JP6908653B2 - 固定化タンパク質及びその使用 - Google Patents
固定化タンパク質及びその使用 Download PDFInfo
- Publication number
- JP6908653B2 JP6908653B2 JP2019095107A JP2019095107A JP6908653B2 JP 6908653 B2 JP6908653 B2 JP 6908653B2 JP 2019095107 A JP2019095107 A JP 2019095107A JP 2019095107 A JP2019095107 A JP 2019095107A JP 6908653 B2 JP6908653 B2 JP 6908653B2
- Authority
- JP
- Japan
- Prior art keywords
- enzyme
- immobilized
- carrier
- cpg
- protein
- 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.)
- Active
Links
- 108010058683 Immobilized Proteins Proteins 0.000 title description 29
- 102000004190 Enzymes Human genes 0.000 claims description 137
- 108090000790 Enzymes Proteins 0.000 claims description 137
- 238000000034 method Methods 0.000 claims description 76
- 229910021645 metal ion Inorganic materials 0.000 claims description 46
- 238000006243 chemical reaction Methods 0.000 claims description 44
- 108010093096 Immobilized Enzymes Proteins 0.000 claims description 31
- 239000011159 matrix material Substances 0.000 claims description 31
- 229920000620 organic polymer Polymers 0.000 claims description 28
- 230000027455 binding Effects 0.000 claims description 21
- -1 polyethylene Polymers 0.000 claims description 16
- 238000006555 catalytic reaction Methods 0.000 claims description 14
- 239000000758 substrate Substances 0.000 claims description 14
- 239000002245 particle Substances 0.000 claims description 12
- KDLHZDBZIXYQEI-UHFFFAOYSA-N Palladium Chemical compound [Pd] KDLHZDBZIXYQEI-UHFFFAOYSA-N 0.000 claims description 11
- 239000002082 metal nanoparticle Substances 0.000 claims description 11
- 239000004793 Polystyrene Substances 0.000 claims description 10
- PXHVJJICTQNCMI-UHFFFAOYSA-N nickel Substances [Ni] PXHVJJICTQNCMI-UHFFFAOYSA-N 0.000 claims description 10
- 238000010523 cascade reaction Methods 0.000 claims description 9
- 229920002223 polystyrene Polymers 0.000 claims description 8
- 239000002105 nanoparticle Substances 0.000 claims description 7
- 229920000193 polymethacrylate Polymers 0.000 claims description 7
- 238000003786 synthesis reaction Methods 0.000 claims description 6
- 239000004743 Polypropylene Substances 0.000 claims description 5
- 239000003960 organic solvent Substances 0.000 claims description 5
- 229920001155 polypropylene Polymers 0.000 claims description 5
- 239000002033 PVDF binder Substances 0.000 claims description 4
- 229920001903 high density polyethylene Polymers 0.000 claims description 4
- 239000004700 high-density polyethylene Substances 0.000 claims description 4
- 229910052763 palladium Inorganic materials 0.000 claims description 4
- 229920001343 polytetrafluoroethylene Polymers 0.000 claims description 4
- 239000004810 polytetrafluoroethylene Substances 0.000 claims description 4
- 229920002981 polyvinylidene fluoride Polymers 0.000 claims description 4
- 238000003541 multi-stage reaction Methods 0.000 claims description 3
- 229920003229 poly(methyl methacrylate) Polymers 0.000 claims description 3
- 239000004926 polymethyl methacrylate Substances 0.000 claims description 3
- 230000007704 transition Effects 0.000 claims description 3
- 229910052723 transition metal Inorganic materials 0.000 claims description 3
- 229910021524 transition metal nanoparticle Inorganic materials 0.000 claims description 3
- 150000003624 transition metals Chemical class 0.000 claims description 3
- 239000004698 Polyethylene Substances 0.000 claims description 2
- 229920010741 Ultra High Molecular Weight Polyethylene (UHMWPE) Polymers 0.000 claims description 2
- 239000010941 cobalt Substances 0.000 claims description 2
- 229910017052 cobalt Inorganic materials 0.000 claims description 2
- GUTLYIVDDKVIGB-UHFFFAOYSA-N cobalt atom Chemical compound [Co] GUTLYIVDDKVIGB-UHFFFAOYSA-N 0.000 claims description 2
- 229910052759 nickel Inorganic materials 0.000 claims description 2
- 229920000573 polyethylene Polymers 0.000 claims description 2
- 230000008569 process Effects 0.000 claims description 2
- 239000000463 material Substances 0.000 description 80
- 108090000623 proteins and genes Proteins 0.000 description 60
- 102000004169 proteins and genes Human genes 0.000 description 59
- 235000018102 proteins Nutrition 0.000 description 58
- 239000000243 solution Substances 0.000 description 35
- 239000000872 buffer Substances 0.000 description 20
- 239000005373 porous glass Substances 0.000 description 19
- 238000002360 preparation method Methods 0.000 description 19
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 18
- 239000012876 carrier material Substances 0.000 description 17
- 108091006055 affinity-tagged proteins Proteins 0.000 description 16
- 229920002704 polyhistidine Polymers 0.000 description 16
- 239000000047 product Substances 0.000 description 16
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 15
- 239000003054 catalyst Substances 0.000 description 14
- 239000000203 mixture Substances 0.000 description 13
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 12
- 150000001412 amines Chemical class 0.000 description 12
- RAXXELZNTBOGNW-UHFFFAOYSA-N imidazole Natural products C1=CNC=N1 RAXXELZNTBOGNW-UHFFFAOYSA-N 0.000 description 12
- 239000011148 porous material Substances 0.000 description 12
- 230000000694 effects Effects 0.000 description 11
- 230000002829 reductive effect Effects 0.000 description 11
- SULYEHHGGXARJS-UHFFFAOYSA-N 2',4'-dihydroxyacetophenone Chemical compound CC(=O)C1=CC=C(O)C=C1O SULYEHHGGXARJS-UHFFFAOYSA-N 0.000 description 10
- YCOZIPAWZNQLMR-UHFFFAOYSA-N pentadecane Chemical compound CCCCCCCCCCCCCCC YCOZIPAWZNQLMR-UHFFFAOYSA-N 0.000 description 10
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 9
- 238000000576 coating method Methods 0.000 description 9
- 125000000524 functional group Chemical group 0.000 description 9
- 239000011521 glass Substances 0.000 description 9
- JKMHFZQWWAIEOD-UHFFFAOYSA-N 2-[4-(2-hydroxyethyl)piperazin-1-yl]ethanesulfonic acid Chemical compound OCC[NH+]1CCN(CCS([O-])(=O)=O)CC1 JKMHFZQWWAIEOD-UHFFFAOYSA-N 0.000 description 8
- 229920008712 Copo Polymers 0.000 description 8
- 125000003277 amino group Chemical group 0.000 description 8
- 239000011942 biocatalyst Substances 0.000 description 8
- 239000011248 coating agent Substances 0.000 description 8
- 230000003100 immobilizing effect Effects 0.000 description 8
- 238000002386 leaching Methods 0.000 description 8
- 229920000642 polymer Polymers 0.000 description 8
- 238000000746 purification Methods 0.000 description 8
- 239000000523 sample Substances 0.000 description 8
- 239000007787 solid Substances 0.000 description 8
- RQEUFEKYXDPUSK-UHFFFAOYSA-N 1-phenylethylamine Chemical compound CC(N)C1=CC=CC=C1 RQEUFEKYXDPUSK-UHFFFAOYSA-N 0.000 description 7
- 239000007995 HEPES buffer Substances 0.000 description 7
- 101001064468 Pseudozyma aphidis (strain ATCC 32657 / CBS 517.83 / DSM 70725 / JCM 10318 / NBRC 10182 / NRRL Y-7954 / St-0401) Lipase A Proteins 0.000 description 7
- 238000002835 absorbance Methods 0.000 description 7
- 230000003197 catalytic effect Effects 0.000 description 7
- 238000001914 filtration Methods 0.000 description 7
- 238000011002 quantification Methods 0.000 description 7
- 239000011541 reaction mixture Substances 0.000 description 7
- WFDIJRYMOXRFFG-UHFFFAOYSA-N Acetic anhydride Chemical compound CC(=O)OC(C)=O WFDIJRYMOXRFFG-UHFFFAOYSA-N 0.000 description 6
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 6
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 6
- BZLVMXJERCGZMT-UHFFFAOYSA-N Methyl tert-butyl ether Chemical compound COC(C)(C)C BZLVMXJERCGZMT-UHFFFAOYSA-N 0.000 description 6
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 6
- YXFVVABEGXRONW-UHFFFAOYSA-N Toluene Chemical compound CC1=CC=CC=C1 YXFVVABEGXRONW-UHFFFAOYSA-N 0.000 description 6
- ZMANZCXQSJIPKH-UHFFFAOYSA-N Triethylamine Chemical compound CCN(CC)CC ZMANZCXQSJIPKH-UHFFFAOYSA-N 0.000 description 6
- 239000007864 aqueous solution Substances 0.000 description 6
- 230000008901 benefit Effects 0.000 description 6
- 239000000969 carrier Substances 0.000 description 6
- 235000019439 ethyl acetate Nutrition 0.000 description 6
- NGVDGCNFYWLIFO-UHFFFAOYSA-N pyridoxal 5'-phosphate Chemical compound CC1=NC=C(COP(O)(O)=O)C(C=O)=C1O NGVDGCNFYWLIFO-UHFFFAOYSA-N 0.000 description 6
- 235000007682 pyridoxal 5'-phosphate Nutrition 0.000 description 6
- 239000011589 pyridoxal 5'-phosphate Substances 0.000 description 6
- 238000003756 stirring Methods 0.000 description 6
- 238000005917 acylation reaction Methods 0.000 description 5
- 150000001299 aldehydes Chemical class 0.000 description 5
- 238000003556 assay Methods 0.000 description 5
- 238000010494 dissociation reaction Methods 0.000 description 5
- 230000005593 dissociations Effects 0.000 description 5
- 150000002466 imines Chemical class 0.000 description 5
- 238000011534 incubation Methods 0.000 description 5
- 229930027945 nicotinamide-adenine dinucleotide Natural products 0.000 description 5
- 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 5
- KLAKIAVEMQMVBT-UHFFFAOYSA-N p-hydroxy-phenacyl alcohol Natural products OCC(=O)C1=CC=C(O)C=C1 KLAKIAVEMQMVBT-UHFFFAOYSA-N 0.000 description 5
- 229920006395 saturated elastomer Polymers 0.000 description 5
- 239000012279 sodium borohydride Substances 0.000 description 5
- 229910000033 sodium borohydride Inorganic materials 0.000 description 5
- 238000005406 washing Methods 0.000 description 5
- DVLFYONBTKHTER-UHFFFAOYSA-N 3-(N-morpholino)propanesulfonic acid Chemical compound OS(=O)(=O)CCCN1CCOCC1 DVLFYONBTKHTER-UHFFFAOYSA-N 0.000 description 4
- 101000774761 Archaeoglobus fulgidus (strain ATCC 49558 / DSM 4304 / JCM 9628 / NBRC 100126 / VC-16) Alanine dehydrogenase Proteins 0.000 description 4
- 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 4
- CDBYLPFSWZWCQE-UHFFFAOYSA-L Sodium Carbonate Chemical compound [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 description 4
- 230000010933 acylation Effects 0.000 description 4
- 239000012062 aqueous buffer Substances 0.000 description 4
- 229920001577 copolymer Polymers 0.000 description 4
- 238000012258 culturing Methods 0.000 description 4
- 238000005516 engineering process Methods 0.000 description 4
- 230000002255 enzymatic effect Effects 0.000 description 4
- MTZQAGJQAFMTAQ-UHFFFAOYSA-N ethyl benzoate Chemical compound CCOC(=O)C1=CC=CC=C1 MTZQAGJQAFMTAQ-UHFFFAOYSA-N 0.000 description 4
- 229940071106 ethylenediaminetetraacetate Drugs 0.000 description 4
- FVTCRASFADXXNN-SCRDCRAPSA-N flavin mononucleotide Chemical compound OP(=O)(O)OC[C@@H](O)[C@@H](O)[C@@H](O)CN1C=2C=C(C)C(C)=CC=2N=C2C1=NC(=O)NC2=O FVTCRASFADXXNN-SCRDCRAPSA-N 0.000 description 4
- FVTCRASFADXXNN-UHFFFAOYSA-N flavin mononucleotide Natural products OP(=O)(O)OCC(O)C(O)C(O)CN1C=2C=C(C)C(C)=CC=2N=C2C1=NC(=O)NC2=O FVTCRASFADXXNN-UHFFFAOYSA-N 0.000 description 4
- 239000011768 flavin mononucleotide Substances 0.000 description 4
- 239000002638 heterogeneous catalyst Substances 0.000 description 4
- 229910052751 metal Inorganic materials 0.000 description 4
- 239000002184 metal Substances 0.000 description 4
- 229920003023 plastic Polymers 0.000 description 4
- 239000004033 plastic Substances 0.000 description 4
- 238000006722 reduction reaction Methods 0.000 description 4
- 235000019231 riboflavin-5'-phosphate Nutrition 0.000 description 4
- 239000000377 silicon dioxide Substances 0.000 description 4
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 4
- 239000011343 solid material Substances 0.000 description 4
- 239000002904 solvent Substances 0.000 description 4
- WAPNOHKVXSQRPX-UHFFFAOYSA-N 1-phenylethanol Chemical compound CC(O)C1=CC=CC=C1 WAPNOHKVXSQRPX-UHFFFAOYSA-N 0.000 description 3
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 3
- 235000014469 Bacillus subtilis Nutrition 0.000 description 3
- 108010031797 Candida antarctica lipase B Proteins 0.000 description 3
- 108010025076 Holoenzymes 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
- 101710098556 Lipase A Proteins 0.000 description 3
- 101710098554 Lipase B Proteins 0.000 description 3
- 101710099648 Lysosomal acid lipase/cholesteryl ester hydrolase Proteins 0.000 description 3
- 102100026001 Lysosomal acid lipase/cholesteryl ester hydrolase Human genes 0.000 description 3
- 239000007993 MOPS buffer Substances 0.000 description 3
- 241001661345 Moesziomyces antarcticus Species 0.000 description 3
- 102220469850 Protein argonaute-3_W104A_mutation Human genes 0.000 description 3
- 108010008281 Recombinant Fusion Proteins Proteins 0.000 description 3
- 102000007056 Recombinant Fusion Proteins Human genes 0.000 description 3
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- XPNGNIFUDRPBFJ-UHFFFAOYSA-N alpha-methylbenzylalcohol Natural products CC1=CC=CC=C1CO XPNGNIFUDRPBFJ-UHFFFAOYSA-N 0.000 description 3
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 3
- 230000002210 biocatalytic effect Effects 0.000 description 3
- 230000015572 biosynthetic process Effects 0.000 description 3
- 239000003638 chemical reducing agent Substances 0.000 description 3
- 238000009826 distribution Methods 0.000 description 3
- 125000003700 epoxy group Chemical group 0.000 description 3
- 238000000605 extraction Methods 0.000 description 3
- 150000004820 halides Chemical class 0.000 description 3
- 125000000487 histidyl group Chemical group [H]N([H])C(C(=O)O*)C([H])([H])C1=C([H])N([H])C([H])=N1 0.000 description 3
- 230000003993 interaction Effects 0.000 description 3
- BPHPUYQFMNQIOC-NXRLNHOXSA-N isopropyl beta-D-thiogalactopyranoside Chemical compound CC(C)S[C@@H]1O[C@H](CO)[C@H](O)[C@H](O)[C@H]1O BPHPUYQFMNQIOC-NXRLNHOXSA-N 0.000 description 3
- 235000019421 lipase Nutrition 0.000 description 3
- 238000005259 measurement Methods 0.000 description 3
- 238000002156 mixing Methods 0.000 description 3
- 239000012074 organic phase Substances 0.000 description 3
- 230000003647 oxidation Effects 0.000 description 3
- 238000007254 oxidation reaction Methods 0.000 description 3
- 239000001301 oxygen Substances 0.000 description 3
- 229910052760 oxygen Inorganic materials 0.000 description 3
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 3
- 230000009467 reduction Effects 0.000 description 3
- SCPYDCQAZCOKTP-UHFFFAOYSA-N silanol Chemical compound [SiH3]O SCPYDCQAZCOKTP-UHFFFAOYSA-N 0.000 description 3
- 239000011780 sodium chloride Substances 0.000 description 3
- 239000006228 supernatant Substances 0.000 description 3
- 238000005891 transamination reaction Methods 0.000 description 3
- DSSYKIVIOFKYAU-XCBNKYQSSA-N (R)-camphor Chemical compound C1C[C@@]2(C)C(=O)C[C@@H]1C2(C)C DSSYKIVIOFKYAU-XCBNKYQSSA-N 0.000 description 2
- 229930007886 (R)-camphor Natural products 0.000 description 2
- KWOLFJPFCHCOCG-UHFFFAOYSA-N Acetophenone Chemical compound CC(=O)C1=CC=CC=C1 KWOLFJPFCHCOCG-UHFFFAOYSA-N 0.000 description 2
- 108010031025 Alanine Dehydrogenase Proteins 0.000 description 2
- 102000007698 Alcohol dehydrogenase Human genes 0.000 description 2
- 108010021809 Alcohol dehydrogenase Proteins 0.000 description 2
- 244000063299 Bacillus subtilis Species 0.000 description 2
- 241000588879 Chromobacterium violaceum Species 0.000 description 2
- QNAYBMKLOCPYGJ-UHFFFAOYSA-N D-alpha-Ala Natural products CC([NH3+])C([O-])=O QNAYBMKLOCPYGJ-UHFFFAOYSA-N 0.000 description 2
- 241000588724 Escherichia coli Species 0.000 description 2
- 101710157404 Flavin reductase Proteins 0.000 description 2
- 102100027944 Flavin reductase (NADPH) Human genes 0.000 description 2
- 102000005720 Glutathione transferase Human genes 0.000 description 2
- 108010070675 Glutathione transferase Proteins 0.000 description 2
- OAKJQQAXSVQMHS-UHFFFAOYSA-N Hydrazine Chemical compound NN OAKJQQAXSVQMHS-UHFFFAOYSA-N 0.000 description 2
- FFOPEPMHKILNIT-UHFFFAOYSA-N Isopropyl butyrate Chemical compound CCCC(=O)OC(C)C FFOPEPMHKILNIT-UHFFFAOYSA-N 0.000 description 2
- QNAYBMKLOCPYGJ-UWTATZPHSA-N L-Alanine Natural products C[C@@H](N)C(O)=O QNAYBMKLOCPYGJ-UWTATZPHSA-N 0.000 description 2
- QNAYBMKLOCPYGJ-REOHCLBHSA-N L-alanine Chemical compound C[C@H](N)C(O)=O QNAYBMKLOCPYGJ-REOHCLBHSA-N 0.000 description 2
- 101710175625 Maltose/maltodextrin-binding periplasmic protein Proteins 0.000 description 2
- BAPJBEWLBFYGME-UHFFFAOYSA-N Methyl acrylate Chemical compound COC(=O)C=C BAPJBEWLBFYGME-UHFFFAOYSA-N 0.000 description 2
- 101150003085 Pdcl gene Proteins 0.000 description 2
- 241000589776 Pseudomonas putida Species 0.000 description 2
- PPBRXRYQALVLMV-UHFFFAOYSA-N Styrene Chemical compound C=CC1=CC=CC=C1 PPBRXRYQALVLMV-UHFFFAOYSA-N 0.000 description 2
- 229960003767 alanine Drugs 0.000 description 2
- 150000001298 alcohols Chemical class 0.000 description 2
- 230000001588 bifunctional effect Effects 0.000 description 2
- 239000012496 blank sample Substances 0.000 description 2
- 239000007853 buffer solution Substances 0.000 description 2
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 2
- 150000001735 carboxylic acids Chemical group 0.000 description 2
- 238000004113 cell culture Methods 0.000 description 2
- 238000005119 centrifugation Methods 0.000 description 2
- 230000009920 chelation Effects 0.000 description 2
- 238000005356 chiral GC Methods 0.000 description 2
- 102000021178 chitin binding proteins Human genes 0.000 description 2
- 108091011157 chitin binding proteins Proteins 0.000 description 2
- 229910001429 cobalt ion Inorganic materials 0.000 description 2
- XLJKHNWPARRRJB-UHFFFAOYSA-N cobalt(2+) Chemical compound [Co+2] XLJKHNWPARRRJB-UHFFFAOYSA-N 0.000 description 2
- 239000005515 coenzyme Substances 0.000 description 2
- 230000000052 comparative effect Effects 0.000 description 2
- 239000012228 culture supernatant Substances 0.000 description 2
- 238000013461 design Methods 0.000 description 2
- 238000001035 drying Methods 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 239000012634 fragment Substances 0.000 description 2
- 238000004817 gas chromatography Methods 0.000 description 2
- HNDVDQJCIGZPNO-UHFFFAOYSA-N histidine Natural products OC(=O)C(N)CC1=CN=CN1 HNDVDQJCIGZPNO-UHFFFAOYSA-N 0.000 description 2
- 238000011065 in-situ storage Methods 0.000 description 2
- 229910052742 iron Inorganic materials 0.000 description 2
- 238000002955 isolation Methods 0.000 description 2
- 150000002576 ketones Chemical class 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 239000012280 lithium aluminium hydride Substances 0.000 description 2
- 239000002609 medium Substances 0.000 description 2
- 239000000178 monomer Substances 0.000 description 2
- 238000006053 organic reaction Methods 0.000 description 2
- 239000012071 phase Substances 0.000 description 2
- 239000008363 phosphate buffer Substances 0.000 description 2
- 229920005553 polystyrene-acrylate Polymers 0.000 description 2
- 108090000765 processed proteins & peptides Proteins 0.000 description 2
- 150000003839 salts Chemical class 0.000 description 2
- 238000000926 separation method Methods 0.000 description 2
- 125000005372 silanol group Chemical group 0.000 description 2
- 229910000029 sodium carbonate Inorganic materials 0.000 description 2
- BEOOHQFXGBMRKU-UHFFFAOYSA-N sodium cyanoborohydride Chemical compound [Na+].[B-]C#N BEOOHQFXGBMRKU-UHFFFAOYSA-N 0.000 description 2
- 239000012064 sodium phosphate buffer Substances 0.000 description 2
- DAEPDZWVDSPTHF-UHFFFAOYSA-M sodium pyruvate Chemical compound [Na+].CC(=O)C([O-])=O DAEPDZWVDSPTHF-UHFFFAOYSA-M 0.000 description 2
- 238000001179 sorption measurement Methods 0.000 description 2
- 230000008961 swelling Effects 0.000 description 2
- 238000012546 transfer Methods 0.000 description 2
- WYTZZXDRDKSJID-UHFFFAOYSA-N (3-aminopropyl)triethoxysilane Chemical compound CCO[Si](OCC)(OCC)CCCN WYTZZXDRDKSJID-UHFFFAOYSA-N 0.000 description 1
- GGKADXREVJTZMF-UHFFFAOYSA-N 1-Phenylethyl butyrate Chemical compound CCCC(=O)OC(C)C1=CC=CC=C1 GGKADXREVJTZMF-UHFFFAOYSA-N 0.000 description 1
- IKYFHRVPKIFGMH-UHFFFAOYSA-N 1-phenoxypropan-2-amine Chemical compound CC(N)COC1=CC=CC=C1 IKYFHRVPKIFGMH-UHFFFAOYSA-N 0.000 description 1
- QWAVNXZAQASOML-UHFFFAOYSA-N 1-phenoxypropan-2-one Chemical compound CC(=O)COC1=CC=CC=C1 QWAVNXZAQASOML-UHFFFAOYSA-N 0.000 description 1
- MLONYBFKXHEPCD-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol Chemical compound OCC(N)(CO)CO.OCC(N)(CO)CO MLONYBFKXHEPCD-UHFFFAOYSA-N 0.000 description 1
- DVGKRPYUFRZAQW-UHFFFAOYSA-N 3 prime Natural products CC(=O)NC1OC(CC(O)C1C(O)C(O)CO)(OC2C(O)C(CO)OC(OC3C(O)C(O)C(O)OC3CO)C2O)C(=O)O DVGKRPYUFRZAQW-UHFFFAOYSA-N 0.000 description 1
- NLHHRLWOUZZQLW-UHFFFAOYSA-N Acrylonitrile Chemical compound C=CC#N NLHHRLWOUZZQLW-UHFFFAOYSA-N 0.000 description 1
- 108090001008 Avidin Proteins 0.000 description 1
- 101710197852 Baeyer-Villiger monooxygenase Proteins 0.000 description 1
- 238000006220 Baeyer-Villiger oxidation reaction Methods 0.000 description 1
- 238000009010 Bradford assay Methods 0.000 description 1
- 241000222120 Candida <Saccharomycetales> Species 0.000 description 1
- XDTMQSROBMDMFD-UHFFFAOYSA-N Cyclohexane Chemical compound C1CCCCC1 XDTMQSROBMDMFD-UHFFFAOYSA-N 0.000 description 1
- 108020004414 DNA Proteins 0.000 description 1
- 101710137307 FAD-containing monooxygenase EthA Proteins 0.000 description 1
- 108090000698 Formate Dehydrogenases Proteins 0.000 description 1
- SXRSQZLOMIGNAQ-UHFFFAOYSA-N Glutaraldehyde Chemical compound O=CCCCC=O SXRSQZLOMIGNAQ-UHFFFAOYSA-N 0.000 description 1
- 108010093488 His-His-His-His-His-His Proteins 0.000 description 1
- 102000004157 Hydrolases Human genes 0.000 description 1
- 108090000604 Hydrolases Proteins 0.000 description 1
- 102000004195 Isomerases Human genes 0.000 description 1
- 108090000769 Isomerases Proteins 0.000 description 1
- 102000003960 Ligases Human genes 0.000 description 1
- 108090000364 Ligases Proteins 0.000 description 1
- 239000006391 Luria-Bertani Medium Substances 0.000 description 1
- 102000004317 Lyases Human genes 0.000 description 1
- 108090000856 Lyases Proteins 0.000 description 1
- CERQOIWHTDAKMF-UHFFFAOYSA-M Methacrylate Chemical compound CC(=C)C([O-])=O CERQOIWHTDAKMF-UHFFFAOYSA-M 0.000 description 1
- CERQOIWHTDAKMF-UHFFFAOYSA-N Methacrylic acid Chemical compound CC(=C)C(O)=O CERQOIWHTDAKMF-UHFFFAOYSA-N 0.000 description 1
- VVQNEPGJFQJSBK-UHFFFAOYSA-N Methyl methacrylate Chemical compound COC(=O)C(C)=C VVQNEPGJFQJSBK-UHFFFAOYSA-N 0.000 description 1
- 102000008109 Mixed Function Oxygenases Human genes 0.000 description 1
- 108010074633 Mixed Function Oxygenases Proteins 0.000 description 1
- 102000010909 Monoamine Oxidase Human genes 0.000 description 1
- 108010062431 Monoamine oxidase Proteins 0.000 description 1
- 102000004316 Oxidoreductases Human genes 0.000 description 1
- 108090000854 Oxidoreductases Proteins 0.000 description 1
- 101150053185 P450 gene Proteins 0.000 description 1
- 108091005804 Peptidases Proteins 0.000 description 1
- 239000004365 Protease Substances 0.000 description 1
- 108020004511 Recombinant DNA Proteins 0.000 description 1
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 description 1
- 229910008051 Si-OH Inorganic materials 0.000 description 1
- BLRPTPMANUNPDV-UHFFFAOYSA-N Silane Chemical compound [SiH4] BLRPTPMANUNPDV-UHFFFAOYSA-N 0.000 description 1
- XUIMIQQOPSSXEZ-UHFFFAOYSA-N Silicon Chemical group [Si] XUIMIQQOPSSXEZ-UHFFFAOYSA-N 0.000 description 1
- 229910006358 Si—OH Inorganic materials 0.000 description 1
- 108010090804 Streptavidin Proteins 0.000 description 1
- 102000004357 Transferases Human genes 0.000 description 1
- 108090000992 Transferases Proteins 0.000 description 1
- 239000007983 Tris buffer Substances 0.000 description 1
- 238000011481 absorbance measurement Methods 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 125000002252 acyl group Chemical group 0.000 description 1
- 238000001042 affinity chromatography Methods 0.000 description 1
- 239000011543 agarose gel Substances 0.000 description 1
- 150000003973 alkyl amines Chemical class 0.000 description 1
- 125000000539 amino acid group Chemical group 0.000 description 1
- 239000008346 aqueous phase Substances 0.000 description 1
- 230000008033 biological extinction Effects 0.000 description 1
- 230000008827 biological function Effects 0.000 description 1
- 150000001732 carboxylic acid derivatives Chemical class 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 239000013522 chelant Substances 0.000 description 1
- 150000004697 chelate complex Chemical class 0.000 description 1
- 230000002860 competitive effect Effects 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 229920006037 cross link polymer Polymers 0.000 description 1
- 239000003431 cross linking reagent Substances 0.000 description 1
- 239000012045 crude solution Substances 0.000 description 1
- XUJNEKJLAYXESH-UHFFFAOYSA-N cysteine Natural products SCC(N)C(O)=O XUJNEKJLAYXESH-UHFFFAOYSA-N 0.000 description 1
- 235000018417 cysteine Nutrition 0.000 description 1
- DEZRYPDIMOWBDS-UHFFFAOYSA-N dcm dichloromethane Chemical compound ClCCl.ClCCl DEZRYPDIMOWBDS-UHFFFAOYSA-N 0.000 description 1
- 238000001212 derivatisation Methods 0.000 description 1
- 230000000368 destabilizing effect Effects 0.000 description 1
- 239000003599 detergent Substances 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- YMWUJEATGCHHMB-UHFFFAOYSA-N dichloromethane Natural products ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 1
- 238000009792 diffusion process Methods 0.000 description 1
- 239000012149 elution buffer Substances 0.000 description 1
- 238000006047 enzymatic hydrolysis reaction Methods 0.000 description 1
- MEGHWIAOTJPCHQ-UHFFFAOYSA-N ethenyl butanoate Chemical compound CCCC(=O)OC=C MEGHWIAOTJPCHQ-UHFFFAOYSA-N 0.000 description 1
- OJCSPXHYDFONPU-UHFFFAOYSA-N etoac etoac Chemical compound CCOC(C)=O.CCOC(C)=O OJCSPXHYDFONPU-UHFFFAOYSA-N 0.000 description 1
- 238000011049 filling Methods 0.000 description 1
- 238000003818 flash chromatography Methods 0.000 description 1
- 229940013640 flavin mononucleotide Drugs 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 230000004927 fusion Effects 0.000 description 1
- 108020001507 fusion proteins Proteins 0.000 description 1
- 102000037865 fusion proteins Human genes 0.000 description 1
- 238000007429 general method Methods 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 239000012052 hydrophilic carrier Substances 0.000 description 1
- 239000012051 hydrophobic carrier Substances 0.000 description 1
- 230000002209 hydrophobic effect Effects 0.000 description 1
- 239000012535 impurity Substances 0.000 description 1
- 230000002779 inactivation Effects 0.000 description 1
- 230000003834 intracellular effect Effects 0.000 description 1
- DEGPIRUPAKWDBU-UHFFFAOYSA-N isoindole-1,3-dione;sodium Chemical compound [Na].C1=CC=C2C(=O)NC(=O)C2=C1 DEGPIRUPAKWDBU-UHFFFAOYSA-N 0.000 description 1
- 238000006317 isomerization reaction Methods 0.000 description 1
- SBUJHOSQTJFQJX-NOAMYHISSA-N kanamycin Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CN)O[C@@H]1O[C@H]1[C@H](O)[C@@H](O[C@@H]2[C@@H]([C@@H](N)[C@H](O)[C@@H](CO)O2)O)[C@H](N)C[C@@H]1N SBUJHOSQTJFQJX-NOAMYHISSA-N 0.000 description 1
- 229930027917 kanamycin Natural products 0.000 description 1
- 229960000318 kanamycin Drugs 0.000 description 1
- 229930182823 kanamycin A Natural products 0.000 description 1
- 239000003446 ligand Substances 0.000 description 1
- 239000007791 liquid phase Substances 0.000 description 1
- 238000011068 loading method Methods 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 125000003588 lysine group Chemical group [H]N([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])(N([H])[H])C(*)=O 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- IEXZQIBYHMBUJQ-SNVBAGLBSA-N n-[(1r)-1-phenylethyl]butanamide Chemical compound CCCC(=O)N[C@H](C)C1=CC=CC=C1 IEXZQIBYHMBUJQ-SNVBAGLBSA-N 0.000 description 1
- 229950006238 nadide Drugs 0.000 description 1
- 230000009871 nonspecific binding Effects 0.000 description 1
- 231100000252 nontoxic Toxicity 0.000 description 1
- 230000003000 nontoxic effect Effects 0.000 description 1
- 239000011368 organic material Substances 0.000 description 1
- 125000002524 organometallic group Chemical group 0.000 description 1
- 230000002018 overexpression Effects 0.000 description 1
- 125000001997 phenyl group Chemical group [H]C1=C([H])C([H])=C(*)C([H])=C1[H] 0.000 description 1
- 229920002239 polyacrylonitrile Polymers 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 230000002035 prolonged effect Effects 0.000 description 1
- 238000000164 protein isolation Methods 0.000 description 1
- 238000001742 protein purification Methods 0.000 description 1
- 229960001327 pyridoxal phosphate Drugs 0.000 description 1
- 230000006340 racemization Effects 0.000 description 1
- 238000009790 rate-determining step (RDS) Methods 0.000 description 1
- 239000000376 reactant Substances 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 238000004064 recycling Methods 0.000 description 1
- 230000008929 regeneration Effects 0.000 description 1
- 238000011069 regeneration method Methods 0.000 description 1
- 230000000717 retained effect Effects 0.000 description 1
- 230000002441 reversible effect Effects 0.000 description 1
- 238000013341 scale-up Methods 0.000 description 1
- 238000004062 sedimentation Methods 0.000 description 1
- 229910000077 silane Inorganic materials 0.000 description 1
- 229910052710 silicon Inorganic materials 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 239000001488 sodium phosphate Substances 0.000 description 1
- 229910000162 sodium phosphate Inorganic materials 0.000 description 1
- 229940054269 sodium pyruvate Drugs 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 230000009870 specific binding Effects 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 description 1
- RYFMWSXOAZQYPI-UHFFFAOYSA-K trisodium phosphate Chemical compound [Na+].[Na+].[Na+].[O-]P([O-])([O-])=O RYFMWSXOAZQYPI-UHFFFAOYSA-K 0.000 description 1
- 239000002699 waste material Substances 0.000 description 1
Images
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J31/00—Catalysts comprising hydrides, coordination complexes or organic compounds
- B01J31/003—Catalysts comprising hydrides, coordination complexes or organic compounds containing enzymes
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/14—Extraction; Separation; Purification
- C07K1/16—Extraction; Separation; Purification by chromatography
- C07K1/22—Affinity chromatography or related techniques based upon selective absorption processes
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K17/00—Carrier-bound or immobilised peptides; Preparation thereof
- C07K17/02—Peptides being immobilised on, or in, an organic carrier
- C07K17/06—Peptides being immobilised on, or in, an organic carrier attached to the carrier via a bridging agent
-
- 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
- C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
- C12N11/02—Enzymes or microbial cells immobilised on or in an organic carrier
- C12N11/08—Enzymes or microbial cells immobilised on or in an organic carrier the carrier being a synthetic polymer
- C12N11/082—Enzymes or microbial cells immobilised on or in an organic carrier the carrier being a synthetic polymer obtained by reactions only involving carbon-to-carbon unsaturated bonds
-
- 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
- C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
- C12N11/02—Enzymes or microbial cells immobilised on or in an organic carrier
- C12N11/08—Enzymes or microbial cells immobilised on or in an organic carrier the carrier being a synthetic polymer
- C12N11/082—Enzymes or microbial cells immobilised on or in an organic carrier the carrier being a synthetic polymer obtained by reactions only involving carbon-to-carbon unsaturated bonds
- C12N11/087—Acrylic polymers
-
- 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
- C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
- C12N11/14—Enzymes or microbial cells immobilised on or in an inorganic carrier
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P13/00—Preparation of nitrogen-containing organic compounds
- C12P13/001—Amines; Imines
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P13/00—Preparation of nitrogen-containing organic compounds
- C12P13/02—Amides, e.g. chloramphenicol or polyamides; Imides or polyimides; Urethanes, i.e. compounds comprising N-C=O structural element or polyurethanes
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P17/00—Preparation of heterocyclic carbon compounds with only O, N, S, Se or Te as ring hetero atoms
- C12P17/02—Oxygen as only ring hetero atoms
- C12P17/06—Oxygen as only ring hetero atoms containing a six-membered hetero ring, e.g. fluorescein
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P7/00—Preparation of oxygen-containing organic compounds
- C12P7/62—Carboxylic acid esters
-
- 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
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
- G01N33/54353—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals with ligand attached to the carrier via a chemical coupling agent
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
- G01N33/544—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals the carrier being organic
- G01N33/545—Synthetic resin
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
- G01N33/551—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals the carrier being inorganic
- G01N33/552—Glass or silica
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
- G01N33/551—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals the carrier being inorganic
- G01N33/553—Metal or metal coated
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/573—Immunoassay; Biospecific binding assay; Materials therefor for enzymes or isoenzymes
-
- 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
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P20/00—Technologies relating to chemical industry
- Y02P20/50—Improvements relating to the production of bulk chemicals
Landscapes
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- Genetics & Genomics (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Immunology (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Biomedical Technology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Microbiology (AREA)
- Biotechnology (AREA)
- Molecular Biology (AREA)
- Urology & Nephrology (AREA)
- Hematology (AREA)
- General Engineering & Computer Science (AREA)
- Medicinal Chemistry (AREA)
- Analytical Chemistry (AREA)
- Physics & Mathematics (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Food Science & Technology (AREA)
- Cell Biology (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Biophysics (AREA)
- Inorganic Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Materials Engineering (AREA)
- Immobilizing And Processing Of Enzymes And Microorganisms (AREA)
- Peptides Or Proteins (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
Description
(a)制御多孔性ガラス(CPG);
(b)ハイブリッド制御多孔性ガラス(ハイブリッドCPG);及び
(c)多孔質有機ポリマー
からなる群から選択され、少なくとも1つのタンパク質がアフィニティタグを含有し、アフィニティマトリックスとの特異的アフィニティ結合を介して担体上に固定化されている固定化タンパク質材料に関する。
(a)制御多孔性ガラス(CPG);
(b)ハイブリッド制御多孔性ガラス(ハイブリッドCPG);及び
(c)多孔質有機ポリマー
からなる群から選択され、タンパク質が、アフィニティマトリックスとの特異的アフィニティ結合を介して担体上に固定化される担体に関する。
i)アフィニティマトリックスが付着している担体材料を含む担体上にアフィニティタグ付きタンパク質を固定化する工程であって、前記担体材料が、
(a)制御多孔性ガラス(CPG);
(b)ハイブリッド制御多孔性ガラス(ハイブリッドCPG);及び
(c)多孔質有機ポリマー
からなる群から選択される工程;並びに
ii)任意選択で、固定化タンパク質材料を水又は適切な緩衝液で洗浄する工程
を含む方法に関する。
i)アミノ基を含有する担体材料を用意する工程であって、前記担体材料が、
(a)制御多孔性ガラス(CPG);
(b)ハイブリッド制御多孔性ガラス(ハイブリッドCPG);及び
(c)多孔質有機ポリマー
からなる群から選択される工程;
ii)担体材料を2.4-ジヒドロキシアセトフェノンと反応させ、それにより、ジヒドロキシフェニル基を担体材料に連結する工程;
iii)ポリヒスチジンタグ付きタンパク質に結合する能力があるキレート複合体を、ジヒドロキシフェニル基と金属イオンとの間で形成する工程;並びに
iv)前記金属イオンを含む担体材料に、ポリヒスチジンタグ付き酵素を結合させる工程
を含む方法を提供する。
i)アフィニティマトリックスが付着している担体材料を含む担体上にアフィニティタグ付きタンパク質を固定化する工程であって、前記担体材料が、
(a)制御多孔性ガラス(CPG);
(b)ハイブリッド制御多孔性ガラス(ハイブリッドCPG);及び
(c)多孔質有機ポリマー
からなる群から選択される工程;
ii)任意選択で、固定化タンパク質材料を水又は適切な緩衝液で洗浄する工程;並びに
iii)精製タンパク質をアフィニティマトリックスから解離させる工程
を含む方法に関する。
用語「アフィニティタグ」は、組換えタンパク質に付着し、かつマトリックス上に固定化された特定の基と結合することができる、有機又は有機金属分子、タンパク質断片等の限定された基を指す。アフィニティタグの例は、ポリヒスチジンタグ、グルタチオンS-トランスフェラーゼ(GST)タグ、キチン結合タンパク質(CBP)タグ、マルトース結合タンパク質(MBP)タグ、FLAGタグ、アビジンタグ、又はストレプトアビジンタグである。アフィニティタグはまた、融合タグと呼ばれる場合もある。
aw 水分活性
AlaDH 枯草菌(B. subtilis)由来のアラニンデヒドロゲナーゼ
CalA カンジダ・アンタルクティカリパーゼA(別名シュードジマ・アンタルクティカ(Pseudozyma antarctica)リパーゼA)
CalB カンジダ・アンタルクティカリパーゼB(別名シュードジマ・アンタルクティカリパーゼB)
DCM ジクロロメタン
2,5-DKCMO シュードモナス・プチダ(Pseudomonas putida)由来2,5-ジケトカンファン
equiv. 当量
EtOAc 酢酸エチル
FMN フラビンモノヌクレオチド
FRE 大腸菌(E. coli)由来フラビンリダクターゼ
GC ガスクロマトグラフィー
HEPES 4-(2-ヒドロキシエチル)ピペラジン-1-エタンスルホン酸
IPTG イソプロピル-β-D-チオガラクトピラノシド
min 分
MOPS 3-(N-モルホリノ)プロパンスルホン酸
MTBE メチルtert-ブチルエーテル(又はtert-ブチルメチルエーテル)
NADH ニコチンアミドアデニンジヌクレオチド(還元型)
PLP ピリドキサール5-リン酸
SDS-PAGE ドデシル硫酸ナトリウム-ポリアクリルアミドゲル電気泳動
Tris トリス(ヒドロキシメチル)アミノメタン
ω-TA クロモバクテリウム・ビオラセウム(Chromobacterium violaceum)由来ω-トランスアミナーゼ
CPG材料をPrime Synthesis, Inc. (Aston、PA、USA)から入手した。以下の制御多孔性ガラス材料を実施例に用いた:
・LCAA CPGは、長鎖アルキルアミンで誘導体化されたCPGである(CPG-0502-N12)。
・コポリ-HybCPGアミン(以下においては、「HybCPGコポ」とも呼ばれる)は、インサイチュで1:1アクリロニトリル:塩化ビニルベンジルから形成され、かつ架橋されたコポリマーでコーティングされたハイブリッドCPGである。
・VBC HybCPG-アミン(以下において、「HybCPG VBC」とも呼ばれる)は、インサイチュで塩化ビニルベンジルモノマーから形成され、かつ架橋されたポリマーでコーティングされたハイブリッドCPGである。
コーティング剤の(コ)ポリマーは、WO 2009/005631に記載されているように、二官能性アミンで架橋されている。その後、アミン基は、フタルイミドナトリウムとの反応、続いてヒドラジンでの脱フタロイルにより、クロロポリマーコーティング剤上へ導入される。
・「中膨張度ポリスチレン」(3-Prime LLC、USAから購入;部品番号04-02-03-32)、アミノ官能化ポリスチレン支持体。ポアサイズ約1000Å(非常に広い分布)、粒子サイズ100μm、アミン負荷222μmol/g
・「低膨張度メタクリレートコポリマー」(SPRIN Technologies S.p.A.、Italyから購入;部品番号1A02BN)、アミン官能化メタクリレート支持体。ポア直径不明、粒子サイズ100〜300μm、アミン負荷270μmol/g。
ポリヒスチジンタグ付き酵素の固定化及び精製のためのキレートCPG担体の調製
所望のタイプのアミノ-CPG(5g)を、メタノール(200mL)中2,4-ジヒドロキシアセトフェノン(CPGのアミノ官能基の1.5当量)で、連続撹拌しながら60分間、処理した。形成されたイミンを、水素化ホウ素ナトリウム(4当量)の逐次添加により、連続撹拌しながら60分間、還元した。この固体材料を濾過し、飽和炭酸ナトリウム水溶液、水、その後、エタノールですすぎ、その後、80℃で2時間、乾燥させた。その後、この粒子を、CoCl2の飽和水溶液(100mL)中に浸漬した。濾過、並びに水及びエタノールでのすすぎ後、この粒子を80℃で2時間、乾燥させた。異なるキレートCPG担体の性質を表1に示す。
ポリヒスチジンタグ付き酵素の固定化及び精製のためのキレート多孔質ポリスチレン及びポリメタクリレート担体の調製
洗浄し(水/エタノール1:1、400mL)、かつ乾燥させた(濾過後、減圧16時間)、所望のタイプ(下記参照)のアミノ官能化多孔質有機ポリマー粒子(2g)を、メタノール(50mL)中2,4-ジヒドロキシアセトフェノン(プラスチックのアミノ官能基の1.5当量)で、連続撹拌しながら60分間、処理した。形成されたイミンを、水素化ホウ素ナトリウム(4当量)の逐次添加により、連続撹拌しながら60分間、還元した。この固体材料を濾過し、飽和炭酸ナトリウム水溶液、水、その後、エタノールですすぎ、その後、25℃で16時間、減圧中、乾燥させた。
ポリヒスチジンタグ付き酵素のキレート担体上への固定化
CalA又はCalBを含有する細胞培養上清を、緩衝化せずに用いた。ω-TAの細胞可溶化物を、HEPES緩衝液(50mM、500mM NaCl、pH8.3)中に細胞を再懸濁することによりにより調製した。界面活性剤(BugBuster(商標)10X、Novagen)の添加後、細胞片を遠心分離により除去した。キレートCPG担体を、その可溶化物又は上清中に浸漬し、続いて、オービタルシェーカー(150rpm)上で撹拌した。固定化中、溶液のブラッドフォード分析される試料によって、タンパク質濃度の減少が止まったら、キレートCPG担体の結合及び飽和の完了が確認された。活性アッセイもまた、CPG担体の濾過による除去後、溶液に実施した。その後、固定化調製物を、緩衝液(CalA及びCalBについてはMOPS(50mM、pH7.4);ω-TAについてはHEPES(上記参照))ですすぎ、減圧下で16時間、乾燥させた。
ω-TA-CPGの触媒としての使用
実施例3において調製されたω-TA-CPGを、フェノキシ-2-プロパノンのエナンチオ特異的アミノ基転移における触媒として適用した:
固定化CalA及びCalBの触媒としての使用
実施例3において調製されたCalA-CPG、CalB-CPG、CalB-ポリスチレン、及びCalB-ポリメタクリレートを、1-フェニルエタノールのエナンチオ選択的アシル化における触媒として適用した(速度論的分割):
- Accurel(登録商標)(多孔質ポリプロピレン粉末)上に固定化されたCalB
- 非修飾型(すなわち、実施例1に従って処理されていない)のアミノ-HybCPGコポ上に固定化されたCalB
3つの異なる酵素を含むカスケード-CPG(「BV-カスケード-CPG」)の調製
2,5-DKCMO、FRE、及びAlaDHの細胞可溶化物を、リン酸ナトリウム緩衝液(50mM、500mM NaCl、pH7.5)中での細胞再懸濁、及びBugBuster(商標)10Xの添加により調製した。遠心分離し細胞片を除去した後、キレートCo2+ CPG担体を、同体積の3つの細胞可溶化物の混合物中に浸漬し、続いて、オービタルシェーカー(150rpm)上で撹拌した。固定化中、溶液のブラッドフォード分析される試料によって、タンパク質濃度の減少が止まったら、CPG担体の結合及び飽和の完了が確認された。活性アッセイもまた、CPGの濾過による除去後、溶液に実施した。その後、固定化調製物を、リン酸ナトリウム緩衝液(上記参照)ですすぎ、その後、減圧下で16時間、乾燥させた。
触媒としてBV-カスケード-CPGを用いる酵素的カスケード反応
実施例6において調製されたカスケード-CPGを、(+)-カンファーのBaeyer-Villiger酸化における触媒として適用した:
CalB及びPd-ナノ粒子を含むカスケード-CPG(「Pd-CalB-CPG」)の調製
アミノ-CPG(5g)を、水(200mL)中Pd(TFA)2(CPGのアミノ官能基の0.5当量)の溶液中に浸漬し、この混合物を、連続して10分間、撹拌した。その後、NaBH4(7当量)を加え、この混合物を更に30分間、撹拌した。その後、2,4-ジヒドロキシアセトフェノン(0.5当量)を加え、その後、この混合物を30分間、撹拌した。この固体材料を濾過し、水で入念にすすぎ、その後、CoCl2の飽和水溶液(100mL)中に浸漬した。濾過及び水での洗浄後、固体材料を、(精製された、又は上清由来の)ポリヒスチジンタグ付きCalBの溶液中に浸漬し、30分間、撹拌した。その後、その固定化調製物を濾過し、緩衝液(20mM MOPS、pH7.4)ですすぎ、その後、減圧下で16時間、乾燥させた。
Pd-CalB-CPGを用いるカスケード反応
実施例8において調製された材料を、1-フェニルエチルアミンのエナンチオ選択的アシル化(速度論的分割)における触媒として適用した:
A.金属イオン浸出の決定
所定の条件において担体から浸出する金属イオンの量を決定するために、CPG及びHybCPGに基づき、かつCo2+か又はFe3+のいずれかを含有する担体を、水性緩衝液中での長時間のインキュベーションに供した。
所定の条件において担体から解離する酵素の量を決定するために、CPG又はHybCPG上のω-TAの固定化調製物を、水性緩衝液中での長時間のインキュベーションに供した。
ポリヒスチジンタグ付きω-TAのHybCPGコポ(Co2+)での精製
50μg/mLカナマイシンを添加した100mL Luria-Bertani培地中でのω-TAのペレット化IPTG誘導化24時間培養からの細胞可溶化物(5.0mL)を、BugBuster(商標)の使用によって調製した。過剰量の補酵素(PLP)を加え、溶液を、37℃で1時間、インキュベートした。その後、過剰な(酵素によって結合されていない)PLPは、PD10カラムを用いてHEPES緩衝液(50mM、500mM NaCl、pH8.2(緩衝液1))に緩衝液交換(2ラウンド)することによって除去し、標的ホロ酵素、天然タンパク質、及び他の不純物を含有する7.0mLの溶液が得られた。
Claims (24)
- 化学合成における酵素触媒反応を触媒するための方法であって、
(a) 担体、及び該担体上に固定化された少なくとも1つの酵素を準備する工程であって、
前記担体が、キレートされた金属イオンを含有するアフィニティマトリックスが付着している多孔質有機ポリマーであり、
前記少なくとも1つの酵素が、ポリヒスチジンタグを含有し、該ポリヒスチジンタグと前記キレートされた金属イオンとの間の特異的アフィニティ結合を介して前記担体上に固定化されている、工程、及び、
(b) 前記固定化された酵素を少なくとも1つの基質と接触させて、これによりに前記反応を触媒する工程、を含む、方法。 - 前記アフィニティマトリックスが、リンカーを介して前記担体の表面に付着している、請求項1に記載の方法。
- 前記多孔質有機ポリマーが、ポリエチレン、超高分子量ポリエチレン(UHMWPE)、高密度ポリエチレン(HDPE)、ポリプロピレン(PP)、ポリテトラフルオロエチレン(PTFE)、ポリビニリデンフルオリド(PVDF)、ポリスチレン、ポリメタクリレート、及びポリ(メチルメタクリレート)からなる群から選択される、請求項1に記載の方法。
- 前記多孔質有機ポリマーがポリスチレンである、請求項1に記載の方法。
- 前記多孔質有機ポリマーがポリメタクリレートである、請求項1に記載の方法。
- 前記多孔質有機ポリマーが、膨張が限られた、多孔質粒子として存在する、請求項1に記載の方法。
- 前記キレートされた金属イオンがFe2+である、請求項1に記載の方法。
- 前記キレートされた金属イオンがFe3+である、請求項1に記載の方法。
- 前記キレートされた金属イオンがCo2+である、請求項1に記載の方法。
- 前記キレートされた金属イオンがNi2+である、請求項1に記載の方法。
- 前記キレートされた金属イオンがCu2+である、請求項1に記載の方法。
- 前記キレートされた金属イオンがZn2+である、請求項1に記載の方法。
- 2つ以上の異なる酵素が前記担体上に固定化され、前記異なる酵素のそれぞれが異なる反応を触媒することができる、請求項1に記載の方法。
- 前記担体が金属ナノ粒子を更に含む、請求項1に記載の方法。
- 前記金属ナノ粒子が遷移金属ナノ粒子である、請求項14に記載の方法。
- 前記金属ナノ粒子が、コバルト、ニッケル、及びパラジウムのナノ粒子からなる群から選択される、請求項14に記載の方法。
- 前記酵素触媒反応が水性条件下で行われる、請求項1に記載の方法。
- 前記酵素触媒反応が有機溶媒中で行われる、請求項1に記載の方法。
- 前記酵素触媒反応が連続フロー反応である、請求項1に記載の方法。
- 前記酵素触媒反応がカスケード反応である、請求項13に記載の方法。
- 第1の酵素のための基質が、第2の酵素のための基質へと変換される、請求項20に記載の方法。
- 基質が第1の酵素によって変換され、かつ前記第1の酵素のための補助因子が第2の酵素によって再生される、請求項20に記載の方法。
- 遷移金属が前記担体に固定化され、かつ前記方法が酵素触媒反応と遷移金属触媒反応を含む多段階反応である、請求項15に記載の方法。
- 前記少なくとも1つの基質が前記酵素触媒反応において生成物に変換される、請求項1に記載の方法。
Applications Claiming Priority (5)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
SE1450105-0 | 2014-01-31 | ||
SE1450105 | 2014-01-31 | ||
SE1450822 | 2014-07-02 | ||
SE1450822-0 | 2014-07-02 | ||
JP2016549481A JP6534161B2 (ja) | 2014-01-31 | 2015-01-30 | 固定化タンパク質及びその使用 |
Related Parent Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP2016549481A Division JP6534161B2 (ja) | 2014-01-31 | 2015-01-30 | 固定化タンパク質及びその使用 |
Publications (2)
Publication Number | Publication Date |
---|---|
JP2019162133A JP2019162133A (ja) | 2019-09-26 |
JP6908653B2 true JP6908653B2 (ja) | 2021-07-28 |
Family
ID=52649088
Family Applications (2)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP2016549481A Active JP6534161B2 (ja) | 2014-01-31 | 2015-01-30 | 固定化タンパク質及びその使用 |
JP2019095107A Active JP6908653B2 (ja) | 2014-01-31 | 2019-05-21 | 固定化タンパク質及びその使用 |
Family Applications Before (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP2016549481A Active JP6534161B2 (ja) | 2014-01-31 | 2015-01-30 | 固定化タンパク質及びその使用 |
Country Status (17)
Country | Link |
---|---|
US (3) | US10793848B2 (ja) |
EP (3) | EP3910337B1 (ja) |
JP (2) | JP6534161B2 (ja) |
KR (2) | KR102465626B1 (ja) |
CN (2) | CN105940104B (ja) |
AU (1) | AU2015211437B2 (ja) |
BR (1) | BR112016017398B1 (ja) |
CA (2) | CA2936690C (ja) |
CL (1) | CL2016001921A1 (ja) |
DK (3) | DK3910337T3 (ja) |
ES (3) | ES2756601T3 (ja) |
FI (1) | FI3910337T3 (ja) |
MX (2) | MX2016009805A (ja) |
PL (3) | PL3910337T3 (ja) |
RU (1) | RU2684619C2 (ja) |
WO (1) | WO2015115993A1 (ja) |
ZA (1) | ZA201605760B (ja) |
Families Citing this family (10)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2017151104A1 (en) * | 2016-02-29 | 2017-09-08 | Hewlett-Packard Development Company, L.P. | Enzymatic sample purification |
CN109022413A (zh) * | 2018-08-10 | 2018-12-18 | 暨南大学 | 一种单胺氧化酶a微反应器及其制备方法和应用 |
CN114599786A (zh) * | 2019-07-17 | 2022-06-07 | 博努莫斯股份有限公司 | 用于生产己糖的固定化酶组合物 |
CN111647561B (zh) * | 2020-05-28 | 2022-12-20 | 大连理工大学 | 纳米抗体在细胞特异性捕获和细胞释放中的应用 |
GB202009513D0 (en) * | 2020-06-22 | 2020-08-05 | Univ Of Stellenbosch | An enzyme-polymer matrix |
CN113061158A (zh) * | 2021-03-30 | 2021-07-02 | 南京工业大学 | 一种利用二价金属离子与组氨酸标签纯化并固定化蛋白的方法及其应用 |
CN113122445A (zh) * | 2021-05-06 | 2021-07-16 | 南京普瑞特生物科技有限公司 | 固定化酶法拆分制备光学纯1-(1-萘基)乙胺的设备及使用方法 |
GB2614570A (en) | 2022-01-10 | 2023-07-12 | Uab Biomatter Designs | C-nucleoside monophosphate synthesis |
WO2023227795A1 (en) | 2022-05-27 | 2023-11-30 | Enginzyme Ab | Biocatalysts for organic synthesis |
WO2023229519A1 (en) | 2022-05-27 | 2023-11-30 | Bunge Sa | Batch process for enzymatic modification of lipids |
Family Cites Families (12)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPS56115727A (en) * | 1980-02-19 | 1981-09-11 | Kuraray Co Ltd | Carrier for immobilizing physiologically active substance |
US4632904A (en) | 1983-12-27 | 1986-12-30 | Ciba Corning Diagnostics Corp. | Immobilized enzyme composites having carriers derivatized with an organotitanate |
US5610274A (en) | 1991-11-20 | 1997-03-11 | Cpg, Inc. | Production and use of magnetic porous inorganic materials |
US6087452A (en) * | 1998-06-02 | 2000-07-11 | University Of Utah | Metal-chelating surfacant |
KR100506646B1 (ko) | 2002-07-22 | 2005-08-03 | 현대모비스 주식회사 | 차량 라디에이터 상부 마운팅 구조 |
US20040115777A1 (en) * | 2002-10-15 | 2004-06-17 | Syngenta Participations Ag | Identification of protein interactions using in vivo post-translationally modified fusion proteins |
EP2170776B1 (en) * | 2007-06-28 | 2018-11-07 | Prime Synthesis, Inc. | Method for preparing a polymer-coated controlled porosity glass particle |
EP2023140A1 (en) | 2007-08-08 | 2009-02-11 | FUJIFILM Corporation | Biosensor chip |
JP2010190891A (ja) | 2009-01-22 | 2010-09-02 | Fujifilm Corp | 担体およびその製造方法並びに抽出操作器具 |
CN102260662B (zh) | 2010-05-24 | 2017-04-05 | 中国科学院过程工程研究所 | 用于固定化酶的载体及其用途和固定有酶的载体 |
SG10201701224UA (en) * | 2012-03-12 | 2017-04-27 | Merck Patent Gmbh | Removal of protein aggregates from biopharmaceutical preparations in a flowthrough mode |
CN102998444B (zh) | 2012-07-15 | 2014-08-06 | 潍坊医学院 | IgG抗体在聚苯乙烯载体表面的立体定向固定方法 |
-
2015
- 2015-01-30 DK DK21184595.3T patent/DK3910337T3/da active
- 2015-01-30 CA CA2936690A patent/CA2936690C/en active Active
- 2015-01-30 CN CN201580005764.2A patent/CN105940104B/zh active Active
- 2015-01-30 ES ES15709372T patent/ES2756601T3/es active Active
- 2015-01-30 FI FIEP21184595.3T patent/FI3910337T3/fi active
- 2015-01-30 MX MX2016009805A patent/MX2016009805A/es unknown
- 2015-01-30 DK DK15709372T patent/DK3100050T3/da active
- 2015-01-30 BR BR112016017398-8A patent/BR112016017398B1/pt active IP Right Grant
- 2015-01-30 ES ES21184595T patent/ES2958266T3/es active Active
- 2015-01-30 JP JP2016549481A patent/JP6534161B2/ja active Active
- 2015-01-30 CA CA3205514A patent/CA3205514A1/en active Pending
- 2015-01-30 EP EP21184595.3A patent/EP3910337B1/en active Active
- 2015-01-30 US US15/112,206 patent/US10793848B2/en active Active
- 2015-01-30 KR KR1020227021766A patent/KR102465626B1/ko active IP Right Grant
- 2015-01-30 ES ES19192953T patent/ES2892319T3/es active Active
- 2015-01-30 KR KR1020167023700A patent/KR102414887B1/ko active IP Right Grant
- 2015-01-30 PL PL21184595.3T patent/PL3910337T3/pl unknown
- 2015-01-30 WO PCT/SE2015/050108 patent/WO2015115993A1/en active Application Filing
- 2015-01-30 DK DK19192953.8T patent/DK3605100T3/da active
- 2015-01-30 PL PL15709372T patent/PL3100050T3/pl unknown
- 2015-01-30 AU AU2015211437A patent/AU2015211437B2/en active Active
- 2015-01-30 RU RU2016134892A patent/RU2684619C2/ru active
- 2015-01-30 CN CN201911330175.8A patent/CN110951720B/zh active Active
- 2015-01-30 EP EP19192953.8A patent/EP3605100B1/en active Active
- 2015-01-30 PL PL19192953T patent/PL3605100T3/pl unknown
- 2015-01-30 EP EP15709372.5A patent/EP3100050B1/en active Active
-
2016
- 2016-07-27 MX MX2022001420A patent/MX2022001420A/es unknown
- 2016-07-28 CL CL2016001921A patent/CL2016001921A1/es unknown
- 2016-08-18 ZA ZA201605760A patent/ZA201605760B/en unknown
-
2019
- 2019-04-15 US US16/383,935 patent/US20190241884A1/en not_active Abandoned
- 2019-05-21 JP JP2019095107A patent/JP6908653B2/ja active Active
-
2021
- 2021-01-19 US US17/152,074 patent/US20210139881A1/en not_active Abandoned
Also Published As
Similar Documents
Publication | Publication Date | Title |
---|---|---|
JP6908653B2 (ja) | 固定化タンパク質及びその使用 | |
Britton et al. | Continuous flow biocatalysis | |
Brena et al. | Immobilization of enzymes: a literature survey | |
Hanefeld et al. | Understanding enzyme immobilisation | |
Cassimjee et al. | A general protein purification and immobilization method on controlled porosity glass: Biocatalytic applications | |
Andrade et al. | Continuous flow synthesis of chiral amines in organic solvents: immobilization of E. coli cells containing both ω-transaminase and PLP | |
Kallenberg et al. | Immobilization of penicillin G acylase: the key to optimum performance | |
T. sriwong et al. | Recent advances in enzyme immobilization utilizing nanotechnology for biocatalysis | |
Quaglia et al. | His-tagged horse liver alcohol dehydrogenase: immobilization and application in the bio-based enantioselective synthesis of (S)-arylpropanols | |
Coloma et al. | Immobilisation and flow chemistry: tools for implementing biocatalysis | |
Zhang et al. | Progress and perspective of enzyme immobilization on zeolite crystal materials | |
Mao et al. | Flow‐through enzymatic reactors using polymer monoliths: from motivation to application | |
Basso et al. | Optimization of metal affinity ketoreductase immobilization for application in batch and flow processes | |
Khanam et al. | Recent advances in immobilized ω-transaminase for chiral amine synthesis | |
Peper et al. | Immobilization and characterization of benzoylformate decarboxylase from Pseudomonas putida on spherical silica carrier | |
Tang et al. | Control of the activity and enantioselectivity in biocatalyzed procedures: immobilization, medium engineering, and protein engineering | |
RU2650668C1 (ru) | Способ получения гетерогенного биокатализатора на основе липазы дрожжей Candida antarctica фракции В | |
Päiviö | Lipase and omega-transaminase catalysis in preparation of alcohol and amine enantiomers |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
A621 | Written request for application examination |
Free format text: JAPANESE INTERMEDIATE CODE: A621 Effective date: 20190527 |
|
A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20200622 |
|
A601 | Written request for extension of time |
Free format text: JAPANESE INTERMEDIATE CODE: A601 Effective date: 20200918 |
|
A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20201222 |
|
TRDD | Decision of grant or rejection written | ||
A01 | Written decision to grant a patent or to grant a registration (utility model) |
Free format text: JAPANESE INTERMEDIATE CODE: A01 Effective date: 20210607 |
|
A61 | First payment of annual fees (during grant procedure) |
Free format text: JAPANESE INTERMEDIATE CODE: A61 Effective date: 20210701 |
|
R150 | Certificate of patent or registration of utility model |
Ref document number: 6908653 Country of ref document: JP Free format text: JAPANESE INTERMEDIATE CODE: R150 |