CN104684958B - Isoprene oligomer, polyisoprene, method, rubber composition and the pneumatic tire for manufacturing isoprene oligomer and polyisoprene - Google Patents
Isoprene oligomer, polyisoprene, method, rubber composition and the pneumatic tire for manufacturing isoprene oligomer and polyisoprene Download PDFInfo
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- CN104684958B CN104684958B CN201380045290.5A CN201380045290A CN104684958B CN 104684958 B CN104684958 B CN 104684958B CN 201380045290 A CN201380045290 A CN 201380045290A CN 104684958 B CN104684958 B CN 104684958B
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- polyisoprene
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- isoprene oligomer
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- RRHGJUQNOFWUDK-UHFFFAOYSA-N Isoprene Chemical compound CC(=C)C=C RRHGJUQNOFWUDK-UHFFFAOYSA-N 0.000 title claims abstract description 752
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- 239000005060 rubber Substances 0.000 title claims abstract description 116
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- 238000004519 manufacturing process Methods 0.000 title claims description 55
- -1 method Polymers 0.000 title claims description 43
- 238000000034 method Methods 0.000 title claims description 41
- 150000001875 compounds Chemical class 0.000 claims abstract description 105
- 230000015572 biosynthetic process Effects 0.000 claims abstract description 51
- 238000003786 synthesis reaction Methods 0.000 claims abstract description 36
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 claims abstract description 25
- 125000004429 atom Chemical group 0.000 claims description 98
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- CBIDRCWHNCKSTO-UHFFFAOYSA-N prenyl diphosphate Chemical compound CC(C)=CCO[P@](O)(=O)OP(O)(O)=O CBIDRCWHNCKSTO-UHFFFAOYSA-N 0.000 claims description 74
- 230000000694 effects Effects 0.000 claims description 56
- 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 33
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- 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
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- 125000000999 tert-butyl group Chemical group [H]C([H])([H])C(*)(C([H])([H])[H])C([H])([H])[H] 0.000 description 2
- 210000004885 white matter Anatomy 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
- DSAYAFZWRDYBQY-UHFFFAOYSA-N 2,5-dimethylhexa-1,5-diene Chemical group CC(=C)CCC(C)=C DSAYAFZWRDYBQY-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
- OSDWBNJEKMUWAV-UHFFFAOYSA-N Allyl chloride Chemical compound ClCC=C OSDWBNJEKMUWAV-UHFFFAOYSA-N 0.000 description 1
- 239000001729 Ammonium fumarate Substances 0.000 description 1
- 229920000856 Amylose Polymers 0.000 description 1
- 241000219194 Arabidopsis Species 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
- OMPJBNCRMGITSC-UHFFFAOYSA-N Benzoylperoxide Chemical group C=1C=CC=CC=1C(=O)OOC(=O)C1=CC=CC=C1 OMPJBNCRMGITSC-UHFFFAOYSA-N 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-JGWLITMVSA-N D-glucitol Chemical compound OC[C@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-JGWLITMVSA-N 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 240000008168 Ficus benjamina 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
- UPZUXIGZPJBAPH-UHFFFAOYSA-N OP(O)(=O)OP(=O)(O)O.C=CC=CC Chemical compound OP(O)(=O)OP(=O)(O)O.C=CC=CC UPZUXIGZPJBAPH-UHFFFAOYSA-N 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- 229930182555 Penicillin Natural products 0.000 description 1
- JGSARLDLIJGVTE-MBNYWOFBSA-N Penicillin G Chemical compound N([C@H]1[C@H]2SC([C@@H](N2C1=O)C(O)=O)(C)C)C(=O)CC1=CC=CC=C1 JGSARLDLIJGVTE-MBNYWOFBSA-N 0.000 description 1
- 239000001888 Peptone Substances 0.000 description 1
- 108010080698 Peptones Proteins 0.000 description 1
- 229920005683 SIBR Polymers 0.000 description 1
- 101710165017 Short-chain Z-isoprenyl diphosphate synthase Proteins 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
- 240000001949 Taraxacum officinale Species 0.000 description 1
- 235000006754 Taraxacum officinale Nutrition 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
- GCNLQHANGFOQKY-UHFFFAOYSA-N [C+4].[O-2].[O-2].[Ti+4] Chemical compound [C+4].[O-2].[O-2].[Ti+4] GCNLQHANGFOQKY-UHFFFAOYSA-N 0.000 description 1
- DWOPFQQGLMXOKL-BYPYZUCNSA-N [C@H]1(CC=C1)P(OP(=O)O)(=O)O Chemical compound [C@H]1(CC=C1)P(OP(=O)O)(=O)O DWOPFQQGLMXOKL-BYPYZUCNSA-N 0.000 description 1
- CCQOIBGABQRPJK-UHFFFAOYSA-N [hydroxy(methyl)phosphoryl]oxy-methylphosphinic acid Chemical compound CP(O)(=O)OP(C)(O)=O CCQOIBGABQRPJK-UHFFFAOYSA-N 0.000 description 1
- 239000006061 abrasive grain Substances 0.000 description 1
- 238000001042 affinity chromatography Methods 0.000 description 1
- 150000001298 alcohols Chemical class 0.000 description 1
- 150000001335 aliphatic alkanes Chemical class 0.000 description 1
- 235000019270 ammonium chloride Nutrition 0.000 description 1
- 235000019297 ammonium fumarate Nutrition 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
- 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
- 210000000988 bone and bone Anatomy 0.000 description 1
- 230000003139 buffering effect Effects 0.000 description 1
- MTAZNLWOLGHBHU-UHFFFAOYSA-N butadiene-styrene rubber Chemical compound C=CC=C.C=CC1=CC=CC=C1 MTAZNLWOLGHBHU-UHFFFAOYSA-N 0.000 description 1
- 229910000019 calcium carbonate Inorganic materials 0.000 description 1
- 229940041514 candida albicans extract Drugs 0.000 description 1
- 150000001732 carboxylic acid derivatives Chemical class 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 238000005660 chlorination reaction Methods 0.000 description 1
- 239000004927 clay Substances 0.000 description 1
- 239000005515 coenzyme Substances 0.000 description 1
- 238000013329 compounding Methods 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
- 229920003244 diene elastomer Polymers 0.000 description 1
- 238000010494 dissociation reaction Methods 0.000 description 1
- 230000005593 dissociations Effects 0.000 description 1
- 239000012154 double-distilled water Substances 0.000 description 1
- 239000000428 dust Substances 0.000 description 1
- 238000005516 engineering process Methods 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
- 239000004744 fabric Substances 0.000 description 1
- 229940043259 farnesol Drugs 0.000 description 1
- 229930002886 farnesol Natural products 0.000 description 1
- 238000011049 filling Methods 0.000 description 1
- 239000000706 filtrate Substances 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 235000019253 formic acid Nutrition 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
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 description 1
- 239000010931 gold Substances 0.000 description 1
- 229910052737 gold Inorganic materials 0.000 description 1
- 150000004795 grignard reagents Chemical class 0.000 description 1
- 238000000227 grinding Methods 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 230000020169 heat generation 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
- 239000003112 inhibitor Substances 0.000 description 1
- 229910001410 inorganic ion Inorganic materials 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
- 238000011068 loading method Methods 0.000 description 1
- 238000012423 maintenance Methods 0.000 description 1
- 235000013372 meat Nutrition 0.000 description 1
- 229910052751 metal Inorganic materials 0.000 description 1
- STEPQTYSZVCJPV-UHFFFAOYSA-N metazachlor Chemical compound CC1=CC=CC(C)=C1N(C(=O)CCl)CN1N=CC=C1 STEPQTYSZVCJPV-UHFFFAOYSA-N 0.000 description 1
- 125000001434 methanylylidene group Chemical group [H]C#[*] 0.000 description 1
- 125000000325 methylidene group Chemical group [H]C([H])=* 0.000 description 1
- SNVLJLYUUXKWOJ-UHFFFAOYSA-N methylidenecarbene Chemical group C=[C] SNVLJLYUUXKWOJ-UHFFFAOYSA-N 0.000 description 1
- GRVDJDISBSALJP-UHFFFAOYSA-N methyloxidanyl Chemical group [O]C GRVDJDISBSALJP-UHFFFAOYSA-N 0.000 description 1
- 125000002816 methylsulfanyl group Chemical group [H]C([H])([H])S[*] 0.000 description 1
- 238000005065 mining Methods 0.000 description 1
- 238000001668 nucleic acid synthesis Methods 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 150000007524 organic acids Chemical class 0.000 description 1
- 239000005416 organic matter Substances 0.000 description 1
- 230000001151 other effect Effects 0.000 description 1
- VDGJOQCBCPGFFD-UHFFFAOYSA-N oxygen(2-) silicon(4+) titanium(4+) Chemical compound [Si+4].[O-2].[O-2].[Ti+4] VDGJOQCBCPGFFD-UHFFFAOYSA-N 0.000 description 1
- 239000012188 paraffin wax Substances 0.000 description 1
- 229940049954 penicillin Drugs 0.000 description 1
- PMJHHCWVYXUKFD-UHFFFAOYSA-N pentadiene group Chemical group C=CC=CC PMJHHCWVYXUKFD-UHFFFAOYSA-N 0.000 description 1
- 235000019319 peptone Nutrition 0.000 description 1
- 238000005373 pervaporation Methods 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 1
- 239000010452 phosphate Substances 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
- 150000003013 phosphoric acid derivatives Chemical class 0.000 description 1
- 125000004437 phosphorous atom Chemical group 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
- 230000000704 physical effect Effects 0.000 description 1
- 229920001084 poly(chloroprene) Polymers 0.000 description 1
- 238000006116 polymerization reaction Methods 0.000 description 1
- 239000011698 potassium fluoride Substances 0.000 description 1
- 239000004323 potassium nitrate Substances 0.000 description 1
- 235000010333 potassium nitrate Nutrition 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 125000001844 prenyl group Chemical group [H]C([*])([H])C([H])=C(C([H])([H])[H])C([H])([H])[H] 0.000 description 1
- 235000019260 propionic acid Nutrition 0.000 description 1
- IUVKMZGDUIUOCP-BTNSXGMBSA-N quinbolone Chemical compound O([C@H]1CC[C@H]2[C@H]3[C@@H]([C@]4(C=CC(=O)C=C4CC3)C)CC[C@@]21C)C1=CCCC1 IUVKMZGDUIUOCP-BTNSXGMBSA-N 0.000 description 1
- 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
- 239000005720 sucrose Substances 0.000 description 1
- BBDNZMUIQBRBJH-UHFFFAOYSA-N sulfurochloridic acid;toluene Chemical compound OS(Cl)(=O)=O.CC1=CC=CC=C1 BBDNZMUIQBRBJH-UHFFFAOYSA-N 0.000 description 1
- 230000002194 synthesizing effect Effects 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
- 238000012549 training Methods 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
- 238000012546 transfer Methods 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 class CCO[Si](OCC)(OCC)CCCSSSSCCC[Si](OCC)(OCC)OCC VTHOKNTVYKTUPI-UHFFFAOYSA-N 0.000 description 1
- WVLBCYQITXONBZ-UHFFFAOYSA-N trimethyl phosphate Chemical compound COP(=O)(OC)OC WVLBCYQITXONBZ-UHFFFAOYSA-N 0.000 description 1
- 238000002525 ultrasonication Methods 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
- 229940088594 vitamin Drugs 0.000 description 1
- 229930003231 vitamin Natural products 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)
- General Health & Medical Sciences (AREA)
- Molecular Biology (AREA)
- Biochemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Mechanical Engineering (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 object of the present invention is to provide the isoprene oligomer and polyisoprene that a kind of molecular skeleton is modified.The present invention also aims to provide a kind of rubber composition comprising the isoprene oligomer and/or the polyisoprene, and one kind includes the pneumatic tire that tyre element (for example, tyre surface or sidewall) is formed by the rubber composition.The present invention relates to a kind of isoprene oligomer, the compound synthesis represented by its pi-allyl diphosphonic acid and following formula (Y) represented by lower formula (X).In formula (X), n represents 1 to 10 integer.In formula (Y), R represents the group beyond methyl.
Description
Technical field
The present invention relates to isoprene oligomer, polyisoprene, manufacture isoprene oligomer and polyisoprene
Method, the composition containing isoprene oligomer and/or polyisoprene rubber and filled by what the rubber composition was formed
Air filled tyre.
Background technology
For a long time, people be directed to providing rubber originally with property outside rubber also with new property produce
Product, according to purposes, import filler of different shapes or other components that different materials make in rubber composition, needed for acquisition
The property wanted.For example, in automobile tire, people are attempted by importing silica or carbon black etc. in the rubber phase of organic matter
Filler, to improve the properties such as wearability, low-heat-generation or wet grip performance.
It is conventionally used in mixed fillers etc. in the rubber phases such as rubber composition, modified rubber (modified dienopolymer)
Method, to increase the compatibility between these components, further improves low-heat-generation, wet grip performance etc..These modified rubbers
By making the rubber molecule in rubber phase be reacted with the compound for example containing nitrogen-containing group, chlorine sulfenyl, and in rubber point
The functional group (for example, referenced patent document 1 and 2) that there is compatibility with filler is introduced in son.
Also, it is known that isoprene oligomer and polyisoprene have the diphosphonic acid of high reaction activity in its chain end
Base etc..Therefore, the method for modifying of traditional isoprene oligomer or polyisoprene is related to above-mentioned high reaction activity end
Diphosphonic acid base and the reaction of the compound containing functional group.As described above, traditionally isoprene oligomer and poly- isoamyl two
Alkene is mainly modified in the end of molecule.On the other hand, in conventional methods where, by high reaction activity end diphosphonic acid base or
The skeleton of the influence of its analog, isoprene oligomer and polyisoprene cannot be fully modified.In addition, traditionally,
Modification to natural rubber is only capable of by carrying out chemistry to the latex collected from Para rubber tree (Hevea brasiliensis)
Handle to complete.It is thus impossible to by analysis be confirmed whether strand it is actual be modified and substitute have it is what kind of
Functional group.
Citation list
Patent document
Patent document 1：JP 2000-001573A
Patent document 2：JP 2000-001575A
The content of the invention
Technical problem
Present invention aim to address the above problem, and a kind of isoprene oligomer of molecular skeleton modification is provided and is gathered
Isoprene.The present invention also aims to provide a kind of rubber comprising the isoprene oligomer and/or the polyisoprene
Glue composition, and provide a kind of comprising the inflated wheel that tyre element (for example, tyre surface or sidewall) is formed by the rubber composition
Tire.
Solution to the problem
The present invention relates to a kind of isoprene oligomer, its pi-allyl diphosphonic acid and formula (Y) institute table represented by formula (X)
The compound synthesis shown：
Wherein, n represents 1 to 10 integer；
Wherein, R represents the group beyond methyl.
It is preferred that what pi-allyl diphosphonic acid of the isoprene oligomer represented by the formula (X), the formula (Y) represented
Compound and prenyl diphosphate synthesis.
It is preferred that pi-allyl diphosphonic acid of the pi-allyl diphosphonic acid represented by formula (X-1) represented by the formula (X)：
Wherein, n represents 1 to 10 integer, and at least one atom or atom contained in the II parts in formula (X-1)
Group is substituted by other (i.e. different) atoms or atomic group, and the atom or atomic group contained by the III parts in formula (X-1) are not
Substituted by other atoms or atomic group.
The synthesis is preferably carried out by using the enzyme with prenyltransferases activity.
The invention further relates to the isoprene oligomer represented by a kind of formula (Z-1) or represented by formula (Z-2), formula (Z-1)
Or contained at least one atom or atomic group is substituted by other atoms or atomic group in the v parts in formula (Z-2).
Wherein, the n in formula (Z-1) represents 1 to 10 integer；M represents 1 to 30 integer；Y represents hydroxyl, formoxyl, carboxylic
Base, alkoxycarbonyl, alkoxy carbonyl or OPP；
Wherein, the n in formula (Z-2) represents 1 to 10 integer；M represents 1 to 30 integer；Y represents hydroxyl, formoxyl, carboxylic
Base, alkoxycarbonyl, alkoxy carbonyl or OPP.
Contained at least one atom or atomic group are by other atoms in iv parts in preferred formula (Z-1) or formula (Z-2)
Or atomic group substitution.
The invention further relates to a kind of method for manufacturing isoprene oligomer, it includes the pi-allyl represented by formula (X)
Compound synthesis isoprene oligomer represented by diphosphonic acid and formula (Y)：
Wherein, n represents 1 to 10 integer；
Wherein, R represents the group beyond methyl.
In the method for manufacture isoprene oligomer, preferably described isoprene oligomer is represented by the formula (X)
Pi-allyl diphosphonic acid, the formula (Y) represent compound and prenyl diphosphate synthesis.
It is preferred that pi-allyl diphosphonic acid of the pi-allyl diphosphonic acid represented by formula (X-1) represented by the formula (X)：
Wherein, n represents 1 to 10 integer, and at least one atom or atom contained in the II parts in formula (X-1)
Group is substituted by other atoms or atomic group, and contained atom or atomic group be not by other atoms in the III parts in formula (X-1)
Or atomic group substitution.
The synthesis is preferably carried out by using the enzyme with prenyltransferases activity.
The invention further relates to a kind of polyisoprene, its pi-allyl diphosphonic acid and formula (Y) represented by formula (X) are represented
Compound synthesis：
Wherein, n represents 1 to 10 integer；
Wherein, R represents the group beyond methyl.
It is preferred that pi-allyl diphosphonic acid, the chemical combination of the formula (Y) expression of the polyisoprene represented by the formula (X)
Thing and prenyl diphosphate synthesis.
It is preferred that pi-allyl diphosphonic acid of the pi-allyl diphosphonic acid represented by formula (X-1) represented by the formula (X)：
Wherein, n represents 1 to 10 integer, and at least one atom or atom contained in the II parts in formula (X-1)
Group is substituted by other atoms or atomic group, and contained atom or atomic group be not by other atoms in the III parts in formula (X-1)
Or atomic group substitution.
The invention further relates to a kind of polyisoprene, it is represented by the isoprene oligomer, at least one formula (Y)
Compound and prenyl diphosphate synthesis.
The invention further relates to a kind of formula (ZZ-1) is represented or formula (ZZ-2) represented by polyisoprene, formula (ZZ-1) or
Contained at least one atom or atomic group are substituted by other atoms or atomic group in v parts in formula (ZZ-2),
Wherein, the n in formula (ZZ-1) represents 1 to 10 integer；Q represents 30 to 40000 integer；Y represents hydroxyl, formyl
Base, carboxyl, alkoxycarbonyl, alkoxy carbonyl or OPP；
Wherein, the n in formula (ZZ-2) represents 1 to 10 integer；Q represents 30 to 40000 integer；Y represents hydroxyl, formyl
Base, carboxyl, alkoxycarbonyl, alkoxy carbonyl or OPP.
Contained at least one atom or atomic group are former by other in iv parts in preferred formula (ZZ-1) or formula (ZZ-2)
Son or atomic group substitution.
The invention further relates to a kind of method for manufacturing polyisoprene, it is included by the isoprene oligomer, at least
Compound and prenyl diphosphate represented by a kind of formula (Y) synthesize the polyisoprene.
The invention further relates to a kind of rubber composition, and it includes in the isoprene oligomer and the polyisoprene
At least one.
The present invention relates to a kind of pneumatic tire formed by the rubber composition.
Invention Beneficial Effect
The isoprene oligomer of the present invention is the pi-allyl diphosphonic acid represented by formula (X) and the change represented by formula (Y)
The isoprene oligomer of compound synthesis, or it is the isoprene oligomer by formula (Z-1) or formula (Z-2) expression, wherein,
Contained at least one atom or atomic group are substituted by other atoms or atomic group in v parts in formula (Z-1) or formula (Z-2).
In addition, the polyisoprene of the present invention is by the pi-allyl diphosphonic acid represented by formula (X) and the conjunction of the compound represented by formula (Y)
Into polyisoprene；Compound and/or iso-amylene either represented by above-mentioned isoprene oligomer, a kind of formula (Y)
The polyisoprene of base diphosphonic acid synthesis；Polyisoprene either represented by the formula (ZZ-1) or represented by formula (ZZ-2),
Contained at least one atom or atomic group are taken by other atoms or atomic group in v parts in formula (ZZ-1) or formula (ZZ-2)
Generation.Therefore, the molecule (rubber molecule) of isoprene oligomer of the invention and the polyisoprene of the present invention is respectively provided with modification
Skeleton, thus its there is outstanding compatibility with filler (such as silica).Therefore, it is if the isoprene of the present invention is low
Polymers and/or the polyisoprene of the present invention are used for rubber composition, and the rubber molecule obtained in rubber composition can be with filling out
Material height mixes.For example, the rubber composition therefore provided has outstanding low-heat-generation and wearability.Further,
If for example, in the tyre element (for example, tyre surface or sidewall) that above-mentioned rubber composition is used for, can provide with outstanding
The pneumatic tire of low-heat-generation and wearability.
In addition, the method for manufacture isoprene oligomer and the method for manufacturing polyisoprene of the present invention of the present invention,
The impregnable skeleton of diphosphonic acid base fully modified isoprene oligomer in high reaction activity end can be respectively provided and gathered different
Pentadiene.
Further, because isoprene oligomer provided by the present invention and polyisoprene are represented using formula (Y)
Compound obtain, add functional group or other groups with known structure in the compound represented by the formula (Y), can
To positively identify the functional group (modified group) in isoprene oligomer or polyisoprene.
Embodiment
In artificial synthesized (biosynthesis) of rubber molecule (polyisoprene), enzyme can be used (for example, isoprene
Based transferase) act on starting material (for example, farnesyl diphosphonic acid (FPP)) and monomer (for example, prenyl diphosphate)
In mixture, producing the addition polymerization on starting material has the isoprene oligomer of 1 to 30 isoprene unit.Then,
These latex components of isoprene oligomer further with the enzyme containing addition polymerizable prenyl diphosphate are mixed.
It is known the result is that being formed in the polyisoprene that many polyisoprene units are connected with oligomer.
As described above, form rubber molecule must use native enzyme in the step of continuously link monomer on starting material
The addition polymerization of catalysis.
This mean that the starting material of the synthesis of rubber molecule (polyisoprene) and monomer need be can use enzyme into
The material of row catalytic reaction.Therefore, the starting material of rubber molecule (polyisoprene) material and the structure of monomer be may be used as
It is restricted.Particularly because the limit caused by the enzyme of prenyltransferases and energy addition polymerization prenyl diphosphate
System, monomer are defined as naturally occurring prenyl diphosphate.
Its result cause the structure design flexibility of the rubber molecule (polyisoprene) of artificial synthesized (biosynthesis) by
Limitation, hence in so that being difficult to provide the flexibility in enough MOLECULE DESIGNs, carrys out the performance not available for additional natural rubber.
For these reasons, the rubber molecule (polyisoprene) of functional group or its analog is imported with order to prepare,
For example, traditionally, as described above, people for example, by by the end diphosphonic acid group of high reaction activity or its analog with containing
The compound reaction of functional group, to cause modification.Especially, the modification to natural rubber is only capable of by from Para rubber tree
The latex that (Hevea brasiliensis) is collected is chemically treated to complete.It is thus impossible to carrying out confirmation by analysis is
No strand is actual, which to be modified and substitute, what kind of functional group.
Conversely, the present invention is had found based on following：By using the prenyl diphosphate of partially modified structure as monomer,
Isoprene oligomer or polyisoprene are manufactured, the isoprene oligomer with functionalization skeleton or poly- isoamyl can be manufactured
Diene.Further, because isoprene oligomer provided by the present invention and polyisoprene use prenyl diphosphate
Obtain, add functional group or other groups with known structure in the prenyl diphosphate, can positively identify
The functional group (modified group) in isoprene oligomer or polyisoprene.
Especially, the present invention is had found based on following：If maintain the structure of naturally occurring monomer prenyl diphosphate
In 3- positions methyl beyond structure it is constant, can also or even during by required group to substitute positioned at 3- methyl
Using the monomer prenyltransferases of enzyme or naturally occurring energy addition are generated by being used as naturally occurring oligomer
It polymerize the polymer generation enzyme of prenyl diphosphate, manufactures isoprene oligomer or polyisoprene.Its reason is not complete
Portion is verified, but the enzyme that can be presumably due to prenyltransferases and energy addition polymerization prenyl diphosphate is attracted to
In structure outside the methyl of the 3- positions of prenyl diphosphate monomer, while these enzymes are relatively unwise to the structure of 3- methyl
Sense.
Based on these discoveries, can provide skeleton has the isoprene oligomer and polyisoprene of required property,
Therefore isoprene oligomer and polyisoprene with additional functions can be provided.
More particularly, in naturally occurring biosynthesis, for example, prenyl diphosphate (IPP) can continuously be gathered
Close on dimethylallyl diphosphate (DMAPP), thus produce geranyl diphosphonic acid (GPP), farnesyl diphosphonic acid (FPP),
Geranyl geranyl diphosphate (GGPP), geranyl farnesyl diphosphonic acid (GFPP), it is as follows.
Conversely, the present invention uses the R-IPP obtained with the methyl of the 3- positions of required group (- R) substitution IPP, with
Instead of prenyl diphosphate (IPP).In this case, continuously R-IPP can be aggregated on dimethylallyl diphosphate,
Thus geranyl diphosphonic acid (R-GPP), farnesyl diphosphonic acid (R-FPP), the framework modification of framework modification of framework modification are produced
Geranyl geranyl diphosphate (R-GGPP), the geranyl farnesyl diphosphonic acid (R-GFPP) of framework modification, it is as follows.
Further, if R-IPP and IPP used at the same time, in other words, and with R-IPP and IPP, by R-IPP and IPP
Continuously it is aggregated on dimethylallyl diphosphate, the geranyl diphosphonic acid (R- of framework modification as follows can be produced
GPP), geranyl geranyl diphosphate (R-GGPP), the skeleton of the farnesyl diphosphonic acid (R-FPP) of framework modification, framework modification
Modified geranyl farnesyl diphosphonic acid (R-GFPP).Therefore, tool can be manufactured with the ratio after adjusting and with R-IPP and IPP
The isoprene oligomer of property is needed.
Herein, the modification of the skeleton of molecule (rubber molecule) means with required functional group substitution molecule (rubber point
Son) structure (especially, with the 3- positions corresponding part of methyl of IPP) in IPP in skeleton, or mean to use other
The structure in IPP in structure (structure beyond methyl) substitution molecule (rubber molecule) skeleton is (especially, with IPP's
The 3- corresponding parts of methyl).
(isoprene oligomer)
A kind of pi-allyl diphosphonic acid of oligomer represented by formula (X) and the compound (R- represented by formula (Y) of the present invention
IPP) (biosynthesis) is synthesized：
Wherein, n represents 1 to 10 integer；
Wherein, R represents the group beyond methyl.
Herein, term " OPP (OPP groups) " means diphosphonic acid group (group represented by following formula (A-1)), and it has
There are 3 hydroxyls being bonded with phosphorus atoms.When its in aqueous when, dissociation occurs for some or all of hydroxyl (for example, OPP becomes
For the group as represented by following formula (A-2)).The intension of term " OPP " herein includes above-mentioned partly or entirely hydroxyl and solves
From group.
The isoprene oligomer of the present invention has a structure close with natural rubber, and with rubber molecule height phase
Hold.In addition, because the isoprene oligomer of the present invention has modified skeletal, it can send out with filler (for example, silica)
Raw strong interaction.Therefore, because the isoprene oligomer of the present invention and rubber highly compatible, while can be with filler
Strong interaction occurs for (for example, silica), if the isoprene oligomer is used for rubber composition, can obtain phase
The rubber composition mixed than traditional composition, its rubber molecule with filler higher degree.For example, thus the rubber composition may be used
To strengthen low-heat-generation, wet grip performance, wearability, elongation at break and fracture tensile strength.
The skeleton of the isoprene oligomer of the present invention includes polar group or its analog.Thus, filler is (for example, two
Silica) situation of dispersion performance when being only present in chain end higher than polar group or its analog.Thus, for example, low life
Hot, wet grip performance, wearability, the increase of the improvement of elongation at break and fracture tensile strength.
The isoprene oligomer of the present invention also has outstanding antimicrobial acivity.Speculate this is because the present invention's is different
Pentadiene oligomer has the structure different from naturally occurring usually isoprene oligomer, therefore, have and for example suppress thin
The enzyme and coenzyme of bacterium, suppress nucleic acid synthesis, suppress cell membrane synthesis, the synthesis for suppressing cytoplasma membrane, infringement cell membrane and infringement
Cytoplasma membrane and other effects.
First, the pi-allyl diphosphonic acid represented by formula (X) is illustrated：
Wherein, n represents 1 to 10 integer.
In formula (X), n represents 1 to 10 integer (being preferably 1 to 4, more preferably 1 to 3).
The example of pi-allyl diphosphonic acid represented by formula (X) includes：Dimethylallyl diphosphate (DMAPP), geranyl
Diphosphonic acid (GPP), farnesyl diphosphonic acid (FPP), geranyl geranyl diphosphate (GGPP), geranyl farnesyl diphosphonic acid
(GFPP).Wherein, preferably DMAPP, GPP and FPP, because they can be as the bottom of polytype prenyltransferases
Thing.
In the pi-allyl diphosphonic acid represented by formula (X), diphosphonic acid group is bonded with isoprene unit.Formula (X) institute table
The pi-allyl diphosphonic acid shown can be derived by being changed the pi-allyl diphosphonic acid of (modification) to part isoprene unit
Thing.
The present inventor just uses pi-allyl diphosphonic acid derivative to manufacture isoprene oligomer or poly- different as starting material
The method of pentadiene proposed patent application (JP2012-036360A).The patent application is summarized as follows.
As described above, it is that can be carried out with enzyme that the starting material and monomer of the synthesis of rubber molecule (polyisoprene), which need,
The material of catalytic reaction.Which has limited the starting material of material and the knot of monomer that may be used as rubber molecule (polyisoprene)
Structure.Particularly because limitation caused by the enzyme of manufacture oligomer, starting material are restricted to naturally occurring dimethallyl
Base diphosphonic acid, geranyl diphosphonic acid, farnesyl diphosphonic acid, geranyl geranyl diphosphate and the like.
On the contrary, the present inventor is through widely studied discovery, by using by such as structure by partially modified two phosphorus of farnesyl
Acid etc. is used as starting material, manufactures isoprene oligomer or polyisoprene, can manufacture the chain end with functionalization
Isoprene oligomer or polyisoprene.
Particularly, the inventors discovered that, if maintaining the following formula in the starting materials such as naturally occurring farnesyl diphosphonic acid
(I) structure of the I parts in, the other parts beyond I parts introduce required structure, can also use and be used as day
So the prenyltransferases of existing oligomer generation enzyme manufacture isoprene oligomer.Its reason is not visited all
It is bright, but can be presumably due in the structure for the I parts that prenyltransferases are attracted in the formula (I) of starting material,
Structure relative insensitivity of these enzymes to other parts at the same time.
Found based on these, the isoprene oligomer and poly- isoamyl two of the chain end with required property can be provided
Alkene, it is, therefore, possible to provide while the script property of isoprene oligomer or polyisoprene is not weakened, be attached with it is a variety of
The isoprene oligomer and polyisoprene of function.
Therefore, in the present invention, by using the prenyltransferases of naturally occurring oligomer generation enzyme, by formula
(Y) compound of (R-IPP) is aggregated on starting material, which is while the I part-structures of the formula that maintains (I)
The pi-allyl diphosphonic acid for the formula (X) being modified, can obtain and be not only skeleton and chain end is (from pi-allyl diphosphonic acid
Structure division) the isoprene oligomer that is also modified.
In particular it is preferred to pi-allyl two phosphorus of the pi-allyl diphosphonic acid represented by formula (X-1) represented by the above-mentioned formula (X)
Acid：
Wherein, n represent 1 to 10 integer (n in formula (X-1) by with it is same in formula (X) in a manner of be defined), and
Contained at least one atom or atomic group are substituted by other atoms or atomic group in II parts in formula (X-1), in formula (X-1)
III parts in contained atom or atomic group do not substituted by other atoms or atomic group.
The result is that a kind of not only isoprene oligomer with modified skeletal, also with modified chain end is manufactured.
Because for the isoprene oligomer not only with modified skeletal, also with modified chain end, it can be with filler (for example, dioxy
SiClx) preferably interact, if the isoprene oligomer is used for rubber composition, the building rubber compound obtained
Rubber molecule in thing can compare traditional composition, it can carry out higher degree with filler and mix.For example, rubber composition by
This can provide the low-heat-generation further enhanced, wet grip performance, wearability, elongation at break and fracture tensile strength.
Herein, the modification of the end of the molecule (rubber molecule) is it is meant that molecule (rubber molecule) end comes from formula
(X) the predetermined part of the pi-allyl diphosphonic acid structure represented by is substituted by required functional group, or means
Tied from the predetermined part of the pi-allyl diphosphonic acid structure represented by formula (X) by other molecule (rubber molecule) end
Structure substitutes.
Contained atom or atomic group (that is, atom or original before substitution in II parts or III parts in formula (X-1)
Son group) example include hydrogen atom, methyl, methylene, carbon atom and methine.
The example of other atoms includes nitrogen-atoms, oxygen atom, sulphur atom, silicon atom and carbon atom.Wherein, preferably nitrogen is former
Son, because it is with strong intermolecular force, therefore can have strong interaction with enzyme or cell membrane, also with filler (for example,
Silica) there is strong interaction.
Other described atomic groups can be nitrogen-containing group, oxy radical, sulfur-containing group, silicon-containing group, carbon-containing group or its
Analog.Its example includes：Acetoxyl group, alkoxy (preferably C1-C3 alkoxies, more preferably methoxyl group), hydroxyl, aryl are (excellent
Select phenyl), alkyl (preferably C1-C5 alkyl, more preferably ethyl and the tert-butyl group), acetyl group, N- alkyl-acetylamino (wherein, alkane
Base preferably has 1 to 5 carbon atom) and azido.
From the angle of anti-microbe ability, preferably N- alkyl-acetylamino (more preferably N- methyl-acetamidos and
N- butyl-acetylamino) and azido, because nitrogen-atoms has strong intermolecular force, therefore can be with enzyme or cell membrane
Strong interaction occurs.Because its strong interaction with filler (for example, silica), it is also preferred that alkoxy and hydroxyl
Base.
The example for the pi-allyl diphosphonic acid (pi-allyl diphosphonic acid derivative) that part isoprene unit is modified includes following
Compound represented by formula (A) to (S).Wherein, the compound of preferred formula (B), (C), (D), (E), (F), (K), (L) and (R),
More preferably formula (B) and the compound of (C), because they have the stronger and interaction of filler (for example, silica), and
And it can more effectively improve low-heat-generation, wet grip performance, wearability, elongation at break and fracture tensile strength.Based on excellent
Elegant anti-microbe ability, it is also preferred that the structure of the structure of the formula (G) to (Q), more preferably formula (K), (L) and (Q).
Technical staff can according to the method disclosed in JP2012-036360A, for example, from dimethylallyl diphosphate,
Geranyl diphosphonic acid, farnesyl diphosphonic acid, geranyl geranyl diphosphate, geraniol, farnesol, trans-Geranylgeraniol or its class
Like thing, the pi-allyl diphosphonic acid derivative represented by formula (A) to (S).
Then, the compound (R-IPP) represented by formula (Y) is illustrated：
Wherein, R represents the group beyond methyl.
In formula (Y), R can be any group beyond methyl.Its example include nitrogen-containing group, oxy radical, sulfur-containing group,
Silicon-containing group and carbon-containing group (in addition to methyl).Preference includes acetoxyl group, alkoxy, hydroxyl, aryl, alkyl and (removes methyl
Outside), acetyl group, N- acetyl group-acetylamino, azido, amino and sulfydryl.Wherein, preferably nitrogen-containing group, oxy radical, contain
Methylthio group, silicon-containing group and carbon-containing group (in addition to methyl), more preferably alkyl (especially, the alkyl of C2-C6), aryl are (especially
Ground, phenyl), sulfydryl, hydroxyl and amino, further preferred sulfydryl and hydroxyl, the reason is that they can more effectively improve it is low
Heat generation, wet grip performance, elongation at break and fracture tensile strength.
Technical staff can be according to the method illustrated in such as embodiment, the compound represented by formula (Y).
The addition isoprene on the pi-allyl diphosphonic acid starting material of formula (X) in the isoprene oligomer of the present invention
The quantity of unit preferably 1 to 30, more preferably 1 to 20, further preferred 1 to 10.
For example, the concrete example of the isoprene oligomer of the present invention is formula (Z-1) or the isoprene represented by formula (Z-2)
Contained at least one atom or atomic group are by other atoms or atom in v parts in oligomer, formula (Z-1) or formula (Z-2)
Group's substitution.
Wherein, the n in formula (Z-1) represents 1 to 10 integer；M represents 1 to 30 integer；Y represents hydroxyl, formoxyl, carboxylic
Base, alkoxycarbonyl, alkoxy carbonyl or OPP；
Wherein, the n in formula (Z-2) represents 1 to 10 integer；M represents 1 to 30 integer；Y represents hydroxyl, formoxyl, carboxylic
Base, alkoxycarbonyl, alkoxy carbonyl or OPP.
Contained atom or atomic group and other described atoms or atomic group in v parts in formula (Z-1) or formula (Z-2)
Can be the atom or atomic group as described in formula (X-1).
Iv parts in formula (Z-1) and (Z-2) are the knots from the pi-allyl diphosphonic acid starting material represented by formula (X)
Structure.Therefore, the n in formula (Z-1) and (Z-2) is defined in a manner of identical with the n in formula (X).
Pi-allyl diphosphonic acid starting material of the v parts represented by adding to formula (X) in formula (Z-1) and (Z-2) it is different
Pentadiene unit forms.Therefore, the m in formula (Z-1) and (Z-2) is originated with the above-mentioned pi-allyl diphosphonic acid for adding to formula (X)
The quantity of the isoprene unit of substrate is corresponding.
Strong interaction due to outstanding antimicrobial properties and with filler (for example, silica), formula (Z-1)
The preferred OPP of Y, hydroxyl or carboxyl in (Z-2).
As described above, the present invention is had found based on following：If maintain the methyl of the 3- positions in prenyl diphosphate structure
Structure in addition is constant, even if the methyl of 3- is substituted by required group, can also use it by naturally occurring enzyme,
Manufacture isoprene oligomer or polyisoprene.Therefore, formula (Z-1) or isoprene oligomer represented by formula (Z-2) are excellent
Elect that formula (Z-1-1) is represented or the isoprene oligomer as represented by formula (Z-2-1) as：
Wherein, n, m and Y in formula (Z-1-1) are defined in a manner of identical with n, m and Y in formula (Z-1), R with
Defined with the same way of the R in formula (Y)；
Wherein, n, m and Y in formula (Z-2-1) are defined in a manner of identical with n, m and Y in formula (Z-2), R with
Defined with the same way of the R in formula (Y)；
As described above, the it is also preferred that material that skeleton and chain end are modified at the same time.It is therefore preferable that formula (Z-1) or formula (Z-2)
At least one atom or atomic group contained by iv parts in (preferred formula (Z-1-1) or formula (Z-2-1)) is by other atoms or original
The substitution of son group.
In iv parts in formula (Z-1), formula (Z-2), formula (Z-1-1) or formula (Z-2-1) contained atom or atomic group with
And other described atoms or atomic group can be as described in formulas (X-1).
Further, as described above, it is preferred to which the pi-allyl diphosphonic acid represented by formula (X) is the allyl represented by formula (X-1)
Base diphosphonic acid, and at least one atom or atomic group contained in the II parts in formula (X-1) are by other atoms or atomic group
Substitute, contained atom or atomic group are not substituted by other atoms or atomic group in the III parts in formula (X-1).Therefore, close
Iv parts in the formula (Z-1) as represented by following formula (Z), formula (Z-2), formula (Z-1-1) and formula (Z-2-1), in preferred formula (Z)
Iv-II parts in contained at least one atom or atomic group substituted by other atoms or atomic group, in formula (Z)
Contained atom or atomic group are not substituted by other atoms or atomic group in iv-III parts.
In iv-II parts or iv-III parts in formula (Z) contained atom or atomic group and other described atoms or
Atomic group can be the atom or atomic group as described in formula (X-1).
The concrete example for the iv parts that chain end is also modified can include the structure as represented by following formula (a) to (s).Its
In, preferred formula (b), (c), (d), (e), (f), (k), the structure of (l) and (r), the more preferably structure of formula (b) and (c), because it
Have stronger with the interaction of filler (for example, silica), and can more effectively improve low-heat-generation, wet grab
Ground performance, wearability, elongation at break and fracture tensile strength.Due to outstanding anti-microbe ability, it is also preferred that formula (g) is extremely
(q) structure of structure, more preferably formula (k), (l) and (q).
(method of manufacture isoprene oligomer)
Pi-allyl diphosphonic acid of the isoprene oligomer represented by formula (X) and the chemical combination represented by formula (Y) of the present invention
Thing is synthesized (biosynthesis).
Compound synthesis (biosynthesis) as the pi-allyl diphosphonic acid represented by formula (X) and represented by formula (Y)
One example of the method for isoprene oligomer of the present invention, includes the use of the enzyme with prenyltransferases activity.It is special
Not, in the presence of the enzyme with prenyltransferases activity, pi-allyl diphosphonic acid represented by formula (X) with can be in alkene
Compound reaction represented by (Y) of the monomer polymerizeing in propyl group diphosphonic acid formula.Pi-allyl diphosphonic acid and formula represented by formula (X)
(Y) compound represented by can be the composition of single compound or multiple compounds respectively.It is, for example, possible to use formula
(Y) composition of the multiple compounds of formula (Y) different R in is low to provide the isoprene with required property
Polymers.
In the present invention, it can be only formula (Y) (R-IPP) to polymerize in the monomer on the pi-allyl diphosphonic acid represented by formula (X)
Represented compound.In this case, can be with the isoprene oligomer shown in fabrication scheme 2.Alternatively, polymerization is in formula (X)
Monomer on represented pi-allyl diphosphonic acid can be the compound and prenyl diphosphate represented by formula (Y) (R-IPP)
(IPP) combination.In this case, can be with the isoprene oligomer shown in fabrication scheme 3.
Herein, term means with catalysis allyl substrate (pi-allyl two " with prenyltransferases activity "
Phosphoric acid) and the condensation reaction of prenyl diphosphate to synthesize the new pi-allyl diphosphonic acid of the isoprene unit with addition
Active enzyme, therefore, can be catalyzed by prenyl diphosphate continuously with allyl substrate (pi-allyl diphosphonic acid) link
Reaction.
As described above, when the methyl for substituting the 3- positions in prenyl diphosphate structure with required group and remain different
The structure in addition to the methyl of 3- of pentenyl diphosphonic acid is constant and obtains the compound of formula (Y) (R-IPP), can be by making
It is oligomeric that isoprene is manufactured with the compound and using such as naturally occurring enzyme of enzyme with prenyltransferases activity
Thing.
As the enzyme with prenyltransferases activity, it has been found that polytype enzyme.
The enzyme (isoprene unit of each new addition has cis-configuration) of Z- isoprene chains, which can be extended, to be included：
Nine prenyl diphosphate synthases of Z- (Ishii, K.et al., (1986) Biochem., J., 233,773.), 11 is different
Amylene diphosphate synthase (Takahashi, I. and Ogura, K., (1982) J.Biochem., 92,1527.；Keenman,
M.V. and Allen, C.M., (1974) Arch.Biochem.Biophys., 161,375.), Z- farnesyl diphosphate synthases
(Identification of a short(C-15)chain Z-isoprenyl diphosphate synthase and a
homologous long(C-50)chain isoprenyl diphosphate synthase in Mycobacterium
Tuberculosis (short chain (C-15) farnesyl diphosphate synthase and homologous long-chain (C-50) isoamyl two in tubercle bacillus
The identification of alkene diphosphate synthase), Schulbach, MC., et al.JOURNAL OF BIOLOGICAL CHEMISTRY, 275
(30), 22876-22881 (2000)), and the long alcohol radical diphosphate synthase of dehydrogenation ((Identification of human
The knowledge of the long alcohol radical diphosphate synthase gene of dehydrodolichyl diphosphate synthase gene mankind's dehydrogenations
Not), Endo, Shota.et al, Biochimica et Biophysica Acta (BBA), 1625 (3), (2003) p.291-
295.)。
Furthermore it is possible to extend the enzyme of E- isoprene chains (isoprene unit of each new addition has anti-configuration)
Including：It is farnesyl diphosphate synthase, geranyl geranyl diphosphate synzyme, six isoprene diphosphate synthases, seven different
Pentadiene diphosphate synthase, eight isoprene diphosphate synthases, ten isoprene diphosphate synthases.
(anti-configuration is cis for the maximum number for the isoprene unit that can be formed and the direction of isoprene chain elongation
Configuration) depend on specific enzyme.Therefore, can according to the target numbers and isoprene chain elongation direction of isoprene unit come
Select enzyme.In the present invention, the isoprene chain of isoprene oligomer upper in any direction can extend that (anti-configuration is cis
Configuration).In other words, for example, the isoprene oligomer of the present invention can be that all isoprene units are connected with anti-configuration
The isoprene oligomer (for example, isoprene oligomer of formula (Z-1)) of knot, isoprene unit are with trans-cis configuration
The isoprene oligomer (for example, isoprene oligomer of formula (Z-2)) or isoprene unit of link are with trans-suitable
The isoprene oligomer that formula-anti-configuration links.
Tellurian existing every kind of biology is provided with the enzyme with prenyltransferases activity.Possess and carry isoamyl
For example, micrococcus luteus B-P26 (Micrococcus luteus B- for the biology of the enzyme of dialkylene transferase active
P26), Escherichia coli (Escherichia coli), saccharomyces cerevisiae (Saccharomyces cerevisiae), arabidopsis
(Arabidopsis thaliana), Para rubber tree (Hevea brasiliensis), periploca sepium (Periploca
Sepium), bacillus stearothermophilus (Bachillus Stearothermophilus), sulfolobus acidocaldarius
(Sulfolobus acidocaldarius (ATCC49426)), homo sapiens (Homo sapiens), sonchus oleraceus (Sonchus
Oleracers L.), Western dandelion (Taraxacum officinale) and sunflower (Helianthus annuus).
The script substrate (starting material) of enzyme with prenyltransferases activity is pi-allyl diphosphonic acid.Then,
Because pi-allyl diphosphonic acid derivative inhibitor as the enzyme originally, under many circumstances, if by pi-allyl diphosphonic acid
Derivative is used as starting material, relatively low (especially, the formula of enzymatic activity which shows on the pi-allyl diphosphonic acid derivative
Compound represented by (G) to (Q)).For these reasons, when using feelings of the pi-allyl diphosphonic acid derivative as starting material
Under condition, the variant enzyme that enzymatic activity improves can be used for pi-allyl diphosphonic acid derivative., can in the case of using variant enzyme
To prepare by technique for gene engineering the biology (transformant) that converts, to express variant enzyme.Especially, technical staff can root
According to the method disclosed in JP2012-036360A, the variation for the enzymatic activity for improving pi-allyl diphosphonic acid derivative is easily prepared
Enzyme.
The isoprene oligomer of the present invention can pass through in the presence of the enzyme with prenyltransferases activity
Pi-allyl diphosphonic acid represented by formula (X) reacts and prepares with the compound represented by formula (Y).
Herein, term means that there are following substances " in the presence of the enzyme with prenyltransferases activity "
In the case of：
Above-mentioned biological culture, the separated organism from the culture, the processed material of the organism, or from institute
State culture or the enzyme of organism purifying；Or
For expressing (turning and the biology converted by technique for gene engineering for the enzyme with prenyltransferases activity
Beggar) culture, the separated organism from the culture, the processed material of the organism, or from the described culture
Or the enzyme of organism purifying；Or its analog.
Technical staff can be by known technique for gene engineering, and preparing to be used to express has isoprene group-transfer
The enzyme of enzymatic activity and the biology being converted.
The biology, can be by being cultivated acquisition by above-mentioned biological organism in appropriate culture medium.These culture mediums
As long as can make the biology breed, be not particularly limited, can be containing common carbon source, nitrogen source, inorganic ions and
The organic nutrition source being added as needed on is generally incubated base.
Can be the arbitrary carbon source that above-mentioned biology can utilize for example, as carbon source.Its specific example, can make
With the carbohydrate of glucose, fructose, maltose, amylose and sucrose etc., the alcohols of D-sorbite, ethanol, glycerine etc., rich horse
The carbohydrate of the organic acid of acid, citric acid, acetic acid, propionic acid etc. and their salt, paraffin etc., and theirs are mixed
Compound.
As the example of nitrogen source, include having for the inorganic ammonium salt of ammonium sulfate, ammonium chloride etc., ammonium fumarate, ammonium citrate etc.
The organonitrogen compound of the nitrate of the ammonium salt of machine acid, sodium nitrate, potassium nitrate etc., peptone, yeast extract, meat extract, corn pulp etc.,
And their mixture.
Can suitably it mix and using the other nutrient sources being generally incubated used in base, for example, inorganic salts, micro gold
Belong to salt, vitamins, hormone etc..
Condition of culture is also not particularly limited.For example, under aerobic conditions, can be in the model that suitable control pH is 5 to 8
Enclose, in the case that temperature is 10 to 60 DEG C of scope, carry out 12 to 480 it is small when or so culture.
The biological culture can be for example, by the biology of above-mentioned condition of culture culture and the training of acquisition
Nutrient solution, or biological (organism) is separated and the culturing filtrate (culture supernatant) of acquisition from the nutrient solution by filtering etc..This
Outside, separated organism can be for example, being obtained by filtering or centrifuging etc. to separate from nutrient solution from above-mentioned culture
The organism (biology) obtained.
The processed material of the organism can be for example, will be obtained from above-mentioned culture after separated organism homogenizing processing
The organism homogenate obtained, or the organism for biology obtain after ultrasonication are homogenized.
The enzyme purified from the culture or the organism is for example, to existing in the culture or the organism
Enzyme saltoutd, enzyme obtained from purification process known to ion-exchange chromatography, affinity chromatography or gel filtration chromatography etc..It is pure
The purity for changing enzyme is not particularly limited.
The isoprene oligomer of the present invention can pass through in the presence of the enzyme with prenyltransferases activity
Pi-allyl diphosphonic acid represented by formula (X) reacts and prepares with the compound represented by formula (Y).Specifically, for example, this is anti-
Should be by adding the life in the solution containing the compound represented by the pi-allyl diphosphonic acid represented by formula (X) and formula (Y)
The culture of thing, the enzyme of purifying or its analog carry out.In addition, reaction temperature can be such as 20 to 60 DEG C, the reaction time can
During being such as 1 to 16 small, pH can be such as 5 to 8.In addition it is also possible to as needed, addition magnesium chloride, surfactant,
2 mercapto ethanol and other materials.
In the isoprene oligomer of the present invention manufactured by the reaction of described above, formula (Z-1), formula (Z-2), formula
(Z-1-1), the Y in formula (Z-2-1) is usually OPP or hydroxyl.OPP comes from IPP or R-IPP.OPP can also be hydrolyzed easily, OPP's
Hydrolysis produces hydroxyl.Here it is why the Y in formula (Z-1), (Z-2), (Z-1-1) or (Z-2-1) is usually OPP or hydroxyl.
In addition, the Y in formula (Z-1) or (Z-2) is the isoprene oligomer of formoxyl, for example, can pass through oxidation-type
(Z-1) or the Y in (Z-2) manufactures for the isoprene oligomer of OPP.
In addition, the Y in formula (Z-1) or (Z-2) is the isoprene oligomer of carboxyl, for example, can pass through oxidation-type (Z-
1) or the Y in (Z-2) manufactures for the isoprene oligomer of OPP.
In addition, Y in formula (Z-1) or (Z-2) is in the isoprene oligomer of alkoxycarbonyl, formula (Z-1) or (Z-2)
Y be alkoxy carbonyl group isoprene oligomer, for example, the Y in method described above carboxylation formula (Z-1) or (Z-2) can be passed through
For OPP isoprene oligomer and be further esterified the isoprene oligomer after the carboxylation and manufacture.
Organic synthesis and the pi-allyl diphosphonic acid derivative that can select as needed except the compound represented by formula (Y)
Starting material outside, isoprene oligomer of the invention can be manufactured by biosynthesis.Therefore, the present invention account for stone
The exhaustion of oily resource and environmental problem.
(polyisoprene)
Then, the polyisoprene of the present invention is illustrated.Alkene of the polyisoprene of the present invention represented by formula (X)
Compound represented by propyl group diphosphonic acid and formula (Y) is synthesized (biosynthesis).It polymerize in the pi-allyl two represented by formula (X)
Monomer on phosphoric acid can be the combination of the compound and prenyl diphosphate represented by formula (Y).For example, the present invention's is poly- different
Pentadiene can be by the compound and/or prenyl diphosphate represented by the isoprene oligomer of the present invention and formula (Y)
Synthesized (biosynthesis).For example, the polyisoprene of the present invention can be by unmodified isoprene oligomer and formula
(Y) compound represented by is synthesized (biosynthesis), or represented by unmodified isoprene oligomer, formula (Y)
Compound and prenyl diphosphate synthesized (biosynthesis).
The polyisoprene of the present invention have the structure close with natural rubber and with rubber molecule highly compatible.
The polyisoprene of the present invention also has modified molecular skeleton, therefore it can occur by force with filler (for example, silica)
Interaction.Therefore, because the polyisoprene of the present invention and rubber highly compatible, while can be with filler (for example, dioxy
SiClx) strong interaction occurs, if the polyisoprene is used for rubber composition, can obtain compared to traditional composition,
The rubber composition that its rubber molecule is mixed with filler higher degree.For example, the rubber composition is it is possible thereby to strengthen low-heat-generation
Property, wet grip performance, wearability, elongation at break and fracture tensile strength.
The skeleton of the polyisoprene of the present invention includes polar group or its analog.Thus, filler is (for example, titanium dioxide
Silicon) dispersion performance be higher than situation of the polar group (for example, end diphosphonic acid group) when being only present in chain end.Therefore,
For example, low-heat-generation, wet grip performance, wearability, the increase of the improvement of elongation at break and fracture tensile strength.
In addition, the pi-allyl diphosphonic acid represented by the formula (X) is the pi-allyl diphosphonic acid represented by formula (X-1), and formula
(X-1) contained at least one atom or atomic group are substituted by other atoms or atomic group in the II parts in, in formula (X-1)
In the case that contained atom or atomic group are not substituted by other atoms or atomic group in III parts, not only bone can be obtained
The polyisoprene that frame and chain end are also modified.Because the polyisoprene incessantly there is modified skeleton also to have
There is modified end, it can preferably interact with filler (for example, silica), if by the polyisoprene
For rubber composition, it can obtain compared to traditional composition, the building rubber compound that its rubber molecule is mixed with filler higher degree
Thing.For example, rubber composition is it is possible thereby to provide the low-heat-generation further enhanced, wet grip performance, wearability, extension at break
Rate and fracture tensile strength.
The isoprene list of addition on the pi-allyl diphosphonic acid starting material of formula (X) in the polyisoprene of the present invention
Member quantity preferably 30 to 40000, more preferably 31 to 38000, further preferred 1000 to 35000, particularly preferred 2000 to
30000。
For example, the specific example of the polyisoprene of the present invention is poly- represented by formula (ZZ-1) or represented by formula (ZZ-2)
In v parts in isoprene, formula (ZZ-1) or formula (ZZ-2) contained at least one atom or atomic group by other atoms or
Atomic group substitutes；
Wherein, the n in formula (ZZ-1) represents 1 to 10 integer；Q represents 30 to 40000 integer；Y represents hydroxyl, formyl
Base, carboxyl, alkoxycarbonyl, alkoxy carbonyl or OPP；
Wherein, the n in formula (ZZ-2) represents 1 to 10 integer；Q represents 30 to 40000 integer；Y represents hydroxyl, formyl
Base, carboxyl, alkoxycarbonyl, alkoxy carbonyl or OPP.
Contained atom or atomic group and other described atoms or atom in v parts in formula (ZZ-1) or formula (ZZ-2)
Group can be as described in formula (X-1).
Iv parts in formula (ZZ-1) and (ZZ-2) are the knots from the pi-allyl diphosphonic acid starting material represented by formula (X)
Structure.Therefore, the n in formula (ZZ-1) and (ZZ-2) is defined in a manner of identical with the n in formula (X).
Pi-allyl diphosphonic acid starting material of the v parts represented by adding to formula (X) in formula (ZZ-1) and (ZZ-2)
Isoprene unit forms.Therefore, pi-allyl diphosphonic acid starting bottoms of the q in formula (ZZ-1) and (ZZ-2) with adding to formula (X)
The quantity of the isoprene unit of thing is corresponding.
Due to the strong interaction with filler (for example, silica), preferred OPP of Y in formula (ZZ-1) and (ZZ-2),
Hydroxyl or carboxyl.
As described above, the present invention is had found based on following：If maintain the first of the 3- positions in the structure of prenyl diphosphate
Structure beyond base is constant, even if the methyl of 3- is substituted by required group, can also be passed through using it naturally occurring
Enzyme manufactures isoprene oligomer or polyisoprene.Therefore, formula (ZZ-1) or polyisoprene represented by formula (ZZ-2) are excellent
Elect the polyisoprene represented by formula (ZZ-1-1) or represented by formula (ZZ-2-1) as：
Wherein, n, q and Y in formula (ZZ-1) are defined in a manner of identical with n, q and Y in formula (ZZ-1), R with
The same way definition of R in formula (Y)；
Wherein, n, q and Y in formula (ZZ-2-1) are defined in a manner of identical with n, q and Y in formula (ZZ-2), R with
Defined with the same way of the R in formula (Y).
As described above, it is also preferred that skeleton is at the same time modified chain end incessantly.It is therefore preferable that formula (ZZ-1) or formula
(ZZ-2) at least one atom or atomic group contained by the iv parts in (preferred formula (ZZ-1-1) or formula (ZZ-2-1)) are by other
Atom or atomic group substitution.
Contained atom or atom in iv parts in formula (ZZ-1), formula (ZZ-2), formula (ZZ-1-1) or formula (ZZ-2-1)
Group and other described atoms or atomic group can be as described in formulas (X-1).
Further, as described above, it is preferred to which the pi-allyl diphosphonic acid represented by formula (X) is the allyl represented by formula (X-1)
Base diphosphonic acid, and at least one atom or atomic group contained in the II parts in formula (X-1) are by other atoms or atomic group
Substitute, contained atom or atomic group are not substituted by other atoms or atomic group in the III parts in formula (X-1).Therefore, close
Iv parts in the formula (ZZ-1) as represented by following formula (Z), formula (ZZ-2), formula (ZZ-1-1) and formula (ZZ-2-1), preferred formula
(Z) at least one atom or atomic group contained in iv-II parts are substituted by other atoms or atomic group in, in formula (Z)
Iv-III parts in contained atom or atomic group do not substituted by other atoms or atomic group.
In iv-II parts or iv-III parts in formula (Z) contained atom or atomic group and other described atoms or
Atomic group can be such as the atom or atomic group as described in formula (X-1).
The concrete example for the iv parts that chain end is also modified can include the structure as represented by above-mentioned formula (a) to (s).Its
In, preferred formula (b), (c), (d), (e), (f), (k), the structure of (l) and (r), the more preferably structure of formula (b) and (c), because it
Have stronger with the interaction of filler (for example, silica), and can more effectively improve low-heat-generation, wet grab
Ground performance, wearability, elongation at break and fracture tensile strength.
(method of manufacture polyisoprene)
The polyisoprene of the present invention can be represented by the isoprene oligomer of the present invention and formula (Y) (R-IPP)
Compound and/or prenyl diphosphate (IPP), by synthesize (biosynthesis) polyisoprene method (A) made
Make.It according to the target physical property of the polyisoprene of acquisition, can suitably change the ratio of the monomer of the R-IPP used and IPP
Value, the percentage of one of which monomer can be 0.Alternatively, the polyisoprene of the present invention can be by unmodified isoprene
Compound represented by oligomer and formula (Y), is manufactured by the method (B) for synthesizing (biosynthesis) polyisoprene, or
Compound and prenyl diphosphate represented by unmodified isoprene oligomer, formula (Y), by synthesis, (biology closes
Into) manufacture of the method (C) of polyisoprene.In addition, in these cases, can be according to the object of the polyisoprene of acquisition
Rationality matter, suitably changes the ratio between R-IPP the and IPP monomers used.
Organic synthesis and the pi-allyl diphosphonic acid derivative that can select as needed except the compound represented by formula (Y)
Starting material outside, polyisoprene of the invention can be manufactured by biosynthesis.Therefore, the present invention account for oil money
The exhaustion in source and environmental problem.
All the time, it is known that contain in natural rubber latex (especially, the natural rubber latex from Para rubber tree)
Enzyme and rubber elongation factor (for example, being capable of the enzyme of addition polymerization prenyl diphosphate as described above) or its analog, its
Activity with the condensation reaction between catalysis isoprene oligomer and prenyl diphosphate, therefore, catalysis successively will be different
Pentenyl diphosphonic acid is connected as Z configurations (isoprene unit of new addition is cis-configuration), generation with isoprene oligomer
The reaction of polyisoprene, it is as follows.
Also, it is known that contain enzyme and rubber elongation factor in the natural rubber latex (rubber latex) from some plant
(for example, being capable of the enzyme of addition polymerization prenyl diphosphate as described above) or its analog, it has catalysis isoprene
The activity of condensation reaction between oligomer and prenyl diphosphate, therefore, catalysis successively by prenyl diphosphate with it is different
Pentadiene oligomer is connected as E configuration (isoprene unit of new addition is anti-configuration), generates the reaction of polyisoprene.
As described above, methyl and maintenance when the 3- positions in the structure with required group substitution prenyl diphosphate
The structure in addition to the methyl of 3- of prenyl diphosphate is constant, and obtains the compound of formula (Y) (R-IPP), Ke Yitong
Cross using the compound and using with being catalyzed the active naturally occurring enzyme of above-mentioned reaction, rubber elongation factor or its is similar
Thing forms polyisoprene.Therefore, in the present invention, above-mentioned enzyme, rubber elongation factor or the manufacture of its analog can be used poly-
Isoprene.
In other words, the compound and/or isopentene group two represented by the isoprene oligomer of the present invention and formula (Y)
Phosphoric acid synthesizes the method (A) of the polyisoprene of (biosynthesis) present invention, by unmodified isoprene oligomer and
Compound represented by formula (Y) synthesizes the method (B) of the polyisoprene of (biosynthesis) present invention, by unmodified isoamyl
Compound and prenyl diphosphate represented by diene oligomer, formula (Y) synthesize the poly- isoamyl of (biosynthesis) present invention
The method (C) of diene, can come by using enzyme, rubber elongation factor or its analog contained in natural rubber latex
Complete.The enzyme cloned from natural rubber latex, rubber elongation factor or its analog can also be used for the above method.
Specifically, can be in the presence of enzyme and/or rubber elongation factor, by the isoamyl two of the present invention in method (A)
Alkene oligomer is reacted with the compound represented by formula (Y) and/or prenyl diphosphate.Similarly, in method (B),
Can be in the presence of enzyme and/or rubber elongation factor, by the chemical combination represented by unmodified isoprene oligomer and formula (Y)
Thing is reacted.Similarly, can be in the presence of enzyme and/or rubber elongation factor in method (C), will be unmodified different
Pentadiene oligomer is reacted with the compound represented by formula (Y), prenyl diphosphate.
More specifically, for example, the reaction can be by isoprene oligomer and formula (Y) institute table containing the present invention
In the compound and/or the solution of prenyl diphosphate shown, add natural rubber latex or separated from natural rubber latex
Enzyme, rubber elongation factor or its analog for going out and trigger.In addition, reaction temperature can be such as 10 to 60 DEG C, reaction time
When can be such as 1 to 72 small, pH can be such as 6 to 8.In addition it is also possible to as needed, addition magnesium chloride, surface-active
Agent, 2 mercapto ethanol, potassium fluoride and other materials.Even in the case of using unmodified isoprene oligomer, reaction
It can also be carried out under similarity condition.
In the present invention, the isoprene chain of polyisoprene upper in any direction can extend (anti-configuration or cis structure
Type).In other words, for example, the polyisoprene of the present invention can be all isoprene units gathering with anti-configuration link
Isoprene (for example, isoprene oligomer of formula (ZZ-1)), isoprene unit are linked poly- different with trans-cis configuration
Pentadiene (for example, polyisoprene of formula (Z-2)) or isoprene unit are linked poly- with Trans-cis-anti-configuration
Isoprene.Wherein, the polyisoprene that preferably isoprene unit is linked with trans-cis configuration, because its with it is industrial wide
The structure of the natural rubber from Para rubber tree of general application is identical.
In the polyisoprene of the invention that the reaction by described above manufactures, formula (ZZ-1), formula (ZZ-2), formula (ZZ-
1-1) or the Y in formula (ZZ-2-1) is usually OPP or hydroxyl.OPP comes from IPP or R-IPP.OPP can also be hydrolyzed easily, OPP's
Hydrolysis produces hydroxyl.Here it is why the Y in formula (ZZ-1), (ZZ-2), (ZZ-1-1) or (ZZ-2-1) be usually OPP or
Hydroxyl.
In addition, the Y in formula (ZZ-1) or (ZZ-2) is the polyisoprene of formoxyl, for example, can pass through oxidation-type
(ZZ-1) or the Y in (ZZ-2) manufactures for the polyisoprene of OPP.
In addition, the Y in formula (ZZ-1) or (ZZ-2) is the polyisoprene of carboxyl, for example, can pass through oxidation-type (ZZ-
1) or the Y in (ZZ-2) manufactures for the polyisoprene of OPP.
In addition, Y in formula (ZZ-1) or (ZZ-2) is in the polyisoprene of alkoxycarbonyl, formula (ZZ-1) or (ZZ-2)
Y be alkoxy carbonyl group polyisoprene, for example, can be by the Y in method described above carboxylation formula (ZZ-1) or (ZZ-2)
The polyisoprene of OPP is simultaneously further esterified the polyisoprene after the carboxylation and manufactures.
The source of natural rubber latex is not particularly limited.Its example includes：Para rubber tree (Hevea
Brasiliensis), rubberplant (Ficus elastica), Ficus lyrata (Ficus lyrata), banyan (Ficus
Benjamina), bodhi tree (Ficus religiosa), banyan (Ficus benghalensis), laetarius volemus
(Lactarius volemus), sonchus oleraceus (Sonchus oleracers L.), Western dandelion (Taraxacum
) and sunflower (Helianthus annuus) officinale.Wherein, preferred Para rubber tree, because rubber of its manufacture has
High molecular weight and latex has high rubber content.
Natural rubber latex, for example, can be by marking channel-shaped with knife or its analog on the trunk of Para rubber tree
Wound (this process referred to as " is tapped rubber "), then recycle the natural rubber latex flowed out from cut-off latex dust and obtain.
For example, by centrifuging natural rubber for separated enzyme or rubber elongation factor from natural rubber latex
Glue latex and separated glue clear (Serum), bottom phase (bottom fraction) or rubber phase (rubber fraction).Institute
State in clear glue, bottom phase and rubber phase and contain enzyme, rubber elongation factor or its analog.
(rubber composition)
The rubber composition of the present invention contains the isoprene oligomer of the present invention and/or the polyisoprene of the present invention.
Therefore, rubber composition of the invention has outstanding low-heat-generation, wet grip performance, wearability, elongation at break and fracture
Tensile strength (particularly low-heat-generation and wearability).The polyisoprene of the present invention may be used as rubber components.
In rubber constituent based on 100 mass % meters, the content of polyisoprene of the invention be preferably 20 mass % with
On, more preferably more than 40 mass %, further preferred more than 60 mass %.It can be 100 mass %.
In addition to the polyisoprene of the present invention, it can include as the example for the material that rubber constituent uses：Isoamyl
Diene rubber (IR), natural rubber (NR), butadiene rubber (BR), butadiene-styrene rubber (SBR), styrene isoprene butadiene rubber
The diene series rubbers such as glue (SIBR), neoprene (CR) and acrylonitrile butadiene rubber (NBR).These elastomeric materials can be independent
Use, can also be used in combination of two or more.Wherein, preferably NR, BR and SBR.
When the isoprene oligomer of the present invention is used in rubber composition, mix for isoprene oligomer
The reasons why property is high, rubber constituent is preferably using NR.By and with the present invention isoprene oligomer and NR, can be more suitably
Obtain the effect of the isoprene oligomer of the present invention.
When the isoprene oligomer of the present invention is used in rubber composition, based in 100 mass % of rubber constituent, NR
Content preferably more than 20 mass %, more preferably more than 40 mass %, further preferred more than 60 mass %.It can be 100 matter
Measure %.
Relative to 100 mass parts of rubber constituent, the content of isoprene oligomer of the invention is preferably more than 1 mass parts,
More preferably more than 2 mass parts.When the isoprene oligomer used is less than 1 mass parts, it may not be possible to fully produce its effect.Institute
The content for stating isoprene oligomer is preferably also 20 below mass part, and more preferably 15 is below mass part.Higher than 20 mass parts
During content, intensity and wearability are likely to reduced.
The example of the filler used in the present invention includes：Silica, carbon black, clay and calcium carbonate.
The preferred silica of filler used in the present invention.During using silica, can fully it obtain by using this
Effect obtained from the isoprene oligomer of invention and/or the polyisoprene of the present invention.Arbitrary silica can be used,
Its example includes dry silica (silicic acid anhydride), wet silica (aqueous silicic acid).Preferably wet silica, because
It contains more silanol groups.
It is also preferred that being used as filler using carbon black in the present invention.In this case, can also fully obtain by using this hair
Effect obtained from bright isoprene oligomer and/or the polyisoprene of the present invention.
The manufacture in rubber composition can also suitably be contained in addition to mentioned component, in rubber composition of the invention
In common other compounding agents, for example, silane coupling agent, zinc oxide, stearic acid, various antiaging agents (i.e. antioxidant), soft
Agent (for example, oil), wax, vulcanizing agent (for example, sulphur) and vulcanization accelerator.
Rubber composition of the present invention can be prepared by conventional known method.For example, can be by using
The rubber mill of open roll, Banbury muller etc. is kneaded the component, then vulcanizes, to manufacture rubber composition.
The rubber composition of the present invention is applicable to tyre element (for example, tyre surface, side wall, tread base, casing ply, slow
Rush layer and carcass) with and the like.
(pneumatic tire)
The pneumatic tire of the present invention can be manufactured using the rubber composition according to usual way.Specifically,
It is the shape corresponding to tyre element (for example, tyre surface or side wall) by unvulcanized rubber composition extrusion process, then, passes through
Usual way is shaped with tyre forming machine, then is bonded with other tyre elements, so that unvulcanized tire is formed, by the non-sulphur
Change tire and hot pressing is carried out in vulcanizer, so as to form tire.
Embodiment
With reference to following embodiments, the present invention is described in detail, but the present invention is not limited only to this.
(manufacture embodiment 1)
(the synthesis (compound (R-IPP) represented by formula (Y)) of 3-R-3- cyclobutenyl diphosphonic acid
The use of n-R- aldehyde is starting material synthesising target compound.By Green et al. (M.B.Green, and
W.J.Hickinbotton, J.Chem.Soc.1957,3262) method, in formic acid using dimethylamine n-R- aldehyde alpha-position
The outer methylene (compound represented by following formula (i)) of upper introducing.Then, the compound of acquisition is reduced to 2-R- with lithium aluminium hydride reduction
Allyl alcohol (compound represented by following formula (ii)).Further, by the compound chlorination it is 2-R- with phosphorus chloride in pyridine
Allyl chloride (compound represented by following formula (iii)), then by it in the presence of the Grignard Reagent of fresh synthesis, with titanium dioxide
Carbon reaction obtains carboxylic acid (compound represented by following formula (iv)).Then, the compound is reduced to form alcohol with lithium aluminium hydride reduction
(compound represented by following formula (v)).Then, in pyridine with toluene sulfochloride by the compound carry out tosylation (under
Compound represented by formula (vi)).Then, which is subjected to phosphorylation with trimethyl phosphate in acetonitrile, be derived from
Target product (compound represented by following formula (vii), i.e., the compound (R-IPP) represented by formula (Y)).By using TLC and
Instrumental Analysis (IR and NMR) confirms the intermediate of each synthesis phase and final product.
In the case where R is respectively ethyl, propyl group, butyl, phenyl, sulfydryl, hydroxyl and amino, formula (Y) institute has been synthesized
The compound (R-IPP) of expression.When R is ethyl, propyl group, butyl, phenyl, sulfydryl, hydroxyl and amino, R-IPP points of formula (Y)
R-IPP-A, R-IPP-B, R-IPP-C, R-IPP-D, R-IPP-E, R-IPP-F and R-IPP-G are not denoted as it.
(embodiment 1)
(manufacture (alltrans) of isoprene oligomer)
The isoprene oligomer that whole isoprene units are linked with anti-configuration is prepared, such as formula (Z-1-1-1) institute
Represent.
(preparation of transformant)
First, transformant is prepared.In the preparation of transformant, importing mankind's geranyl geranyl diphosphate synzyme is used
pET15b(pET15b/human-GGPS).The pET15b/human-GGPS by multi-component Science Institute of Northeastern University assistant
Upper rich associate professor (Associate Professor Hiroshi SAGAMI, Institute of Multidisciplinary
Research for Advanced Materials, Tohoku University) provide.
Pass through heat shock Transformed E .coli BL21 (DE3) using pET15b/human-GGPS.The transformant is coated with
After on the LB agar mediums of the ampicillin containing 50 μ g/mL, overnight incubation at 37 DEG C, then, selects transformant.
(manufacture with the protein of prenyltransferases activity)
Obtained E.coli BL21 (DE3)/pET15b/human-GGPS (wild type) is inoculated into and contains 50 equipped with 3mL
In the test tube of the LB culture mediums of the ampicillin of μ g/mL, when shaken cultivation 5 is small at 37 DEG C.In obtained nutrient solution, take
1mL deciles are seeded in the 500mL conical flasks of the LB culture mediums containing 50 μ g/mL ampicillins equipped with 100mL, at 37 DEG C
It is lower by cell oscillation culture 3 it is small when.Then, add IPTG to its concentration be 0.1mmol/L, then by cell oscillation at 30 DEG C
Cultivate 18 it is small when.The nutrient solution is centrifuged, obtains wet thallus.After being homogenized the wet thallus by ultrasonic wave, centrifuge and obtain
Supernatant.The egg with prenyltransferases activity is purified from the supernatant using HisTrap (Amersham companies)
White matter.The SDS-PAGE that is purified by of protein is confirmed.
(preparation of isoprene oligomer)
Prepare containing after purification protein 10mg, 50mM Tris-HCl buffer solutions (pH 7.5), 40mM magnesium chlorides,
25mM 2 mercapto ethanols, 1mM starting materials (geranyl diphosphonic acid (GPP) and 1mM prenyl diphosphates (IPP) or system
Make a kind of reaction solution in R-IPP manufactured in embodiment 1.Carried out in 37 DEG C of water-bath the reaction 1 it is small when.Reaction
After, saturated salt solution 100ml and pentane 500ml is added, after stirring the mixture, is stood.Then, it is dense by evaporating
Contracting supernatant (pentane layer) is extremely dry.Remnant parts are taken to confirm product structure by NMR, it is isoprene oligomer.By
The concrete condition (n, m and R in formula (Z-1-1-1)) of this obtained isoprene oligomer is as shown in table 1.Herein, based on making
The information of starting material and the isoprene chain length measured with TLC, calculate the n and m in formula (Z-1-1-1).In addition, formula
(Z-1-1-1) the R structures in are identified by NMR, TLC and GC-MS.
In addition, monomer IPP, R-IPP-A, R-IPP-B, R-IPP-C, R-IPP-D, R-IPP-E, R-IPP-F are used respectively
With R-IPP-G obtain isoprene oligomer, it is aftermentioned experiment use in be referred to as isoprene oligomer (t- controls),
Isoprene oligomer (t-A), isoprene oligomer (t-B), isoprene oligomer (t-C), isoprene oligomer (t-
D), isoprene oligomer (t-E), isoprene oligomer (t-F) and isoprene oligomer (t-G).It is in addition, different in experiment
Pentadiene oligomer (t-E), isoprene oligomer (t-F) are respectively as isoprene oligomer a and isoprene oligomer e
Add in rubber composition.
[table 1]
(relative activity of monomer)
Make IPP or manufacture embodiment 1 in R-IPP in one kind with geranyl diphosphonic acid (GPP) under the following conditions into
Row reaction.Each R-IPP is represented for the relative activity of GPP with index relative to IPP active (=100).
Prepare containing after purification protein 500ng, 50mM Tris-HCl buffer solutions (pH 7.5), 40mM magnesium chlorides,
25mM 2 mercapto ethanols, 12.5 μM GPP and 50 μM of [1-14C] in IPP or manufacture embodiment 1 in the R-IPP for preparing one
Kind.Carried out in 37 DEG C of water-bath the reaction 1 it is small when.After reaction, counted using liquid scintillation with TLC to measure in each condition
Activity.It is as shown in table 2 relative to the activity (=100) of IPP, the relative activity of each R-IPP.
Isoprene oligomer using with by the way of same in 1 part of embodiment (preparation of isoprene oligomer) into
Row purifying.Then, the concrete condition (n, m and R in formula (Z-1-1-1)) of the isoprene oligomer of acquisition is shown in table 2.
Herein, information based on the starting material used and the isoprene chain length measured with TLC, calculate n in formula (Z-1-1-1) and
m.In addition, the R structures in formula (Z-1-1-1) are identified by NMR, TLC and GC-MS.
[table 2]
Table 1 and 2 the results show that even if using R-IPP, the isoprene oligomer of acquisition also with using IPP when have
Identical molecular weight.The skeleton for obtaining isoprene oligomer is also demonstrated to be modified there occurs the R-IPP with using is corresponding.
(embodiment 2)
(preparation of polyisoprene)
It is oligomeric with the isoprene that anti-configuration links using whole isoprene units as represented by formula (Z-1-1-1)
Thing (all trans) prepares polyisoprene.
Prepare containing 10 μ L latex components, 50mM Tris-HCl buffer solutions (pH 7.5), 25mM magnesium chlorides, 40mM2- mercaptos
Base ethanol, 40mM potassium fluorides, 50 μM of isoprene oligomer and 1mM IPP are manufactured in the R-IPP prepared in embodiment 1
A kind of reaction solution.The reaction 3 days is carried out in 30 DEG C of water-bath.After reaction, molecular weight is measured by GPC.Then, it is based on
The information of the numerical value of molecular weight and the starting material used, calculates the number of the isoprene unit added on starting material GPP
Amount.The results are shown in Table 3.
Latex component used herein is by clear and the glue obtained to obtaining latex progress ultracentrifugation from Para rubber tree.
The isoprene oligomer used is isoamyl two prepared in above-mentioned part (preparation of isoprene oligomer)
Alkene oligomer (t- controls), isoprene oligomer (t-A), isoprene oligomer (t-B), isoprene oligomer (t-C),
Isoprene oligomer (t-D), isoprene oligomer (t-E), isoprene oligomer (t-F) and isoprene oligomer
(t-G) one kind in.
[table 3]
Table 3 the results show that even if using R-IPP, the polyisoprene of acquisition also with using during IPP with identical point
Son amount.Further, the polyisoprene of acquisition is analyzed with NMR, TLC and GC-MS, it is thus identified that oligomeric with isoprene
The situation of thing is similar, and the skeleton of polyisoprene has carried out be modified corresponding with used R-IPP.
(embodiment 3)
(manufacture (Trans-cis) of isoprene oligomer)
The isoprene oligomer that isoprene unit is linked with trans-cis configuration is prepared, such as formula (Z-2-1-1) institute table
Show.
(preparation of transformant)
First, transformant is prepared.In the preparation of transformant, 11 iso-amylene two for importing micrococcus luteus B-P26 are used
The pET22b (pET22b/MLU-UPS) of phosphate synthase is used as dsDNA templates.The pET22b/MLU-UPS is polynary by Northeastern University
Pretty professor (Prof.Tanetoshi Koyama, the Institute of of the ancient mountain kind of material science research institute
Multidisciplinary Research for Advanced Materials, Tohoku University) provide.
Pass through heat shock Transformed E .coli BL21 (DE3) using pET22b/MLU-UPS.The transformant is coated on and is contained
After having on the LB agar mediums of the ampicillin of 50 μ g/mL, overnight incubation at 37 DEG C, then, selects transformant.
(manufacture with the protein of prenyltransferases activity)
Obtained E.coli BL21 (DE3)/pET22b/MLU-UPS is inoculated into the ammonia benzyl for containing 50 μ g/mL equipped with 3mL
In the test tube of the LB culture mediums of penicillin, when shaken cultivation 5 is small at 37 DEG C.In obtained nutrient solution, 1mL deciles are taken to be seeded to
In the 500mL conical flasks of the LB culture mediums containing 50 μ g/mL ampicillins equipped with 100mL, by cell oscillation at 37 DEG C
Cultivate 3 it is small when.Then, add IPTG to its concentration be 0.1mmol/L, then at 30 DEG C by cell oscillation culture 18 it is small when.
The nutrient solution is centrifuged, obtains wet thallus.After being homogenized the wet thallus by ultrasonic wave, centrifuge and obtain supernatant.Use
HisTrap (Amersham companies) purifies the protein with prenyltransferases activity from the supernatant.Protein
The SDS-PAGE that is purified by confirmed.
(preparation of isoprene oligomer)
Prepare containing after purification protein 10mg, 50mM Tris-HCl buffer solutions (pH 7.5), 40mM magnesium chlorides,
40mM Triton X-100,25mM 2 mercapto ethanols, 1mM starting materials (farnesyl diphosphonic acid (FPP) and 1mM IPP or
One kind in the R-IPP prepared in manufacture embodiment 1.Carried out in 37 DEG C of water-bath the reaction 1 it is small when.After reaction, add
Enter saturated salt solution 100ml and n-butyl alcohol 500ml, after stirring the mixture, stood.Then, by being concentrated by evaporation supernatant
Liquid (n-butyl alcohol layer) is extremely dry.Remnant parts are taken to confirm product structure by NMR, it is isoprene oligomer.Obtain
The concrete condition (n, m and R in formula (Z-2-1-1)) of isoprene oligomer is as shown in table 4.Herein, based on the starting used
The information of substrate and the isoprene chain length measured with TLC, calculate the n and m in formula (Z-2-1-1).In addition, formula (Z-2-1-1)
In R structures identified by NMR, TLC and GC.
In addition, monomer IPP, R-IPP-A, R-IPP-B, R-IPP-C, R-IPP-D, R-IPP-E, R-IPP-F are used respectively
With R-IPP-G and the isoprene oligomer that obtains, (tc- pairs of isoprene oligomer is referred to as in aftermentioned experiment use
According to), isoprene oligomer (tc-A), isoprene oligomer (tc-B), isoprene oligomer (tc-C), isoprene it is low
Polymers (tc-D), isoprene oligomer (tc-E), isoprene oligomer (tc-F) and isoprene oligomer (tc-G).This
Outside, in experiment, isoprene oligomer (tc-E) is added in rubber composition as isoprene oligomer (b).In addition, experiment
In, each isoprene oligomer (tc-A), isoprene oligomer (tc-B), isoprene oligomer (tc-C), isoamyl two
Alkene oligomer (tc-D), isoprene oligomer (tc-E), isoprene oligomer (tc-F) and isoprene oligomer (tc-
G) respectively as isoprene oligomer h, isoprene oligomer i, isoprene oligomer j, isoprene oligomer k, different
Pentadiene oligomer l, isoprene oligomer m and isoprene oligomer n are added in rubber composition.
[table 4]
(relative activity of monomer)
Make IPP or manufacture embodiment 1 in R-IPP in one kind with farnesyl diphosphonic acid (FPP) under the following conditions into
Row reaction.Each R-IPP is indicated for the relative activity of FPP with the index relative to IPP active (=100).
Prepare containing after purification protein 500ng, 50mM Tris-HCl buffer solutions (pH 7.5), 40mM magnesium chlorides,
40mM Triton X-100,25mM 2 mercapto ethanols, 12.5 μM FPP and 50 μM of [1-14C] IPP or manufacture embodiment 1
A kind of reaction solution in the R-IPP of middle preparation.Carried out in 37 DEG C of water-bath the reaction 1 it is small when.After reaction, using liquid
Scinticounting and TLC measure activity in each condition.Relative to the activity (=100) of IPP, the relative activity of each R-IPP
As shown in table 5.
Isoprene oligomer using with by the way of same in 3 part of embodiment (preparation of isoprene oligomer) into
Row purifying.Then, the concrete condition (n, m and R in formula (Z-2-1-1)) of the isoprene oligomer of acquisition is shown in table 5.
Herein, information based on the starting material used and the isoprene chain length measured with TLC, calculate n in formula (Z-2-1-1) and
m.In addition, the R structures in formula (Z-2-1-1) are identified by NMR, TLC and GC-MS.
[table 5]
Table 4 and 5 the results show that even if using R-IPP, the isoprene oligomer of acquisition also with using IPP when have
Identical molecular weight.The skeleton for obtaining isoprene oligomer is also demonstrated to be modified there occurs the R-IPP with using is corresponding.
(embodiment 4)
(preparation of polyisoprene)
It is low with the isoprene that trans-cis configuration links using the isoprene unit as represented by formula (Z-2-1-1)
Polymers (Trans-cis) prepares polyisoprene.
Prepare containing 10 μ L latex components, 50mM Tris-HCl buffer solutions (pH 7.5), 25mM magnesium chlorides, 40mM2- mercaptos
Base ethanol, 40mM potassium fluorides, 50 μM of isoprene oligomer and 1mM IPP are manufactured in the R-IPP prepared in embodiment 1
A kind of reaction solution.The reaction 3 days is carried out in 30 DEG C of water-bath.After reaction, molecular weight is measured by GPC.Then, it is based on
The information of the numerical value of molecular weight and the starting material used, calculates the number of the isoprene unit added on starting material FPP
Amount.The result is shown in table 6.
Latex component used herein is by clear and the glue obtained to obtaining latex progress ultracentrifugation from Para rubber tree.
The isoprene oligomer used is isoamyl two prepared in above-mentioned part (preparation of isoprene oligomer)
Alkene oligomer (tc- controls), isoprene oligomer (tc-A), isoprene oligomer (tc-B), isoprene oligomer
(tc-C), isoprene oligomer (tc-D), isoprene oligomer (tc-E), isoprene oligomer (tc-F) and isoamyl two
One kind in alkene oligomer (tc-G).In addition, in subsequent experiment, isoprene oligomer (tc-E), isoamyl two are used
The polyisoprene of alkene oligomer (tc-F) manufacture adds rubber composition respectively as polyisoprene D and polyisoprene A
In.In addition, each use isoprene oligomer (tc-A), isoprene oligomer (tc-B), isoprene oligomer (tc-
C), isoprene oligomer (tc-D), isoprene oligomer (tc-E), isoprene oligomer (tc-F), isoprene are low
The polyisoprene of polymers (tc-G) manufacture is respectively as polyisoprene I, polyisoprene J, polyisoprene K, poly- isoamyl
Diene L, polyisoprene M, polyisoprene N, polyisoprene P, add in rubber composition.
[table 6]
Table 6 the results show that even if using R-IPP, the polyisoprene of acquisition also with using during IPP with identical point
Son amount.Further, the polyisoprene of acquisition is analyzed with NMR, TLC and GC-MS, it is thus identified that oligomeric with isoprene
The situation of thing is similar, and the skeleton of polyisoprene has carried out be modified corresponding with used R-IPP.
(embodiment 5)
(manufacture (all trans) of isoprene oligomer)
The isoprene oligomer that whole isoprene units are linked with anti-configuration is prepared, such as formula (Z-1-1-1) institute
Represent.
(preparation of transformant)
First, transformant is prepared.In the preparation of transformant, closed using bacillus stearothermophilus farnesyl diphosphonic acid is imported
Into the pET15b (pET15b/bacillus-FPS) of enzyme.The pET15b/bacillus-FPS is by Northeastern University's multi-component science
Pretty professor (Prof.Tanetoshi Koyama, the Institute of Multidisciplinary of the ancient mountain kind of research institute
Research for Advanced Materials, Tohoku University) provide.
Pass through heat shock Transformed E .coli BL21 (DE3) using pET15b/bacillus-FPS.The transformant is applied
After cloth is on the LB agar mediums of the ampicillin containing 50 μ g/mL, overnight incubation at 37 DEG C, then, selects transformant.
(manufacture with the protein of prenyltransferases activity)
Obtained E.coli BL21 (DE3)/pET15b/bacillus-FPS (wild type) is inoculated into and contained equipped with 3mL
In the test tube of the LB culture mediums of the ampicillin of 50 μ g/mL, when shaken cultivation 5 is small at 37 DEG C.In obtained nutrient solution, take
1mL deciles are seeded in the 500mL conical flasks of the LB culture mediums containing 50 μ g/mL ampicillins equipped with 100mL, at 37 DEG C
It is lower by cell oscillation culture 3 it is small when.Then, add IPTG to its concentration be 0.1mmol/L, then by cell oscillation at 30 DEG C
Cultivate 18 it is small when.The nutrient solution is centrifuged, obtains wet thallus.After being homogenized the wet thallus by ultrasonic wave, centrifuge and obtain
Supernatant.The egg with prenyltransferases activity is purified from the supernatant using HisTrap (Amersham companies)
White matter.The SDS-PAGE that is purified by of protein is confirmed.
(preparation of isoprene oligomer)
Prepare containing after purification protein 10mg, 50mM Tris-HCl buffer solutions (pH 7.5), 40mM magnesium chlorides,
25mM 2 mercapto ethanols, 1mM starting materials (dimethylallyl diphosphate (DMAPP) and 1mM prenyl diphosphates
(IPP) or in manufacture embodiment 1 a kind of reaction solution in the R-IPP prepared.It is small that the reaction 1 is carried out in 37 DEG C of water-bath
When.After reaction, saturated salt solution 100ml and pentane 500ml is added, after stirring the mixture, is stood.Then, lead to
Pervaporation concentrated supernatant (pentane layer) is extremely dry.Remnant parts are taken to confirm product structure by NMR, it is low for isoprene
Polymers.The concrete condition (n, m and R in formula (Z-1-1-1)) of obtained isoprene oligomer is as shown in table 7.Herein, it is based on
The information of the starting material used and the isoprene chain length measured with TLC, calculate the n and m in formula (Z-1-1-1).In addition, formula
(Z-1-1-1) the R structures in are identified by NMR, TLC and GC-MS.
In addition, monomer IPP, R-IPP-A, R-IPP-B, R-IPP-C, R-IPP-D, R-IPP-E, R-IPP-F are used respectively
With R-IPP-G and the isoprene oligomer that obtains, (t- pairs of isoprene oligomer is referred to as in aftermentioned experiment use
According to -1), isoprene oligomer (t-A-1), isoprene oligomer (t-B-1), isoprene oligomer (t-C-1), isoamyl
Diene oligomer (t-D-1), isoprene oligomer (t-E-1), isoprene oligomer (t-F-1) and isoprene oligomer
(t-G-1)。
[table 7]
(relative activity of monomer)
One kind in the R-IPP for making to prepare in IPP or manufacture embodiment 1 is with dimethyl diphosphonic acid (DMAPP) in following bar
Reacted under part.Each R-IPP is represented for the relative activity of DMAPP with index relative to IPP active (=100).
Prepare containing after purification protein 500ng, 50mM Tris-HCl buffer solutions (pH 7.5), 40mM magnesium chlorides,
25mM 2 mercapto ethanols, 12.5 μM DMAPP and 50 μM of [1-14C] in IPP or manufacture embodiment 1 in the R-IPP for preparing
A kind of reaction solution.Carried out in 37 DEG C of water-bath the reaction 1 it is small when.After reaction, counted using liquid scintillation with TLC to survey
Fixed activity in each condition.It is as shown in table 8 relative to the activity (=100) of IPP, the relative activity of each R-IPP.
Isoprene oligomer using with by the way of same in 5 part of embodiment (preparation of isoprene oligomer) into
Row purifying.Then, the concrete condition (n, m and R in formula (Z-1-1-1)) of the isoprene oligomer of acquisition is shown in table 8.
Herein, information based on the starting material used and the isoprene chain length measured with TLC, calculate n in formula (Z-1-1-1) and
m.In addition, the R structures in formula (Z-1-1-1) are identified by NMR, TLC and GC-MS.
[table 8]
Table 7 and 8 the results show that even if using R-IPP, the isoprene oligomer of acquisition also with using IPP when have
Identical molecular weight.The skeleton for obtaining isoprene oligomer is also demonstrated to be modified there occurs the R-IPP with using is corresponding.
(embodiment 6)
(preparation of polyisoprene)
Then, using the isoamyl two linked such as whole isoprene units represented by formula (Z-1-1-1) with anti-configuration
Alkene oligomer (all trans) prepares polyisoprene.
Prepare containing 10 μ L latex components, 50mM Tris-HCl buffer solutions (pH 7.5), 25mM magnesium chlorides, 40mM2- mercaptos
Base ethanol, 40mM potassium fluorides, 50 μM of isoprene oligomer and 1mM IPP are manufactured in the R-IPP prepared in embodiment 1
A kind of reaction solution.The reaction 3 days is carried out in 30 DEG C of water-bath.After reaction, molecular weight is measured by GPC.Then, it is based on
The information of the numerical value of molecular weight and the starting material used, calculating add to the isoprene unit on starting material DMAPP
Quantity.The results are shown in Table 9.
Latex component used herein is by clear and the glue obtained to obtaining latex progress ultracentrifugation from Para rubber tree.
The isoprene oligomer used is isoamyl two prepared in above-mentioned part (preparation of isoprene oligomer)
Alkene oligomer (t- controls -1), isoprene oligomer (t-A-1), isoprene oligomer (t-B-1), isoprene oligomer
(t-C-1), isoprene oligomer (t-D-1), isoprene oligomer (t-E-1), isoprene oligomer (t-F-1) and different
One kind in pentadiene oligomer (t-G-1).
[table 9]
Table 9 the results show that even if using R-IPP, the polyisoprene of acquisition also with using during IPP with identical point
Son amount.Further, the polyisoprene of acquisition is analyzed with NMR, TLC and GC-MS, it is thus identified that oligomeric with isoprene
The situation of thing is similar, and the skeleton of polyisoprene has carried out be modified corresponding with used R-IPP.
(embodiment 7)
(manufacture (Trans-cis) of isoprene oligomer)
Then, using pi-allyl diphosphonic acid derivative, the isoprene unit as represented by formula (Z-2-1-1) is prepared with anti-
It is that formula-cis-configuration links and not only modified isoprene oligomer (skeleton and last-in-chain(LIC) also occur for skeleton but also chain end
Hold modified isoprene oligomer).
(preparation of pi-allyl diphosphonic acid derivative)
First, synthesis of allyl diphosphonic acid derivative.
(manufacture embodiment 2)
(- ten two carbon of 8- methoxyl group -3,7- dimethyl-(2E, 6E)-dialkylene diphosphonic acid (compound represented by formula (B))
Synthesis)
The use of geraniol is starting material synthesising target compound.With pyridine and acetic anhydride to perfume (or spice) in anhydrous methylene chloride
Leaf-alcohol carries out acetylation, obtains acetic acid esters (compound represented by following formula (bi)) (yield 95%).Then, by the acetic acid esters
The carbon of 8- carries out selenolite in ethanol, obtains aldehyde body (compound represented by following formula (bii)) (yield 24%).Then,
The aldehyde is subjected to basic hydrolysis with potassium hydroxide, forms alcohol (compound represented by following formula (biii)) (yield 38%).Connect
, the alcohol is handled with imidazoles, tert-butyl diphenyl chlorosilane (TBDPS) in anhydrous methylene chloride, obtain shown in following (biv)
Compound (yield 80%).Then, the compound is reacted with butyl lithium in absolute ether, obtain butyl alcohol (following formula
(bv) compound represented by) (yield 73%).Then, in anhydrous tetrahydro furan, which is converted into sodium salt, therewith
Methyl iodide is added in the sodium salt and by Williamson synthesis, obtain ether body (compound represented by following (bvi)) (yield
95%).Then, the ether is subjected to elimination reaction using four positive ammonium fluorides in anhydrous tetrahydro furan, obtains alcohol (following formula
(bvii) compound represented by) (yield 87%).Then, N- chloro ambers are used below -40 DEG C, in anhydrous methylene chloride solvent
Amber acid imide carries out chlorine substitution with dimethyl sulphide to primary hydroxyl, obtains chloride (compound represented by following (bviii))
(yield 92%).Then, diphosphate is carried out to the chloride with three (tetra-n-butyl ammoniums) hydrogenation diphosphonic acid in anhydrous acetonitrile,
Obtain target product (compound represented by following formula (bix), i.e. compound represented by formula (B)) (yield 26%).
The confirmation of the intermediate and final product of each synthesis phase is carried out using TLC and Instrumental Analysis (IR and NMR).
(manufacture embodiment 3)
(- ten two carbon of 8- hydroxyl -3,7- dimethyl-(2E, 6E)-dialkylene diphosphonic acid (compound represented by formula (C))
Synthesis)
The use of geraniol is starting material synthesising target compound.With pyridine and acetic anhydride to perfume (or spice) in anhydrous methylene chloride
Leaf-alcohol carries out acetylation, obtains acetic acid esters (compound represented by following formula (ci)) (yield 97%).Then, by the acetic acid esters
The carbon of 8- carries out selenolite in ethanol, obtains aldehyde body (compound represented by following formula (cii)) (yield 20%).Then,
The aldehyde is subjected to basic hydrolysis with potassium hydroxide, forms alcohol (compound represented by following formula (ciii)) (yield 42%).Connect
, the alcohol is handled with imidazoles, tert-butyl diphenyl chlorosilane (TBDPS) in anhydrous methylene chloride, obtain shown in following (civ)
Compound (yield 80%).Then, the compound is reacted with butyl lithium in anhydrous ether, obtain butanol body (yield
62%).Then, four positive ammonium fluorides are used in anhydrous tetrahydro furan by the pure carry out elimination reaction, two alcohol (following formulas of acquisition
(cvi) compound represented by) (yield 94%).Then, N- chloro ambers are used below -40 DEG C, in anhydrous methylene chloride solvent
Amber acid imide carries out chlorine substitution with dimethyl sulphide to primary hydroxyl, obtains chloride (compound represented by following (cvii))
(yield 90%).Then, diphosphate is carried out to the chloride with three (tetra-n-butyl ammoniums) hydrogenation diphosphonic acid in anhydrous acetonitrile,
Obtain target product (compound represented by following formula (cviii), i.e. compound represented by formula (C)) (yield 46%).
The confirmation of the intermediate and final product of each synthesis phase is carried out using TLC and Instrumental Analysis (IR and NMR).
(preparation of mutant enzyme)
Using pi-allyl diphosphonic acid derivative as starting material, even if being turned using wild type with prenyl
In the case of the enzyme for moving enzymatic activity, reaction can also be carried out and prepare isoprene oligomer.However, in order to improve reaction efficiency,
It is prepared for improving the variant enzyme to the enzymatic activity of pi-allyl diphosphonic acid derivative.
The reagent used comes from QuickChange site-directed mutagenesis kits (QuickChange Site-Directed
Mutagenesis Kit) (Stratagene companies).Primer is designed, is mutated with allowing to introduce in target site.Mutation is drawn
Thing 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 templates using import from micrococcus luteus B-P26 11 prenyl diphosphate synthases (with
Under, also referred to as wild-type enzyme) pET22b (pET22b/MLU-UPS).The pET22b/MLU-UPS is by Northeastern University's multi-component
Pretty professor (Prof.Tanetoshi Koyama, the Institute of Multidisciplinary of the ancient mountain kind of Science Institute
Research for Advanced Materials, Tohoku University) provide.Mix 10x Pfu polymerases buffering
2 μ L of liquid, dsDNA templates 2-20ng, forward primer 50ng, 0.4 μ L of reverse primer 50ng, dNTPs (every kind of is 2.5mM),
ddH2O totally 20 μ L, Pfu polymerase (2.5U/ μ l) 0.4mL, carry out PCR reactions.The condition of PCR reactions 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, 0.4 μ L Dpn I are added in PCR solution, at 37 DEG C the PCR solution is carried out with Dpn I 1 it is small when
Processing.Using the Dpn I treatment fluids of 1 to 10 μ L, pass through heat shock Transformed E .coli DH5 α.The transformant is coated on
After on the LB agar mediums of ampicillin containing 50 μ g/mL, overnight incubation at 37 DEG C, then, selects transformant.Connect
, cultivate the transformant in the LB culture mediums of the ampicillin containing 50 μ g/mL and stay overnight, pass through from obtained nutrient solution
Alkali SDS legal system prepared plasmids.The mutation of the plasmid is confirmed using sequenator.
(manufacture with the protein of prenyltransferases activity)
Obtained E.coli BL21 (DE3)/pET22b/MLU-UPS (wild and saltant type) is inoculated into and is contained equipped with 3mL
In the test tube for having the LB culture mediums of the ampicillin of 50 μ g/mL, when shaken cultivation 5 is small at 37 DEG C.In obtained nutrient solution,
1mL deciles are taken to be seeded in the 500mL conical flasks of the LB culture mediums containing 50 μ g/mL ampicillins equipped with 100mL, 37
When the shaken cultivation cell 3 is small at DEG C.Then, add IPTG to its concentration be 0.1mmol/L, the then shaken cultivation at 30 DEG C
When the cell 18 is small.The nutrient solution is centrifuged, obtains wet thallus.After being homogenized the wet thallus by ultrasonic wave, centrifugation obtains
Obtain supernatant.Purified using HisTrap (Amersham companies) from the supernatant with prenyltransferases activity
Protein.The SDS-PAGE that is purified by of protein is confirmed.The ammonia of thus obtained variant enzyme N77A and variant enzyme L91D
The SEQ ID NO of base acid sequence such as sequence table：Shown in 5 and 6.
(preparation of isoprene oligomer)
Prepare containing after purification protein 10mg, 50mM Tris-HCl buffer solutions (pH 7.5), 40mM magnesium chlorides,
40mM Triton X-100,25mM 2 mercapto ethanols, 1mM starting materials (farnesyl diphosphonic acid (FPP) or manufacture embodiment 2
With 3 in the R-IPP for preparing in one kind and 1mM IPP or manufacture embodiment 1 in the pi-allyl diphosphonic acid derivative for preparing
A kind of reaction solution.Carried out in 37 DEG C of water-bath the reaction 1 it is small when.After reaction, saturated salt solution 100ml is added
With n-butyl alcohol 500ml, after stirring the mixture, stood.Then, it is extremely dry by being concentrated by evaporation supernatant (n-butyl alcohol layer)
It is dry.Remnant parts are taken to confirm product structure by NMR, it is isoprene oligomer.Obtained isoprene oligomer
Concrete condition (n, m and R in formula (Z-2-1-1)) is as shown in table 10.Herein, information and use based on the starting material used
The isoprene chain length of TLC measure, calculates the n and m in formula (Z-2-1-1).In addition, the R structures in formula (Z-2-1-1) pass through
NMR, TLC and GC-MS are identified.
In addition, by the use of variant enzyme N77A as enzyme, using the compound represented by formula (B) as starting material, respectively using single
The isoprene that body IPP, R-IPP-A, R-IPP-B, R-IPP-C, R-IPP-D, R-IPP-E, R-IPP-F and R-IPP-G are obtained
Oligomer, isoprene oligomer (tc- controls-AB), isoprene oligomer (tc- are referred to as in aftermentioned experiment use
A-AB), isoprene oligomer (tc-B-AB), isoprene oligomer (tc-C-AB), isoprene oligomer (tc-D-
AB), isoprene oligomer (tc-E-AB), isoprene oligomer (tc-F-AB) and isoprene oligomer (tc-G-AB).
In addition, by the use of variant enzyme L91D as enzyme, using the compound represented by formula (B) as starting material, respectively using single
The isoprene that body IPP, R-IPP-A, R-IPP-B, R-IPP-C, R-IPP-D, R-IPP-E, R-IPP-F and R-IPP-G are obtained
Oligomer, isoprene oligomer (tc- controls-DB), isoprene oligomer (tc- are referred to as in aftermentioned experiment use
A-DB), isoprene oligomer (tc-B-DB), isoprene oligomer (tc-C-DB), isoprene oligomer (tc-D-
DB), isoprene oligomer (tc-E-DB), isoprene oligomer (tc-F-DB) and isoprene oligomer (tc-G-DB).
In addition, by the use of variant enzyme N77A as enzyme, using the compound represented by formula (C) as starting material, respectively using single
The isoprene that body IPP, R-IPP-A, R-IPP-B, R-IPP-C, R-IPP-D, R-IPP-E, R-IPP-F and R-IPP-G are obtained
Oligomer, isoprene oligomer (tc- controls-AC), isoprene oligomer (tc- are referred to as in aftermentioned experiment use
A-AC), isoprene oligomer (tc-B-AC), isoprene oligomer (tc-C-AC), isoprene oligomer (tc-D-
AC), isoprene oligomer (tc-E-AC), isoprene oligomer (tc-F-AC) and isoprene oligomer (tc-G-AC).
In addition, by the use of variant enzyme L91D as enzyme, using the compound represented by formula (C) as starting material, respectively using single
The isoprene that body IPP, R-IPP-A, R-IPP-B, R-IPP-C, R-IPP-D, R-IPP-E, R-IPP-F and R-IPP-G are obtained
Oligomer, isoprene oligomer (tc- controls-DC), isoprene oligomer (tc- are referred to as in aftermentioned experiment use
A-DC), isoprene oligomer (tc-B-DC), isoprene oligomer (tc-C-DC), isoprene oligomer (tc-D-
DC), isoprene oligomer (tc-E-DC), isoprene oligomer (tc-F-DC) and isoprene oligomer (tc-G-DC).
In addition, in experiment, each isoprene oligomer (tc-E-AB), isoprene oligomer (tc-F-AB), isoamyl
Diene oligomer (tc-E-DB) and isoprene oligomer (tc-F-DB) are respectively as isoprene oligomer c, isoprene
Oligomer f, isoprene oligomer d, isoprene oligomer g are added in rubber composition.
[table 10]
(relative activity of monomer)
Make IPP or manufacture one kind in embodiment 1 in the R-IPP for preparing, implement with farnesyl diphosphonic acid (FPP) or manufacture
One kind in the pi-allyl diphosphonic acid derivative prepared in example 2 and 3 is reacted under the following conditions.Be respectively relative to FPP and
The activity of IPP, by the relative activity of each pi-allyl diphosphonic acid derivative and each R-IPP with exponential representation, wherein, FPP and
The activity of IPP is set to 100.
Prepare containing after purification protein 500ng, 50mM Tris-HCl buffer solutions (pH7.5), 40mM magnesium chlorides,
The pi-allyl two prepared in 40mM Triton X-100,25mM 2 mercapto ethanols, 12.5 μM of FPP or manufacture embodiment 2 and 3
One kind and 50 μM of [1- in phosphoric acid derivatives14C] a kind of reaction in IPP or manufacture embodiment 1 in the R-IPP for preparing
Solution.Carried out in 37 DEG C of water-bath the reaction 1 it is small when.After reaction, counted using liquid scintillation with TLC to measure in each condition
Under activity.Relative to the activity (=100) of IPP, the relative activity such as institute of table 11 of pi-allyl diphosphonic acid derivative and R-IPP
Show.
Isoprene oligomer is used to carry out with the same manner in 7 part of embodiment (preparation of isoprene oligomer)
Purifying.Then, the concrete condition (n, m and R in formula (Z-2-1-1)) of the isoprene oligomer of acquisition is shown in table 11.
Herein, information based on the starting material used and the isoprene chain length measured with TLC, calculate n in formula (Z-2-1-1) and
m.In addition, the R structures in formula (Z-2-1-1) are identified by NMR, TLC and GC-MS.
[table 11]
Table 10 and 11 the results show that even if low using R-IPP and pi-allyl diphosphonic acid derivative, the isoprene of acquisition
Polymers also with using IPP and FPP when with identical molecular weight.Also demonstrate obtain isoprene oligomer there occurs with use
R-IPP and the corresponding modification of pi-allyl diphosphonic acid derivative.
(embodiment 8)
(preparation of polyisoprene)
Then, polyisoprene is prepared using following isoprene oligomer：Prepared using pi-allyl diphosphonic acid derivative,
As represented by formula (Z-2-1-1), that isoprene unit is linked with trans-cis configuration and not only skeleton but also chain end
Also modified isoprene oligomer (the isoprene oligomer that skeleton and chain end are modified) occurs.
Prepare containing 10 μ L latex components, 50mM Tris-HCl buffer solutions (pH 7.5), 25mM magnesium chlorides, 40mM2- mercaptos
Base ethanol, 40mM potassium fluorides, 50 μM of isoprene oligomer and 1mM IPP are manufactured in the R-IPP prepared in embodiment 1
A kind of reaction solution.The reaction 3 days is carried out in 30 DEG C of water-bath.After reaction, molecular weight is measured by GPC.Then, base
In the numerical value of molecular weight and the information of the starting material used, calculating adds to starting material FPP or pi-allyl diphosphonic acid derives
The quantity of isoprene unit on thing.As a result it is as shown in table 12.
Latex component used herein is by clear and the glue obtained to obtaining latex progress ultracentrifugation from Para rubber tree.
The isoprene oligomer used is isoamyl two prepared in above-mentioned part (preparation of isoprene oligomer)
Alkene oligomer (tc- controls-AB), isoprene oligomer (tc-A-AB), isoprene oligomer (tc-B-AB), isoprene
Oligomer (tc-C-AB), isoprene oligomer (tc-D-AB), isoprene oligomer (tc-E-AB), isoprene are oligomeric
Thing (tc-F-AB), isoprene oligomer (tc-G-AB), isoprene oligomer (tc- controls-DB), isoprene oligomer
(tc-A-DB), isoprene oligomer (tc-B-DB), isoprene oligomer (tc-C-DB), isoprene oligomer (tc-
D-DB), isoprene oligomer (tc-E-DB), isoprene oligomer (tc-F-DB), isoprene oligomer (tc-G-
DB), isoprene oligomer (tc- controls-AC), isoprene oligomer (tc-A-AC), isoprene oligomer (tc-B-
AC), isoprene oligomer (tc-C-AC), isoprene oligomer (tc-D-AC), isoprene oligomer (tc-E-AC),
It is isoprene oligomer (tc-F-AC), isoprene oligomer (tc-G-AC), isoprene oligomer (tc- controls-DC), different
Pentadiene oligomer (tc-A-DC), isoprene oligomer (tc-B-DC), isoprene oligomer (tc-C-DC), isoamyl two
Alkene oligomer (tc-D-DC), isoprene oligomer (tc-E-DC), isoprene oligomer (tc-F-DC) and isoprene are low
One kind in polymers (tc-G-DC).
In aftermentioned experiment, it is each using isoprene oligomer (tc-E-AB), it is isoprene oligomer (tc-F-AB), different
Pentadiene oligomer (tc-E-DB), isoprene oligomer (tc-F-DB), isoprene oligomer (tc-E-AC) and isoamyl two
Alkene oligomer (tc-E-DC) manufacture polyisoprene respectively as polyisoprene E, polyisoprene B, polyisoprene F,
Polyisoprene C, polyisoprene G, polyisoprene H, add in rubber composition.
[table 12]
Table 12 the results show that even if the isoprene oligomer being modified using R-IPP and skeleton and chain end, acquisition
Polyisoprene also with using IPP and unmodified isoprene oligomer when with identical molecular weight.Further, to obtaining
The polyisoprene obtained is analyzed with NMR, TLC and GC-MS, it is thus identified that and it is similar with the situation of isoprene oligomer, gather different
Pentadiene has carried out the corresponding modification of isoprene oligomer with used R-IPP and the modification of skeleton and chain end.
The isoprene oligomer and the chain end of polyisoprene prepared in embodiment is (with formula (Z-1), formula (Z-2), formula
(ZZ-1) or formula (ZZ-2) Y it is corresponding) be hydroxyl or OPP.
Chemical reagent for embodiment 9 to 41 and comparative example 1 to 4 is listed below.
NR：TSR20
BR：BR01 (JSR companies)
Carbon black：DIABLACK (N220) (Mitsubishi Chemical Ind)
Isoprene oligomer a to n：Prepared isoprene oligomer in above-described embodiment
Polyisoprene A to P：Prepared polyisoprene in above-described embodiment
Zinc oxide：Zinc oxide #1 (mining company of Mitsui Metal Co., Ltd.)
Stearic acid：Stearic acid (You companies)
Antiaging agent：NOCRAC 6C (N- (1,3- dimethylbutyls)-N '-diphenyl-para-phenylene diamine) (the emerging chemical works of imperial palace
Industry company)
Wax：SUNNOC waxes (the emerging chemical industrial company of imperial palace)
Sulphur：Powder sulphur (He Jian chemical companies)
Vulcanization accelerator NS：NOCCELER NS (N tert butyl benzothiazole 2 sulfenamide) (the emerging chemical works of imperial palace
Industry company)
Silica：Nipsil AQ (wet silica) (Japanese silica company)
Silane coupling agent：Si266 (double (3- triethoxysilylpropyltetrasulfides) disulphide) (goldschmidt chemical corporation)
Accelerator D PG：NOCCELER D (N, N- diphenylguanidine) (the emerging chemical industrial company of imperial palace)
Formula according to table 13 to 16, is kneaded beyond sulphur and vulcanization accelerator using 1.7L banbury mixers
Material, obtains mixture.Then, mix sulphur and vulcanization accelerator in the mixture, and be kneaded with open roll, obtain not
Vulcanize rubber composition.Using steam vulcanizing pree in pressure 80kgf/cm2, at 150 DEG C to the unvulcanized rubber composition
Vulcanization 30 minutes is carried out, prepares vulcanization rubber composition.
Following evaluations are carried out to the vulcanization rubber composition thus prepared.As a result as shown in table 13 to 16.The benchmark of table 13
It is the formula of comparative example 2 to be formulated as the formula of comparative example 1, the benchmark formula of table 14, and the benchmark formula of table 15 is matching somebody with somebody for comparative example 3
Side, the benchmark formula of table 16 are the formula of comparative example 4.
(test for viscoelastic)
Using viscoplasticity spectrometer (this making of rock institute), in 70 DEG C, strain 2%, the condition of (elongation at initial stage) carries out tan
The measure of δ.Using the tan δ of benchmark formula as 100, by its numerical value with exponential representation.Index is higher, and heat is bigger.Index is 100
Following formula, which is considered as, improves heat resistance (low-heat-generation).In other words, index is lower, and low-heat-generation is better.
(blue Berne abrasion test)
Using blue Berne abrasiometer (this making of rock institute) and in loading 3kg, slip rate 40% and Sha Su 15g/ minutes
Under conditions of, implement abrasion test.The shape of sample is thickness 5mm, diameter 50mm.The use of grinding stone is the GC abrasive grains of granularity #80.
Using the tan δ of benchmark formula as 100 (benchmark), by result of the test with exponential representation.The index is higher, and wearability is better.Index is big
In 100 formula, being considered as improves wearability.
(tension test)
According to JIS K6251 " rubber, vulcanization rubber or thermoplastic elastomer-tension and the definite method of emergent property ",
Implement tension test using No. 3 dumbbell shape samples being made of above-mentioned vulcanization sheet rubber, to measure fracture tensile strength (TB)
(MPa), elongation at break (EB) (%).If rubber composition of the elongation at break less than 480% is used for giant tire, hold
Rubbery chip is also easy to produce, thus needs to improve.Further, since low fracture tensile strength can cause tire to destroy, it is therefore desirable to anti-
Only because of caused by the change of material fracture tensile strength decline.
Table 13 and 15 show, using the present invention isoprene oligomer embodiment show outstanding low-heat-generation,
Wearability and elongation at break.
Table 14 and 16 is shown, outstanding low-heat-generation, wear-resisting is shown using the embodiment of the polyisoprene of the present invention
Property and fracture tensile strength.
(independent text of sequence table)
SEQ ID NO：1：Prepare the forward primer of variant enzyme N77A
SEQ ID NO：2：Prepare the reverse primer of variant enzyme N77A
SEQ ID NO：3：Prepare the forward primer of variant enzyme L91D
SEQ ID NO：4：Prepare the reverse primer of variant enzyme L91D
SEQ ID NO：5：The amino acid sequence of variant enzyme N77A
SEQ ID NO：6：The amino acid sequence of variant enzyme L91D
Claims (15)
1. a kind of polyisoprene, the compound synthesis represented by its pi-allyl diphosphonic acid and formula (Y) represented by formula (X)：
In formula (X), n represents 1 to 10 integer, and part isoprene unit can be modified；
In formula (Y), R represents nitrogen-containing group, oxy radical, sulfur-containing group or silicon-containing group,
In the polyisoprene, on the pi-allyl diphosphonic acid quantity of the isoprene unit of addition for 30 to
40000。
2. polyisoprene according to claim 1, its pi-allyl diphosphonic acid represented by the formula (X), described formula
(Y) compound and the prenyl diphosphate synthesis represented.
3. polyisoprene according to claim 1 or 2, wherein, the pi-allyl diphosphonic acid represented by the formula (X) is formula
(X-1) the pi-allyl diphosphonic acid represented by：
In formula (X-1), n represents 1 to 10 integer, and
Contained at least one atom or atomic group are substituted by different atom or atomic group in II parts in formula (X-1), formula
(X-1) contained atom or atomic group are not substituted by different atom or atomic group in the III parts in.
4. a kind of polyisoprene, it is by isoprene oligomer, and two phosphorus of compound and isopentene group represented by formula (Y)
At least one of acid synthesizes,
Pi-allyl diphosphonic acid of the isoprene oligomer represented by formula (X) and the compound synthesis represented by formula (Y), its
In on the pi-allyl diphosphonic acid isoprene unit of addition quantity be 1 to 30,
In formula (X), n represents 1 to 10 integer, and part isoprene unit can be modified；
In formula (Y), R represents the group beyond methyl；
In the polyisoprene, on the pi-allyl diphosphonic acid quantity of the isoprene unit of addition for 30 to
40000。
5. polyisoprene according to claim 4, wherein, the isoprene oligomer is represented by the formula (X)
Pi-allyl diphosphonic acid, the formula (Y) represent compound and prenyl diphosphate synthesis.
6. polyisoprene according to claim 4 or 5, wherein, the pi-allyl diphosphonic acid represented by the formula (X) is formula
(X-1) the pi-allyl diphosphonic acid represented by：
In formula (X-1), n represents 1 to 10 integer, and
Contained at least one atom or atomic group are substituted by different atom or atomic group in II parts in formula (X-1), formula
(X-1) contained atom or atomic group are not substituted by different atom or atomic group in the III parts in.
7. polyisoprene according to claim 4 or 5, wherein, the synthesis of the isoprene oligomer is using tool
What the enzyme for having prenyltransferases activity carried out.
8. a kind of polyisoprene, it is represented by formula (ZZ-1) or represented by formula (ZZ-2)：
In formula (ZZ-1), n represents 1 to 10 integer；Q represents 30 to 40000 integer；Y represents hydroxyl, formoxyl, carboxyl, alkane
Epoxide carboxyl, alkoxy carbonyl or OPP；At least one atom or atomic group contained by iv parts can be by other atoms or atomic groups
Substitution,
In formula (ZZ-2), n represents 1 to 10 integer；Q represents 30 to 40000 integer；Y represents hydroxyl, formoxyl, carboxyl, alkane
Epoxide carboxyl, alkoxy carbonyl or OPP；At least one atom or atomic group contained by iv parts can be by other atoms or atomic groups
Substitution,
In v parts in the formula (ZZ-1) or the formula (ZZ-2) contained at least one atom or atomic group by nitrogen-atoms,
Oxygen atom, sulphur atom, silicon atom, nitrogen-containing group, oxy radical, sulfur-containing group or silicon-containing group substitution.
9. polyisoprene according to claim 8, wherein, the iv parts in the formula (ZZ-1) or the formula (ZZ-2)
In contained at least one atom or atomic group substituted by different atom or atomic group.
10. a kind of method for manufacturing polyisoprene, it is included by isoprene oligomer, and the chemical combination represented by formula (Y)
At least one of thing and prenyl diphosphate carry out synthetic polyisoprenes,
Pi-allyl diphosphonic acid of the isoprene oligomer represented by formula (X) and the compound synthesis represented by formula (Y), its
In on the pi-allyl diphosphonic acid isoprene unit of addition quantity be 1 to 30,
In formula (X), n represents 1 to 10 integer, and part isoprene unit can be modified；
In formula (Y), R represents nitrogen-containing group, oxy radical, sulfur-containing group or silicon-containing group；；
In the polyisoprene, on the pi-allyl diphosphonic acid quantity of the isoprene unit of addition for 30 to
40000。
11. the method for manufacture polyisoprene according to claim 10, wherein, the isoprene oligomer is by described
The compound and prenyl diphosphate synthesis that pi-allyl diphosphonic acid, the formula (Y) represented by formula (X) represent.
12. the method for the manufacture polyisoprene according to claim 10 or 11, wherein, the alkene represented by the formula (X)
Pi-allyl diphosphonic acid of the propyl group diphosphonic acid represented by formula (X-1)：
In formula (X-1), n represents 1 to 10 integer, and
Contained at least one atom or atomic group are substituted by different atom or atomic group in II parts in formula (X-1), formula
(X-1) contained atom or atomic group are not substituted by different atom or atomic group in the III parts in.
13. the method for the manufacture polyisoprene according to claim 10 or 11, wherein, the isoprene oligomer
Synthesis is to use the enzyme with prenyltransferases activity to carry out.
14. a kind of rubber composition, it contains at least one in the polyisoprene in claim 1~9 described in any one
Kind.
15. a kind of pneumatic tire, it is formed by rubber composition according to claim 14.
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| 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 |
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| 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 |
| FR2914652B1 (en) * | 2007-04-03 | 2013-01-18 | Librophyt | GENES ENCODING Z, Z-FARNESYL DIPHOSPHATE SYNTHASE AND MULTI-PRODUCT SESQUITERPENE SYNTHASE AND USES THEREOF |
| 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 |
| JP5058332B2 (en) * | 2010-07-14 | 2012-10-24 | 住友ゴム工業株式会社 | Isoprene oligomer, polyisoprene, and production methods thereof, rubber composition, and pneumatic tire |
-
2013
- 2013-09-02 CN CN201810340783.6A patent/CN108484663B/en active Active
- 2013-09-02 WO PCT/JP2013/073509 patent/WO2014042027A1/en active Application Filing
- 2013-09-02 MY MYPI2015700468A patent/MY172203A/en unknown
- 2013-09-02 BR BR112015004224A patent/BR112015004224A2/en not_active Application Discontinuation
- 2013-09-02 CN CN201380045290.5A patent/CN104684958B/en active Active
- 2013-09-02 JP JP2014535489A patent/JP6468512B2/en active Active
-
2019
- 2019-01-07 JP JP2019000505A patent/JP6757938B2/en active Active
Non-Patent Citations (2)
| Title |
|---|
| Artificial substrates for undecaprenyl diphosphate synthase from Micrococcus luteus B-P 26;Masahiko Nagaki et al.;《Journal of Molecular Catalysis B:Enzymatic》;20001231;第9卷;第33-38页 * |
| Substrate specificities of medium-prenylchain elongating enzymes, hexaprenyl- and heptaprenyl diphosphate synthases;Masahiko Nagaki et al.;《Journal of Molecular Catalysis B:Enzymatic》;20031231;第22卷;第97-103页 * |
Also Published As
| Publication number | Publication date |
|---|---|
| WO2014042027A1 (en) | 2014-03-20 |
| CN108484663A (en) | 2018-09-04 |
| BR112015004224A2 (en) | 2017-08-08 |
| JP2019090030A (en) | 2019-06-13 |
| CN104684958A (en) | 2015-06-03 |
| JPWO2014042027A1 (en) | 2016-08-18 |
| CN108484663B (en) | 2020-09-18 |
| MY172203A (en) | 2019-11-15 |
| JP6757938B2 (en) | 2020-09-23 |
| JP6468512B2 (en) | 2019-02-13 |
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