¶²ã«æŽ»æ§10Uãæããé µçŽ æº¶æ¶²
0.1mlãæ·»å ãããïŒåºŠèžæºæ°Žã®æ·»å ã«ããã
ããã2.8mlã®æçµå®¹éã«èª¿ç¯ããã25âã§ïŒ
åææž©ä¿æããã
次ãã§ããšããžã³ã¡ãã¹ã«ããšãŒã0.6ÎŒã¢
ã«ããã³ïŒã»ïŒâãžã¯ãã«ããšããŒã«ã€ã³ãã
ãšããŒã«0.6ÎŒã¢ã«ãå«æãã溶液0.2mlã®æ·»å
ã«ããåå¿ãéå§ããããåå èšãçšããŠ
600nïœã§ã®åžå ã枬å®ãããã°ãªã»ãªã³0.01ã
0.05ÎŒã¢ã«ã®ç¯å²ã§çŽç·çãªçµéãèªããã
ãã
ã«ããã³ïŒã»ïŒâãžã¯ãã«ããšããŒã«ã€ã³ãã
ãšããŒã«0.6ÎŒã¢ã«ãå«æãã溶液0.2mlã®æ·»å
ã«ããåå¿ãéå§ããããåå èšãçšããŠ
600nïœã§ã®åžå ã枬å®ãããã°ãªã»ãªã³0.01ã
0.05ÎŒã¢ã«ã®ç¯å²ã§çŽç·çãªçµéãèªããã
ãã
ïŒ æªç¥ã®è©Šæã®æž¬å®
æªç¥ã®ã°ãªã»ãªã³å«éãæããè©ŠæïŒmlã«ã
ãŠãããäžã§ããšããžã³ã¡ãã¹ã«ããšãŒã0.6
ÎŒã¢ã«ããã³ïŒã»ïŒâãžã¯ãã«ããšããŒã«ã€ã³
ãããšããŒã«0.6ÎŒã¢ã«ãå«ã溶液ïŒmlãæ·»ã
ããããªã¹âHClâç·©è¡æ¶²ïŒPH7.5ïŒ0.5mlïŒ100
ÎŒã¢ã«ïŒãå ãããã€25âã§ïŒåææž©ä¿æã
ããæ€åºåå¿ã掻æ§10Uã®é µçŽ 溶液0.1mlã®æ·»
å ã«ããéå§ãããã600nïœã§ã®åžå ã®æžå°
ã枬å®ããã枬å®ããåžå å·®ãèŒæ£æ²ç·ãšæ¯ã¹
ãŠäœ¿çšè©Šæã®ã°ãªã»ãªã³å«éã枬å®ããã
ãŠãããäžã§ããšããžã³ã¡ãã¹ã«ããšãŒã0.6
ÎŒã¢ã«ããã³ïŒã»ïŒâãžã¯ãã«ããšããŒã«ã€ã³
ãããšããŒã«0.6ÎŒã¢ã«ãå«ã溶液ïŒmlãæ·»ã
ããããªã¹âHClâç·©è¡æ¶²ïŒPH7.5ïŒ0.5mlïŒ100
ÎŒã¢ã«ïŒãå ãããã€25âã§ïŒåææž©ä¿æã
ããæ€åºåå¿ã掻æ§10Uã®é µçŽ 溶液0.1mlã®æ·»
å ã«ããéå§ãããã600nïœã§ã®åžå ã®æžå°
ã枬å®ããã枬å®ããåžå å·®ãèŒæ£æ²ç·ãšæ¯ã¹
ãŠäœ¿çšè©Šæã®ã°ãªã»ãªã³å«éã枬å®ããã
äŸ ïŒ
ããšããžã³ã¡ãã¹ã«ããšãŒãããã³ããã©ãŸãª
ãŠã å¡©ãçšããŠã°ãªã»ãªã³ã®å®é æªç¥ã®ã°ãªã»ãªã³å«éãæããè©ŠæïŒmlã«ããŠ
ãããäžã§ããšããžã³ã¡ãã¹ã«ããšãŒã0.1ÎŒã¢
ã«ãïŒâïŒïŒã»ïŒâãžã¡ãã«ãã¢ãŸãªã«âïŒïŒâ
ïŒã»ïŒâãžããšãã«âããã©ãŸãªãŠã ãããã
1.2ÎŒã¢ã«ããã³ããªã¹âHClâç·©è¡æ¶²ïŒ0.1MïŒ
PH7.5ïŒ100ÎŒã¢ã«ãå«æãã溶液ïŒmlãå ããã
ããã25âã§ïŒåææž©ä¿æãããæ¬çºæã«ããé µ
çŽ 1Uãæã€æº¶æ¶²20ÎŒã®æ·»å ã«ããæ€åºåå¿ã
éå§ãããã570nïœã§åžå å€åã枬å®ããã枬
å®ãããåžå å·®ãèŒæ£æ²ç·ïŒããã¯æ¢ç¥ã®ã°ãªã»
ãªã³å«éãæããè©Šæã®æž¬å®ã«ããäœæããããš
ãã§ããïŒãšæ¯ã¹ãŠäœ¿çšãããè©Šæã®ã°ãªã»ãªã³
å«éã枬å®ããã
ãŠã å¡©ãçšããŠã°ãªã»ãªã³ã®å®é æªç¥ã®ã°ãªã»ãªã³å«éãæããè©ŠæïŒmlã«ããŠ
ãããäžã§ããšããžã³ã¡ãã¹ã«ããšãŒã0.1ÎŒã¢
ã«ãïŒâïŒïŒã»ïŒâãžã¡ãã«ãã¢ãŸãªã«âïŒïŒâ
ïŒã»ïŒâãžããšãã«âããã©ãŸãªãŠã ãããã
1.2ÎŒã¢ã«ããã³ããªã¹âHClâç·©è¡æ¶²ïŒ0.1MïŒ
PH7.5ïŒ100ÎŒã¢ã«ãå«æãã溶液ïŒmlãå ããã
ããã25âã§ïŒåææž©ä¿æãããæ¬çºæã«ããé µ
çŽ 1Uãæã€æº¶æ¶²20ÎŒã®æ·»å ã«ããæ€åºåå¿ã
éå§ãããã570nïœã§åžå å€åã枬å®ããã枬
å®ãããåžå å·®ãèŒæ£æ²ç·ïŒããã¯æ¢ç¥ã®ã°ãªã»
ãªã³å«éãæããè©Šæã®æž¬å®ã«ããäœæããããš
ãã§ããïŒãšæ¯ã¹ãŠäœ¿çšãããè©Šæã®ã°ãªã»ãªã³
å«éã枬å®ããã
äŸ ïŒ
ããªã°ãªã»ãªãã®å®é
ããè¡æž
0.02mlã«ããŠãããäžã§ãªãã¿ã³ãã¯
質ãªããŒãŒ100ã200UãŸãã¯ãªããŒãŒããã³ãšã¹
ãã©ãŒãŒãå«æããé µçŽ æº¶æ¶²0.5mlãå ãããã€
25âã§20åææž©ä¿æããã
質ãªããŒãŒ100ã200UãŸãã¯ãªããŒãŒããã³ãšã¹
ãã©ãŒãŒãå«æããé µçŽ æº¶æ¶²0.5mlãå ãããã€
25âã§20åææž©ä¿æããã
次ãã§ããšããžã³ã¡ãã¹ã«ããšãŒã0.6ÎŒã¢
ã«ãïŒã»ïŒâãžã¯ãã«ããšããŒã«ã€ã³ãããšããŒ
ã«0.2ÎŒã¢ã«ããã³ããªã¹âHClâç·©è¡æ¶²ïŒPH
7.5ïŒ100ÎŒã¢ã«ãå«ã溶液ïŒmlãæ·»å ããã次ã
ã§25âã§ïŒåææž©ä¿æããã次ãã§æ€åºåå¿ãæ¬
çºæã«ããã°ãªã»ãªã³ããããã²ããŒãŒ1Uãå«
æãã溶液0.1mlã®æ·»å ã«ããéå§ãããã
ã«ãïŒã»ïŒâãžã¯ãã«ããšããŒã«ã€ã³ãããšããŒ
ã«0.2ÎŒã¢ã«ããã³ããªã¹âHClâç·©è¡æ¶²ïŒPH
7.5ïŒ100ÎŒã¢ã«ãå«ã溶液ïŒmlãæ·»å ããã次ã
ã§25âã§ïŒåææž©ä¿æããã次ãã§æ€åºåå¿ãæ¬
çºæã«ããã°ãªã»ãªã³ããããã²ããŒãŒ1Uãå«
æãã溶液0.1mlã®æ·»å ã«ããéå§ãããã
600nïœã§ã®åžå
æžå°ã枬å®ãããåžå
å·®ãã
äŸïŒã«èšèŒã®æ¹æ³ã§ã°ãªã»ãªã³å«éããããã€ãŠ
ããªã°ãªã»ãªãå«éã枬å®ããããšãã§ãããå¹³
åããªã°ãªã»ãªãå«é110mg/dlãå€æããã
äŸïŒã«èšèŒã®æ¹æ³ã§ã°ãªã»ãªã³å«éããããã€ãŠ
ããªã°ãªã»ãªãå«éã枬å®ããããšãã§ãããå¹³
åããªã°ãªã»ãªãå«é110mg/dlãå€æããã
ããªã°ãªã»ãªãå®éã§ã¯ç²æ€å€æž¬å®ã®å®æœãæš
奚ãããã
奚ãããã
Claims (1)
- ãç¹èš±è«æ±ã®ç¯å²ã ïŒ è³é©PHå€ãPH7.0ã8.5ã§ãããå®å®PHå€ãPH
6.0ã10.0ã§ããã枩床25ã37âã§æé«ã®æŽ»æ§ã
æãããªããµã¡ãŒãã«ãã掻æ§ã¯è»œãæå¶ããã
ãªã³è質ã«ãã掻æ§ã¯çŽ20ïŒ å¢å ããããã«ãã
ãªã³ããã³ãè£æ¬ ååå£ãšããŠæãããç¹ç°çã«
ã°ãªã»ãªã³ãè±æ°ŽçŽ ããŠãžããããã·ã¢ã»ãã³ã«
ãããã®éåé¢ãããïŒã€ã®ãããã³ããã³ïŒã€
ã®é»åã¯NADïŒïŒ°ïŒãäžç·ã«äœ¿çšããã«å¥œé©ãª
å容äœã«ç§»ããããNADïŒïŒ°ïŒâç¬ç«æ§ã°ãªã»ãª
ã³ããããã²ããŒãŒã ïŒ ã°ãªã»ãªã³ãå容äœã®ååšã§ãžããããã·ã¢
ã»ãã³ã«å€ãããç¹èš±è«æ±ã®ç¯å²ç¬¬ïŒé ã«èšèŒã®
NADïŒïŒ°ïŒâç¬ç«æ§ã°ãªã»ãªã³ããããã²ããŒ
ãŒã ïŒ NADïŒïŒ°ïŒâç¬ç«æ§ã°ãªã»ãªã³ããããã²ã
ãŒãŒã補é ããããã®æ¹æ³ã«ãããŠããã®é µçŽ ã
圢æãåŸããããœã€ãã¢ãã¹ãã»ã©ãã¢ãã¯ã¬ã
ã·ãšã©ããšã«ãŠã€ãã¢ãã¢ã¹ãã«ã®ã«ã¹ãããã·
ãªãŠã ããªãŸãã¹ãã¢ã»ããã¯ã¿ãŒãŸãã¯ã°ã«ã³
ããã¯ã¿ãŒå±ã®åŸ®çç©ãå¹å°ã§å¹é€ãããã€é µçŽ
ãå¹é€æ±ããåé¢ããããšãç¹åŸŽãšãããè³é©PH
å€ãPH7.0ã8.5ã§ãããå®å®PHå€ãPH6.0ã10.0ã§
ããã枩床25ã37âã§æé«ã®æŽ»æ§ãæãããªããµ
ã¡ãŒãã«ãã掻æ§ã¯è»œãæå¶ããããªã³è質ã«ã
ã掻æ§ã¯çŽ20ïŒ å¢å ããããã«ãããªã³ããã³ã
è£æ¬ ååå£ãšããŠæãããç¹ç°çã«ã°ãªã»ãªã³ã
è±æ°ŽçŽ ããŠãžããããã·ã¢ã»ãã³ã«ãããã®éå
é¢ãããïŒã€ã®ãããã³ããã³ïŒã€ã®é»åã¯
NADïŒïŒ°ïŒãäžç·ã«äœ¿çšããã«å¥œé©ãªå容äœã«
移ããããNADïŒïŒ°ïŒâç¬ç«æ§ã°ãªã»ãªã³ããã
ãã²ããŒãŒã®è£œæ³ã ïŒ åŸ®çç©ãšããŠã¢ã»ããã¯ã¿ãŒã»ãã·ãªãã¹
DSM2623ãã°ã«ã³ããã¯ã¿ãŒã»ã»ãªãã¹
DSM2622ãŸãã¯ã°ã«ã³ããã¯ã¿ãŒã»ã€ã³ãã¹ã
ãªãŠã¹DSM2621ã䜿çšãããç¹èš±è«æ±ã®ç¯å²ç¬¬
ïŒé èšèŒã®æ¹æ³ã ïŒ ã°ãªã»ãªã³ãå®éããããã®æ¹æ³ã«ãããŠã
NADïŒïŒ°ïŒâç¬ç«æ§ã°ãªã»ãªã³ããããã²ããŒãŒ
ã®ååšãå容äœã®ååšã§ã°ãªã»ãªã³ããžãããã
ã·ã¢ã»ãã³ã«å€ãããã€é žååå容äœã®æžå°ãã
ãã¯éå åå容äœã®å¢å ãããã¯éå åå容äœã®
å¢å ãããã¯ãžããããã·ã¢ã»ãã³ã®å¢å ãå®é
ããããšãç¹åŸŽãšãããã°ãªã»ãªã³ãå®éããã
ãã®æ¹æ³ã ïŒ å容äœãšããŠé žçŽ ã䜿çšãããã€é žçŽ ã®æ¶è²»
éã枬å®ãããç¹èš±è«æ±ã®ç¯å²ç¬¬ïŒé èšèŒã®æ¹
æ³ã ïŒ ç¬¬ïŒå容äœâã¬ããã¯ã¹ç³»ãããïŒã€ã®ã¬ã
ãã¯ã¹æ瀺è¬ãšçµåãããç¹èš±è«æ±ã®ç¯å²ç¬¬ïŒé
èšèŒã®æ¹æ³ã ïŒ å容äœãšããŠããšããžã³ã¡ãã¹ã«ããšãŒã
ãããã€ä»ã®ã¬ããã¯ã¹æ瀺è¬ãšããŠïŒã»ïŒâãž
ã¯ãã«ããšããŒã«ã€ã³ãããšããŒã«ã䜿çšããã
ã€ïŒã»ïŒâãžã¯ãã«ããšããŒã«ã€ã³ãããšããŒã«
ã®åžå äœäžã600nïœã§æž¬å®ãããç¹èš±è«æ±ã®ç¯
å²ç¬¬ïŒé èšèŒã®æ¹æ³ã ïŒ å容äœãšããŠããšããžã³ã¡ãã¹ã«ããšãŒã
ãããã€ä»ã®ã¬ããã¯ã¹æ瀺è¬ãšããŠããã©ãŸãª
ãŠã å¡©ã䜿çšãããã€ãã«ãã¶ã³åœ¢æã枬å æ³ã§
枬å®ãããç¹èš±è«æ±ã®ç¯å²ç¬¬ïŒé èšèŒã®æ¹æ³ã ïŒïŒ ããªã°ãªã»ãªããå®éããããã®æ¹æ³ã«ã
ããŠãããªã°ãªã»ãªããããåããŠã°ãªã»ãªã³ãš
éé¢ã®èèªé žã«ãããã€NADïŒïŒ°ïŒâç¬ç«æ§ã°ãª
ã»ãªã³ããããã²ããŒãŒã®ååšãå容äœã®ååšã§
éé¢ã°ãªã»ãªã³ããžããããã·ã¢ã»ãã³ã«å€ãã
ãã€é žååå容äœã®æžå°ãããã¯éå åå容äœã®
å¢å ãããã¯ãžããããã·ã¢ã»ãã³ã®å¢å ãå®é
ããããšãç¹åŸŽãšãããããªã°ãªã»ãªããå®éã
ãããã®æ¹æ³ã ïŒïŒ 該詊è¬äžã§å Žåã«ããããªã°ãªã»ãªããã
ãåãããŠã°ãªã»ãªã³ãšéé¢èèªé žã«ãªããã°ãª
ã»ãªã³ã奜é©ãªå容äœã®ååšã§ã°ãªã»ãªã³ããã
ãã²ããŒãŒã«ãã€ãŠãžããããã·ã¢ã»ãã³ã«å€æ
ããããã€å容äœã®æžå°ãŸãã¯å¢å ãããã¯ãžã
ãããã·ã¢ã»ãã³ã®å¢å ãå®éããããã°ãªã»ãª
ã³äžŠã³ã«ããªã°ãªã»ãªããå®éããããã®è©Šè¬ã«
ãããŠã該詊è¬ãNADïŒïŒ°ïŒâç¬ç«æ§ã°ãªã»ãªã³
ããããã²ããŒãŒãå«æããããšãç¹åŸŽãšããã
ã°ãªã»ãªã³äžŠã³ã«ããªã°ãªã»ãªããå®éãããã
ã®è©Šè¬ã
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
DE19833311027 DE3311027A1 (de) | 1983-03-25 | 1983-03-25 | Nad(p)-unabhaengige glycerindehydrogenase, verfahren zu deren herstellung sowie deren verwendung zur bestimmung von glycerin und triglyceriden |
DE3311027.1 | 1983-03-25 |
Publications (2)
Publication Number | Publication Date |
---|---|
JPS59183689A JPS59183689A (ja) | 1984-10-18 |
JPS6122952B2 true JPS6122952B2 (ja) | 1986-06-03 |
Family
ID=6194725
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP59054611A Granted JPS59183689A (ja) | 1983-03-25 | 1984-03-23 | ïœïœïŒïœïŒâç¬ç«æ§ã°ãªã»ãªã³ããããã²ãâãŒããã®è£œæ³ããã³ã°ãªã»ãªã³ããã³ããªã°ãªã»ãªããå®éããããã®æ¹æ³ããã³è©Šè¬ |
Country Status (6)
Country | Link |
---|---|
US (1) | US4576913A (ja) |
EP (1) | EP0120440B1 (ja) |
JP (1) | JPS59183689A (ja) |
AT (1) | ATE55410T1 (ja) |
CA (1) | CA1225950A (ja) |
DE (2) | DE3311027A1 (ja) |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2008220367A (ja) * | 2007-02-13 | 2008-09-25 | Cci Corp | ããªãªãŒã«è±æ°ŽçŽ é µçŽ ã®è£œé æ¹æ³ |
Families Citing this family (11)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
DE3826922A1 (de) * | 1988-08-09 | 1990-02-22 | Boehringer Mannheim Gmbh | Verfahren zur kolorimetrischen bestimmung eines analyten mittels enzymatischer oxidation |
US4929545A (en) * | 1989-04-14 | 1990-05-29 | Boehringer Mannheim Corporation | Method and reagent for determination of an analyte via enzymatic means using a ferricyanide/ferric compound system |
ES2138596T3 (es) * | 1991-02-27 | 2000-01-16 | Roche Diagnostics Corp | Estabilizacion de un reactivo que contiene una enzima para la determinacion de un analito. |
WO1992015704A1 (en) * | 1991-02-27 | 1992-09-17 | Boehringer Mannheim Corporation | Improved method and reagent for determination of an analyte |
US5707820A (en) * | 1991-09-19 | 1998-01-13 | Boehringer Mannheim Corporation | Reagent and assay methods including a phenazine-containing indicator |
JP3041840B2 (ja) * | 1992-02-24 | 2000-05-15 | æ±æŽçŽ¡çžŸæ ªåŒäŒç€Ÿ | æ°èŠãªã°ãªã»ããŒã«ããããã²ããŒãŒããã®è£œæ³åã³ãã®çšé |
EP0665896B1 (en) * | 1992-09-28 | 1999-11-17 | Roche Diagnostics Corporation | Stable reagent for ferric ion complex indicator systems |
ES2185647T3 (es) * | 1993-01-28 | 2003-05-01 | Roche Diagnostics Corp | Composiciones utiles en la determinacion anaerobia de analitos. |
ES2284162T3 (es) * | 1995-02-27 | 2007-11-01 | Dsm Ip Assets B.V. | D-sorbitol dehidrogenasa. |
US7115321B2 (en) * | 2002-07-26 | 2006-10-03 | Kimberly-Clark Worldwide, Inc. | Absorbent binder coating |
KR101226264B1 (ko) | 2005-03-29 | 2013-01-25 | ì ê²ê°ìŽì€ ìŒí°ìì ìží°ëŽì ë | ë°ìŽì€ìŒì |
Family Cites Families (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CA647693A (en) * | 1962-08-28 | Baxter Laboratories | Process of microbial fermentation | |
US2948658A (en) * | 1960-02-25 | 1960-08-09 | Baxter Laboratories Inc | Process for producing dihydroxy acetone |
FR2212046A5 (ja) * | 1972-12-25 | 1974-07-19 | Ono Pharmaceutical Co | |
US4223090A (en) * | 1978-07-13 | 1980-09-16 | American Hospital Supply Corporation | Reagents for the enzymatic determination of triglycerides |
JPS5934118B2 (ja) * | 1978-08-31 | 1984-08-20 | ååéé µå·¥æ¥æ ªåŒäŒç€Ÿ | ãã€ãã€ãããã·ã¢ã»ãã³ã®å®éæ³ |
JPS6057830B2 (ja) * | 1979-07-02 | 1985-12-17 | ç§æ å±±ç° | ã°ãªã»ããŒã«è±æ°ŽçŽ é µçŽ ã®è£œé æ³ |
JPS603396B2 (ja) * | 1981-12-26 | 1985-01-28 | æ ªåŒäŒç€Ÿ åä»ååŠç 究æ | ããã©ãŸãªãŠã å¡©ååç©ããã³ãã®ååç©ãçšããè±æ°ŽçŽ é µçŽ ã®åžå å 床å®éæ³ |
-
1983
- 1983-03-25 DE DE19833311027 patent/DE3311027A1/de not_active Withdrawn
-
1984
- 1984-03-20 DE DE8484103042T patent/DE3482892D1/de not_active Expired - Lifetime
- 1984-03-20 EP EP84103042A patent/EP0120440B1/de not_active Expired - Lifetime
- 1984-03-20 AT AT84103042T patent/ATE55410T1/de not_active IP Right Cessation
- 1984-03-23 JP JP59054611A patent/JPS59183689A/ja active Granted
- 1984-03-23 US US06/592,708 patent/US4576913A/en not_active Expired - Lifetime
- 1984-03-23 CA CA000450438A patent/CA1225950A/en not_active Expired
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2008220367A (ja) * | 2007-02-13 | 2008-09-25 | Cci Corp | ããªãªãŒã«è±æ°ŽçŽ é µçŽ ã®è£œé æ¹æ³ |
Also Published As
Publication number | Publication date |
---|---|
CA1225950A (en) | 1987-08-25 |
EP0120440A2 (de) | 1984-10-03 |
ATE55410T1 (de) | 1990-08-15 |
US4576913A (en) | 1986-03-18 |
EP0120440B1 (de) | 1990-08-08 |
JPS59183689A (ja) | 1984-10-18 |
DE3482892D1 (de) | 1990-09-13 |
DE3311027A1 (de) | 1984-09-27 |
EP0120440A3 (en) | 1986-12-03 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
Bryant et al. | Purification and properties of primary and secondary alcohol dehydrogenases from Thermoanaerobacter ethanolicus | |
JPH031949B2 (ja) | ||
WO2011004654A1 (ja) | ã ã³ãŒã«å±ç±æ¥ã®ã°ã«ã³ãŒã¹ããããã²ããŒãŒ | |
JP2008035748A (ja) | ã¢ã¹ãã«ã®ã«ã¹å±èç±æ¥æ°èŠã°ã«ã³ãŒã¹ããããã²ããŒãŒ | |
JPS6122952B2 (ja) | ||
FR2555197A1 (fr) | Nouvelle maltose deshydrogenase, sa production, methode d'analyse l'utilisant et composition utilisable pour cette methode | |
Adachi et al. | 3-Dehydroquinate production by oxidative fermentation and further conversion of 3-dehydroquinate to the intermediates in the shikimate pathway | |
Poels et al. | NAD (P)+âindependent aldehyde dehydrogenase from Pseudomonas testosteroni: A novel type of molybdenumâcontaining hydroxylase | |
JP4511655B2 (ja) | ãœã«ãããŒã«è±æ°ŽçŽ é µçŽ ããããç£çãã埮çç©ããã³ãã®è£œé æ¹æ³ | |
Smolander et al. | Large-scale applicable purification and characterization of a membrane-bound PQQ-dependent aldose dehydrogenase | |
US4636465A (en) | Method and composition for determination of glycerol | |
JP2872983B2 (ja) | ïŒïŒïŒâã¢ã³ãããã°ã«ã·ããŒã«ã®å®éæ³åã³å®éçšè©Šè¬ | |
Ano et al. | Energy metabolism of a unique acetic acid bacterium, Asaia bogorensis, that lacks ethanol oxidation activity | |
US4889801A (en) | 3α-hydroxysteroid oxidase and quantitative analysis of 3α-hydroxysteroid making use of same | |
JP2726826B2 (ja) | é µçŽ ããã³ãã®è£œé æ¹æ³ | |
Boontim et al. | Purification and characterization of D-glucose dehydrogenase from Bacillus thuringiensis M15 | |
JP3819094B2 (ja) | ïŒïŒïŒâã¢ã³ãããã°ã«ã·ããŒã«è±æ°ŽçŽ é µçŽ ããã®è£œé æ³åã³ãããçšããïŒïŒïŒâã¢ã³ãããã°ã«ã·ããŒã«ã®å®éæ³ | |
JPS5935592B2 (ja) | âãã«ã¯ãâã¹ã®å®éæ³ | |
US4550079A (en) | Determination of L(+)-tartrate or D(+)-malate with L(+)-tartrate dehydrogenase | |
JPH0249720B2 (ja) | ||
Takeda et al. | Role of cytochrome c-553 (CO), the second subunit of alcohol dehydrogenase, in the azide-insensitive respiratory chain and in oxidative fermentation of Gluconobacter species | |
JP5311615B2 (ja) | ååŠä¿®é£ŸããªãªãŒã«è±æ°ŽçŽ é µçŽ ããã³ãã®è£œé æ¹æ³ | |
JPS6317432B2 (ja) | ||
JPS6342519B2 (ja) | ||
JP3929576B2 (ja) | ãªã³é žåïŒïŒïŒâã¢ã³ãããã°ã«ã·ããŒã«ã«äœçšããé µçŽ ãç£çãã埮çç©ãåœè©²åŸ®çç©ãç£çããé µçŽ åã³ããã䜿çšãããªã³é žåïŒïŒïŒâã¢ã³ãããã°ã«ã·ããŒã«ã®å®éæ¹æ³ |