CN104684958A - Isoprene oligomer, polyisoprene, method for producing isoprene oligomer and polyisoprene, rubber composition, and pneumatic tire - Google Patents
Isoprene oligomer, polyisoprene, method for producing isoprene oligomer and polyisoprene, rubber composition, and pneumatic tire Download PDFInfo
- Publication number
- CN104684958A CN104684958A CN201380045290.5A CN201380045290A CN104684958A CN 104684958 A CN104684958 A CN 104684958A CN 201380045290 A CN201380045290 A CN 201380045290A CN 104684958 A CN104684958 A CN 104684958A
- Authority
- CN
- China
- Prior art keywords
- formula
- isoprene
- isoprene oligopolymer
- polyisoprene
- represented
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- RRHGJUQNOFWUDK-UHFFFAOYSA-N Isoprene Chemical compound CC(=C)C=C RRHGJUQNOFWUDK-UHFFFAOYSA-N 0.000 title claims abstract description 791
- 229920001195 polyisoprene Polymers 0.000 title claims abstract description 162
- 229920001971 elastomer Polymers 0.000 title claims abstract description 112
- 239000005060 rubber Substances 0.000 title claims abstract description 112
- 238000004519 manufacturing process Methods 0.000 title claims description 57
- 239000000203 mixture Substances 0.000 title abstract description 26
- 150000001875 compounds Chemical class 0.000 claims abstract description 104
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 claims abstract description 30
- 108090000790 Enzymes Proteins 0.000 claims description 93
- 102000004190 Enzymes Human genes 0.000 claims description 93
- CBIDRCWHNCKSTO-UHFFFAOYSA-N prenyl diphosphate Chemical compound CC(C)=CCO[P@](O)(=O)OP(O)(O)=O CBIDRCWHNCKSTO-UHFFFAOYSA-N 0.000 claims description 83
- 230000000694 effects Effects 0.000 claims description 56
- 238000000034 method Methods 0.000 claims description 44
- 238000003786 synthesis reaction Methods 0.000 claims description 37
- 230000015572 biosynthetic process Effects 0.000 claims description 35
- XRQVVFIEYAHKBV-OGYJWPHRSA-N opp protocol Chemical compound CNNCC1=CC=C(C(=O)NC(C)C)C=C1.O=C1C=C[C@]2(C)[C@H]3C(=O)C[C@](C)([C@@](CC4)(O)C(=O)CO)[C@@H]4[C@@H]3CCC2=C1.C([C@H](C[C@]1(C(=O)OC)C=2C(=C3C([C@]45[C@H]([C@@]([C@H](OC(C)=O)[C@]6(CC)C=CCN([C@H]56)CC4)(O)C(=O)OC)N3C=O)=CC=2)OC)C[C@@](C2)(O)CC)N2CCC2=C1NC1=CC=CC=C21 XRQVVFIEYAHKBV-OGYJWPHRSA-N 0.000 claims description 35
- 102000019337 Prenyltransferases Human genes 0.000 claims description 34
- 108050006837 Prenyltransferases Proteins 0.000 claims description 34
- 125000002887 hydroxy group Chemical group [H]O* 0.000 claims description 33
- 125000004453 alkoxycarbonyl group Chemical group 0.000 claims description 26
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 claims description 16
- CFHIDWOYWUOIHU-UHFFFAOYSA-N oxomethyl Chemical compound O=[CH] CFHIDWOYWUOIHU-UHFFFAOYSA-N 0.000 claims description 14
- 229920003051 synthetic elastomer Polymers 0.000 claims description 2
- 230000004048 modification Effects 0.000 abstract description 44
- 238000012986 modification Methods 0.000 abstract description 44
- 238000002156 mixing Methods 0.000 abstract description 7
- 125000000746 allylic group Chemical group 0.000 abstract 1
- XPPKVPWEQAFLFU-UHFFFAOYSA-N diphosphoric acid Chemical compound OP(O)(=O)OP(O)(O)=O XPPKVPWEQAFLFU-UHFFFAOYSA-N 0.000 abstract 1
- 125000004429 atom Chemical group 0.000 description 74
- 238000006243 chemical reaction Methods 0.000 description 58
- 238000002360 preparation method Methods 0.000 description 56
- 239000007858 starting material Substances 0.000 description 55
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 54
- 244000043261 Hevea brasiliensis Species 0.000 description 29
- 239000000178 monomer Substances 0.000 description 29
- TWRXJAOTZQYOKJ-UHFFFAOYSA-L Magnesium chloride Chemical compound [Mg+2].[Cl-].[Cl-] TWRXJAOTZQYOKJ-UHFFFAOYSA-L 0.000 description 28
- 239000000945 filler Substances 0.000 description 27
- 239000000377 silicon dioxide Substances 0.000 description 27
- 235000012239 silicon dioxide Nutrition 0.000 description 25
- 229960001866 silicon dioxide Drugs 0.000 description 23
- DGVVWUTYPXICAM-UHFFFAOYSA-N β‐Mercaptoethanol Chemical compound OCCS DGVVWUTYPXICAM-UHFFFAOYSA-N 0.000 description 20
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 18
- 230000003570 biosynthesizing effect Effects 0.000 description 18
- 229920000126 latex Polymers 0.000 description 18
- 239000004816 latex Substances 0.000 description 17
- 108090000623 proteins and genes Proteins 0.000 description 17
- 239000000463 material Substances 0.000 description 16
- 235000018102 proteins Nutrition 0.000 description 16
- 102000004169 proteins and genes Human genes 0.000 description 16
- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical compound CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 description 15
- -1 Sauerstoffatom Chemical group 0.000 description 15
- 229910001629 magnesium chloride Inorganic materials 0.000 description 14
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 13
- 229910052799 carbon Inorganic materials 0.000 description 13
- 238000002474 experimental method Methods 0.000 description 13
- 230000003993 interaction Effects 0.000 description 13
- 229920003052 natural elastomer Polymers 0.000 description 13
- 229920001194 natural rubber Polymers 0.000 description 13
- 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 12
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 12
- 102000002508 Peptide Elongation Factors Human genes 0.000 description 12
- 108010068204 Peptide Elongation Factors Proteins 0.000 description 12
- 125000005340 bisphosphate group Chemical group 0.000 description 12
- 238000013016 damping Methods 0.000 description 12
- 239000012530 fluid Substances 0.000 description 12
- 125000000524 functional group Chemical group 0.000 description 12
- 229920006173 natural rubber latex Polymers 0.000 description 12
- 235000015097 nutrients Nutrition 0.000 description 12
- 230000009257 reactivity Effects 0.000 description 12
- 239000006228 supernatant Substances 0.000 description 12
- 238000002290 gas chromatography-mass spectrometry Methods 0.000 description 11
- 239000000126 substance Substances 0.000 description 11
- GLZPCOQZEFWAFX-UHFFFAOYSA-N Geraniol Chemical compound CC(C)=CCCC(C)=CCO GLZPCOQZEFWAFX-UHFFFAOYSA-N 0.000 description 10
- 238000005119 centrifugation Methods 0.000 description 10
- 238000012360 testing method Methods 0.000 description 10
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 9
- 241000588724 Escherichia coli Species 0.000 description 9
- NINIDFKCEFEMDL-UHFFFAOYSA-N Sulfur Chemical group [S] NINIDFKCEFEMDL-UHFFFAOYSA-N 0.000 description 9
- KLOHDWPABZXLGI-YWUHCJSESA-M ampicillin sodium Chemical compound [Na+].C1([C@@H](N)C(=O)N[C@H]2[C@H]3SC([C@@H](N3C2=O)C([O-])=O)(C)C)=CC=CC=C1 KLOHDWPABZXLGI-YWUHCJSESA-M 0.000 description 9
- 238000006555 catalytic reaction Methods 0.000 description 9
- 210000004027 cell Anatomy 0.000 description 9
- OINNEUNVOZHBOX-QIRCYJPOSA-N 2-trans,6-trans,10-trans-geranylgeranyl diphosphate Chemical compound CC(C)=CCC\C(C)=C\CC\C(C)=C\CC\C(C)=C\COP(O)(=O)OP(O)(O)=O OINNEUNVOZHBOX-QIRCYJPOSA-N 0.000 description 8
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 8
- OFBQJSOFQDEBGM-UHFFFAOYSA-N Pentane Chemical compound CCCCC OFBQJSOFQDEBGM-UHFFFAOYSA-N 0.000 description 8
- JUJWROOIHBZHMG-UHFFFAOYSA-N Pyridine Chemical compound C1=CC=NC=C1 JUJWROOIHBZHMG-UHFFFAOYSA-N 0.000 description 8
- 238000012644 addition polymerization Methods 0.000 description 8
- 239000000047 product Substances 0.000 description 8
- 230000002194 synthesizing effect Effects 0.000 description 8
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 7
- 102220466384 PRA1 family protein 2_N77A_mutation Human genes 0.000 description 7
- 239000002585 base Substances 0.000 description 7
- 235000002639 sodium chloride Nutrition 0.000 description 7
- 0 CC*(C)CC*CCCC(N1C(C)CC1)=*C=C Chemical compound CC*(C)CC*CCCC(N1C(C)CC1)=*C=C 0.000 description 6
- OINNEUNVOZHBOX-XBQSVVNOSA-N Geranylgeranyl diphosphate Natural products [P@](=O)(OP(=O)(O)O)(OC/C=C(\CC/C=C(\CC/C=C(\CC/C=C(\C)/C)/C)/C)/C)O OINNEUNVOZHBOX-XBQSVVNOSA-N 0.000 description 6
- KWYUFKZDYYNOTN-UHFFFAOYSA-M Potassium hydroxide Chemical compound [OH-].[K+] KWYUFKZDYYNOTN-UHFFFAOYSA-M 0.000 description 6
- WYURNTSHIVDZCO-UHFFFAOYSA-N Tetrahydrofuran Chemical compound C1CCOC1 WYURNTSHIVDZCO-UHFFFAOYSA-N 0.000 description 6
- XLOMVQKBTHCTTD-UHFFFAOYSA-N Zinc monoxide Chemical compound [Zn]=O XLOMVQKBTHCTTD-UHFFFAOYSA-N 0.000 description 6
- 239000003292 glue Substances 0.000 description 6
- QJGQUHMNIGDVPM-UHFFFAOYSA-N nitrogen group Chemical group [N] QJGQUHMNIGDVPM-UHFFFAOYSA-N 0.000 description 6
- 230000010355 oscillation Effects 0.000 description 6
- WFCGYZPOBJZGDF-UHFFFAOYSA-N 2-methylbuta-1,3-diene;phosphono dihydrogen phosphate Chemical compound CC(=C)C=C.OP(O)(=O)OP(O)(O)=O WFCGYZPOBJZGDF-UHFFFAOYSA-N 0.000 description 5
- 239000005792 Geraniol Substances 0.000 description 5
- GLZPCOQZEFWAFX-YFHOEESVSA-N Geraniol Natural products CC(C)=CCC\C(C)=C/CO GLZPCOQZEFWAFX-YFHOEESVSA-N 0.000 description 5
- 239000005864 Sulphur Substances 0.000 description 5
- 125000000217 alkyl group Chemical group 0.000 description 5
- 238000004458 analytical method Methods 0.000 description 5
- 230000000052 comparative effect Effects 0.000 description 5
- 239000000470 constituent Substances 0.000 description 5
- 229940113087 geraniol Drugs 0.000 description 5
- NROKBHXJSPEDAR-UHFFFAOYSA-M potassium fluoride Chemical compound [F-].[K+] NROKBHXJSPEDAR-UHFFFAOYSA-M 0.000 description 5
- 230000008569 process Effects 0.000 description 5
- 244000020551 Helianthus annuus Species 0.000 description 4
- 235000003222 Helianthus annuus Nutrition 0.000 description 4
- 241000191938 Micrococcus luteus Species 0.000 description 4
- MZRVEZGGRBJDDB-UHFFFAOYSA-N N-Butyllithium Chemical class [Li]CCCC MZRVEZGGRBJDDB-UHFFFAOYSA-N 0.000 description 4
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 4
- 150000001299 aldehydes Chemical class 0.000 description 4
- 125000003545 alkoxy group Chemical group 0.000 description 4
- 239000006229 carbon black Substances 0.000 description 4
- 230000021523 carboxylation Effects 0.000 description 4
- 238000006473 carboxylation reaction Methods 0.000 description 4
- 230000008859 change Effects 0.000 description 4
- 239000001177 diphosphate Substances 0.000 description 4
- XPPKVPWEQAFLFU-UHFFFAOYSA-J diphosphate(4-) Chemical compound [O-]P([O-])(=O)OP([O-])([O-])=O XPPKVPWEQAFLFU-UHFFFAOYSA-J 0.000 description 4
- 235000011180 diphosphates Nutrition 0.000 description 4
- 230000002255 enzymatic effect Effects 0.000 description 4
- 235000019439 ethyl acetate Nutrition 0.000 description 4
- 238000001704 evaporation Methods 0.000 description 4
- 230000008020 evaporation Effects 0.000 description 4
- 238000011534 incubation Methods 0.000 description 4
- 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 4
- 238000005567 liquid scintillation counting Methods 0.000 description 4
- 239000006916 nutrient agar Substances 0.000 description 4
- 125000001997 phenyl group Chemical group [H]C1=C([H])C([H])=C(*)C([H])=C1[H] 0.000 description 4
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 4
- 238000001742 protein purification Methods 0.000 description 4
- UMJSCPRVCHMLSP-UHFFFAOYSA-N pyridine Natural products COC1=CC=CN=C1 UMJSCPRVCHMLSP-UHFFFAOYSA-N 0.000 description 4
- 238000011160 research Methods 0.000 description 4
- 229920006395 saturated elastomer Polymers 0.000 description 4
- 230000035939 shock Effects 0.000 description 4
- 239000011780 sodium chloride Substances 0.000 description 4
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 4
- 238000003756 stirring Methods 0.000 description 4
- 239000000758 substrate Substances 0.000 description 4
- MHYGQXWCZAYSLJ-UHFFFAOYSA-N tert-butyl-chloro-diphenylsilane Chemical compound C=1C=CC=CC=1[Si](Cl)(C(C)(C)C)C1=CC=CC=C1 MHYGQXWCZAYSLJ-UHFFFAOYSA-N 0.000 description 4
- GPRLSGONYQIRFK-MNYXATJNSA-N triton Chemical compound [3H+] GPRLSGONYQIRFK-MNYXATJNSA-N 0.000 description 4
- 238000005199 ultracentrifugation Methods 0.000 description 4
- 238000004073 vulcanization Methods 0.000 description 4
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 3
- 102000053602 DNA Human genes 0.000 description 3
- 108020004414 DNA Proteins 0.000 description 3
- 102100035111 Farnesyl pyrophosphate synthase Human genes 0.000 description 3
- 108010026318 Geranyltranstransferase Proteins 0.000 description 3
- 229920000459 Nitrile rubber Polymers 0.000 description 3
- 239000005062 Polybutadiene Substances 0.000 description 3
- XUIMIQQOPSSXEZ-UHFFFAOYSA-N Silicon Chemical compound [Si] XUIMIQQOPSSXEZ-UHFFFAOYSA-N 0.000 description 3
- 235000021355 Stearic acid Nutrition 0.000 description 3
- 150000001298 alcohols Chemical class 0.000 description 3
- 125000003275 alpha amino acid group Chemical group 0.000 description 3
- 125000003118 aryl group Chemical group 0.000 description 3
- 125000000484 butyl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])C([H])([H])[H] 0.000 description 3
- 150000001721 carbon Chemical group 0.000 description 3
- 239000000460 chlorine Substances 0.000 description 3
- 229910052801 chlorine Inorganic materials 0.000 description 3
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 description 3
- 238000006482 condensation reaction Methods 0.000 description 3
- 238000013461 design Methods 0.000 description 3
- 125000001495 ethyl group Chemical group [H]C([H])([H])C([H])([H])* 0.000 description 3
- 239000012467 final product Substances 0.000 description 3
- 238000010353 genetic engineering Methods 0.000 description 3
- 230000006872 improvement Effects 0.000 description 3
- 125000004433 nitrogen atom Chemical group N* 0.000 description 3
- QIQXTHQIDYTFRH-UHFFFAOYSA-N octadecanoic acid Chemical compound CCCCCCCCCCCCCCCCCC(O)=O QIQXTHQIDYTFRH-UHFFFAOYSA-N 0.000 description 3
- OQCDKBAXFALNLD-UHFFFAOYSA-N octadecanoic acid Natural products CCCCCCCC(C)CCCCCCCCC(O)=O OQCDKBAXFALNLD-UHFFFAOYSA-N 0.000 description 3
- 150000003839 salts Chemical class 0.000 description 3
- 229910052710 silicon Inorganic materials 0.000 description 3
- 239000010703 silicon Substances 0.000 description 3
- 239000008117 stearic acid Substances 0.000 description 3
- 229920003048 styrene butadiene rubber Polymers 0.000 description 3
- 239000011593 sulfur Substances 0.000 description 3
- 229910052717 sulfur Inorganic materials 0.000 description 3
- 230000001629 suppression Effects 0.000 description 3
- YLQBMQCUIZJEEH-UHFFFAOYSA-N tetrahydrofuran Natural products C=1C=COC=1 YLQBMQCUIZJEEH-UHFFFAOYSA-N 0.000 description 3
- 239000001993 wax Substances 0.000 description 3
- 239000011787 zinc oxide Substances 0.000 description 3
- 125000003903 2-propenyl group Chemical group [H]C([*])([H])C([H])=C([H])[H] 0.000 description 2
- YKAYSCGASBCKPU-UHFFFAOYSA-N 3-bromo-1-chloropyrrolidine-2,5-dione Chemical compound ClN1C(=O)CC(Br)C1=O YKAYSCGASBCKPU-UHFFFAOYSA-N 0.000 description 2
- WFDIJRYMOXRFFG-UHFFFAOYSA-N Acetic anhydride Chemical compound CC(=O)OC(C)=O WFDIJRYMOXRFFG-UHFFFAOYSA-N 0.000 description 2
- NLXLAEXVIDQMFP-UHFFFAOYSA-N Ammonia chloride Chemical compound [NH4+].[Cl-] NLXLAEXVIDQMFP-UHFFFAOYSA-N 0.000 description 2
- KAKZBPTYRLMSJV-UHFFFAOYSA-N Butadiene Chemical group C=CC=C KAKZBPTYRLMSJV-UHFFFAOYSA-N 0.000 description 2
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 description 2
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 2
- ZAMOUSCENKQFHK-UHFFFAOYSA-N Chlorine atom Chemical compound [Cl] ZAMOUSCENKQFHK-UHFFFAOYSA-N 0.000 description 2
- QMMFVYPAHWMCMS-UHFFFAOYSA-N Dimethyl sulfide Chemical compound CSC QMMFVYPAHWMCMS-UHFFFAOYSA-N 0.000 description 2
- ROSDSFDQCJNGOL-UHFFFAOYSA-N Dimethylamine Chemical compound CNC ROSDSFDQCJNGOL-UHFFFAOYSA-N 0.000 description 2
- LYCAIKOWRPUZTN-UHFFFAOYSA-N Ethylene glycol Chemical compound OCCO LYCAIKOWRPUZTN-UHFFFAOYSA-N 0.000 description 2
- 240000003537 Ficus benghalensis Species 0.000 description 2
- 244000286663 Ficus elastica Species 0.000 description 2
- 241000983593 Ficus lyrata Species 0.000 description 2
- VZCYOOQTPOCHFL-OWOJBTEDSA-N Fumaric acid Chemical compound OC(=O)\C=C\C(O)=O VZCYOOQTPOCHFL-OWOJBTEDSA-N 0.000 description 2
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 2
- 241000282414 Homo sapiens Species 0.000 description 2
- 241001406327 Periploca sepium Species 0.000 description 2
- 229920005683 SIBR Polymers 0.000 description 2
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 2
- 239000006087 Silane Coupling Agent Substances 0.000 description 2
- 241000488874 Sonchus Species 0.000 description 2
- 244000113428 Sonchus oleraceus Species 0.000 description 2
- 235000006745 Sonchus oleraceus Nutrition 0.000 description 2
- 239000002174 Styrene-butadiene Substances 0.000 description 2
- 241000205098 Sulfolobus acidocaldarius Species 0.000 description 2
- 241000245665 Taraxacum Species 0.000 description 2
- 240000001949 Taraxacum officinale Species 0.000 description 2
- 235000006754 Taraxacum officinale Nutrition 0.000 description 2
- 235000005187 Taraxacum officinale ssp. officinale Nutrition 0.000 description 2
- 125000002777 acetyl group Chemical group [H]C([H])([H])C(*)=O 0.000 description 2
- 239000013543 active substance Substances 0.000 description 2
- 238000005904 alkaline hydrolysis reaction Methods 0.000 description 2
- LDDQLRUQCUTJBB-UHFFFAOYSA-N ammonium fluoride Chemical class [NH4+].[F-] LDDQLRUQCUTJBB-UHFFFAOYSA-N 0.000 description 2
- 230000000845 anti-microbial effect Effects 0.000 description 2
- 150000001720 carbohydrates Chemical class 0.000 description 2
- 235000014633 carbohydrates Nutrition 0.000 description 2
- 210000000170 cell membrane Anatomy 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 230000004087 circulation Effects 0.000 description 2
- 238000012790 confirmation Methods 0.000 description 2
- 238000006356 dehydrogenation reaction Methods 0.000 description 2
- XBDQKXXYIPTUBI-UHFFFAOYSA-N dimethylselenoniopropionate Natural products CCC(O)=O XBDQKXXYIPTUBI-UHFFFAOYSA-N 0.000 description 2
- 150000002031 dolichols Chemical class 0.000 description 2
- 230000007613 environmental effect Effects 0.000 description 2
- 230000032050 esterification Effects 0.000 description 2
- 238000005886 esterification reaction Methods 0.000 description 2
- 230000006870 function Effects 0.000 description 2
- 238000007306 functionalization reaction Methods 0.000 description 2
- 238000005984 hydrogenation reaction Methods 0.000 description 2
- 230000007062 hydrolysis Effects 0.000 description 2
- 238000006460 hydrolysis reaction Methods 0.000 description 2
- 150000002460 imidazoles Chemical class 0.000 description 2
- NUHSROFQTUXZQQ-UHFFFAOYSA-N isopentenyl diphosphate Chemical compound CC(=C)CCO[P@](O)(=O)OP(O)(O)=O NUHSROFQTUXZQQ-UHFFFAOYSA-N 0.000 description 2
- 239000012280 lithium aluminium hydride Substances 0.000 description 2
- 238000012423 maintenance Methods 0.000 description 2
- HZVOZRGWRWCICA-UHFFFAOYSA-N methanediyl Chemical compound [CH2] HZVOZRGWRWCICA-UHFFFAOYSA-N 0.000 description 2
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 description 2
- 230000007935 neutral effect Effects 0.000 description 2
- 239000003208 petroleum Substances 0.000 description 2
- 239000013612 plasmid Substances 0.000 description 2
- 229920001084 poly(chloroprene) Polymers 0.000 description 2
- FGIUAXJPYTZDNR-UHFFFAOYSA-N potassium nitrate Chemical compound [K+].[O-][N+]([O-])=O FGIUAXJPYTZDNR-UHFFFAOYSA-N 0.000 description 2
- 239000000843 powder Substances 0.000 description 2
- 125000001436 propyl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])[H] 0.000 description 2
- 239000003223 protective agent Substances 0.000 description 2
- 230000035484 reaction time Effects 0.000 description 2
- 239000011435 rock Substances 0.000 description 2
- RMAQACBXLXPBSY-UHFFFAOYSA-N silicic acid Chemical compound O[Si](O)(O)O RMAQACBXLXPBSY-UHFFFAOYSA-N 0.000 description 2
- 238000002741 site-directed mutagenesis Methods 0.000 description 2
- VWDWKYIASSYTQR-UHFFFAOYSA-N sodium nitrate Chemical compound [Na+].[O-][N+]([O-])=O VWDWKYIASSYTQR-UHFFFAOYSA-N 0.000 description 2
- 159000000000 sodium salts Chemical class 0.000 description 2
- 239000002904 solvent Substances 0.000 description 2
- 238000005987 sulfurization reaction Methods 0.000 description 2
- DZLFLBLQUQXARW-UHFFFAOYSA-N tetrabutylammonium Chemical compound CCCC[N+](CCCC)(CCCC)CCCC DZLFLBLQUQXARW-UHFFFAOYSA-N 0.000 description 2
- 239000004636 vulcanized rubber Substances 0.000 description 2
- CRDAMVZIKSXKFV-FBXUGWQNSA-N (2-cis,6-cis)-farnesol Chemical compound CC(C)=CCC\C(C)=C/CC\C(C)=C/CO CRDAMVZIKSXKFV-FBXUGWQNSA-N 0.000 description 1
- 239000000260 (2E,6E)-3,7,11-trimethyldodeca-2,6,10-trien-1-ol Substances 0.000 description 1
- 101710116641 (2Z,6E)-farnesyl diphosphate synthase Proteins 0.000 description 1
- 125000006527 (C1-C5) alkyl group Chemical group 0.000 description 1
- OJISWRZIEWCUBN-QIRCYJPOSA-N (E,E,E)-geranylgeraniol Chemical compound CC(C)=CCC\C(C)=C\CC\C(C)=C\CC\C(C)=C\CO OJISWRZIEWCUBN-QIRCYJPOSA-N 0.000 description 1
- MFHNAXHSHOCFEC-UHFFFAOYSA-N 1,1-diphenylguanidine Chemical compound C=1C=CC=CC=1N(C(=N)N)C1=CC=CC=C1 MFHNAXHSHOCFEC-UHFFFAOYSA-N 0.000 description 1
- OWRCNXZUPFZXOS-UHFFFAOYSA-N 1,3-diphenylguanidine Chemical compound C=1C=CC=CC=1NC(=N)NC1=CC=CC=C1 OWRCNXZUPFZXOS-UHFFFAOYSA-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
- HDECRAPHCDXMIJ-UHFFFAOYSA-N 2-methylbenzenesulfonyl chloride Chemical compound CC1=CC=CC=C1S(Cl)(=O)=O HDECRAPHCDXMIJ-UHFFFAOYSA-N 0.000 description 1
- OSWFIVFLDKOXQC-UHFFFAOYSA-N 4-(3-methoxyphenyl)aniline Chemical compound COC1=CC=CC(C=2C=CC(N)=CC=2)=C1 OSWFIVFLDKOXQC-UHFFFAOYSA-N 0.000 description 1
- 239000001729 Ammonium fumarate Substances 0.000 description 1
- 229920000856 Amylose Polymers 0.000 description 1
- 241000219195 Arabidopsis thaliana Species 0.000 description 1
- 241000193830 Bacillus <bacterium> Species 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- VZHHNDCSESIXJW-UHFFFAOYSA-N C(=CC(C)=C)OP(=O)(O)OP(=O)(O)O Chemical compound C(=CC(C)=C)OP(=O)(O)OP(=O)(O)O VZHHNDCSESIXJW-UHFFFAOYSA-N 0.000 description 1
- 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 1
- 241000196324 Embryophyta Species 0.000 description 1
- 240000008168 Ficus benjamina Species 0.000 description 1
- 244000292839 Ficus religiosa Species 0.000 description 1
- 229930091371 Fructose Natural products 0.000 description 1
- 239000005715 Fructose Substances 0.000 description 1
- RFSUNEUAIZKAJO-ARQDHWQXSA-N Fructose Chemical compound OC[C@H]1O[C@](O)(CO)[C@@H](O)[C@@H]1O RFSUNEUAIZKAJO-ARQDHWQXSA-N 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- 239000007818 Grignard reagent Substances 0.000 description 1
- 241001237908 Lactarius volemus Species 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
- 241000187479 Mycobacterium tuberculosis Species 0.000 description 1
- 229910002651 NO3 Inorganic materials 0.000 description 1
- NHNBFGGVMKEFGY-UHFFFAOYSA-N Nitrate Chemical compound [O-][N+]([O-])=O NHNBFGGVMKEFGY-UHFFFAOYSA-N 0.000 description 1
- 239000001888 Peptone Substances 0.000 description 1
- 108010080698 Peptones Proteins 0.000 description 1
- 108010002747 Pfu DNA polymerase Proteins 0.000 description 1
- 235000014680 Saccharomyces cerevisiae Nutrition 0.000 description 1
- 101710165017 Short-chain Z-isoprenyl diphosphate synthase Proteins 0.000 description 1
- 229920002472 Starch Polymers 0.000 description 1
- 229930006000 Sucrose Natural products 0.000 description 1
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 1
- 240000007313 Tilia cordata Species 0.000 description 1
- 238000006959 Williamson synthesis reaction Methods 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
- 239000006061 abrasive grain Substances 0.000 description 1
- 238000001042 affinity chromatography Methods 0.000 description 1
- 239000003513 alkali Substances 0.000 description 1
- 235000019270 ammonium chloride Nutrition 0.000 description 1
- 235000019297 ammonium fumarate Nutrition 0.000 description 1
- 150000003863 ammonium salts Chemical class 0.000 description 1
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 description 1
- 229910052921 ammonium sulfate Inorganic materials 0.000 description 1
- 235000011130 ammonium sulphate Nutrition 0.000 description 1
- 230000000844 anti-bacterial effect Effects 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- 230000003078 antioxidant effect Effects 0.000 description 1
- 150000004945 aromatic hydrocarbons Chemical class 0.000 description 1
- CKKXWJDFFQPBQL-SEPHDYHBSA-N azane;(e)-but-2-enedioic acid Chemical compound N.N.OC(=O)\C=C\C(O)=O CKKXWJDFFQPBQL-SEPHDYHBSA-N 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 1
- GUBGYTABKSRVRQ-QUYVBRFLSA-N beta-maltose Chemical compound OC[C@H]1O[C@H](O[C@H]2[C@H](O)[C@@H](O)[C@H](O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@@H]1O GUBGYTABKSRVRQ-QUYVBRFLSA-N 0.000 description 1
- 229910000019 calcium carbonate Inorganic materials 0.000 description 1
- 229940041514 candida albicans extract Drugs 0.000 description 1
- 239000001569 carbon dioxide Substances 0.000 description 1
- 229910002092 carbon dioxide Inorganic materials 0.000 description 1
- 150000001732 carboxylic acid derivatives Chemical class 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 238000005660 chlorination reaction Methods 0.000 description 1
- 239000004927 clay Substances 0.000 description 1
- 238000010367 cloning Methods 0.000 description 1
- 239000005515 coenzyme Substances 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 235000005822 corn Nutrition 0.000 description 1
- 239000012228 culture supernatant Substances 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- IOJUPLGTWVMSFF-UHFFFAOYSA-N cyclobenzothiazole Natural products C1=CC=C2SC=NC2=C1 IOJUPLGTWVMSFF-UHFFFAOYSA-N 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 108010040223 dehydrodolichyl diphosphate synthetase Proteins 0.000 description 1
- 150000001993 dienes Chemical class 0.000 description 1
- 235000012489 doughnuts Nutrition 0.000 description 1
- 239000000428 dust Substances 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 150000002170 ethers Chemical class 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- 238000001125 extrusion Methods 0.000 description 1
- 229940043259 farnesol Drugs 0.000 description 1
- 229930002886 farnesol Natural products 0.000 description 1
- 239000000706 filtrate Substances 0.000 description 1
- 235000019253 formic acid Nutrition 0.000 description 1
- 239000001530 fumaric acid Substances 0.000 description 1
- 238000001641 gel filtration chromatography Methods 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 235000011187 glycerol Nutrition 0.000 description 1
- 150000004795 grignard reagents Chemical class 0.000 description 1
- 238000000227 grinding Methods 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 239000005556 hormone Substances 0.000 description 1
- 229940088597 hormone Drugs 0.000 description 1
- 238000007731 hot pressing Methods 0.000 description 1
- 125000004435 hydrogen atom Chemical group [H]* 0.000 description 1
- WGCNASOHLSPBMP-UHFFFAOYSA-N hydroxyacetaldehyde Natural products OCC=O WGCNASOHLSPBMP-UHFFFAOYSA-N 0.000 description 1
- 230000008676 import Effects 0.000 description 1
- 239000003112 inhibitor Substances 0.000 description 1
- 229910052500 inorganic mineral Inorganic materials 0.000 description 1
- 230000000968 intestinal effect Effects 0.000 description 1
- INQOMBQAUSQDDS-UHFFFAOYSA-N iodomethane Chemical compound IC INQOMBQAUSQDDS-UHFFFAOYSA-N 0.000 description 1
- 238000004255 ion exchange chromatography Methods 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- 238000011068 loading method Methods 0.000 description 1
- 235000013372 meat Nutrition 0.000 description 1
- 229910052751 metal Inorganic materials 0.000 description 1
- 125000001434 methanylylidene group Chemical group [H]C#[*] 0.000 description 1
- QYQRNKAKVJOLKF-UHFFFAOYSA-N methyl dihydrogen phosphate Chemical compound COP(O)(O)=O.COP(O)(O)=O QYQRNKAKVJOLKF-UHFFFAOYSA-N 0.000 description 1
- GRVDJDISBSALJP-UHFFFAOYSA-N methyloxidanyl Chemical group [O]C GRVDJDISBSALJP-UHFFFAOYSA-N 0.000 description 1
- 235000010755 mineral Nutrition 0.000 description 1
- 239000011707 mineral Substances 0.000 description 1
- 238000005065 mining Methods 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 102000039446 nucleic acids Human genes 0.000 description 1
- 108020004707 nucleic acids Proteins 0.000 description 1
- 150000007523 nucleic acids Chemical class 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 239000003921 oil Substances 0.000 description 1
- 150000007524 organic acids Chemical class 0.000 description 1
- 150000002897 organic nitrogen compounds Chemical class 0.000 description 1
- 239000012188 paraffin wax Substances 0.000 description 1
- 235000019319 peptone Nutrition 0.000 description 1
- VWFJDQUYCIWHTN-ONUHYBHYSA-N phosphono [(2Z)-3,7,11-trimethyldodeca-2,6,10-trienyl] hydrogen phosphate Chemical compound C(\C=C(\C)/CCC=C(C)CCC=C(C)C)OP(=O)(O)OP(=O)(O)O VWFJDQUYCIWHTN-ONUHYBHYSA-N 0.000 description 1
- 229910052698 phosphorus Inorganic materials 0.000 description 1
- FAIAAWCVCHQXDN-UHFFFAOYSA-N phosphorus trichloride Chemical compound ClP(Cl)Cl FAIAAWCVCHQXDN-UHFFFAOYSA-N 0.000 description 1
- 230000026731 phosphorylation Effects 0.000 description 1
- 238000006366 phosphorylation reaction Methods 0.000 description 1
- 238000006116 polymerization reaction Methods 0.000 description 1
- 239000004323 potassium nitrate Substances 0.000 description 1
- 235000010333 potassium nitrate Nutrition 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 235000019260 propionic acid Nutrition 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- IUVKMZGDUIUOCP-BTNSXGMBSA-N quinbolone Chemical compound O([C@H]1CC[C@H]2[C@H]3[C@@H]([C@]4(C=CC(=O)C=C4CC3)C)CC[C@@]21C)C1=CCCC1 IUVKMZGDUIUOCP-BTNSXGMBSA-N 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 238000010079 rubber tapping Methods 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 210000002966 serum Anatomy 0.000 description 1
- 125000005372 silanol group Chemical group 0.000 description 1
- 239000004317 sodium nitrate Substances 0.000 description 1
- 235000010344 sodium nitrate Nutrition 0.000 description 1
- 229940001516 sodium nitrate Drugs 0.000 description 1
- 239000000600 sorbitol Substances 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
- 239000004575 stone Substances 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- 125000000999 tert-butyl group Chemical group [H]C([H])([H])C(*)(C([H])([H])[H])C([H])([H])[H] 0.000 description 1
- 229920001169 thermoplastic Polymers 0.000 description 1
- 239000004416 thermosoftening plastic Substances 0.000 description 1
- 238000007070 tosylation reaction Methods 0.000 description 1
- 229910021654 trace metal Inorganic materials 0.000 description 1
- CRDAMVZIKSXKFV-UHFFFAOYSA-N trans-Farnesol Natural products CC(C)=CCCC(C)=CCCC(C)=CCO CRDAMVZIKSXKFV-UHFFFAOYSA-N 0.000 description 1
- VZCYOOQTPOCHFL-UHFFFAOYSA-N trans-butenedioic acid Natural products OC(=O)C=CC(O)=O VZCYOOQTPOCHFL-UHFFFAOYSA-N 0.000 description 1
- YWYZEGXAUVWDED-UHFFFAOYSA-N triammonium citrate Chemical compound [NH4+].[NH4+].[NH4+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O YWYZEGXAUVWDED-UHFFFAOYSA-N 0.000 description 1
- VTHOKNTVYKTUPI-UHFFFAOYSA-N triethoxy-[3-(3-triethoxysilylpropyltetrasulfanyl)propyl]silane Chemical compound CCO[Si](OCC)(OCC)CCCSSSSCCC[Si](OCC)(OCC)OCC VTHOKNTVYKTUPI-UHFFFAOYSA-N 0.000 description 1
- CYTQBVOFDCPGCX-UHFFFAOYSA-N trimethyl phosphite Chemical compound COP(OC)OC CYTQBVOFDCPGCX-UHFFFAOYSA-N 0.000 description 1
- 238000002525 ultrasonication Methods 0.000 description 1
- 108010028731 undecaprenyl pyrophosphate synthetase Proteins 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
- 229940088594 vitamin Drugs 0.000 description 1
- 229930003231 vitamin Natural products 0.000 description 1
- 230000003313 weakening effect Effects 0.000 description 1
- 239000012138 yeast extract Substances 0.000 description 1
Classifications
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B60—VEHICLES IN GENERAL
- B60C—VEHICLE TYRES; TYRE INFLATION; TYRE CHANGING; CONNECTING VALVES TO INFLATABLE ELASTIC BODIES IN GENERAL; DEVICES OR ARRANGEMENTS RELATED TO TYRES
- B60C1/00—Tyres characterised by the chemical composition or the physical arrangement or mixture of the composition
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
- C07F9/00—Compounds containing elements of Groups 5 or 15 of the Periodic Table
- C07F9/02—Phosphorus compounds
- C07F9/06—Phosphorus compounds without P—C bonds
- C07F9/08—Esters of oxyacids of phosphorus
- C07F9/09—Esters of phosphoric acids
- C07F9/113—Esters of phosphoric acids with unsaturated acyclic alcohols
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08F—MACROMOLECULAR COMPOUNDS OBTAINED BY REACTIONS ONLY INVOLVING CARBON-TO-CARBON UNSATURATED BONDS
- C08F36/00—Homopolymers and copolymers of compounds having one or more unsaturated aliphatic radicals, at least one having two or more carbon-to-carbon double bonds
- C08F36/02—Homopolymers and copolymers of compounds having one or more unsaturated aliphatic radicals, at least one having two or more carbon-to-carbon double bonds the radical having only two carbon-to-carbon double bonds
- C08F36/04—Homopolymers and copolymers of compounds having one or more unsaturated aliphatic radicals, at least one having two or more carbon-to-carbon double bonds the radical having only two carbon-to-carbon double bonds conjugated
- C08F36/08—Isoprene
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08G—MACROMOLECULAR COMPOUNDS OBTAINED OTHERWISE THAN BY REACTIONS ONLY INVOLVING UNSATURATED CARBON-TO-CARBON BONDS
- C08G61/00—Macromolecular compounds obtained by reactions forming a carbon-to-carbon link in the main chain of the macromolecule
- C08G61/02—Macromolecular compounds containing only carbon atoms in the main chain of the macromolecule, e.g. polyxylylenes
- C08G61/04—Macromolecular compounds containing only carbon atoms in the main chain of the macromolecule, e.g. polyxylylenes only aliphatic carbon atoms
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08L—COMPOSITIONS OF MACROMOLECULAR COMPOUNDS
- C08L15/00—Compositions of rubber derivatives
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08L—COMPOSITIONS OF MACROMOLECULAR COMPOUNDS
- C08L9/00—Compositions of homopolymers or copolymers of conjugated diene hydrocarbons
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08G—MACROMOLECULAR COMPOUNDS OBTAINED OTHERWISE THAN BY REACTIONS ONLY INVOLVING UNSATURATED CARBON-TO-CARBON BONDS
- C08G2261/00—Macromolecular compounds obtained by reactions forming a carbon-to-carbon link in the main chain of the macromolecule
- C08G2261/30—Monomer units or repeat units incorporating structural elements in the main chain
- C08G2261/33—Monomer units or repeat units incorporating structural elements in the main chain incorporating non-aromatic structural elements in the main chain
- C08G2261/332—Monomer units or repeat units incorporating structural elements in the main chain incorporating non-aromatic structural elements in the main chain containing only carbon atoms
- C08G2261/3327—Monomer units or repeat units incorporating structural elements in the main chain incorporating non-aromatic structural elements in the main chain containing only carbon atoms alkene-based
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Polymers & Plastics (AREA)
- Medicinal Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Mechanical Engineering (AREA)
- Engineering & Computer Science (AREA)
- Molecular Biology (AREA)
- General Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Compositions Of Macromolecular Compounds (AREA)
- Addition Polymer Or Copolymer, Post-Treatments, Or Chemical Modifications (AREA)
- Tires In General (AREA)
- Polyoxymethylene Polymers And Polymers With Carbon-To-Carbon Bonds (AREA)
- Enzymes And Modification Thereof (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The purpose of the present invention is to provide an isoprene oligomer and polyisoprene in which a modification has been added to the main-chain moiety of the molecule. Also provided are a rubber composition obtained by blending the isoprene oligomer and/or polyisoprene, and a pneumatic tire in which the rubber composition is used for the members of the tire (for example, the tread or sidewalls). The present invention pertains to an isoprene oligomer obtained by being synthesized from an allylic diphosphoric acid represented by formula (X), and a compound represented by formula (Y). (In formula (X), n represents an integer 1 to 10. In formula (Y), R represents a group other than a methyl group.)
Description
Technical field
The pneumatic tyre that the present invention relates to the method for isoprene oligopolymer, polyisoprene, manufacture isoprene oligopolymer and polyisoprene, composition containing isoprene oligopolymer and/or polyisoprene rubber and formed by this rubber compound.
Background technology
For a long time, people are devoted to be provided in the rubber product also outside character that rubber has originally with new property, according to purposes, import difform filler or other components that differing materials makes, the character required for acquisition in rubber combination.Such as, in doughnut, people attempt by importing the filler such as silicon-dioxide or carbon black in organic rubber phase, improve the character such as wear resistance, low-heat-generation or wet earth-catching property.
Be conventionally used in the method for mixed fillers in the rubber phase such as rubber combination, modified rubber (modified dienopolymer) etc., increase the affinity between these components, improve low-heat-generation, wet earth-catching property etc. further.These modified rubbers react with the compound such as containing nitrogen-containing group, chlorine sulfenyl by making the rubber molecule in rubber phase, and in rubber molecule, introduce the functional group (such as, referenced patent document 1 and 2) with filler with affinity.
Meanwhile, known isoprene oligopolymer and polyisoprene have the bisphosphate base etc. of high reaction activity at its chain end.Therefore, traditional isoprene oligopolymer or the method for modifying of polyisoprene relate to the bisphosphate base of above-mentioned high reaction activity end and the reaction of the compound containing functional group.As mentioned above, isoprene oligopolymer and polyisoprene mainly carry out modification at the end of molecule traditionally.On the other hand, in conventional methods where, by the bisphosphate base of high reaction activity end or the impact of its analogue, the skeleton of isoprene oligopolymer and polyisoprene can not by modification fully.In addition, traditionally, the modification of natural rubber has only been come by carrying out chemical treatment to the latex collected from Para rubber tree (Hevea brasiliensis).What therefore, do not undertaken being confirmed whether that molecular chain is actual by analysis be modified and replace the functional group having type.
Citing document list
Patent documentation
Patent documentation 1:JP 2000-001573A
Patent documentation 2:JP 2000-001575A
Summary of the invention
Technical problem
The object of the invention is to solve the problem, and a kind of isoprene oligopolymer and polyisoprene of molecular skeleton modification are provided.The present invention also aims to provide a kind of rubber combination comprising this isoprene oligopolymer and/or this polyisoprene, and provide one to comprise the pneumatic tyre being formed tyre element (such as, tyre surface or sidewall) by this rubber combination.
The method of dealing with problems
The present invention relates to a kind of isoprene oligopolymer, its allyl group bisphosphate represented by formula (X) and the compou nd synthesis represented by formula (Y):
Wherein, n represents the integer of 1 to 10;
Wherein, R represents the group beyond methyl.
The compound that the allyl group bisphosphate of preferred described isoprene oligopolymer represented by described formula (X), described formula (Y) represent and prenyl diphosphate synthesis.
The allyl group bisphosphate of allyl group bisphosphate represented by formula (X-1) preferably described in this represented by formula (X):
Wherein, n represents the integer of 1 to 10, and at least one atom contained in the II part in formula (X-1) or atomic group are replaced by other (namely different) atoms or atomic group, and atom contained in the III part in formula (X-1) or atomic group are not all replaced by other atoms or atomic group.
Described synthesis is carried out preferably by using the enzyme with prenyltransferases activity.
The invention still further relates to the represented or isoprene oligopolymer represented by formula (Z-2) of a kind of formula (Z-1), in the v part in formula (Z-1) or formula (Z-2), at least one contained atom or atomic group are replaced by other atoms or atomic group.
Wherein, the n in formula (Z-1) represents the integer of 1 to 10; M represents the integer of 1 to 30; Y represents hydroxyl, formyl radical, carboxyl, alkoxycarbonyl, alkoxy carbonyl or OPP;
Wherein, the n in formula (Z-2) represents the integer of 1 to 10; M represents the integer of 1 to 30; Y represents hydroxyl, formyl radical, carboxyl, alkoxycarbonyl, alkoxy carbonyl or OPP.
At least one atom contained in iv part in preferred formula (Z-1) or formula (Z-2) or atomic group are replaced by other atoms or atomic group.
The invention still further relates to a kind of method manufacturing isoprene oligopolymer, it comprises the allyl group bisphosphate represented by formula (X) and the compou nd synthesis isoprene oligopolymer represented by formula (Y):
Wherein, n represents the integer of 1 to 10;
Wherein, R represents the group beyond methyl.
In the method manufacturing isoprene oligopolymer, the compound that the allyl group bisphosphate of preferred described isoprene oligopolymer represented by described formula (X), described formula (Y) represent and prenyl diphosphate synthesis.
The allyl group bisphosphate of allyl group bisphosphate represented by formula (X-1) preferably described in this represented by formula (X):
Wherein, n represents the integer of 1 to 10, and at least one atom contained in the II part in formula (X-1) or atomic group are replaced by other atoms or atomic group, atom contained in the III part in formula (X-1) or atomic group are not all replaced by other atoms or atomic group.
Described synthesis is carried out preferably by using the enzyme with prenyltransferases activity.
The invention still further relates to a kind of polyisoprene, its allyl group bisphosphate represented by formula (X) and the compou nd synthesis represented by formula (Y):
Wherein, n represents the integer of 1 to 10;
Wherein, R represents the group beyond methyl.
The compound that the allyl group bisphosphate of preferred described polyisoprene represented by described formula (X), described formula (Y) represent and prenyl diphosphate synthesis.
The allyl group bisphosphate of allyl group bisphosphate represented by formula (X-1) preferably described in this represented by formula (X):
Wherein, n represents the integer of 1 to 10, and at least one atom contained in the II part in formula (X-1) or atomic group are replaced by other atoms or atomic group, atom contained in the III part in formula (X-1) or atomic group are not all replaced by other atoms or atomic group.
The invention still further relates to a kind of polyisoprene, its compound represented by described isoprene oligopolymer, at least one formula (Y) and prenyl diphosphate synthesis.
The invention still further relates to the polyisoprene represented by a kind of formula (ZZ-1) or represented by formula (ZZ-2), at least one atom contained in v part in formula (ZZ-1) or formula (ZZ-2) or atomic group are replaced by other atoms or atomic group
Wherein, the n in formula (ZZ-1) represents the integer of 1 to 10; Q represents the integer of 30 to 40000; Y represents hydroxyl, formyl radical, carboxyl, alkoxycarbonyl, alkoxy carbonyl or OPP;
Wherein, the n in formula (ZZ-2) represents the integer of 1 to 10; Q represents the integer of 30 to 40000; Y represents hydroxyl, formyl radical, carboxyl, alkoxycarbonyl, alkoxy carbonyl or OPP.
At least one atom contained in iv part in preferred formula (ZZ-1) or formula (ZZ-2) or atomic group are replaced by other atoms or atomic group.
The invention still further relates to a kind of method manufacturing polyisoprene, it comprises compound represented by described isoprene oligopolymer, at least one formula (Y) and prenyl diphosphate synthesizes this polyisoprene.
The invention still further relates to a kind of rubber combination, it comprises at least one in described isoprene oligopolymer and described polyisoprene.
The present invention relates to a kind of pneumatic tyre formed by described rubber combination.
Invention Beneficial Effect
Isoprene oligopolymer of the present invention is the isoprene oligopolymer of the allyl group bisphosphate represented by formula (X) and the compou nd synthesis represented by formula (Y), or it is the isoprene oligopolymer represented by formula (Z-1) or formula (Z-2), wherein, at least one atom contained in the v part in formula (Z-1) or formula (Z-2) or atomic group are replaced by other atoms or atomic group.In addition, polyisoprene of the present invention is the polyisoprene by the allyl group bisphosphate represented by formula (X) and the compou nd synthesis represented by formula (Y); Or the polyisoprene of compound represented by above-mentioned isoprene oligopolymer, a kind of formula (Y) and/or prenyl diphosphate synthesis; Or represented by formula (ZZ-1) or represented by formula (ZZ-2) polyisoprene, in the v part in formula (ZZ-1) or formula (ZZ-2), at least one contained atom or atomic group are replaced by other atoms or atomic group.Therefore, the molecule (rubber molecule) of isoprene oligopolymer of the present invention and polyisoprene of the present invention all has the skeleton of modification, and itself and filler (as silicon-dioxide) have outstanding affinity thus.Therefore, if isoprene oligopolymer of the present invention and/or polyisoprene of the present invention are used for rubber combination, the rubber molecule obtained in rubber combination can mix with packed height.Such as, therefore provided rubber combination has outstanding low-heat-generation and wear resistance.Such as, further, if in the tyre element that above-mentioned rubber combination is used for (such as, tyre surface or sidewall), can provide and there is outstanding low-heat-generation and the pneumatic tyre of wear resistance.
In addition, the method for manufacture isoprene oligopolymer of the present invention and the method for manufacture polyisoprene of the present invention, can provide isoprene oligopolymer and the polyisoprene of the abundant modification of the impregnable skeleton of high reaction activity end bisphosphate base respectively.
Further, because isoprene oligopolymer provided by the present invention and polyisoprene use the compound represented by formula (Y) to obtain, add in compound represented by this formula (Y) have functional group or other there is the group of known structure, this functional group (modified group) in isoprene oligopolymer or polyisoprene can be identified clearly.
Embodiment
In the synthetic (biosynthesizing) of rubber molecule (polyisoprene), enzyme can be used (such as, prenyltransferases) act on starting material (such as, farnesyl bisphosphate (FPP)) and monomer is (such as, prenyl diphosphate) mixture in, produce the isoprene oligopolymer that addition polymerization on starting material has 1 to 30 isoprene unit.Then, by these isoprene oligopolymer further with containing can the latex component of enzyme of addition polymerization prenyl diphosphate mix.Knownly consequently be formed in polyisoprene oligopolymer being connected with many polyisoprene units.
As described above, the step linking monomer on starting material continuously forming rubber molecule must use natural enzymatic addition polymerization.
This just means that the starting material of the synthesis of rubber molecule (polyisoprene) and monomer needs are the materials that can carry out catalyzed reaction with enzyme.Therefore, the structure of the starting material and monomer that can be used as rubber molecule (polyisoprene) material is restricted.Especially, because the restriction caused by enzyme of prenyltransferases and energy addition polymerization prenyl diphosphate, monomer is restricted to naturally occurring prenyl diphosphate.
Its result causes the structure design limited flexibility system of the rubber molecule (polyisoprene) of synthetic (biosynthesizing), therefore make to be difficult to provide the handiness in enough molecular designing, carry out the performance not available for additional natural rubber.
For these reasons, in order to prepare the rubber molecule (polyisoprene) being imported with functional group or its analogue, such as, traditionally, as described above, people are by such as reacting the end bisphosphate group of high reaction activity or its analogue, to cause modification with the compound containing functional group.Especially, the modification of natural rubber has only been come by carrying out chemical treatment to the latex collected from Para rubber tree (Hevea brasiliensis).What therefore, do not undertaken being confirmed whether that molecular chain is actual by analysis be modified and replace the functional group having type.
Otherwise, the present invention is based on following discovery: by using the prenyl diphosphate of partially modified structure as monomer, manufacturing isoprene oligopolymer or polyisoprene, the isoprene oligopolymer or polyisoprene with functionalization skeleton can be manufactured.Further, because isoprene oligopolymer provided by the present invention and polyisoprene use prenyl diphosphate to obtain, add in this prenyl diphosphate have functional group or other there is the group of known structure, this functional group (modified group) in isoprene oligopolymer or polyisoprene can be identified clearly.
Especially, the present invention is based on following discovery: if the structure beyond the methyl maintaining the 3-position in the structure of naturally occurring monomer prenyl diphosphate is constant, even by required group replace be positioned at 3-position methyl time, also this monomer can be used to generate enzyme by the polymkeric substance of the prenyltransferases or naturally occurring energy addition polymerization prenyl diphosphate that generate enzyme as naturally occurring oligopolymer, manufacture isoprene oligopolymer or polyisoprene.Its reason is not all verified, but can infer due to prenyltransferases and can the enzyme of addition polymerization prenyl diphosphate be attracted to the 3-position of prenyl diphosphate monomer methyl outside structure on, these enzymes are to the structure relative insensitivity of 3-position methyl simultaneously.
Find based on these, the isoprene oligopolymer that skeleton can be provided to have required character and polyisoprene, therefore can provide the isoprene oligopolymer and polyisoprene with additional functions.
More particularly, in naturally occurring biosynthesizing, such as, prenyl diphosphate (IPP) can be aggregated on dimethylallyl diphosphate (DMAPP) continuously, produce geranyl bisphosphate (GPP), farnesyl bisphosphate (FPP), geranyl geranyl diphosphate (GGPP), geranyl farnesyl bisphosphate (GFPP) thus, as follows.
Otherwise the present invention uses and replaces the methyl of the 3-position of IPP and the R-IPP that obtains, to replace prenyl diphosphate (IPP) with required group (-R).In this situation, can continuously R-IPP be aggregated on dimethylallyl diphosphate, produce the geranyl bisphosphate (R-GPP) of framework modification, the farnesyl bisphosphate (R-FPP) of framework modification, the geranyl geranyl diphosphate (R-GGPP) of framework modification, the geranyl farnesyl bisphosphate (R-GFPP) of framework modification thus, as follows.
Further, if R-IPP and IPP is used simultaneously, in other words, and with R-IPP and IPP, R-IPP and IPP is aggregated on dimethylallyl diphosphate continuously, the geranyl bisphosphate (R-GPP) of framework modification as follows, the farnesyl bisphosphate (R-FPP) of framework modification, the geranyl geranyl diphosphate (R-GGPP) of framework modification, the geranyl farnesyl bisphosphate (R-GFPP) of framework modification can be produced.Therefore, the isoprene oligopolymer with required character can be manufactured with R-IPP and IPP with the ratio after regulating.
Herein, the modification of the skeleton of molecule (rubber molecule) mean with required functional group replace in molecule (rubber molecule) skeleton from the structure in IPP (especially, the part corresponding with the 3-position methyl of IPP), or mean with other structures (structure beyond methyl) replace in molecule (rubber molecule) skeleton from the structure (especially, corresponding with the 3-position methyl of IPP part) in IPP.
(isoprene oligopolymer)
The allyl group bisphosphate of a kind of oligopolymer of the present invention represented by formula (X) and the synthesis of the compound (R-IPP) represented by formula (Y) (biosynthesizing):
Wherein, n represents the integer of 1 to 10;
Wherein, R represents the group beyond methyl.
Herein, term " OPP (OPP group) " means bisphosphate group (group represented by following formula (A-1)), and its have 3 with the hydroxyl of phosphorus atom bonding.When its in aqueous time, some or all of hydroxyl dissociates (such as, OPP becomes as shown in the formula the group represented by (A-2)).The intension of term " OPP " herein comprises the group that above-mentioned part or all of hydroxyl dissociates.
Isoprene oligopolymer of the present invention has the akin structure with natural rubber, and with rubber molecule highly compatible.In addition, because isoprene oligopolymer of the present invention has modified skeletal, can there is strong interaction with filler (such as, silicon-dioxide) in it.Therefore, due to isoprene oligopolymer of the present invention and rubber highly compatible, simultaneously can with filler (such as, silicon-dioxide) there is strong interaction, if this isoprene oligopolymer is used for rubber combination, can obtain and compare traditional composition, the rubber combination that its rubber molecule mixes with filler higher degree.Such as, this rubber combination can strengthen low-heat-generation, wet earth-catching property, wear resistance, elongation at break and fracture tensile strength thus.
The skeleton of isoprene oligopolymer of the present invention comprises polar group or its analogue.Thus, the situation when dispersing property of filler (such as, silicon-dioxide) only appears at chain end higher than polar group or its analogue.Therefore, such as, the improvement effect of low-heat-generation, wet earth-catching property, wear resistance, elongation at break and fracture tensile strength increases.
Isoprene oligopolymer of the present invention also has outstanding antimicrobial acivity.Infer this is because isoprene oligopolymer of the present invention has the structure different from naturally occurring usual isoprene oligopolymer, therefore, there is the effects such as the enzyme of such as anti-bacteria and coenzyme, the synthesis of suppression nucleic acid, the synthesis of T suppression cell film, the synthesis of T suppression cell plasma membrane, infringement cytolemma and infringement cytoplasmic membrane.
First, the allyl group bisphosphate represented by formula (X) is described:
Wherein, n represents the integer of 1 to 10.
In formula (X), n represents the integer (be preferably 1 to 4, be more preferably 1 to 3) of 1 to 10.
The example of the allyl group bisphosphate represented by formula (X) comprises: dimethylallyl diphosphate (DMAPP), geranyl bisphosphate (GPP), farnesyl bisphosphate (FPP), geranyl geranyl diphosphate (GGPP), geranyl farnesyl bisphosphate (GFPP).Wherein, preferred DMAPP, GPP and FPP, because they can as the substrate of polytype prenyltransferases.
In the allyl group bisphosphate represented by formula (X), bisphosphate group and isoprene unit bonding.Allyl group bisphosphate represented by formula (X) can be the allyl group bisphosphate derivative by changing (modification) to part isoprene unit.
The present inventor proposed patent application (JP2012-036360A) with regard to using allyl group bisphosphate derivative as the method for starting material manufacture isoprene oligopolymer or polyisoprene.This patent application is summarized as follows.
As mentioned above, the starting material of the synthesis of rubber molecule (polyisoprene) and monomer need to be the material that can carry out catalyzed reaction with enzyme.Which has limited the starting material of material and the structure of monomer that can be used as rubber molecule (polyisoprene).Especially, because the restriction that the enzyme manufacturing oligopolymer brings, starting material is restricted to naturally occurring dimethylallyl diphosphate, geranyl bisphosphate, farnesyl bisphosphate, geranyl geranyl diphosphate and analogue thereof.
On the contrary, the present inventor is through extensively studying discovery, by use using such as structure by partially modified farnesyl bisphosphate etc. as starting material, manufacture isoprene oligopolymer or polyisoprene, isoprene oligopolymer or the polyisoprene of the chain end with functionalization can be manufactured.
Especially, the present inventor finds, if maintain the structure of the I part in the following formula (I) in the starting material such as naturally occurring farnesyl bisphosphate, even if the structure required for the introducing of other parts beyond I part, naturally occurring oligopolymer also can be used as to generate the prenyltransferases of enzyme to manufacture isoprene oligopolymer.Its reason is not all verified, but can infer that these enzymes are to the structure relative insensitivity of other parts simultaneously because prenyltransferases is attracted in the structure of the I part in the formula (I) of starting material.
Find based on these, isoprene oligopolymer and the polyisoprene of the chain end with required character can be provided, therefore, while can be provided in the script character not weakening isoprene oligopolymer or polyisoprene, isoprene oligopolymer and the polyisoprene of several functions is attached with.
Therefore, in the present invention, by the prenyltransferases using naturally occurring oligopolymer to generate enzyme, by the compound polymerization of formula (Y) (R-IPP) on starting material, this starting material is the allyl group bisphosphate of the formula (X) of carrying out modification while the I part-structure maintaining formula (I), can obtain and be not only the isoprene oligopolymer that skeleton and chain end (structure division from allyl group bisphosphate) have also carried out modification.
Especially, preferred this allyl group bisphosphate allyl group bisphosphate represented by formula (X-1) represented by above-mentioned formula (X):
Wherein, n represents the integer (n in formula (X-1) defines in the mode same with formula (X)) of 1 to 10, and at least one atom contained in the II part in formula (X-1) or atomic group are replaced by other atoms or atomic group, atom contained in the III part in formula (X-1) or atomic group are not all replaced by other atoms or atomic group.
Consequently manufacture a kind of isoprene oligopolymer that not only there is modified skeletal, also there is modification chain end.Because this isoprene oligopolymer not only has modified skeletal, also has modification chain end, its can with filler (such as, silicon-dioxide) interact better, if this isoprene oligopolymer is used for rubber combination, rubber molecule in the rubber combination obtained can compare traditional composition, and itself and filler can carry out higher degree and mix.Such as, rubber combination can provide the low-heat-generation strengthened further, wet earth-catching property, wear resistance, elongation at break and fracture tensile strength thus.
Herein, the modification of the end of described molecule (rubber molecule) means, the predetermined part from the allyl group bisphosphate structure represented by formula (X) of molecule (rubber molecule) end is replaced by required functional group, or means that the predetermined part from the allyl group bisphosphate structure represented by formula (X) of molecule (rubber molecule) end is replaced by other structures.
In II part in formula (X-1) or III part, the example of contained atom or atomic group (that is, the atom before replacement or atomic group) comprises hydrogen atom, methyl, methylene radical, carbon atom and methyne.
The example of other atoms comprises nitrogen-atoms, Sauerstoffatom, sulphur atom, Siliciumatom and carbon atom.Wherein, preferred nitrogen atom, because it has strong Intermolecular Forces, therefore can have strong interaction with enzyme or cytolemma, also have strong interaction with filler (such as, silicon-dioxide).
Other atomic groups described can be nitrogen-containing group, oxy radical, sulfur-containing group, silicon-containing group, carbon-containing group or its analogue.Its example comprises: acetoxyl group, alkoxyl group (preferred C1-C3 alkoxyl group; more preferably methoxyl group), hydroxyl, aryl (preferred phenyl), alkyl (preferred C1-C5 alkyl; more preferably ethyl and the tertiary butyl), ethanoyl, N-alkyl-kharophen (wherein, alkyl preferably has 1 to 5 carbon atom) and azido-.
From the angle of anti-microbe ability, preferred N-alkyl-kharophen (more preferably N-methyl-acetamido and N-butyl-kharophen) and azido-, because nitrogen-atoms has strong Intermolecular Forces, therefore strong interaction can be there is with enzyme or cytolemma.Because the strong interaction of itself and filler (such as, silicon-dioxide), also preferably alkoxyl group and hydroxyl.
The example of the allyl group bisphosphate (allyl group bisphosphate derivative) of part isoprene unit modification comprises the compound represented by following formula (A) to (S).Wherein, the compound of preferred formula (B), (C), (D), (E), (F), (K), (L) and (R), more preferably the compound of formula (B) and (C), because they have stronger with filler (such as, silicon-dioxide) interaction, and more effectively can improve low-heat-generation, wet earth-catching property, wear resistance, elongation at break and fracture tensile strength.Based on outstanding anti-microbe ability, the also structure of preferred this formula (G) to (Q), the more preferably structure of formula (K), (L) and (Q).
Technician can according to the method disclosed in JP2012-036360A, such as, from dimethylallyl diphosphate, geranyl bisphosphate, farnesyl bisphosphate, geranyl geranyl diphosphate, Geraniol, farnesol, trans-Geranylgeraniol or its analogue, the allyl group bisphosphate derivative represented by preparation formula (A) to (S).
Then, the compound (R-IPP) represented by formula (Y) is described:
Wherein, R represents the group beyond methyl.
In formula (Y), R can be any group beyond methyl.Its example comprises nitrogen-containing group, oxy radical, sulfur-containing group, silicon-containing group and carbon-containing group (except methyl).Preference comprises acetoxyl group, alkoxyl group, hydroxyl, aryl, alkyl (except methyl), ethanoyl, N-ethanoyl-kharophen, azido-, amino and sulfydryl.Wherein, preferred nitrogen-containing group, oxy radical, sulfur-containing group, silicon-containing group and carbon-containing group (except methyl), more preferably alkyl (especially, the alkyl of C2-C6), aryl (especially, phenyl), sulfydryl, hydroxyl and amino, preferred sulfydryl and hydroxyl further, its reason is that they more effectively can improve low-heat-generation, wet earth-catching property, elongation at break and fracture tensile strength.
Technician can according to method illustrated in such as embodiment, the compound represented by preparation formula (Y).
The quantity preferably 1 to 30 of addition isoprene unit on the allyl group bisphosphate starting material of formula (X) in isoprene oligopolymer of the present invention, more preferably 1 to 20, further preferably 1 to 10.
Such as, the concrete example of isoprene oligopolymer of the present invention is formula (Z-1) or the isoprene oligopolymer represented by formula (Z-2), and at least one atom contained in the v part in formula (Z-1) or formula (Z-2) or atomic group are replaced by other atoms or atomic group.
Wherein, the n in formula (Z-1) represents the integer of 1 to 10; M represents the integer of 1 to 30; Y represents hydroxyl, formyl radical, carboxyl, alkoxycarbonyl, alkoxy carbonyl or OPP;
Wherein, the n in formula (Z-2) represents the integer of 1 to 10; M represents the integer of 1 to 30; Y represents hydroxyl, formyl radical, carboxyl, alkoxycarbonyl, alkoxy carbonyl or OPP.
Atom contained in v part in formula (Z-1) or formula (Z-2) or atomic group and other atoms described or atomic group can be such as formula the atom described in (X-1) or atomic group.
Iv part in formula (Z-1) and (Z-2) is the structure from the allyl group bisphosphate starting material represented by formula (X).Therefore, formula (Z-1) defines in the mode identical with the n in formula (X) with the n in (Z-2).
V part in formula (Z-1) and (Z-2) is made up of the isoprene unit of the allyl group bisphosphate starting material added to represented by formula (X).Therefore, formula (Z-1) is corresponding with the quantity of the isoprene unit of the allyl group bisphosphate starting material of the above-mentioned formula that adds to (X) with the m in (Z-2).
Due to outstanding antimicrobial properties and the strong interaction with filler (such as, silicon-dioxide), the preferred OPP of Y, hydroxyl or carboxyl in formula (Z-1) and (Z-2).
As mentioned above, the present invention is based on following discovery: if the structure beyond the methyl of the 3-position in maintenance prenyl diphosphate structure is constant, even if the methyl of 3-position is replaced by required group, it also can be used to pass through naturally occurring enzyme, manufacture isoprene oligopolymer or polyisoprene.Therefore, formula (Z-1) or the isoprene oligopolymer represented by formula (Z-2) are preferably represented by formula (Z-1-1) or such as formula the isoprene oligopolymer represented by (Z-2-1):
Wherein, n, m in formula (Z-1-1) define in the mode identical with Y with n, the m in formula (Z-1) with Y, and R is to define with the same way of the R in formula (Y);
Wherein, n, m in formula (Z-2-1) define in the mode identical with Y with n, the m in formula (Z-2) with Y, and R is to define with the same way of the R in formula (Y);
As mentioned above, the also preferred material that is simultaneously modified of skeleton and chain end.Therefore, at least one atom contained by iv part in preferred formula (Z-1) or formula (Z-2) (preferred formula (Z-1-1) or formula (Z-2-1)) or atomic group are replaced by other atoms or atomic group.
Atom contained in iv part in formula (Z-1), formula (Z-2), formula (Z-1-1) or formula (Z-2-1) or atomic group and other atoms described or atomic group can such as formula described in (X-1).
Further, as mentioned above, allyl group bisphosphate represented by preferred formula (X) is the allyl group bisphosphate represented by formula (X-1), and at least one atom contained in the II part in formula (X-1) or atomic group are replaced by other atoms or atomic group, atom contained in the III part in formula (X-1) or atomic group are not all replaced by other atoms or atomic group.Therefore, about as shown in the formula the iv part in the formula (Z-1) represented by (Z), formula (Z-2), formula (Z-1-1) and formula (Z-2-1), at least one atom contained in iv-II part in preferred formula (Z) or atomic group are replaced by other atoms or atomic group, and atom contained in the iv-III part in formula (Z) or atomic group are not all replaced by other atoms or atomic group.
In iv-II part in formula (Z) or iv-III part, contained atom or atomic group and other atoms described or atomic group can for such as formula the atom described in (X-1) or atomic groups.
The concrete example of the iv part that chain end is also modified can comprise as shown in the formula the structure represented by (a) to (s).Wherein, the structure of preferred formula (b), (c), (d), (e), (f), (k), (l) and (r), more preferably the structure of formula (b) and (c), because they have stronger with filler (such as, silicon-dioxide) interaction, and more effectively can improve low-heat-generation, wet earth-catching property, wear resistance, elongation at break and fracture tensile strength.Due to outstanding anti-microbe ability, the also structure of preferred formula (g) to (q), the more preferably structure of formula (k), (l) and (q).
(manufacturing the method for isoprene oligopolymer)
The allyl group bisphosphate of isoprene oligopolymer of the present invention represented by formula (X) and the compound represented by formula (Y) carry out synthesizing (biosynthesizing).
As an example of the method for the isoprene oligopolymer of the present invention of the allyl group bisphosphate represented by formula (X) and the compou nd synthesis (biosynthesizing) represented by formula (Y), comprise the enzyme using and there is prenyltransferases activity.Especially, under the existence of enzyme with prenyltransferases activity, the compound represented by (Y) of the allyl group bisphosphate represented by formula (X) and the monomer that can be polymerized in allyl group bisphosphate formula reacts.Allyl group bisphosphate represented by formula (X) and the compound represented by formula (Y) can be independent compound or the composition of multiple compounds respectively.Such as, the composition of the multiple compounds of the formula (Y) that the R in formula (Y) can be used different, to provide the oligopolymer of the isoprene with required character.
In the present invention, the monomer be polymerized on the allyl group bisphosphate represented by formula (X) can be only the compound represented by formula (Y) (R-IPP).In this situation, can the isoprene oligopolymer shown in fabrication scheme 2.Or the monomer be polymerized on the allyl group bisphosphate represented by formula (X) can be the combination of compound represented by formula (Y) (R-IPP) and prenyl diphosphate (IPP).In this situation, can the isoprene oligopolymer shown in fabrication scheme 3.
Herein, term " has prenyltransferases activity " and means that the condensation reaction with catalysis allyl group substrate (allyl group bisphosphate) and prenyl diphosphate has the enzyme of the activity of the new allyl group bisphosphate of the isoprene unit of addition with synthesis, therefore, can catalysis reaction that prenyl diphosphate is linked with allyl group substrate (allyl group bisphosphate) continuously.
As mentioned above, when the methyl replacing the 3-position in prenyl diphosphate structure with required group and to maintain the structure except the methyl of 3-position except of prenyl diphosphate constant and obtain the compound of formula (Y) (R-IPP), can by using this compound and adopting the naturally occurring enzyme manufacture isoprene oligopolymer such as the enzyme as having prenyltransferases activity.
As the enzyme with prenyltransferases activity, have been found that polytype enzyme.
The enzyme (isoprene unit of each new addition has cis-configuration) that can extend Z-isoprene chain comprises: Z-nine prenyl diphosphate synthase (Ishii, K.et al., (1986) Biochem., J., 233,773.), 11 isopentenyl diphosphate synthetic enzyme (Takahashi, I. and Ogura, K., (1982) J.Biochem., 92,1527., Keenman, and Allen M.V., C.M., (1974) Arch.Biochem.Biophys., 161, 375.), Z-farnesyl diphosphate synthase (Identification of a short (C-15) chain Z-isoprenyl diphosphate synthase and a homologous long (C-50) chain isoprenyl diphosphate synthase in Mycobacterium tuberculosis (identification of long-chain (C-50) the isoprene diphosphate synthase of short chain (C-15) the farnesyl diphosphate synthase in tubercule bacillus and homology), Schulbach, MC., et al.JOURNAL OF BIOLOGICAL CHEMISTRY, 275 (30), 22876-22881 (2000)), and dehydrogenation dolichol base diphosphate synthase ((identification of Identification of human dehydrodolichyl diphosphate synthase gene mankind dehydrogenation dolichol base diphosphate synthase gene), Endo, Shota.et al, Biochimica et Biophysica Acta (BBA), 1625 (3), (2003) p.291-295.).
In addition, the enzyme (isoprene unit of each new addition has transconfiguration) that can extend E-isoprene chain comprises: farnesyl diphosphate synthase, geranyl geranyl diphosphate synthetic enzyme, six isoprene diphosphate synthases, seven isoprene diphosphate synthases, eight isoprene diphosphate synthases, ten isoprene diphosphate synthases.
Concrete enzyme is depended in the maximum number of the isoprene unit that can be formed and the direction (transconfiguration or cis-configuration) of isoprene chain extension.Therefore, enzyme can be selected according to the target numbers of isoprene unit and isoprene chain extension direction.In the present invention, the isoprene chain of isoprene oligopolymer above in any direction can extend (transconfiguration or cis-configuration).In other words, such as, isoprene oligopolymer of the present invention can be all isoprene units all with transconfiguration link isoprene oligopolymer (such as, the isoprene oligopolymer of formula (Z-1)), the isoprene oligopolymer that links with Trans-cis-transconfiguration of the isoprene oligopolymer (such as, the isoprene oligopolymer of formula (Z-2)) that links with trans-cis configuration of isoprene unit or isoprene unit.
Often kind of biology of tellurian existence all possesses the enzyme with prenyltransferases activity.The biology possessed with the enzyme of prenyltransferases activity can be enumerated such as, micrococcus luteus B-P26 (Micrococcus luteus B-P26), intestinal bacteria (Escherichia coli), yeast saccharomyces cerevisiae (Saccharomyces cerevisiae), Arabidopis thaliana (Arabidopsis thaliana), Para rubber tree (Hevea brasiliensis), periploca sepium (Periploca sepium), bacstearothermophilus (Bachillus Stearothermophilus), sulfolobus acidocaldarius (Sulfolobus acidocaldarius (ATCC49426)), homo sapiens (Homo sapiens), sonchus oleraceus (Sonchus oleracers L.), the West taraxacum (Taraxacum officinale) and Sunflower Receptacle (Helianthus annuus).
The script substrate (starting material) with the enzyme of prenyltransferases activity is allyl group bisphosphate.Then, because allyl group bisphosphate derivative is originally as the inhibitor of described enzyme, under many circumstances, if allyl group bisphosphate derivative is used as starting material, the enzymatic activity that this enzyme shows on this allyl group bisphosphate derivative lower (compound especially, represented by formula (G) to (Q)).For these reasons, when using allyl group bisphosphate derivative as starting material, the variant enzyme that enzymatic activity improves can be used for allyl group bisphosphate derivative.When using variant enzyme, the biology (transformant) transformed by genetic engineering technique can be prepared, to express variant enzyme.Especially, technician can according to the method disclosed in JP2012-036360A, and easily preparation improves the variant enzyme of the enzymic activity of allyl group bisphosphate derivative.
Isoprene oligopolymer of the present invention can under the existence of enzyme with prenyltransferases activity, and the allyl group bisphosphate represented by through type (X) and the compound represented by formula (Y) react and prepare.
Herein, term " under the existence of enzyme with prenyltransferases activity " means when there is following substances:
The culture of above-mentioned biology, the organism be separated from described cultivation, the handled thing of described organism, or from the enzyme of described culture or organism purifying; Or
For expressing the culture of the biology (transformant) transformed by genetic engineering technique of the enzyme with prenyltransferases activity, the organism be separated from described cultivation, the handled thing of described organism, or from the enzyme of this culture described or organism purifying; Or its analogue.
Technician can pass through known genetic engineering technique, is converted for the preparation of expression has the enzyme of prenyltransferases activity the biology obtained.
The organism of above-mentioned biology, can by cultivating acquisition by this biology in suitable substratum.As long as these substratum can make this biology breed, being not particularly limited, can be containing common carbon source, nitrogenous source, mineral ion and the usual substratum in organotrophy source that adds as required.
Such as, as carbon source, it can be the arbitrary carbon source that above-mentioned biology can utilize.Its concrete example, the carbohydrate of glucose, fructose, maltose, amylose starch and sucrose etc. can be used, the alcohols of Sorbitol Powder, ethanol, glycerine etc., the organic acid of fumaric acid, citric acid, acetic acid, propionic acid etc. and their salt, the carbohydrate of paraffin etc., and their mixture.
As the example of nitrogenous source, comprise the inorganic ammonium salt of ammonium sulfate, ammonium chloride etc., the organic acid ammonium salt of ammonium fumarate, ammonium citrate etc., the nitrate of SODIUMNITRATE, saltpetre etc., the organic nitrogen compound of peptone, yeast extract, meat extract, corn steep liquor etc., and their mixture.
Suitably can mix and adopt other nutrition source used in usual substratum, such as, inorganic salts, trace metal salts, vitamins, hormone etc.
Culture condition is also not particularly limited.Such as, under aerobic conditions, can be the scope of 5 to 8 at suitable control pH, temperature is when being the scope of 10 to 60 DEG C, carry out the cultivation of 12 to 480 hours.
The culture of described biology can be such as, the described biology cultivated by above-mentioned culture condition and the nutrient solution that obtains, or by filter wait from this nutrient solution separating bio (organism) acquisition culturing filtrate (culture supernatant).In addition, the organism be separated from above-mentioned culture can be such as, the organism (biology) obtained from nutrient solution separation by filter or centrifugation etc.
The handled thing of described organism can be such as, by the organism homogenate obtained after the organism homogenizing process be separated from above-mentioned culture, or the organism homogenate obtained after ultrasonication is carried out to biology.
Be such as from the enzyme of described culture or described organism purifying, the enzyme existed in described culture or described organism saltoutd, the known purification process such as ion-exchange chromatography, affinity chromatography or gel filtration chromatography and the enzyme obtained.The purity of purifying enzyme is not particularly limited.
Isoprene oligopolymer of the present invention can under the existence of enzyme with prenyltransferases activity, and the allyl group bisphosphate represented by through type (X) and the compound represented by formula (Y) react and prepare.Particularly, such as, this reaction is undertaken by adding the culture of described biology, the enzyme of purifying or its analogue in the solution containing the allyl group bisphosphate represented by formula (X) and the compound represented by formula (Y).In addition, temperature of reaction can be such as 20 to 60 DEG C, the reaction times can be such as 1 to 16 hour, pH can be such as 5 to 8.In addition, also as required, magnesium chloride, tensio-active agent, 2 mercapto ethanol and other materials can be added.
In the isoprene oligopolymer of the present invention that the reaction by above-mentioned explanation manufactures, the Y in formula (Z-1), formula (Z-2), formula (Z-1-1), formula (Z-2-1) is generally OPP or hydroxyl.OPP is from IPP or R-IPP.OPP also can be hydrolyzed easily, and the hydrolysis of OPP produces hydroxyl.The Y that Here it is why in formula (Z-1), (Z-2), (Z-1-1) or (Z-2-1) is generally OPP or hydroxyl.
In addition, the Y in formula (Z-1) or (Z-2) is the isoprene oligopolymer of formyl radical, such as, can be that the isoprene oligopolymer of OPP manufactures by the Y in oxidation-type (Z-1) or (Z-2).
In addition, the Y in formula (Z-1) or (Z-2) is the isoprene oligopolymer of carboxyl, such as, can be that the isoprene oligopolymer of OPP manufactures by the Y in oxidation-type (Z-1) or (Z-2).
In addition, the isoprene oligopolymer that Y in formula (Z-1) or (Z-2) is the isoprene oligopolymer of alkoxycarbonyl, Y in formula (Z-1) or (Z-2) is carbalkoxy, such as, can by taking the Y in aforesaid method carboxylation formula (Z-1) or (Z-2) as the isoprene oligopolymer of OPP and the isoprene oligopolymer further after this carboxylation of esterification manufactures.
Except the organic synthesis of the compound represented by formula (Y) and the starting material of the allyl group bisphosphate derivative that can select as required, isoprene oligopolymer of the present invention can be manufactured by biosynthesizing.Therefore, the present invention account for exhaustion and the environmental problem of petroleum resources.
(polyisoprene)
Then, polyisoprene of the present invention is described.The allyl group bisphosphate of polyisoprene of the present invention represented by formula (X) and the compound represented by formula (Y) carry out synthesizing (biosynthesizing).The monomer be polymerized on the allyl group bisphosphate represented by formula (X) can be the combination of compound represented by formula (Y) and prenyl diphosphate.Such as, polyisoprene of the present invention can compound represented by isoprene oligopolymer of the present invention and formula (Y) and/or prenyl diphosphate carry out synthesizing (biosynthesizing).Such as, polyisoprene of the present invention can carry out synthesizing (biosynthesizing) by the compound represented by unmodified isoprene oligopolymer and formula (Y), or compound represented by unmodified isoprene oligopolymer, formula (Y) and prenyl diphosphate carry out synthesizing (biosynthesizing).
Polyisoprene of the present invention there is the akin structure with natural rubber and with rubber molecule highly compatible.Polyisoprene of the present invention also has the molecular skeleton of modification, and therefore it with filler (such as, silicon-dioxide), strong interaction can occur.Therefore, due to polyisoprene of the present invention and rubber highly compatible, simultaneously can with filler (such as, silicon-dioxide) there is strong interaction, if this polyisoprene is used for rubber combination, can obtain and compare traditional composition, the rubber combination that its rubber molecule mixes with filler higher degree.Such as, this rubber combination can strengthen low-heat-generation, wet earth-catching property, wear resistance, elongation at break and fracture tensile strength thus.
The skeleton of polyisoprene of the present invention comprises polar group or its analogue.Thus, the situation when dispersing property of filler (such as, silicon-dioxide) only appears at chain end higher than polar group (such as, end bisphosphate group) etc.Therefore, such as, the improvement effect of low-heat-generation, wet earth-catching property, wear resistance, elongation at break and fracture tensile strength increases.
In addition, when the allyl group bisphosphate that the allyl group bisphosphate represented by formula (X) is represented by formula (X-1), and at least one atom contained in the II part in formula (X-1) or atomic group are replaced by other atoms or atomic group, when atom contained in III part in formula (X-1) or atomic group are not all replaced by other atoms or atomic group, not only skeleton but also chain end can be obtained and also carried out the polyisoprene of modification.Because the skeleton that described polyisoprene has modification incessantly also has the end of modification, its can with filler (such as, silicon-dioxide) interact better, if this polyisoprene is used for rubber combination, can obtain and compare traditional composition, the rubber combination that its rubber molecule mixes with filler higher degree.Such as, rubber combination can provide the low-heat-generation strengthened further, wet earth-catching property, wear resistance, elongation at break and fracture tensile strength thus.
The quantity preferably 30 to 40000 of the isoprene unit of addition on the allyl group bisphosphate starting material of formula (X) in polyisoprene of the present invention, more preferably 31 to 38000, further preferably 1000 to 35000, particularly preferably 2000 to 30000.
Such as, the object lesson of polyisoprene of the present invention is the represented or polyisoprene represented by formula (ZZ-2) of formula (ZZ-1), and in the v part in formula (ZZ-1) or formula (ZZ-2), at least one contained atom or atomic group are replaced by other atoms or atomic group;
Wherein, the n in formula (ZZ-1) represents the integer of 1 to 10; Q represents the integer of 30 to 40000; Y represents hydroxyl, formyl radical, carboxyl, alkoxycarbonyl, alkoxy carbonyl or OPP;
Wherein, the n in formula (ZZ-2) represents the integer of 1 to 10; Q represents the integer of 30 to 40000; Y represents hydroxyl, formyl radical, carboxyl, alkoxycarbonyl, alkoxy carbonyl or OPP.
Atom contained in v part in formula (ZZ-1) or formula (ZZ-2) or atomic group and other atoms described or atomic group can such as formula described in (X-1).
Iv part in formula (ZZ-1) and (ZZ-2) is the structure from the allyl group bisphosphate starting material represented by formula (X).Therefore, formula (ZZ-1) defines in the mode identical with the n in formula (X) with the n in (ZZ-2).
V part in formula (ZZ-1) and (ZZ-2) is made up of the isoprene unit of the allyl group bisphosphate starting material added to represented by formula (X).Therefore, formula (ZZ-1) and the q in (ZZ-2) are corresponding with the quantity of isoprene unit of the allyl group bisphosphate starting material adding to formula (X).
Due to the strong interaction with filler (such as, silicon-dioxide), the preferred OPP of Y, hydroxyl or carboxyl in formula (ZZ-1) and (ZZ-2).
As mentioned above, the present invention is based on following discovery: if the structure beyond the methyl of the 3-position in the structure of maintenance prenyl diphosphate is constant, even if the methyl of 3-position is replaced by required group, it also can be used to pass through naturally occurring enzyme and to manufacture isoprene oligopolymer or polyisoprene.Therefore, formula (ZZ-1) or the polyisoprene represented by formula (ZZ-2) are preferably the polyisoprene represented by formula (ZZ-1-1) or represented by formula (ZZ-2-1):
Wherein, n, q in formula (ZZ-1) define in the mode identical with Y with n, the q in formula (ZZ-1) with Y, and R is to define with the same way of the R in formula (Y);
Wherein, n, q in formula (ZZ-2-1) define in the mode identical with Y with n, the q in formula (ZZ-2) with Y, and R is to define with the same way of the R in formula (Y).
As mentioned above, also preferably more than to chain end, modification is carried out to skeleton simultaneously.Therefore, at least one atom contained by iv part in preferred formula (ZZ-1) or formula (ZZ-2) (preferred formula (ZZ-1-1) or formula (ZZ-2-1)) or atomic group are replaced by other atoms or atomic group.
Atom contained in iv part in formula (ZZ-1), formula (ZZ-2), formula (ZZ-1-1) or formula (ZZ-2-1) or atomic group and other atoms described or atomic group can such as formula described in (X-1).
Further, as mentioned above, allyl group bisphosphate represented by preferred formula (X) is the allyl group bisphosphate represented by formula (X-1), and at least one atom contained in the II part in formula (X-1) or atomic group are replaced by other atoms or atomic group, atom contained in the III part in formula (X-1) or atomic group are not all replaced by other atoms or atomic group.Therefore, about as shown in the formula the iv part in the formula (ZZ-1) represented by (Z), formula (ZZ-2), formula (ZZ-1-1) and formula (ZZ-2-1), at least one atom contained in iv-II part in preferred formula (Z) or atomic group are replaced by other atoms or atomic group, and atom contained in the iv-III part in formula (Z) or atomic group are not all replaced by other atoms or atomic group.
In iv-II part in formula (Z) or iv-III part, contained atom or atomic group and other atoms described or atomic group can such as formula the atom described in (X-1) or atomic groups.
The concrete example of the iv part that chain end is also modified can comprise the structure represented by above-mentioned formula (a) to (s).Wherein, the structure of preferred formula (b), (c), (d), (e), (f), (k), (l) and (r), more preferably the structure of formula (b) and (c), because they have stronger with filler (such as, silicon-dioxide) interaction, and more effectively can improve low-heat-generation, wet earth-catching property, wear resistance, elongation at break and fracture tensile strength.
(manufacturing the method for polyisoprene)
Polyisoprene of the present invention can compound represented by isoprene oligopolymer of the present invention and formula (Y) (R-IPP) and/or prenyl diphosphate (IPP), is manufactured by the method (A) of synthesizing (biosynthesizing) polyisoprene.Can according to the target physical character of the polyisoprene obtained, be suitable for the ratio of the monomer of R-IPP and IPP changing use, wherein a kind of percentage ratio of monomer can be 0.Or, polyisoprene of the present invention can compound represented by unmodified isoprene oligopolymer and formula (Y), manufactured by the method (B) of synthesizing (biosynthesizing) polyisoprene, or the compound represented by unmodified isoprene oligopolymer, formula (Y) and prenyl diphosphate, manufactured by the method (C) of synthesizing (biosynthesizing) polyisoprene.In addition, in these cases, according to the target physical character of the polyisoprene obtained, the ratio changed between R-IPP and the IPP monomer of use can be suitable for.
Except the organic synthesis of the compound represented by formula (Y) and the starting material of the allyl group bisphosphate derivative that can select as required, polyisoprene of the present invention can be manufactured by biosynthesizing.Therefore, the present invention account for exhaustion and the environmental problem of petroleum resources.
All the time, known natural rubber latex (especially, natural rubber latex from Para rubber tree) in containing enzyme and rubber elongation factor (such as, as above can the enzyme of addition polymerization prenyl diphosphate) or its analogue, it has the activity of the condensation reaction between catalysis isoprene oligopolymer and prenyl diphosphate, therefore, prenyl diphosphate is connected for Z configuration (isoprene unit of new addition is cis-configuration), the reaction generating polyisoprene by catalysis successively with isoprene oligopolymer, as follows.
Simultaneously, in the known natural rubber latex from some plant (rubber latex) containing enzyme and rubber elongation factor (such as, as above can the enzyme of addition polymerization prenyl diphosphate) or its analogue, it has the activity of the condensation reaction between catalysis isoprene oligopolymer and prenyl diphosphate, therefore, prenyl diphosphate is connected with isoprene oligopolymer for E (isoprene unit of new addition is transconfiguration), the reaction generating polyisoprene by catalysis successively.
As mentioned above, when the methyl replacing the 3-position in the structure of prenyl diphosphate with required group and to maintain the structure except the methyl of 3-position of prenyl diphosphate constant, and obtain the compound of formula (Y) (R-IPP), can by using this compound and adopting naturally occurring enzyme, rubber elongation factor or its analogue with the activity of the above-mentioned reaction of catalysis to form polyisoprene.Therefore, in the present invention, above-mentioned enzyme, rubber elongation factor or its analogue can be used to manufacture polyisoprene.
In other words, compound represented by isoprene oligopolymer of the present invention and formula (Y) and/or prenyl diphosphate synthesize the method (A) of (biosynthesizing) polyisoprene of the present invention, compound represented by unmodified isoprene oligopolymer and formula (Y) synthesizes the method (B) of (biosynthesizing) polyisoprene of the present invention, by unmodified isoprene oligopolymer, compound represented by formula (Y) and prenyl diphosphate synthesize the method (C) of (biosynthesizing) polyisoprene of the present invention, can by the enzyme such as using natural rubber latex Ruzhong to contain, rubber elongation factor or its analogue have come.Also the enzyme of cloning from natural rubber latex Ruzhong, rubber elongation factor or its analogue can be used for aforesaid method.
Particularly, in method (A), under the existence of enzyme and/or rubber elongation factor, the compound represented by isoprene oligopolymer of the present invention and formula (Y) and/or prenyl diphosphate can be reacted.Similarly, in method (B), under the existence of enzyme and/or rubber elongation factor, the compound represented by unmodified isoprene oligopolymer and formula (Y) can be reacted.Similarly, in method (C), under the existence of enzyme and/or rubber elongation factor, the compound represented by unmodified isoprene oligopolymer and formula (Y), prenyl diphosphate can be reacted.
More specifically, such as, described reaction by the solution containing the compound represented by isoprene oligopolymer of the present invention and formula (Y) and/or prenyl diphosphate, can be added natural rubber latex or causes from the isolated enzyme in natural rubber latex Ruzhong, rubber elongation factor or its analogue.In addition, temperature of reaction can be such as 10 to 60 DEG C, the reaction times can be such as 1 to 72 hour, pH can be such as 6 to 8.In addition, also as required, magnesium chloride, tensio-active agent, 2 mercapto ethanol, Potassium monofluoride and other materials can be added.Even when using unmodified isoprene oligopolymer, reaction also can be carried out under similarity condition.
In the present invention, the isoprene chain of polyisoprene above in any direction can extend (transconfiguration or cis-configuration).In other words, such as, polyisoprene of the present invention can be all isoprene units all with transconfiguration link polyisoprene (such as, the isoprene oligopolymer of formula (ZZ-1)), the polyisoprene that links with Trans-cis-transconfiguration of the polyisoprene (such as, the polyisoprene of formula (Z-2)) that links with trans-cis configuration of isoprene unit or isoprene unit.Wherein, the polyisoprene that preferred isoprene unit links with trans-cis configuration, because it is identical with the structure of the natural rubber from Para rubber tree of industrial widespread use.
At the polyisoprene of the present invention that the reaction by above-mentioned explanation manufactures, the Y in formula (ZZ-1), formula (ZZ-2), formula (ZZ-1-1) or formula (ZZ-2-1) is generally OPP or hydroxyl.OPP is from IPP or R-IPP.OPP also can be hydrolyzed easily, and the hydrolysis of OPP produces hydroxyl.The Y that Here it is why in formula (ZZ-1), (ZZ-2), (ZZ-1-1) or (ZZ-2-1) is generally OPP or hydroxyl.
In addition, the Y in formula (ZZ-1) or (ZZ-2) is the polyisoprene of formyl radical, such as, can be that the polyisoprene of OPP manufactures by the Y in oxidation-type (ZZ-1) or (ZZ-2).
In addition, the Y in formula (ZZ-1) or (ZZ-2) is the polyisoprene of carboxyl, such as, can be that the polyisoprene of OPP manufactures by the Y in oxidation-type (ZZ-1) or (ZZ-2).
In addition, Y in formula (ZZ-1) or (ZZ-2) is the polyisoprene of alkoxycarbonyl, Y in formula (ZZ-1) or (ZZ-2) is the polyisoprene of carbalkoxy, such as, can by taking the Y in aforesaid method carboxylation formula (ZZ-1) or (ZZ-2) as the polyisoprene of OPP and the polyisoprene further after this carboxylation of esterification manufactures.
The source of natural rubber latex is not particularly limited.Its example comprises: Para rubber tree (Hevea brasiliensis), rubberplant (Ficus elastica), Root of Fiddleleaf Fig (Ficus lyrata), banyan (Ficus benjamina), linden (Ficus religiosa), banian (Ficus benghalensis), laetarius volemus (Lactarius volemus), sonchus oleraceus (Sonchus oleracers L.), the West taraxacum (Taraxacum officinale) and Sunflower Receptacle (Helianthus annuus).Wherein, preferred Para rubber tree, the rubber because of its manufacture has high molecular and latex has high rubber content.
Natural rubber latex, such as, can by marking the wound (this process is called " rubber tapping ") of channel-shaped on the trunk of Para rubber tree with cutter or its analogue, then reclaim the natural rubber latex that flows out from cut-off latex dust and obtain.
The enzyme be separated from natural rubber latex Ruzhong or rubber elongation factor can be enumerated such as, the glue be separated by centrifugation natural rubber latex clear (Serum), bottom phase (bottom fraction) or rubber phase (rubber fraction).Described glue is clear, in bottom phase and rubber phase containing enzyme, rubber elongation factor or its analogue.
(rubber combination)
Rubber combination of the present invention contains isoprene oligopolymer of the present invention and/or polyisoprene of the present invention.Therefore, rubber combination of the present invention has outstanding low-heat-generation, wet earth-catching property, wear resistance, elongation at break and fracture tensile strength (particularly low-heat-generation and wear resistance).Polyisoprene of the present invention can be used as rubber components.
In rubber constituent based on 100 quality % meters, the content of polyisoprene of the present invention is preferably more than 20 quality %, more preferably more than 40 quality %, further preferred more than 60 quality %.It can be 100 quality %.
Except polyisoprene of the present invention, the example of the material that can use as rubber constituent comprises: the diene series rubbers such as synthetic polyisoprene (IR), natural rubber (NR), divinyl rubber (BR), styrene-butadiene rubber(SBR) (SBR), styrene isoprene butadiene rubber (SIBR), chloroprene rubber (CR) and acrylonitrile butadiene rubber (NBR).These elastomeric materials can be used alone, and also can be used in combination of two or more.Wherein, preferred NR, BR and SBR.
When being used in rubber combination by isoprene oligopolymer of the present invention, for the reason high with the intermiscibility of isoprene oligopolymer, rubber constituent preferably uses NR.By and with isoprene oligopolymer of the present invention and NR, more suitably can obtain the effect of isoprene oligopolymer of the present invention.
When isoprene oligopolymer of the present invention is used in rubber combination, based in rubber constituent 100 quality %, content preferably more than the 20 quality % of NR, more preferably more than 40 quality %, further preferred more than 60 quality %.It can be 100 quality %.
Relative to rubber constituent 100 mass parts, the content of isoprene oligopolymer of the present invention is preferably more than 1 mass parts, more preferably more than 2 mass parts.When the isoprene oligopolymer used is less than 1 mass parts, may not fully produce its effect.The content of described isoprene oligopolymer is also preferably below 20 mass parts, more preferably below 15 mass parts.During content higher than 20 mass parts, intensity and wear resistance may be reduced.
The example of the filler used in the present invention comprises: silicon-dioxide, carbon black, clay and calcium carbonate.
The preferred silicon-dioxide of filler used in the present invention.When using silicon-dioxide, fully can obtain the effect obtained by using isoprene oligopolymer of the present invention and/or polyisoprene of the present invention.Can use arbitrary silicon-dioxide, its example comprises dry silica (anhydrous silicic acid), wet silica (moisture silicic acid).Be preferably wet silica, because it contains more silanol group.
Also preferably use carbon black as filler in the present invention.In this case, also can fully obtain by using isoprene oligopolymer of the present invention and/or polyisoprene of the present invention and the effect obtained.
Except mentioned component, can also suitably containing other Synergist S-421 95 conventional in the manufacture of rubber combination in rubber combination of the present invention, such as, silane coupling agent, zinc oxide, stearic acid, various protective agent (i.e. antioxidant), tenderizer are (such as, oil), wax, vulcanizing agent (such as, sulphur) and vulcanization accelerator.
Rubber combination of the present invention can be prepared by conventional known method.Such as, can by using the mixing described component of rubber mill of open roll, Banbury muller etc., then sulfuration, to manufacture rubber combination.
Rubber combination of the present invention is applicable to tyre element (such as, tyre surface, sidewall, tread base, cord body, buffer layer and carcass) and its analogue.
(pneumatic tyre)
Pneumatic tyre of the present invention can use described rubber combination to manufacture according to usual method.Particularly, by unvulcanized rubber combination extrusion processing for corresponding to tyre element (such as, tyre surface or sidewall) shape, then, be shaped by usual method tyre forming machine, then fit with other tyre element, thus form unvulcanized tire, this unvulcanized tire is carried out hot pressing in vulcanizer, thus forms tire.
Embodiment
With reference to following examples, the present invention is described in detail, but the present invention is not limited only to this.
(manufacturing embodiment 1)
(the synthesis (compound (R-IPP) represented by formula (Y)) of 3-R-3-butenyl bisphosphate
N-R-aldehyde is used to be starting material synthesising target compound.By the people such as Green (M.B.Green, and W.J.Hickinbotton, J.Chem.Soc.1957,3262) method, uses dimethylamine to introduce outer methylene radical (compound represented by following formula (i)) on the alpha-position of n-R-aldehyde in formic acid.Then, with lithium aluminum hydride, the compound of acquisition is reduced to 2-R-vinyl carbinol (compound represented by following formula (ii)).Further, be 2-R-chlorallylene (compound represented by following formula (iii)) with phosphorus chloride by this compound chlorination in pyridine, again by it under the existence of the Grignard reagent of fresh synthesis, obtain carboxylic acid (compound represented by following formula (iv)) with carbon dioxide reaction.Then, with lithium aluminum hydride, the reduction of this compound is formed alcohol body (compound represented by following formula (v)).Then, in pyridine, with toluene sulfonyl chloride, this compound is carried out tosylation (compound represented by following formula (vi)).Then, this compound is carried out phosphorylation with trimethyl phosphite 99 in acetonitrile, obtain target product (compound represented by following formula (vii), the compound (R-IPP) namely represented by formula (Y)) thus.By the intermediate and the final product that use TLC and instrumental analysis (IR and NMR) to confirm each synthesis phase.
When R is respectively ethyl, propyl group, butyl, phenyl, sulfydryl, hydroxyl and amino, synthesize the compound (R-IPP) represented by formula (Y).When R is ethyl, propyl group, butyl, phenyl, sulfydryl, hydroxyl and amino, the R-IPP of formula (Y) is denoted as R-IPP-A, R-IPP-B, R-IPP-C, R-IPP-D, R-IPP-E, R-IPP-F and R-IPP-G respectively.
(embodiment 1)
(manufacture (alltrans) of isoprene oligopolymer)
Prepare whole isoprene unit all with the isoprene oligopolymer that transconfiguration links, represented by (Z-1-1-1).
(preparation of transformant)
First, transformant is prepared.In the preparation of transformant, use the pET15b (pET15b/human-GGPS) importing mankind's geranyl geranyl diphosphate synthetic enzyme.This pET15b/human-GGPS is by associate professor (Associate Professor Hiroshi SAGAMI rich in the assistant of multi-component Science Institute of Northeastern University, Institute of Multidisciplinary Research for Advanced Materials, Tohoku University) provide.
Use pET15b/human-GGPS by heat shock Transformed E .coli BL21 (DE3).After on the LB nutrient agar this transformant being coated on the penbritin containing 50 μ g/mL, overnight incubation at 37 DEG C, then, selects transformant.
(there is the manufacture of the protein of prenyltransferases activity)
The E.coli BL21 (DE3) obtained/pET15b/human-GGPS (wild-type) is inoculated into and 3mL is housed contains in the test tube of the LB substratum of the penbritin of 50 μ g/mL, shaking culture 5 hours at 37 DEG C.In the nutrient solution obtained, get 1mL decile and be seeded in the 500mL Erlenmeyer flask of the LB substratum containing 50 μ g/mL penbritins that 100mL is housed, at 37 DEG C, cell oscillation is cultivated 3 hours.Then, adding IPTG is 0.1mmol/L to its concentration, then at 30 DEG C, cell oscillation is cultivated 18 hours.This nutrient solution of centrifugation, obtains wet thallus.After this wet thallus of ultrasonic wave homogenizing, centrifugation obtains supernatant liquor.HisTrap (Amersham company) purifying from this supernatant liquor is used to have the protein of prenyltransferases activity.Protein purification is confirmed by SDS-PAGE.
(preparation of isoprene oligopolymer)
Preparation is containing protein 10mg, 50mM Tris-HCl damping fluid (pH 7.5) after purifying, 40mM magnesium chloride, 25mM 2 mercapto ethanol, 1mM starting material (geranyl bisphosphate (GPP) and 1mM prenyl diphosphate (IPP) or a kind of reaction soln manufactured in R-IPP manufactured in embodiment 1.This reaction 1 hour is carried out in the water-bath of 37 DEG C.After reaction terminates, add saturated aqueous common salt 100ml and pentane 500ml, after stirring this mixture, left standstill.Then, extremely dry by evaporation concentration supernatant liquor (pentane layer).Get remnant parts and confirm product structure by NMR, it is isoprene oligopolymer.The particular case (n, m and R in formula (Z-1-1-1)) of the isoprene oligopolymer obtained thus is as shown in table 1.Herein, based on the information of the starting material used and the isoprene chain length by TLC mensuration, n and m in formula (Z-1-1-1) is calculated.In addition, the R structure in formula (Z-1-1-1) is identified by NMR, TLC and GC-MS.
In addition, use the isoprene oligopolymer of monomer I PP, R-IPP-A, R-IPP-B, R-IPP-C, R-IPP-D, R-IPP-E, R-IPP-F and R-IPP-G acquisition respectively, in aftermentioned experiment uses, be called isoprene oligopolymer (t-contrast), isoprene oligopolymer (t-A), isoprene oligopolymer (t-B), isoprene oligopolymer (t-C), isoprene oligopolymer (t-D), isoprene oligopolymer (t-E), isoprene oligopolymer (t-F) and isoprene oligopolymer (t-G).In addition, in experiment, isoprene oligopolymer (t-E), isoprene oligopolymer (t-F) add in rubber combination respectively as isoprene oligopolymer a and isoprene oligopolymer e.
[table 1]
(relative reactivity of monomer)
The one making IPP or manufacture in the R-IPP in embodiment 1 and geranyl bisphosphate (GPP) react under the following conditions.Each R-IPP for the relative reactivity of GPP to represent relative to the index of IPP activity (=100).
Preparation is containing protein 500ng, 50mM Tris-HCl damping fluid (pH 7.5) after purifying, 40mM magnesium chloride, 25mM the 2 mercapto ethanol, [1-of 12.5 μMs GPP and 50 μM
14c] IPP or the one that manufactures in the R-IPP of preparation in embodiment 1.This reaction 1 hour is carried out in the water-bath of 37 DEG C.After reaction, employing liquid scintillation counting(LSC) and TLC measure activity in each condition.Relative to the activity (=100) of IPP, the relative reactivity of each R-IPP is as shown in table 2.
Isoprene oligopolymer adopts and carries out purifying with the same mode in embodiment 1 part (preparation of isoprene oligopolymer).Then, the particular case (n, m and R in formula (Z-1-1-1)) of the isoprene oligopolymer obtained is shown in table 2.Herein, based on the information of the starting material used and the isoprene chain length by TLC mensuration, n and m in formula (Z-1-1-1) is calculated.In addition, the R structure in formula (Z-1-1-1) is identified by NMR, TLC and GC-MS.
[table 2]
Table 1 and 2 result display, even if use R-IPP, the isoprene oligopolymer of acquisition also has identical molecular weight with during use IPP.Also demonstrate the skeleton obtaining isoprene oligopolymer and there occurs the modification corresponding to the R-IPP used.
(embodiment 2)
(preparation of polyisoprene)
The isoprene oligopolymer (all trans) linked with transconfiguration such as formula the whole isoprene units represented by (Z-1-1-1) is used to prepare polyisoprene.
A kind of reaction soln in the R-IPP of preparation containing preparation in 10 μ L latex component, 50mM Tris-HCl damping fluid (pH 7.5), 25mM magnesium chloride, 40mM2-mercaptoethanol, 40mM Potassium monofluoride, 50 μMs of isoprene oligopolymer and 1mM IPP or manufacture embodiment 1.This reaction 3 days is carried out in the water-bath of 30 DEG C.After reaction, by GPC determining molecular weight.Then, based on the information of the numerical value of molecular weight and the starting material of use, the quantity of the isoprene unit added on starting material GPP is calculated.Result is as shown in table 3.
Latex component used herein is by carrying out ultracentrifugation and the glue obtained is clear to obtaining latex from Para rubber tree.
The isoprene oligopolymer used is the one in isoprene oligopolymer (t-contrast), isoprene oligopolymer (t-A), isoprene oligopolymer (t-B), isoprene oligopolymer (t-C), isoprene oligopolymer (t-D), isoprene oligopolymer (t-E), isoprene oligopolymer (t-F) and isoprene oligopolymer (t-G) prepared in above-mentioned part (preparation of isoprene oligopolymer).
[table 3]
The result display of table 3, even if use R-IPP, the polyisoprene of acquisition also has identical molecular weight with during use IPP.Further, analyze polyisoprene NMR, TLC and the GC-MS obtained, confirm with the situation of isoprene oligopolymer similar, the skeleton of polyisoprene has carried out the modification corresponding to used R-IPP.
(embodiment 3)
(manufacture (Trans-cis) of isoprene oligopolymer)
Prepare the isoprene oligopolymer that isoprene unit links with trans-cis configuration, represented by (Z-2-1-1).
(preparation of transformant)
First, transformant is prepared.In the preparation of transformant, use the pET22b (pET22b/MLU-UPS) of the undecaprenyl-diphosphate synthase importing micrococcus luteus B-P26 as dsDNA template.This pET22b/MLU-UPS is by ancient mountain kind professor person of outstanding talent (the Prof.Tanetoshi Koyama of multi-component Science Institute of Northeastern University, Institute of Multidisciplinary Research for Advanced Materials, Tohoku University) provide.
Use pET22b/MLU-UPS by heat shock Transformed E .coli BL21 (DE3).After on the LB nutrient agar this transformant being coated on the penbritin containing 50 μ g/mL, overnight incubation at 37 DEG C, then, selects transformant.
(there is the manufacture of the protein of prenyltransferases activity)
E.coli BL21 (the DE3)/pET22b/MLU-UPS obtained is inoculated into and 3mL is housed contains in the test tube of the LB substratum of the penbritin of 50 μ g/mL, shaking culture 5 hours at 37 DEG C.In the nutrient solution obtained, get 1mL decile and be seeded in the 500mL Erlenmeyer flask of the LB substratum containing 50 μ g/mL penbritins that 100mL is housed, at 37 DEG C, cell oscillation is cultivated 3 hours.Then, adding IPTG is 0.1mmol/L to its concentration, then at 30 DEG C, cell oscillation is cultivated 18 hours.This nutrient solution of centrifugation, obtains wet thallus.After this wet thallus of ultrasonic wave homogenizing, centrifugation obtains supernatant liquor.HisTrap (Amersham company) purifying from this supernatant liquor is used to have the protein of prenyltransferases activity.Protein purification is confirmed by SDS-PAGE.
(preparation of isoprene oligopolymer)
Preparation is containing protein 10mg, 50mM Tris-HCl damping fluid (pH 7.5) after purifying, 40mM magnesium chloride, 40mM Triton X-100,25mM 2 mercapto ethanol, 1mM starting material (farnesyl bisphosphate (FPP) and 1mM IPP or the one manufactured in the R-IPP of preparation in embodiment 1.This reaction 1 hour is carried out in the water-bath of 37 DEG C.After reaction terminates, add saturated aqueous common salt 100ml and n-butyl alcohol 500ml, after stirring this mixture, left standstill.Then, extremely dry by evaporation concentration supernatant liquor (n-butyl alcohol layer).Get remnant parts and confirm product structure by NMR, it is isoprene oligopolymer.The particular case (n, m and R in formula (Z-2-1-1)) of the isoprene oligopolymer obtained is as shown in table 4.Herein, based on the information of the starting material used and the isoprene chain length by TLC mensuration, n and m in formula (Z-2-1-1) is calculated.In addition, the R structure in formula (Z-2-1-1) is identified by NMR, TLC and GC.
In addition, use monomer I PP, R-IPP-A, R-IPP-B, R-IPP-C, R-IPP-D, R-IPP-E, R-IPP-F and R-IPP-G and the isoprene oligopolymer that obtains respectively, in aftermentioned experiment use, be called isoprene oligopolymer (tc-contrast), isoprene oligopolymer (tc-A), isoprene oligopolymer (tc-B), isoprene oligopolymer (tc-C), isoprene oligopolymer (tc-D), isoprene oligopolymer (tc-E), isoprene oligopolymer (tc-F) and isoprene oligopolymer (tc-G).In addition, in experiment, isoprene oligopolymer (tc-E) adds in rubber combination as isoprene oligopolymer (b).In addition, in experiment, each isoprene oligopolymer (tc-A), isoprene oligopolymer (tc-B), isoprene oligopolymer (tc-C), isoprene oligopolymer (tc-D), isoprene oligopolymer (tc-E), isoprene oligopolymer (tc-F) and isoprene oligopolymer (tc-G) add in rubber combination respectively as isoprene oligopolymer h, isoprene oligopolymer i, isoprene oligopolymer j, isoprene oligopolymer k, isoprene oligopolymer l, isoprene oligopolymer m and isoprene oligopolymer n.
[table 4]
(relative reactivity of monomer)
The one making IPP or manufacture in the R-IPP in embodiment 1 and farnesyl bisphosphate (FPP) react under the following conditions.Each R-IPP for the relative reactivity of FPP to represent relative to the index of IPP activity (=100).
Preparation is containing protein 500ng, 50mM Tris-HCl damping fluid (pH 7.5) after purifying, 40mM magnesium chloride, 40mM Triton X-100,25mM 2 mercapto ethanol, [1-of 12.5 μMs FPP and 50 μM
14c] IPP or a kind of reaction soln of manufacturing in the R-IPP of preparation in embodiment 1.This reaction 1 hour is carried out in the water-bath of 37 DEG C.After reaction, employing liquid scintillation counting(LSC) and TLC measure activity in each condition.Relative to the activity (=100) of IPP, the relative reactivity of each R-IPP is as shown in table 5.
Isoprene oligopolymer adopts and carries out purifying with the same mode in embodiment 3 part (preparation of isoprene oligopolymer).Then, the particular case (n, m and R in formula (Z-2-1-1)) of the isoprene oligopolymer obtained is shown in table 5.Herein, based on the information of the starting material used and the isoprene chain length by TLC mensuration, n and m in formula (Z-2-1-1) is calculated.In addition, the R structure in formula (Z-2-1-1) is identified by NMR, TLC and GC-MS.
[table 5]
Table 4 and 5 result display, even if use R-IPP, the isoprene oligopolymer of acquisition also has identical molecular weight with during use IPP.Also demonstrate the skeleton obtaining isoprene oligopolymer and there occurs the modification corresponding to the R-IPP used.
(embodiment 4)
(preparation of polyisoprene)
The isoprene oligopolymer (Trans-cis) linked with trans-cis configuration such as formula the isoprene unit represented by (Z-2-1-1) is used to prepare polyisoprene.
A kind of reaction soln in the R-IPP of preparation containing preparation in 10 μ L latex component, 50mM Tris-HCl damping fluid (pH 7.5), 25mM magnesium chloride, 40mM2-mercaptoethanol, 40mM Potassium monofluoride, 50 μMs of isoprene oligopolymer and 1mM IPP or manufacture embodiment 1.This reaction 3 days is carried out in the water-bath of 30 DEG C.After reaction, by GPC determining molecular weight.Then, based on the information of the numerical value of molecular weight and the starting material of use, the quantity of the isoprene unit added on starting material FPP is calculated.Result is presented in table 6.
Latex component used herein is by carrying out ultracentrifugation and the glue obtained is clear to obtaining latex from Para rubber tree.
The isoprene oligopolymer used is the one in isoprene oligopolymer (tc-contrast), isoprene oligopolymer (tc-A), isoprene oligopolymer (tc-B), isoprene oligopolymer (tc-C), isoprene oligopolymer (tc-D), isoprene oligopolymer (tc-E), isoprene oligopolymer (tc-F) and isoprene oligopolymer (tc-G) prepared in above-mentioned part (preparation of isoprene oligopolymer).In addition, in experiment subsequently, the polyisoprene that use isoprene oligopolymer (tc-E), isoprene oligopolymer (tc-F) manufacture adds in rubber combination respectively as polyisoprene D and polyisoprene A.In addition, each polyisoprene using isoprene oligopolymer (tc-A), isoprene oligopolymer (tc-B), isoprene oligopolymer (tc-C), isoprene oligopolymer (tc-D), isoprene oligopolymer (tc-E), isoprene oligopolymer (tc-F), isoprene oligopolymer (tc-G) to manufacture, respectively as polyisoprene I, polyisoprene J, polyisoprene K, polyisoprene L, polyisoprene M, polyisoprene N, polyisoprene P, adds in rubber combination.
[table 6]
The result display of table 6, even if use R-IPP, the polyisoprene of acquisition also has identical molecular weight with during use IPP.Further, analyze polyisoprene NMR, TLC and the GC-MS obtained, confirm with the situation of isoprene oligopolymer similar, the skeleton of polyisoprene has carried out the modification corresponding to used R-IPP.
(embodiment 5)
(manufacture (all trans) of isoprene oligopolymer)
Prepare whole isoprene unit all with the isoprene oligopolymer that transconfiguration links, represented by (Z-1-1-1).
(preparation of transformant)
First, transformant is prepared.In the preparation of transformant, use the pET15b (pET15b/bacillus-FPS) importing bacstearothermophilus farnesyl diphosphate synthase.This pET15b/bacillus-FPS is by ancient mountain kind professor person of outstanding talent (the Prof.Tanetoshi Koyama of multi-component Science Institute of Northeastern University, Institute of Multidisciplinary Research for Advanced Materials, Tohoku University) provide.
Use pET15b/bacillus-FPS by heat shock Transformed E .coli BL21 (DE3).After on the LB nutrient agar this transformant being coated on the penbritin containing 50 μ g/mL, overnight incubation at 37 DEG C, then, selects transformant.
(there is the manufacture of the protein of prenyltransferases activity)
The E.coli BL21 (DE3) obtained/pET15b/bacillus-FPS (wild-type) is inoculated into and 3mL is housed contains in the test tube of the LB substratum of the penbritin of 50 μ g/mL, shaking culture 5 hours at 37 DEG C.In the nutrient solution obtained, get 1mL decile and be seeded in the 500mL Erlenmeyer flask of the LB substratum containing 50 μ g/mL penbritins that 100mL is housed, at 37 DEG C, cell oscillation is cultivated 3 hours.Then, adding IPTG is 0.1mmol/L to its concentration, then at 30 DEG C, cell oscillation is cultivated 18 hours.This nutrient solution of centrifugation, obtains wet thallus.After this wet thallus of ultrasonic wave homogenizing, centrifugation obtains supernatant liquor.HisTrap (Amersham company) purifying from this supernatant liquor is used to have the protein of prenyltransferases activity.Protein purification is confirmed by SDS-PAGE.
(preparation of isoprene oligopolymer)
Preparation is containing protein 10mg, 50mM Tris-HCl damping fluid (pH 7.5) after purifying, 40mM magnesium chloride, 25mM 2 mercapto ethanol, 1mM starting material (dimethylallyl diphosphate (DMAPP) and 1mM prenyl diphosphate (IPP) or a kind of reaction soln manufactured in the R-IPP of preparation in embodiment 1.This reaction 1 hour is carried out in the water-bath of 37 DEG C.After reaction terminates, add saturated aqueous common salt 100ml and pentane 500ml, after stirring this mixture, left standstill.Then, extremely dry by evaporation concentration supernatant liquor (pentane layer).Get remnant parts and confirm product structure by NMR, it is isoprene oligopolymer.The particular case (n, m and R in formula (Z-1-1-1)) of the isoprene oligopolymer obtained is as shown in table 7.Herein, based on the information of the starting material used and the isoprene chain length by TLC mensuration, n and m in formula (Z-1-1-1) is calculated.In addition, the R structure in formula (Z-1-1-1) is identified by NMR, TLC and GC-MS.
In addition, use monomer I PP respectively, R-IPP-A, R-IPP-B, R-IPP-C, R-IPP-D, R-IPP-E, R-IPP-F and R-IPP-G and the isoprene oligopolymer obtained, isoprene oligopolymer (t-contrast-1) is called in aftermentioned experiment uses, isoprene oligopolymer (t-A-1), isoprene oligopolymer (t-B-1), isoprene oligopolymer (t-C-1), isoprene oligopolymer (t-D-1), isoprene oligopolymer (t-E-1), isoprene oligopolymer (t-F-1) and isoprene oligopolymer (t-G-1).
[table 7]
(relative reactivity of monomer)
IPP or the one manufactured in the R-IPP of preparation in embodiment 1 and dimethyl bisphosphate (DMAPP) are reacted under the following conditions.Each R-IPP for the relative reactivity of DMAPP to represent relative to the index of IPP activity (=100).
Preparation is containing protein 500ng, 50mM Tris-HCl damping fluid (pH 7.5) after purifying, 40mM magnesium chloride, 25mM the 2 mercapto ethanol, [1-of 12.5 μMs DMAPP and 50 μM
14c] IPP or a kind of reaction soln of manufacturing in the R-IPP of preparation in embodiment 1.This reaction 1 hour is carried out in the water-bath of 37 DEG C.After reaction, employing liquid scintillation counting(LSC) and TLC measure activity in each condition.Relative to the activity (=100) of IPP, the relative reactivity of each R-IPP is as shown in table 8.
Isoprene oligopolymer adopts and carries out purifying with the same mode in embodiment 5 part (preparation of isoprene oligopolymer).Then, the particular case (n, m and R in formula (Z-1-1-1)) of the isoprene oligopolymer obtained is shown in table 8.Herein, based on the information of the starting material used and the isoprene chain length by TLC mensuration, n and m in formula (Z-1-1-1) is calculated.In addition, the R structure in formula (Z-1-1-1) is identified by NMR, TLC and GC-MS.
[table 8]
Table 7 and 8 result display, even if use R-IPP, the isoprene oligopolymer of acquisition also has identical molecular weight with during use IPP.Also demonstrate the skeleton obtaining isoprene oligopolymer and there occurs the modification corresponding to the R-IPP used.
(embodiment 6)
(preparation of polyisoprene)
Then, the isoprene oligopolymer (all trans) linked with transconfiguration such as formula the whole isoprene units represented by (Z-1-1-1) is used to prepare polyisoprene.
A kind of reaction soln in the R-IPP of preparation containing preparation in 10 μ L latex component, 50mM Tris-HCl damping fluid (pH 7.5), 25mM magnesium chloride, 40mM2-mercaptoethanol, 40mM Potassium monofluoride, 50 μMs of isoprene oligopolymer and 1mM IPP or manufacture embodiment 1.This reaction 3 days is carried out in the water-bath of 30 DEG C.After reaction, by GPC determining molecular weight.Then, based on the information of the numerical value of molecular weight and the starting material of use, the quantity of the isoprene unit added on starting material DMAPP is calculated.Result is as shown in table 9.
Latex component used herein is by carrying out ultracentrifugation and the glue obtained is clear to obtaining latex from Para rubber tree.
The isoprene oligopolymer used is the isoprene oligopolymer (t-contrast-1) prepared by above-mentioned part (preparation of isoprene oligopolymer), the one in isoprene oligopolymer (t-A-1), isoprene oligopolymer (t-B-1), isoprene oligopolymer (t-C-1), isoprene oligopolymer (t-D-1), isoprene oligopolymer (t-E-1), isoprene oligopolymer (t-F-1) and isoprene oligopolymer (t-G-1).
[table 9]
The result display of table 9, even if use R-IPP, the polyisoprene of acquisition also has identical molecular weight with during use IPP.Further, analyze polyisoprene NMR, TLC and the GC-MS obtained, confirm with the situation of isoprene oligopolymer similar, the skeleton of polyisoprene has carried out the modification corresponding to used R-IPP.
(embodiment 7)
(manufacture (Trans-cis) of isoprene oligopolymer)
Then, use allyl group bisphosphate derivative, prepare such as formula the isoprene unit represented by (Z-2-1-1) with trans-cis configuration link and not only also there is the isoprene oligopolymer (the isoprene oligopolymer of skeleton and chain end modification) of modification in skeleton but also chain end.
(preparation of allyl group bisphosphate derivative)
First, synthesis of allyl bisphosphate derivative.
(manufacturing embodiment 2)
(synthesis of 8-methoxyl group-3,7-dimethyl-ten two carbon-(2E, 6E)-dialkylene bisphosphate (compound represented by formula (B)))
Use Geraniol is starting material synthesising target compound.In anhydrous methylene chloride, with pyridine and diacetyl oxide, acetylize is carried out to Geraniol, obtain acetic ester (compound represented by following formula (bi)) (yield 95%).Then, the carbon of the 8-position of this acetic ester is carried out selenolite in ethanol, obtain aldehyde body (compound represented by following formula (bii)) (yield 24%).Then, with potassium hydroxide, this aldehyde is carried out alkaline hydrolysis, form alcohol body (compound represented by following formula (biii)) (yield 38%).Then, in anhydrous methylene chloride, process this alcohol with imidazoles, tert-butyl diphenyl chlorosilane (TBDPS), obtain the compound (yield 80%) shown in following (biv).Then, in anhydrous ether, make this compound and butyllithium react, obtain butyl alcohol body (compound represented by following formula (bv)) (yield 73%).Then, in anhydrous tetrahydro furan, be sodium salt by this converting compounds, in this sodium salt, add methyl-iodide thereupon and by Williamson synthesis, obtain ether body (compound represented by following (bvi)) (yield 95%).Then, in anhydrous tetrahydro furan, use four positive Neutral ammonium fluorides that this ether is carried out eliminative reaction, obtain alcohol body (compound represented by following formula (bvii)) (yield 87%).Then, below-40 DEG C, in anhydrous methylene chloride solvent, with N-chloro bromosuccinimide and dimethyl sulphide, chlorine replacement is carried out to primary hydroxyl, obtain muriate (compound represented by following (bviii)) (yield 92%).Then, in anhydrous acetonitrile, with three (tetra-n-butyl ammonium) hydrogenation bisphosphate, diphosphate is carried out to this muriate, obtain target product (compound represented by following formula (bix), the compound namely represented by formula (B)) (yield 26%).
TLC and instrumental analysis (IR and NMR) is used to carry out the intermediate of each synthesis phase and the confirmation of final product.
(manufacturing embodiment 3)
(synthesis of 8-hydroxyl-3,7-dimethyl-ten two carbon-(2E, 6E)-dialkylene bisphosphate (compound represented by formula (C)))
Use Geraniol is starting material synthesising target compound.In anhydrous methylene chloride, with pyridine and diacetyl oxide, acetylize is carried out to Geraniol, obtain acetic ester (compound represented by following formula (ci)) (yield 97%).Then, the carbon of the 8-position of this acetic ester is carried out selenolite in ethanol, obtain aldehyde body (compound represented by following formula (cii)) (yield 20%).Then, with potassium hydroxide, this aldehyde is carried out alkaline hydrolysis, form alcohol body (compound represented by following formula (ciii)) (yield 42%).Then, in anhydrous methylene chloride, process this alcohol with imidazoles, tert-butyl diphenyl chlorosilane (TBDPS), obtain the compound (yield 80%) shown in following (civ).Then, in anhydrous diethyl ether, make this compound and butyllithium react, obtain butanols body (yield 62%).Then, in anhydrous tetrahydro furan, four positive Neutral ammonium fluorides are used this purely to be carried out eliminative reaction, acquisition glycol body (compound represented by following formula (cvi)) (yield 94%).Then, below-40 DEG C, in anhydrous methylene chloride solvent, with N-chloro bromosuccinimide and dimethyl sulphide, chlorine replacement is carried out to primary hydroxyl, obtain muriate (compound represented by following (cvii)) (yield 90%).Then, in anhydrous acetonitrile, with three (tetra-n-butyl ammonium) hydrogenation bisphosphate, diphosphate is carried out to this muriate, obtain target product (compound represented by following formula (cviii), the compound namely represented by formula (C)) (yield 46%).
TLC and instrumental analysis (IR and NMR) is used to carry out the intermediate of each synthesis phase and the confirmation of final product.
(preparation of mutant enzyme)
Use allyl group bisphosphate derivative as starting material, though when use wild-type there is the enzyme of prenyltransferases activity, also can carry out reaction and prepare isoprene oligopolymer.But, in order to improve reaction efficiency, prepare the variant enzyme improved the enzymatic activity of allyl group bisphosphate derivative.
The reagent used is from QuickChange site-directed mutagenesis kit (QuickChange Site-Directed Mutagenesis Kit) (Stratagene company).Design primer, to allow to introduce sudden change at target site.The primer of sudden change is purchased from company of Institute of Health on Nutriology (MEDICAL & BIOLOGICAL LABORATORIES CO., LTD) (manufacturer: IDT).The primer of design is as follows.
-prepare the primer of variant enzyme N77A
Forward primer 5'-act gaa gca tgg tct cgt cct aaa g-3'(SEQ ID NO:1)
Reverse primer 5'-gag acc atg ctt cag ttg aaa atg c-3'(SEQ ID NO:2)
-prepare the primer of variant enzyme L91D
Forward primer 5'-gat gaa aga tcc ggg tga ttt ttt aa-3'(SEQ ID NO:3)
Reverse primer 5'-cac ccg gat ctt tca tca agt aat ta-3'(SEQ ID NO:4)
DsDNA template uses the pET22b (pET22b/MLU-UPS) imported from the 11 prenyl diphosphate synthases (hereinafter also referred to as wild-type enzyme) of micrococcus luteus B-P26.This pET22b/MLU-UPS is by ancient mountain kind professor person of outstanding talent (the Prof.Tanetoshi Koyama of multi-component Science Institute of Northeastern University, Institute of Multidisciplinary Research for Advanced Materials, Tohoku University) provide.Mixing 10x Pfu polymerase buffer 2 μ L, dsDNA template 2-20ng, forward primer 50ng, reverse primer 50ng, dNTPs 0.4 μ L (often kind is 2.5mM), ddH
2o is totally 20 μ L, Pfu polysaccharases (2.5U/ μ l) 0.4mL, carries out PCR reaction.The condition of PCR reaction is as follows:
95 DEG C, 30 seconds, a circulation;
95 DEG C, 30 seconds/55 DEG C, 1 minute/68 DEG C, 8 minutes, 15 circulations.
After PCR, in PCR solution, add 0.4 μ L Dpn I, this PCR solution is carried out to the process of 1 hour with Dpn I at 37 DEG C.Use the Dpn I treatment solution of 1 to 10 μ L, by heat shock Transformed E .coli DH5 α.After on the LB nutrient agar this transformant being coated on the penbritin containing 50 μ g/mL, overnight incubation at 37 DEG C, then, selects transformant.Then, in the LB substratum of the penbritin containing 50 μ g/mL, cultivate this transformant to spend the night, by alkali SDS legal system prepared plasmid from the nutrient solution obtained.Sequenator is used to confirm the sudden change of this plasmid.
(there is the manufacture of the protein of prenyltransferases activity)
E.coli BL21 (the DE3)/pET22b/MLU-UPS (wild and saltant type) obtained is inoculated into and 3mL is housed contains in the test tube of the LB substratum of the penbritin of 50 μ g/mL, shaking culture 5 hours at 37 DEG C.In the nutrient solution obtained, get 1mL decile and be seeded in the 500mL Erlenmeyer flask of the LB substratum containing 50 μ g/mL penbritins that 100mL is housed, this cell of shaking culture 3 hours at 37 DEG C.Then, adding IPTG is 0.1mmol/L to its concentration, then this cell of shaking culture 18 hours at 30 DEG C.This nutrient solution of centrifugation, obtains wet thallus.After this wet thallus of ultrasonic wave homogenizing, centrifugation obtains supernatant liquor.HisTrap (Amersham company) purifying from this supernatant liquor is used to have the protein of prenyltransferases activity.Protein purification is confirmed by SDS-PAGE.The aminoacid sequence of thus obtained variant enzyme N77A and variant enzyme L91D is as shown in the SEQ ID NO:5 and 6 of sequence table.
(preparation of isoprene oligopolymer)
Preparation is containing protein 10mg, 50mM Tris-HCl damping fluid (pH 7.5) after purifying, 40mM magnesium chloride, 40mM Triton X-100,25mM 2 mercapto ethanol, 1mM starting material (farnesyl bisphosphate (FPP) or a kind of and 1mM IPP manufactured in the allyl group bisphosphate derivative of preparation in embodiment 2 and 3 or a kind of reaction soln manufactured in the R-IPP of preparation in embodiment 1.This reaction 1 hour is carried out in the water-bath of 37 DEG C.After reaction terminates, add saturated aqueous common salt 100ml and n-butyl alcohol 500ml, after stirring this mixture, left standstill.Then, extremely dry by evaporation concentration supernatant liquor (n-butyl alcohol layer).Get remnant parts and confirm product structure by NMR, it is isoprene oligopolymer.The particular case (n, m and R in formula (Z-2-1-1)) of the isoprene oligopolymer obtained is as shown in table 10.Herein, based on the information of the starting material used and the isoprene chain length by TLC mensuration, n and m in formula (Z-2-1-1) is calculated.In addition, the R structure in formula (Z-2-1-1) is identified by NMR, TLC and GC-MS.
In addition, with variant enzyme N77A as enzyme, using the compound represented by formula (B) as starting material, use monomer I PP respectively, R-IPP-A, R-IPP-B, R-IPP-C, R-IPP-D, R-IPP-E, the isoprene oligopolymer that R-IPP-F and R-IPP-G obtains, isoprene oligopolymer (tc-contrast-AB) is called in aftermentioned experiment uses, isoprene oligopolymer (tc-A-AB), isoprene oligopolymer (tc-B-AB), isoprene oligopolymer (tc-C-AB), isoprene oligopolymer (tc-D-AB), isoprene oligopolymer (tc-E-AB), isoprene oligopolymer (tc-F-AB) and isoprene oligopolymer (tc-G-AB).
In addition, with variant enzyme L91D as enzyme, using the compound represented by formula (B) as starting material, use monomer I PP respectively, R-IPP-A, R-IPP-B, R-IPP-C, R-IPP-D, R-IPP-E, the isoprene oligopolymer that R-IPP-F and R-IPP-G obtains, isoprene oligopolymer (tc-contrast-DB) is called in aftermentioned experiment uses, isoprene oligopolymer (tc-A-DB), isoprene oligopolymer (tc-B-DB), isoprene oligopolymer (tc-C-DB), isoprene oligopolymer (tc-D-DB), isoprene oligopolymer (tc-E-DB), isoprene oligopolymer (tc-F-DB) and isoprene oligopolymer (tc-G-DB).
In addition, with variant enzyme N77A as enzyme, using the compound represented by formula (C) as starting material, use monomer I PP respectively, R-IPP-A, R-IPP-B, R-IPP-C, R-IPP-D, R-IPP-E, the isoprene oligopolymer that R-IPP-F and R-IPP-G obtains, isoprene oligopolymer (tc-contrast-AC) is called in aftermentioned experiment uses, isoprene oligopolymer (tc-A-AC), isoprene oligopolymer (tc-B-AC), isoprene oligopolymer (tc-C-AC), isoprene oligopolymer (tc-D-AC), isoprene oligopolymer (tc-E-AC), isoprene oligopolymer (tc-F-AC) and isoprene oligopolymer (tc-G-AC).
In addition, with variant enzyme L91D as enzyme, using the compound represented by formula (C) as starting material, use monomer I PP respectively, R-IPP-A, R-IPP-B, R-IPP-C, R-IPP-D, R-IPP-E, the isoprene oligopolymer that R-IPP-F and R-IPP-G obtains, isoprene oligopolymer (tc-contrast-DC) is called in aftermentioned experiment uses, isoprene oligopolymer (tc-A-DC), isoprene oligopolymer (tc-B-DC), isoprene oligopolymer (tc-C-DC), isoprene oligopolymer (tc-D-DC), isoprene oligopolymer (tc-E-DC), isoprene oligopolymer (tc-F-DC) and isoprene oligopolymer (tc-G-DC).
In addition, in experiment, each isoprene oligopolymer (tc-E-AB), isoprene oligopolymer (tc-F-AB), isoprene oligopolymer (tc-E-DB) and isoprene oligopolymer (tc-F-DB) add in rubber combination respectively as isoprene oligopolymer c, isoprene oligopolymer f, isoprene oligopolymer d, isoprene oligopolymer g.
[table 10]
(relative reactivity of monomer)
The one making IPP or manufacture in the R-IPP of preparation in embodiment 1, the one in the allyl group bisphosphate derivative prepared in embodiment 2 and 3 with farnesyl bisphosphate (FPP) or manufacture is reacted under the following conditions.Respectively relative to the activity of FPP and IPP, by the relative reactivity of each allyl group bisphosphate derivative and each R-IPP with exponential representation, wherein, the activity of FPP and IPP is set to 100.
Preparation is containing protein 500ng, 50mM Tris-HCl damping fluid (pH7.5) after purifying, 40mM magnesium chloride, 40mM Triton X-100,25mM 2 mercapto ethanol, 12.5 μMs of FPP or the one manufactured in the allyl group bisphosphate derivative of preparation in embodiment 2 and 3 and 50 μMs of [1-
14c] IPP or a kind of reaction soln of manufacturing in the R-IPP of preparation in embodiment 1.This reaction 1 hour is carried out in the water-bath of 37 DEG C.After reaction, employing liquid scintillation counting(LSC) and TLC measure activity in each condition.Relative to the activity (=100) of IPP, the relative reactivity of allyl group bisphosphate derivative and R-IPP is as shown in table 11.
Isoprene oligopolymer adopts and carries out purifying with the same manner in embodiment 7 part (preparation of isoprene oligopolymer).Then, the particular case (n, m and R in formula (Z-2-1-1)) of the isoprene oligopolymer obtained is shown in table 11.Herein, based on the information of the starting material used and the isoprene chain length by TLC mensuration, n and m in formula (Z-2-1-1) is calculated.In addition, the R structure in formula (Z-2-1-1) is identified by NMR, TLC and GC-MS.
[table 11]
Table 10 and 11 result display, even if use R-IPP and allyl group bisphosphate derivative, the isoprene oligopolymer of acquisition also has identical molecular weight with during use IPP and FPP.Also demonstrate acquisition isoprene oligopolymer and there occurs the modification corresponding with the R-IPP used and allyl group bisphosphate derivative.
(embodiment 8)
(preparation of polyisoprene)
Then, use following isoprene oligopolymer to prepare polyisoprene: to use the preparation of allyl group bisphosphate derivative, such as formula represented by (Z-2-1-1), isoprene unit with trans-cis configuration link and not only also there is the isoprene oligopolymer (the isoprene oligopolymer of skeleton and chain end modification) of modification in skeleton but also chain end.
A kind of reaction soln in the R-IPP of preparation containing preparation in 10 μ L latex component, 50mM Tris-HCl damping fluid (pH 7.5), 25mM magnesium chloride, 40mM2-mercaptoethanol, 40mM Potassium monofluoride, 50 μMs of isoprene oligopolymer and 1mM IPP or manufacture embodiment 1.This reaction 3 days is carried out in the water-bath of 30 DEG C.After reaction, by GPC determining molecular weight.Then, based on the information of the numerical value of molecular weight and the starting material of use, the quantity of the isoprene unit added on starting material FPP or allyl group bisphosphate derivative is calculated.Result is as shown in table 12.
Latex component used herein is by carrying out ultracentrifugation and the glue obtained is clear to obtaining latex from Para rubber tree.
The isoprene oligopolymer used is isoprene oligopolymer (tc-contrast-AB) prepared in above-mentioned part (preparation of isoprene oligopolymer), isoprene oligopolymer (tc-A-AB), isoprene oligopolymer (tc-B-AB), isoprene oligopolymer (tc-C-AB), isoprene oligopolymer (tc-D-AB), isoprene oligopolymer (tc-E-AB), isoprene oligopolymer (tc-F-AB), isoprene oligopolymer (tc-G-AB), isoprene oligopolymer (tc-contrast-DB), isoprene oligopolymer (tc-A-DB), isoprene oligopolymer (tc-B-DB), isoprene oligopolymer (tc-C-DB), isoprene oligopolymer (tc-D-DB), isoprene oligopolymer (tc-E-DB), isoprene oligopolymer (tc-F-DB), isoprene oligopolymer (tc-G-DB), isoprene oligopolymer (tc-contrast-AC), isoprene oligopolymer (tc-A-AC), isoprene oligopolymer (tc-B-AC), isoprene oligopolymer (tc-C-AC), isoprene oligopolymer (tc-D-AC), isoprene oligopolymer (tc-E-AC), isoprene oligopolymer (tc-F-AC), isoprene oligopolymer (tc-G-AC), isoprene oligopolymer (tc-contrast-DC), isoprene oligopolymer (tc-A-DC), isoprene oligopolymer (tc-B-DC), isoprene oligopolymer (tc-C-DC), isoprene oligopolymer (tc-D-DC), isoprene oligopolymer (tc-E-DC), one in isoprene oligopolymer (tc-F-DC) and isoprene oligopolymer (tc-G-DC).
In aftermentioned experiment, each polyisoprene using isoprene oligopolymer (tc-E-AB), isoprene oligopolymer (tc-F-AB), isoprene oligopolymer (tc-E-DB), isoprene oligopolymer (tc-F-DB), isoprene oligopolymer (tc-E-AC) and isoprene oligopolymer (tc-E-DC) to manufacture, respectively as polyisoprene E, polyisoprene B, polyisoprene F, polyisoprene C, polyisoprene G, polyisoprene H, adds in rubber combination.
[table 12]
The result display of table 12, even if use the isoprene oligopolymer of R-IPP and skeleton and chain end modification, the polyisoprene of acquisition also has identical molecular weight with when use IPP and unmodified isoprene oligopolymer.Further, polyisoprene NMR, TLC and GC-MS of obtaining are analyzed, confirm with the situation of isoprene oligopolymer similar, polyisoprene has carried out the modification corresponding with the isoprene oligopolymer of used R-IPP and skeleton and chain end modification.
The chain end (corresponding to the Y of formula (Z-1), formula (Z-2), formula (ZZ-1) or formula (ZZ-2)) of the isoprene oligopolymer prepared in embodiment and polyisoprene is hydroxyl or OPP.
For embodiment 9 to 41 and comparative example 1 to 4 chemical reagent as follows listed by.
NR：TSR20
BR:BR01 (JSR company)
Carbon black: DIABLACK (N220) (Mitsubishi Chemical Ind)
Isoprene oligopolymer a to n: isoprene oligopolymer prepared in above-described embodiment
Polyisoprene A to P: polyisoprene prepared in above-described embodiment
Zinc oxide: zinc oxide #1 (mining company of Mitsui Metal Co., Ltd.)
Stearic acid: stearic acid (You company)
Protective agent: NOCRAC 6C (N-(1,3-dimethylbutyl)-N '-diphenyl-para-phenylene diamine) (the emerging chemical industrial company of imperial palace)
Wax: SUNNOC wax (the emerging chemical industrial company of imperial palace)
Sulphur: powder sulphur (He Jian chemical company)
Vulcanization accelerator NS: NOCCELER NS (N tert butyl benzothiazole 2 sulfenamide) (the emerging chemical industrial company of imperial palace)
Silicon-dioxide: Nipsil AQ (wet silica) (Japanese silicon-dioxide company)
Silane coupling agent: Si266 (two (3-triethoxysilylpropyltetrasulfide) disulphide) (goldschmidt chemical corporation)
Accelerator D PG:NOCCELER D (N, N-diphenylguanidine) (the emerging chemical industrial company of imperial palace)
According to the formula shown by table 13 to 16, use the material beyond the mixing sulphur of 1.7L banbury mixers and vulcanization accelerator, obtain mixing thing.Then, mix sulphur and vulcanization accelerator in this mixing thing, and undertaken mixing by open roll, obtain unvulcanized rubber composition.Use steam pressurized vulcanizer at pressure 80kgf/cm
2, sulfuration is carried out 30 minutes to this unvulcanized rubber composition at 150 DEG C, prepare vulcanizate compositions.
Following evaluation is carried out to the vulcanizate compositions prepared thus.Result is as shown in table 13 to 16.The benchmark formula of table 13 is the formula of comparative example 1, and the benchmark formula of table 14 is the formula of comparative example 2, and the benchmark formula of table 15 is the formula of comparative example 3, and the benchmark formula of table 16 is the formula of comparative example 4.
(test for viscoelastic)
Use visco-elasticity spectrograph (this making of rock institute) 70 DEG C, strain 2% time (initial stage elongation) condition carry out the mensuration of tan δ.With the tan δ of benchmark formula for 100, by its numerical value with exponential representation.Index is higher, and heat-dissipating is larger.Index be less than 100 formula be considered as improving heat-dissipating resistance (low-heat-generation).In other words, index is lower, and low-heat-generation is better.
(blue Berne wear test)
Use blue Berne wear testing machine (this making of rock institute) and under the condition of loading 3kg, slip rate 40% and Sha Su 15g/ minute, implement wear test.The shape of sample is thick 5mm, diameter 50mm.Use grinding stone is the GC abrasive grain of granularity #80.Be 100 (benchmark) with the tan δ of benchmark formula, by test-results with exponential representation.This index is higher, and wear resistance is better.The formula that index is greater than 100, is considered as improving wear resistance.
(tension test)
According to JIS K6251 " defining method of rubber, vulcanized rubber or thermoplastic elastomer-tensile stress and emergent property ", No. 3 dumbbell shape samples be made up of above-mentioned vulcanized rubber sheet are used to implement tension test, to measure fracture tensile strength (TB) (MPa), elongation at break (EB) (%).If when rubber combination elongation at break being less than 480% is used for outsize tire, easily produce rubbery chip, thus need improvement.In addition, because low fracture tensile strength can cause tire to destroy, the decline of the fracture tensile strength preventing from causing because of the change of material is therefore needed.
Table 13 and 15 displays, use the embodiment of isoprene oligopolymer of the present invention to show outstanding low-heat-generation, wear resistance and elongation at break.
Table 14 and 16 displays, use the embodiment of polyisoprene of the present invention to show outstanding low-heat-generation, wear resistance and fracture tensile strength.
(the independent text of sequence table)
SEQ ID NO:1: the forward primer preparing variant enzyme N77A
SEQ ID NO:2: the reverse primer preparing variant enzyme N77A
SEQ ID NO:3: the forward primer preparing variant enzyme L91D
SEQ ID NO:4: the reverse primer preparing variant enzyme L91D
SEQ ID NO:5: the aminoacid sequence of variant enzyme N77A
SEQ ID NO:6: the aminoacid sequence of variant enzyme L91D
Claims (19)
1. an isoprene oligopolymer, its allyl group bisphosphate represented by formula (X) and the compou nd synthesis represented by formula (Y):
In formula (X), n represents the integer of 1 to 10;
In formula (Y), R represents the group beyond methyl.
2. isoprene oligopolymer according to claim 1, the compound that its allyl group bisphosphate represented by described formula (X), described formula (Y) represent and prenyl diphosphate synthesis.
3. isoprene oligopolymer according to claim 1 and 2, wherein, the allyl group bisphosphate of the allyl group bisphosphate represented by described formula (X) represented by formula (X-1):
Wherein, the n in formula (X-1) represents the integer of 1 to 10, and
At least one atom contained in II part in formula (X-1) or atomic group are replaced by different atoms or atomic group, and atom contained in the III part in formula (X-1) or atomic group are not all replaced by different atoms or atomic group.
4. isoprene oligopolymer as claimed in any of claims 1 to 3, wherein, described synthesis uses the enzyme with prenyltransferases activity to carry out.
5. an isoprene oligopolymer, it is represented by formula (Z-1) or represented by formula (Z-2):
In formula (Z-1), n represents the integer of 1 to 10; M represents the integer of 1 to 30; Y represents hydroxyl, formyl radical, carboxyl, alkoxycarbonyl, alkoxy carbonyl or OPP;
In formula (Z-2), n represents the integer of 1 to 10; M represents the integer of 1 to 30; Y represents hydroxyl, formyl radical, carboxyl, alkoxycarbonyl, alkoxy carbonyl or OPP,
At least one atom contained in v part in described formula (Z-1) or described formula (Z-2) or atomic group are replaced by different atoms or atomic group.
6. isoprene oligopolymer according to claim 5, wherein, at least one atom contained in the iv part in described formula (Z-1) or described formula (Z-2) or atomic group are replaced by different atoms or atomic group.
7. manufacture a method for isoprene oligopolymer, it comprises the allyl group bisphosphate represented by formula (X) and the compound represented by formula (Y) to synthesize isoprene oligopolymer:
In formula (X), n represents the integer of 1 to 10;
In formula (Y), R represents the group beyond methyl.
8. the method for manufacture isoprene oligopolymer according to claim 7, wherein, the compound that it comprises allyl group bisphosphate represented by described formula (X), described formula (Y) represents and prenyl diphosphate are to synthesize isoprene oligopolymer.
9. the method for the manufacture isoprene oligopolymer according to claim 7 or 8, wherein, the allyl group bisphosphate of the allyl group bisphosphate represented by described formula (X) represented by formula (X-1):
In formula (X-1), n represents the integer of 1 to 10, and
At least one atom contained in II part in formula (X-1) or atomic group are replaced by different atoms or atomic group, and atom contained in the III part in formula (X-1) or atomic group are not all replaced by different atoms or atomic group.
10. according to the method for the manufacture isoprene oligopolymer in claim 7 to 9 described in any one, wherein, described synthesis uses the enzyme with prenyltransferases activity to carry out.
11. 1 kinds of polyisoprene, its allyl group bisphosphate represented by formula (X) and the compou nd synthesis represented by formula (Y):
In formula (X), n represents the integer of 1 to 10;
In formula (Y), R represents the group beyond methyl.
12. polyisoprene according to claim 11, the compound that its allyl group bisphosphate represented by described formula (X), described formula (Y) represent and prenyl diphosphate synthesis.
13. polyisoprene according to claim 11 or 12, wherein, the allyl group bisphosphate of the allyl group bisphosphate represented by described formula (X) represented by formula (X-1):
In formula (X-1), n represents the integer of 1 to 10, and
At least one atom contained in II part in formula (X-1) or atomic group are replaced by different atoms or atomic group, and atom contained in the III part in formula (X-1) or atomic group are not all replaced by different atoms or atomic group.
14. 1 kinds of polyisoprene, its compound represented by formula (Y) described in isoprene oligopolymer as claimed in any of claims 1 to 6, at least one and prenyl diphosphate synthesis.
15. 1 kinds of polyisoprene, it is represented by formula (ZZ-1) or represented by formula (ZZ-2):
In formula (ZZ-1), n represents the integer of 1 to 10; Q represents the integer of 30 to 40000; Y represents hydroxyl, formyl radical, carboxyl, alkoxycarbonyl, alkoxy carbonyl or OPP;
In formula (ZZ-2), n represents the integer of 1 to 10; Q represents the integer of 30 to 40000; Y represents hydroxyl, formyl radical, carboxyl, alkoxycarbonyl, alkoxy carbonyl or OPP,
At least one atom contained in v part in described formula (ZZ-1) or described formula (ZZ-2) or atomic group are replaced by different atoms or atomic group.
16. polyisoprene according to claim 15, wherein, at least one atom contained in the iv part in described formula (ZZ-1) or described formula (ZZ-2) or atomic group are replaced by different atoms or atomic group.
17. 1 kinds of methods manufacturing polyisoprene, it comprises compound represented by formula (Y) described in isoprene oligopolymer as claimed in any of claims 1 to 6, at least one and prenyl diphosphate carrys out synthetic polyisoprenes.
18. 1 kinds of rubber combinations, it is containing the isoprene oligopolymer in good grounds claim 1 ~ 6 described in any one with according at least one in the polyisoprene in claim 11 ~ 16 described in any one.
19. 1 kinds of pneumatic tyres, it is formed by rubber combination according to claim 18.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810340783.6A CN108484663B (en) | 2012-09-12 | 2013-09-02 | Isoprene oligomer and method for producing isoprene oligomer |
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2012-200884 | 2012-09-12 | ||
| JP2012200884 | 2012-09-12 | ||
| PCT/JP2013/073509 WO2014042027A1 (en) | 2012-09-12 | 2013-09-02 | Isoprene oligomer, polyisoprene, method for producing isoprene oligomer and polyisoprene, rubber composition, and pneumatic tire |
Related Child Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201810340783.6A Division CN108484663B (en) | 2012-09-12 | 2013-09-02 | Isoprene oligomer and method for producing isoprene oligomer |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN104684958A true CN104684958A (en) | 2015-06-03 |
| CN104684958B CN104684958B (en) | 2018-05-15 |
Family
ID=50278144
Family Applications (2)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201810340783.6A Active CN108484663B (en) | 2012-09-12 | 2013-09-02 | Isoprene oligomer and method for producing isoprene oligomer |
| CN201380045290.5A Active CN104684958B (en) | 2012-09-12 | 2013-09-02 | Isoprene oligomer, polyisoprene, method, rubber composition and the pneumatic tire for manufacturing isoprene oligomer and polyisoprene |
Family Applications Before (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201810340783.6A Active CN108484663B (en) | 2012-09-12 | 2013-09-02 | Isoprene oligomer and method for producing isoprene oligomer |
Country Status (5)
| Country | Link |
|---|---|
| JP (2) | JP6468512B2 (en) |
| CN (2) | CN108484663B (en) |
| BR (1) | BR112015004224A2 (en) |
| MY (1) | MY172203A (en) |
| WO (1) | WO2014042027A1 (en) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP6531304B2 (en) * | 2014-05-28 | 2019-06-19 | 住友ゴム工業株式会社 | Isoprene oligomer, polyisoprene, method for producing the same, rubber composition, and pneumatic tire |
| JP7209718B2 (en) * | 2018-06-20 | 2023-01-20 | エルジー・ケム・リミテッド | Modified polymerization initiator and method for producing the same |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101688191A (en) * | 2007-04-03 | 2010-03-31 | 菲力普莫里斯产品股份公司 | Genes encoding z,z-farnesyl diphosphate synthase and a sesquiterpene synthase with multiple products and uses thereof |
| WO2012008298A1 (en) * | 2010-07-14 | 2012-01-19 | 住友ゴム工業株式会社 | Isoprene oligomer, polyisoprene, processes for producing these materials, rubber composition, and pneumatic tire |
Family Cites Families (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2714402B2 (en) * | 1988-07-13 | 1998-02-16 | 日清製粉株式会社 | Cancer metastasis inhibitor |
| JP2960183B2 (en) * | 1991-03-12 | 1999-10-06 | エーザイ株式会社 | Novel terpene derivative and method for producing the same |
| JP3888397B2 (en) * | 1996-01-31 | 2007-02-28 | 株式会社日清製粉グループ本社 | Isoprene derivatives |
| JPH10316715A (en) * | 1997-05-20 | 1998-12-02 | Jsr Corp | Aqueous dispersion of epoxy-modified polymer |
| KR100302100B1 (en) * | 1999-05-03 | 2001-11-22 | 박찬구 | Recombinant Microorganisms Expressing Small Rubber Particle-Associated Protein |
| JP2003238603A (en) * | 2002-02-20 | 2003-08-27 | Hiroshi Okamoto | Modified rubber |
| JP4197706B2 (en) * | 2005-06-07 | 2008-12-17 | ユニケミカル株式会社 | Solid polymer electrolyte membrane obtained by copolymerizing phosphoric acid group-containing unsaturated monomer and unsaturated liquid oligomer, and fuel cell using the same |
| JP5035871B2 (en) * | 2005-09-16 | 2012-09-26 | 株式会社ブリヂストン | Gene group of prenyltransferase of Para rubber tree |
| JP5645366B2 (en) * | 2009-02-25 | 2014-12-24 | 日立造船株式会社 | Long chain trans prenyl diphosphate synthase gene |
| JP2011089066A (en) * | 2009-10-23 | 2011-05-06 | Bridgestone Corp | Tire |
-
2013
- 2013-09-02 JP JP2014535489A patent/JP6468512B2/en active Active
- 2013-09-02 BR BR112015004224A patent/BR112015004224A2/en not_active Application Discontinuation
- 2013-09-02 CN CN201810340783.6A patent/CN108484663B/en active Active
- 2013-09-02 MY MYPI2015700468A patent/MY172203A/en unknown
- 2013-09-02 CN CN201380045290.5A patent/CN104684958B/en active Active
- 2013-09-02 WO PCT/JP2013/073509 patent/WO2014042027A1/en active Application Filing
-
2019
- 2019-01-07 JP JP2019000505A patent/JP6757938B2/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101688191A (en) * | 2007-04-03 | 2010-03-31 | 菲力普莫里斯产品股份公司 | Genes encoding z,z-farnesyl diphosphate synthase and a sesquiterpene synthase with multiple products and uses thereof |
| WO2012008298A1 (en) * | 2010-07-14 | 2012-01-19 | 住友ゴム工業株式会社 | Isoprene oligomer, polyisoprene, processes for producing these materials, rubber composition, and pneumatic tire |
Non-Patent Citations (2)
| Title |
|---|
| MASAHIKO NAGAKI ET AL.: "Artificial substrates for undecaprenyl diphosphate synthase from Micrococcus luteus B-P 26", 《JOURNAL OF MOLECULAR CATALYSIS B:ENZYMATIC》 * |
| MASAHIKO NAGAKI ET AL.: "Substrate specificities of medium-prenylchain elongating enzymes, hexaprenyl- and heptaprenyl diphosphate synthases", 《JOURNAL OF MOLECULAR CATALYSIS B:ENZYMATIC》 * |
Also Published As
| Publication number | Publication date |
|---|---|
| JPWO2014042027A1 (en) | 2016-08-18 |
| CN104684958B (en) | 2018-05-15 |
| BR112015004224A2 (en) | 2017-08-08 |
| JP2019090030A (en) | 2019-06-13 |
| MY172203A (en) | 2019-11-15 |
| JP6757938B2 (en) | 2020-09-23 |
| JP6468512B2 (en) | 2019-02-13 |
| CN108484663B (en) | 2020-09-18 |
| CN108484663A (en) | 2018-09-04 |
| WO2014042027A1 (en) | 2014-03-20 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| JP5058332B2 (en) | Isoprene oligomer, polyisoprene, and production methods thereof, rubber composition, and pneumatic tire | |
| CN102770418B (en) | The vinyl modifier composition improved and the technique utilizing said composition | |
| US20100190885A1 (en) | Tire with rubber component containing silica and use of combination of blocked and unblocked alkoxyorganomercaptosilane coupling agents | |
| JP5574562B2 (en) | Rubber composition for tire and pneumatic tire using the same | |
| CN102796292B (en) | High-elongation wear-resistant oil-resistant rubber sole and preparation method thereof | |
| CN105175801B (en) | Truck tyre or bus tire | |
| JPH09118785A (en) | Elastomeric composition useful as tire tread | |
| US6465560B1 (en) | Tire with tread of spatially defined elastomer composition | |
| CN109071708A (en) | Polymer-modified manufacturing method, polymer-modified, rubber composition and tire | |
| CN102731856A (en) | Butadiene rubber and chloroprene rubber mixed rubber sealing ring | |
| JP6757938B2 (en) | Isoprene oligomers, polyisoprene, and methods of making them, rubber compositions, and pneumatic tires. | |
| CN107903458A (en) | One kind filling environment-friendly type white carbon styrene-butadiene rubbers for tread rubber and preparation method | |
| JP5992749B2 (en) | Rubber composition for tire and pneumatic tire | |
| JP5670970B2 (en) | Rubber composition for tire and pneumatic tire | |
| WO2018116726A1 (en) | Method for producing trans-polyisoprenoid, vector, transgenic plant, method for producing pneumatic tire and method for producing rubber product | |
| EP2966098B1 (en) | Modified conjugated diene polymer, method for producing same, and rubber composition using same | |
| CA2233823A1 (en) | Hydrosilated crystalline high trans polybutadiene with silica filler, tires having a base of said composition and method of preparing same | |
| CN109384972B (en) | Rubber composition for shoe sole, vulcanized rubber, and preparation method and application thereof | |
| CN115286862B (en) | Soft light sole material and preparation method thereof | |
| JP6531304B2 (en) | Isoprene oligomer, polyisoprene, method for producing the same, rubber composition, and pneumatic tire | |
| CN102443217A (en) | Inner tube of vehicle tire | |
| US10245888B2 (en) | Rubber composition for tires and pneumatic tire | |
| CN109384975B (en) | Rubber composition for shoe sole, vulcanized rubber, and preparation method and application thereof | |
| WO2021177074A1 (en) | Method for producing polyisoprenoid, vector, transformed plant, method for producing pneumatic tire, and method for producing rubber product | |
| CN116333384B (en) | Wear-resistant heat-resistant high-elastic rubber outsole |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |