JPWO2017104722A1 - スクロース資化性を有するpha生産微生物、及び該微生物を用いたphaの製造方法 - Google Patents
スクロース資化性を有するpha生産微生物、及び該微生物を用いたphaの製造方法 Download PDFInfo
- Publication number
- JPWO2017104722A1 JPWO2017104722A1 JP2017556109A JP2017556109A JPWO2017104722A1 JP WO2017104722 A1 JPWO2017104722 A1 JP WO2017104722A1 JP 2017556109 A JP2017556109 A JP 2017556109A JP 2017556109 A JP2017556109 A JP 2017556109A JP WO2017104722 A1 JPWO2017104722 A1 JP WO2017104722A1
- Authority
- JP
- Japan
- Prior art keywords
- gene
- sucrose
- pha
- strain
- microorganism
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 229930006000 Sucrose Natural products 0.000 title claims abstract description 94
- 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 title claims abstract description 94
- 239000005720 sucrose Substances 0.000 title claims abstract description 94
- 238000004519 manufacturing process Methods 0.000 title claims abstract description 85
- 244000005700 microbiome Species 0.000 title claims abstract description 81
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 114
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims abstract description 42
- 229910052799 carbon Inorganic materials 0.000 claims abstract description 42
- 108010010718 poly(3-hydroxyalkanoic acid) synthase Proteins 0.000 claims abstract description 38
- 101710117283 Sucrose permease Proteins 0.000 claims abstract description 19
- 101710173142 Beta-fructofuranosidase, cell wall isozyme Proteins 0.000 claims abstract description 18
- 230000000694 effects Effects 0.000 claims abstract description 14
- 238000012258 culturing Methods 0.000 claims abstract description 13
- 108090000765 processed proteins & peptides Proteins 0.000 claims abstract description 11
- 229920001184 polypeptide Polymers 0.000 claims abstract description 9
- 102000004196 processed proteins & peptides Human genes 0.000 claims abstract description 9
- FWMNVWWHGCHHJJ-SKKKGAJSSA-N 4-amino-1-[(2r)-6-amino-2-[[(2r)-2-[[(2r)-2-[[(2r)-2-amino-3-phenylpropanoyl]amino]-3-phenylpropanoyl]amino]-4-methylpentanoyl]amino]hexanoyl]piperidine-4-carboxylic acid Chemical compound C([C@H](C(=O)N[C@H](CC(C)C)C(=O)N[C@H](CCCCN)C(=O)N1CCC(N)(CC1)C(O)=O)NC(=O)[C@H](N)CC=1C=CC=CC=1)C1=CC=CC=C1 FWMNVWWHGCHHJJ-SKKKGAJSSA-N 0.000 claims abstract description 3
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 33
- 239000008103 glucose Substances 0.000 claims description 32
- 150000001413 amino acids Chemical group 0.000 claims description 30
- 108010011384 acyl-CoA dehydrogenase (NADP+) Proteins 0.000 claims description 10
- 230000014509 gene expression Effects 0.000 claims description 9
- 108030003420 Ethylmalonyl-CoA decarboxylases Proteins 0.000 claims description 6
- 108091000039 acetoacetyl-CoA reductase Proteins 0.000 claims description 5
- 230000002194 synthesizing effect Effects 0.000 claims description 5
- 238000000034 method Methods 0.000 abstract description 29
- 125000003275 alpha amino acid group Chemical group 0.000 abstract 4
- 239000001963 growth medium Substances 0.000 abstract 1
- 239000002609 medium Substances 0.000 description 62
- 108020004414 DNA Proteins 0.000 description 57
- 108020004705 Codon Proteins 0.000 description 41
- 239000013600 plasmid vector Substances 0.000 description 41
- 239000013612 plasmid Substances 0.000 description 36
- 239000000203 mixture Substances 0.000 description 33
- 239000012634 fragment Substances 0.000 description 31
- 210000000349 chromosome Anatomy 0.000 description 28
- 210000004027 cell Anatomy 0.000 description 26
- 108091081024 Start codon Proteins 0.000 description 25
- 230000000052 comparative effect Effects 0.000 description 23
- 229930091371 Fructose Natural products 0.000 description 22
- 239000005715 Fructose Substances 0.000 description 22
- 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 22
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 18
- 238000012217 deletion Methods 0.000 description 18
- 230000037430 deletion Effects 0.000 description 18
- 101710108497 p-hydroxybenzoate hydroxylase Proteins 0.000 description 17
- 241000498271 Necator Species 0.000 description 16
- 235000000346 sugar Nutrition 0.000 description 15
- 229920001817 Agar Polymers 0.000 description 14
- 239000008272 agar Substances 0.000 description 14
- 101150108672 ccr gene Proteins 0.000 description 13
- 101150027065 nagE gene Proteins 0.000 description 12
- 239000006916 nutrient agar Substances 0.000 description 12
- 238000002360 preparation method Methods 0.000 description 12
- 241000607516 Aeromonas caviae Species 0.000 description 11
- 108090000790 Enzymes Proteins 0.000 description 11
- 230000012010 growth Effects 0.000 description 11
- 108091008146 restriction endonucleases Proteins 0.000 description 11
- 102000004190 Enzymes Human genes 0.000 description 10
- 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 10
- 101150113739 nagR gene Proteins 0.000 description 10
- 101150104605 phaZ2 gene Proteins 0.000 description 10
- 241000894006 Bacteria Species 0.000 description 9
- 238000003780 insertion Methods 0.000 description 9
- 230000037431 insertion Effects 0.000 description 9
- 239000000243 solution Substances 0.000 description 9
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 8
- 229930027917 kanamycin Natural products 0.000 description 8
- SBUJHOSQTJFQJX-NOAMYHISSA-N kanamycin Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CN)O[C@@H]1O[C@H]1[C@H](O)[C@@H](O[C@@H]2[C@@H]([C@@H](N)[C@H](O)[C@@H](CO)O2)O)[C@H](N)C[C@@H]1N SBUJHOSQTJFQJX-NOAMYHISSA-N 0.000 description 8
- 229960000318 kanamycin Drugs 0.000 description 8
- 229930182823 kanamycin A Natural products 0.000 description 8
- 239000000178 monomer Substances 0.000 description 8
- 101150051591 phaZ1 gene Proteins 0.000 description 8
- 239000013598 vector Substances 0.000 description 8
- 241000588724 Escherichia coli Species 0.000 description 7
- 101150018392 cscA gene Proteins 0.000 description 7
- 235000015097 nutrients Nutrition 0.000 description 7
- 101150048611 phaC gene Proteins 0.000 description 7
- 101150097421 phaJ gene Proteins 0.000 description 7
- 238000011144 upstream manufacturing Methods 0.000 description 7
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 6
- 230000015572 biosynthetic process Effects 0.000 description 6
- 101150055897 bktB gene Proteins 0.000 description 6
- 229920001577 copolymer Polymers 0.000 description 6
- 230000000813 microbial effect Effects 0.000 description 6
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 description 6
- 239000013587 production medium Substances 0.000 description 6
- 235000002639 sodium chloride Nutrition 0.000 description 6
- 238000003786 synthesis reaction Methods 0.000 description 6
- 238000012546 transfer Methods 0.000 description 6
- 102000012410 DNA Ligases Human genes 0.000 description 5
- 108010061982 DNA Ligases Proteins 0.000 description 5
- 101100061504 Escherichia coli cscB gene Proteins 0.000 description 5
- 239000002253 acid Substances 0.000 description 5
- 238000005119 centrifugation Methods 0.000 description 5
- 239000013611 chromosomal DNA Substances 0.000 description 5
- 230000021615 conjugation Effects 0.000 description 5
- 238000010353 genetic engineering Methods 0.000 description 5
- 229920001519 homopolymer Polymers 0.000 description 5
- 101150046540 phaA gene Proteins 0.000 description 5
- 241000607534 Aeromonas Species 0.000 description 4
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 4
- 238000004458 analytical method Methods 0.000 description 4
- 125000004432 carbon atom Chemical group C* 0.000 description 4
- 230000008859 change Effects 0.000 description 4
- 238000000855 fermentation Methods 0.000 description 4
- 230000004151 fermentation Effects 0.000 description 4
- 230000006801 homologous recombination Effects 0.000 description 4
- 238000002744 homologous recombination Methods 0.000 description 4
- 235000013379 molasses Nutrition 0.000 description 4
- 239000002773 nucleotide Substances 0.000 description 4
- 125000003729 nucleotide group Chemical group 0.000 description 4
- 229920000642 polymer Polymers 0.000 description 4
- 239000011780 sodium chloride Substances 0.000 description 4
- 238000006467 substitution reaction Methods 0.000 description 4
- 238000012360 testing method Methods 0.000 description 4
- 101710175935 (R)-specific enoyl-CoA hydratase Proteins 0.000 description 3
- WHBMMWSBFZVSSR-UHFFFAOYSA-N 3-hydroxybutyric acid Chemical compound CC(O)CC(O)=O WHBMMWSBFZVSSR-UHFFFAOYSA-N 0.000 description 3
- 241000588722 Escherichia Species 0.000 description 3
- 241001131785 Escherichia coli HB101 Species 0.000 description 3
- 102100029106 Ethylmalonyl-CoA decarboxylase Human genes 0.000 description 3
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 3
- 101100463818 Pseudomonas oleovorans phaC1 gene Proteins 0.000 description 3
- 241000232299 Ralstonia Species 0.000 description 3
- 101100297400 Rhizobium meliloti (strain 1021) phaAB gene Proteins 0.000 description 3
- LFVGISIMTYGQHF-UHFFFAOYSA-N ammonium dihydrogen phosphate Chemical compound [NH4+].OP(O)([O-])=O LFVGISIMTYGQHF-UHFFFAOYSA-N 0.000 description 3
- 229910000387 ammonium dihydrogen phosphate Inorganic materials 0.000 description 3
- 239000007864 aqueous solution Substances 0.000 description 3
- CRFNGMNYKDXRTN-CITAKDKDSA-N butyryl-CoA Chemical compound O[C@@H]1[C@H](OP(O)(O)=O)[C@@H](COP(O)(=O)OP(O)(=O)OCC(C)(C)[C@@H](O)C(=O)NCCC(=O)NCCSC(=O)CCC)O[C@H]1N1C2=NC=NC(N)=C2N=C1 CRFNGMNYKDXRTN-CITAKDKDSA-N 0.000 description 3
- 230000010261 cell growth Effects 0.000 description 3
- 238000006243 chemical reaction Methods 0.000 description 3
- 235000014113 dietary fatty acids Nutrition 0.000 description 3
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 description 3
- -1 emd Proteins 0.000 description 3
- 229930195729 fatty acid Natural products 0.000 description 3
- 239000000194 fatty acid Substances 0.000 description 3
- 150000004665 fatty acids Chemical class 0.000 description 3
- WRUGWIBCXHJTDG-UHFFFAOYSA-L magnesium sulfate heptahydrate Chemical compound O.O.O.O.O.O.O.[Mg+2].[O-]S([O-])(=O)=O WRUGWIBCXHJTDG-UHFFFAOYSA-L 0.000 description 3
- 229940061634 magnesium sulfate heptahydrate Drugs 0.000 description 3
- 230000007246 mechanism Effects 0.000 description 3
- 235000019837 monoammonium phosphate Nutrition 0.000 description 3
- 239000003921 oil Substances 0.000 description 3
- 235000019198 oils Nutrition 0.000 description 3
- 108010093591 phosphoenolpyruvate-sucrose phosphotransferase Proteins 0.000 description 3
- 239000001509 sodium citrate Substances 0.000 description 3
- NLJMYIDDQXHKNR-UHFFFAOYSA-K sodium citrate Chemical compound O.O.[Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O NLJMYIDDQXHKNR-UHFFFAOYSA-K 0.000 description 3
- NDPLAKGOSZHTPH-UHFFFAOYSA-N 3-hydroxyoctanoic acid Chemical compound CCCCCC(O)CC(O)=O NDPLAKGOSZHTPH-UHFFFAOYSA-N 0.000 description 2
- REKYPYSUBKSCAT-UHFFFAOYSA-N 3-hydroxypentanoic acid Chemical compound CCC(O)CC(O)=O REKYPYSUBKSCAT-UHFFFAOYSA-N 0.000 description 2
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 2
- NLXLAEXVIDQMFP-UHFFFAOYSA-N Ammonia chloride Chemical compound [NH4+].[Cl-] NLXLAEXVIDQMFP-UHFFFAOYSA-N 0.000 description 2
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 description 2
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 2
- 241001528480 Cupriavidus Species 0.000 description 2
- 101150111184 Emd gene Proteins 0.000 description 2
- 101100459630 Escherichia coli (strain K12) nagC gene Proteins 0.000 description 2
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 2
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 2
- 108091000080 Phosphotransferase Proteins 0.000 description 2
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical compound [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 description 2
- 108700005078 Synthetic Genes Proteins 0.000 description 2
- ZSLZBFCDCINBPY-ZSJPKINUSA-N acetyl-CoA Chemical compound O[C@@H]1[C@H](OP(O)(O)=O)[C@@H](COP(O)(=O)OP(O)(=O)OCC(C)(C)[C@@H](O)C(=O)NCCC(=O)NCCSC(=O)C)O[C@H]1N1C2=NC=NC(N)=C2N=C1 ZSLZBFCDCINBPY-ZSJPKINUSA-N 0.000 description 2
- 210000004507 artificial chromosome Anatomy 0.000 description 2
- 230000001580 bacterial effect Effects 0.000 description 2
- 230000005540 biological transmission Effects 0.000 description 2
- 229940041514 candida albicans extract Drugs 0.000 description 2
- 238000003776 cleavage reaction Methods 0.000 description 2
- 101150075169 cscB gene Proteins 0.000 description 2
- POULHZVOKOAJMA-UHFFFAOYSA-N dodecanoic acid Chemical compound CCCCCCCCCCCC(O)=O POULHZVOKOAJMA-UHFFFAOYSA-N 0.000 description 2
- 239000000284 extract Substances 0.000 description 2
- 238000001914 filtration Methods 0.000 description 2
- 239000007789 gas Substances 0.000 description 2
- 238000004817 gas chromatography Methods 0.000 description 2
- IPCSVZSSVZVIGE-UHFFFAOYSA-N hexadecanoic acid Chemical compound CCCCCCCCCCCCCCCC(O)=O IPCSVZSSVZVIGE-UHFFFAOYSA-N 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 238000010369 molecular cloning Methods 0.000 description 2
- 229910052757 nitrogen Inorganic materials 0.000 description 2
- 239000003960 organic solvent Substances 0.000 description 2
- 238000007254 oxidation reaction Methods 0.000 description 2
- 101150032462 phaC1 gene Proteins 0.000 description 2
- 238000006116 polymerization reaction Methods 0.000 description 2
- 239000002994 raw material Substances 0.000 description 2
- 230000006798 recombination Effects 0.000 description 2
- 101150025220 sacB gene Proteins 0.000 description 2
- 150000003839 salts Chemical class 0.000 description 2
- 230000007017 scission Effects 0.000 description 2
- 239000011734 sodium Substances 0.000 description 2
- 229910052708 sodium Inorganic materials 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 239000012138 yeast extract Substances 0.000 description 2
- MTCFGRXMJLQNBG-REOHCLBHSA-N (2S)-2-Amino-3-hydroxypropansäure Chemical compound OC[C@H](N)C(O)=O MTCFGRXMJLQNBG-REOHCLBHSA-N 0.000 description 1
- WRIDQFICGBMAFQ-UHFFFAOYSA-N (E)-8-Octadecenoic acid Natural products CCCCCCCCCC=CCCCCCCC(O)=O WRIDQFICGBMAFQ-UHFFFAOYSA-N 0.000 description 1
- LQJBNNIYVWPHFW-UHFFFAOYSA-N 20:1omega9c fatty acid Natural products CCCCCCCCCCC=CCCCCCCCC(O)=O LQJBNNIYVWPHFW-UHFFFAOYSA-N 0.000 description 1
- HPMGFDVTYHWBAG-UHFFFAOYSA-N 3-hydroxyhexanoic acid Chemical compound CCCC(O)CC(O)=O HPMGFDVTYHWBAG-UHFFFAOYSA-N 0.000 description 1
- 102100026105 3-ketoacyl-CoA thiolase, mitochondrial Human genes 0.000 description 1
- SJZRECIVHVDYJC-UHFFFAOYSA-N 4-hydroxybutyric acid Chemical compound OCCCC(O)=O SJZRECIVHVDYJC-UHFFFAOYSA-N 0.000 description 1
- 229940006015 4-hydroxybutyric acid Drugs 0.000 description 1
- 101150039504 6 gene Proteins 0.000 description 1
- QSBYPNXLFMSGKH-UHFFFAOYSA-N 9-Heptadecensaeure Natural products CCCCCCCC=CCCCCCCCC(O)=O QSBYPNXLFMSGKH-UHFFFAOYSA-N 0.000 description 1
- 108010003902 Acetyl-CoA C-acyltransferase Proteins 0.000 description 1
- 239000004254 Ammonium phosphate Substances 0.000 description 1
- 244000063299 Bacillus subtilis Species 0.000 description 1
- 235000014469 Bacillus subtilis Nutrition 0.000 description 1
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 description 1
- 101100326160 Cupriavidus necator (strain ATCC 17699 / DSM 428 / KCTC 22496 / NCIMB 10442 / H16 / Stanier 337) bktB gene Proteins 0.000 description 1
- 101100506339 Cupriavidus necator (strain ATCC 17699 / DSM 428 / KCTC 22496 / NCIMB 10442 / H16 / Stanier 337) phaZ2 gene Proteins 0.000 description 1
- NBSCHQHZLSJFNQ-GASJEMHNSA-N D-Glucose 6-phosphate Chemical compound OC1O[C@H](COP(O)(O)=O)[C@@H](O)[C@H](O)[C@H]1O NBSCHQHZLSJFNQ-GASJEMHNSA-N 0.000 description 1
- ZZZCUOFIHGPKAK-UHFFFAOYSA-N D-erythro-ascorbic acid Natural products OCC1OC(=O)C(O)=C1O ZZZCUOFIHGPKAK-UHFFFAOYSA-N 0.000 description 1
- ZAFNJMIOTHYJRJ-UHFFFAOYSA-N Diisopropyl ether Chemical compound CC(C)OC(C)C ZAFNJMIOTHYJRJ-UHFFFAOYSA-N 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 241001646716 Escherichia coli K-12 Species 0.000 description 1
- 241000901842 Escherichia coli W Species 0.000 description 1
- VUGZQVCBBBEZQE-VRQRJWBYSA-N Ethylmalonyl-CoA Chemical compound O[C@@H]1[C@H](OP(O)(O)=O)[C@@H](COP(O)(=O)OP(O)(=O)OCC(C)(C)C(O)C(=O)NCCC(=O)NCCSC(=O)C(C(O)=O)CC)O[C@H]1N1C2=NC=NC(N)=C2N=C1 VUGZQVCBBBEZQE-VRQRJWBYSA-N 0.000 description 1
- 102000018711 Facilitative Glucose Transport Proteins Human genes 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- 206010064571 Gene mutation Diseases 0.000 description 1
- 108700039691 Genetic Promoter Regions Proteins 0.000 description 1
- VFRROHXSMXFLSN-UHFFFAOYSA-N Glc6P Natural products OP(=O)(O)OCC(O)C(O)C(O)C(O)C=O VFRROHXSMXFLSN-UHFFFAOYSA-N 0.000 description 1
- 108010021582 Glucokinase Proteins 0.000 description 1
- 108091052347 Glucose transporter family Proteins 0.000 description 1
- 239000004471 Glycine Substances 0.000 description 1
- 241000011814 Ichthyobodo necator Species 0.000 description 1
- 241000221089 Jatropha Species 0.000 description 1
- ONIBWKKTOPOVIA-BYPYZUCNSA-N L-Proline Chemical compound OC(=O)[C@@H]1CCCN1 ONIBWKKTOPOVIA-BYPYZUCNSA-N 0.000 description 1
- QNAYBMKLOCPYGJ-REOHCLBHSA-N L-alanine Chemical compound C[C@H](N)C(O)=O QNAYBMKLOCPYGJ-REOHCLBHSA-N 0.000 description 1
- AYFVYJQAPQTCCC-GBXIJSLDSA-N L-threonine Chemical compound C[C@@H](O)[C@H](N)C(O)=O AYFVYJQAPQTCCC-GBXIJSLDSA-N 0.000 description 1
- 239000005639 Lauric acid Substances 0.000 description 1
- 108010085747 Methylmalonyl-CoA Decarboxylase Proteins 0.000 description 1
- OVRNDRQMDRJTHS-UHFFFAOYSA-N N-acelyl-D-glucosamine Natural products CC(=O)NC1C(O)OC(CO)C(O)C1O OVRNDRQMDRJTHS-UHFFFAOYSA-N 0.000 description 1
- OVRNDRQMDRJTHS-FMDGEEDCSA-N N-acetyl-beta-D-glucosamine Chemical compound CC(=O)N[C@H]1[C@H](O)O[C@H](CO)[C@@H](O)[C@@H]1O OVRNDRQMDRJTHS-FMDGEEDCSA-N 0.000 description 1
- MBLBDJOUHNCFQT-LXGUWJNJSA-N N-acetylglucosamine Natural products CC(=O)N[C@@H](C=O)[C@@H](O)[C@H](O)[C@H](O)CO MBLBDJOUHNCFQT-LXGUWJNJSA-N 0.000 description 1
- 238000005481 NMR spectroscopy Methods 0.000 description 1
- 108091028043 Nucleic acid sequence Proteins 0.000 description 1
- 239000005642 Oleic acid Substances 0.000 description 1
- ZQPPMHVWECSIRJ-UHFFFAOYSA-N Oleic acid Natural products CCCCCCCCC=CCCCCCCCC(O)=O ZQPPMHVWECSIRJ-UHFFFAOYSA-N 0.000 description 1
- 108090000854 Oxidoreductases Proteins 0.000 description 1
- 102000004316 Oxidoreductases Human genes 0.000 description 1
- 235000019482 Palm oil Nutrition 0.000 description 1
- 235000021314 Palmitic acid Nutrition 0.000 description 1
- 239000001888 Peptone Substances 0.000 description 1
- 108010080698 Peptones Proteins 0.000 description 1
- 239000002202 Polyethylene glycol Substances 0.000 description 1
- ONIBWKKTOPOVIA-UHFFFAOYSA-N Proline Natural products OC(=O)C1CCCN1 ONIBWKKTOPOVIA-UHFFFAOYSA-N 0.000 description 1
- 241000589516 Pseudomonas Species 0.000 description 1
- 235000019484 Rapeseed oil Nutrition 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- MTCFGRXMJLQNBG-UHFFFAOYSA-N Serine Natural products OCC(N)C(O)=O MTCFGRXMJLQNBG-UHFFFAOYSA-N 0.000 description 1
- 235000021355 Stearic acid Nutrition 0.000 description 1
- 101100309436 Streptococcus mutans serotype c (strain ATCC 700610 / UA159) ftf gene Proteins 0.000 description 1
- WQQSIXKPRAUZJL-UGDNZRGBSA-N Sucrose 6-phosphate Natural products 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](COP(O)(O)=O)O1 WQQSIXKPRAUZJL-UGDNZRGBSA-N 0.000 description 1
- 108020005038 Terminator Codon Proteins 0.000 description 1
- AYFVYJQAPQTCCC-UHFFFAOYSA-N Threonine Natural products CC(O)C(N)C(O)=O AYFVYJQAPQTCCC-UHFFFAOYSA-N 0.000 description 1
- 239000004473 Threonine Substances 0.000 description 1
- 108091023040 Transcription factor Proteins 0.000 description 1
- 102000040945 Transcription factor Human genes 0.000 description 1
- 229930003451 Vitamin B1 Natural products 0.000 description 1
- 229930003779 Vitamin B12 Natural products 0.000 description 1
- 229930003268 Vitamin C Natural products 0.000 description 1
- OJFDKHTZOUZBOS-CITAKDKDSA-N acetoacetyl-CoA Chemical compound O[C@@H]1[C@H](OP(O)(O)=O)[C@@H](COP(O)(=O)OP(O)(=O)OCC(C)(C)[C@@H](O)C(=O)NCCC(=O)NCCSC(=O)CC(=O)C)O[C@H]1N1C2=NC=NC(N)=C2N=C1 OJFDKHTZOUZBOS-CITAKDKDSA-N 0.000 description 1
- 150000007513 acids Chemical class 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 235000004279 alanine Nutrition 0.000 description 1
- 125000000217 alkyl group Chemical group 0.000 description 1
- 235000001014 amino acid Nutrition 0.000 description 1
- 229910021529 ammonia Inorganic materials 0.000 description 1
- 235000019270 ammonium chloride Nutrition 0.000 description 1
- 229910000148 ammonium phosphate Inorganic materials 0.000 description 1
- 235000019289 ammonium phosphates Nutrition 0.000 description 1
- 150000003863 ammonium salts Chemical class 0.000 description 1
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 description 1
- 229910052921 ammonium sulfate Inorganic materials 0.000 description 1
- 235000011130 ammonium sulphate Nutrition 0.000 description 1
- 239000000560 biocompatible material Substances 0.000 description 1
- 239000001110 calcium chloride Substances 0.000 description 1
- 229910001628 calcium chloride Inorganic materials 0.000 description 1
- 238000003965 capillary gas chromatography Methods 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 235000014633 carbohydrates Nutrition 0.000 description 1
- 239000001569 carbon dioxide Substances 0.000 description 1
- 229910002092 carbon dioxide Inorganic materials 0.000 description 1
- 239000012159 carrier gas Substances 0.000 description 1
- 230000003197 catalytic effect Effects 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 238000010367 cloning Methods 0.000 description 1
- AGVAZMGAQJOSFJ-WZHZPDAFSA-M cobalt(2+);[(2r,3s,4r,5s)-5-(5,6-dimethylbenzimidazol-1-yl)-4-hydroxy-2-(hydroxymethyl)oxolan-3-yl] [(2r)-1-[3-[(1r,2r,3r,4z,7s,9z,12s,13s,14z,17s,18s,19r)-2,13,18-tris(2-amino-2-oxoethyl)-7,12,17-tris(3-amino-3-oxopropyl)-3,5,8,8,13,15,18,19-octamethyl-2 Chemical compound [Co+2].N#[C-].[N-]([C@@H]1[C@H](CC(N)=O)[C@@]2(C)CCC(=O)NC[C@@H](C)OP(O)(=O)O[C@H]3[C@H]([C@H](O[C@@H]3CO)N3C4=CC(C)=C(C)C=C4N=C3)O)\C2=C(C)/C([C@H](C\2(C)C)CCC(N)=O)=N/C/2=C\C([C@H]([C@@]/2(CC(N)=O)C)CCC(N)=O)=N\C\2=C(C)/C2=N[C@]1(C)[C@@](C)(CC(N)=O)[C@@H]2CCC(N)=O AGVAZMGAQJOSFJ-WZHZPDAFSA-M 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 235000005687 corn oil Nutrition 0.000 description 1
- 239000002285 corn oil Substances 0.000 description 1
- KFWWCMJSYSSPSK-PAXLJYGASA-N crotonoyl-CoA Chemical compound O[C@@H]1[C@H](OP(O)(O)=O)[C@@H](COP(O)(=O)OP(O)(=O)OCC(C)(C)[C@@H](O)C(=O)NCCC(=O)NCCSC(=O)/C=C/C)O[C@H]1N1C2=NC=NC(N)=C2N=C1 KFWWCMJSYSSPSK-PAXLJYGASA-N 0.000 description 1
- 239000010779 crude oil Substances 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 238000006114 decarboxylation reaction Methods 0.000 description 1
- 238000000354 decomposition reaction Methods 0.000 description 1
- 239000007857 degradation product Substances 0.000 description 1
- 230000000593 degrading effect Effects 0.000 description 1
- MNNHAPBLZZVQHP-UHFFFAOYSA-N diammonium hydrogen phosphate Chemical compound [NH4+].[NH4+].OP([O-])([O-])=O MNNHAPBLZZVQHP-UHFFFAOYSA-N 0.000 description 1
- 150000002016 disaccharides Chemical class 0.000 description 1
- BNIILDVGGAEEIG-UHFFFAOYSA-L disodium hydrogen phosphate Chemical compound [Na+].[Na+].OP([O-])([O-])=O BNIILDVGGAEEIG-UHFFFAOYSA-L 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 238000004520 electroporation Methods 0.000 description 1
- 230000008030 elimination Effects 0.000 description 1
- 238000003379 elimination reaction Methods 0.000 description 1
- 230000001747 exhibiting effect Effects 0.000 description 1
- 239000013604 expression vector Substances 0.000 description 1
- 239000000706 filtrate Substances 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 238000005227 gel permeation chromatography Methods 0.000 description 1
- 238000012215 gene cloning Methods 0.000 description 1
- 230000004190 glucose uptake Effects 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 230000003301 hydrolyzing effect Effects 0.000 description 1
- 230000010354 integration Effects 0.000 description 1
- QXJSBBXBKPUZAA-UHFFFAOYSA-N isooleic acid Natural products CCCCCCCC=CCCCCCCCCC(O)=O QXJSBBXBKPUZAA-UHFFFAOYSA-N 0.000 description 1
- GVALZJMUIHGIMD-UHFFFAOYSA-H magnesium phosphate Chemical compound [Mg+2].[Mg+2].[Mg+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O GVALZJMUIHGIMD-UHFFFAOYSA-H 0.000 description 1
- 239000004137 magnesium phosphate Substances 0.000 description 1
- 229960002261 magnesium phosphate Drugs 0.000 description 1
- 229910000157 magnesium phosphate Inorganic materials 0.000 description 1
- 235000010994 magnesium phosphates Nutrition 0.000 description 1
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 1
- 235000019341 magnesium sulphate Nutrition 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 235000013372 meat Nutrition 0.000 description 1
- 230000002503 metabolic effect Effects 0.000 description 1
- 238000012269 metabolic engineering Methods 0.000 description 1
- WCYAALZQFZMMOM-UHFFFAOYSA-N methanol;sulfuric acid Chemical compound OC.OS(O)(=O)=O WCYAALZQFZMMOM-UHFFFAOYSA-N 0.000 description 1
- 150000004702 methyl esters Chemical class 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 1
- 235000019796 monopotassium phosphate Nutrition 0.000 description 1
- 230000035772 mutation Effects 0.000 description 1
- WQEPLUUGTLDZJY-UHFFFAOYSA-N n-Pentadecanoic acid Natural products CCCCCCCCCCCCCCC(O)=O WQEPLUUGTLDZJY-UHFFFAOYSA-N 0.000 description 1
- 229950006780 n-acetylglucosamine Drugs 0.000 description 1
- QIQXTHQIDYTFRH-UHFFFAOYSA-N octadecanoic acid Chemical compound CCCCCCCCCCCCCCCCCC(O)=O QIQXTHQIDYTFRH-UHFFFAOYSA-N 0.000 description 1
- OQCDKBAXFALNLD-UHFFFAOYSA-N octadecanoic acid Natural products CCCCCCCC(C)CCCCCCCCC(O)=O OQCDKBAXFALNLD-UHFFFAOYSA-N 0.000 description 1
- 235000014593 oils and fats Nutrition 0.000 description 1
- ZQPPMHVWECSIRJ-KTKRTIGZSA-N oleic acid Chemical compound CCCCCCCC\C=C/CCCCCCCC(O)=O ZQPPMHVWECSIRJ-KTKRTIGZSA-N 0.000 description 1
- 235000021313 oleic acid Nutrition 0.000 description 1
- 239000004006 olive oil Substances 0.000 description 1
- 235000008390 olive oil Nutrition 0.000 description 1
- 229920000620 organic polymer Polymers 0.000 description 1
- 239000003346 palm kernel oil Substances 0.000 description 1
- 235000019865 palm kernel oil Nutrition 0.000 description 1
- 239000002540 palm oil Substances 0.000 description 1
- 230000037361 pathway Effects 0.000 description 1
- 235000019319 peptone Nutrition 0.000 description 1
- 229930029653 phosphoenolpyruvate Natural products 0.000 description 1
- DTBNBXWJWCWCIK-UHFFFAOYSA-N phosphoenolpyruvic acid Chemical compound OC(=O)C(=C)OP(O)(O)=O DTBNBXWJWCWCIK-UHFFFAOYSA-N 0.000 description 1
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 description 1
- 102000020233 phosphotransferase Human genes 0.000 description 1
- 230000029553 photosynthesis Effects 0.000 description 1
- 238000010672 photosynthesis Methods 0.000 description 1
- 230000000704 physical effect Effects 0.000 description 1
- 229920000728 polyester Polymers 0.000 description 1
- 229920001223 polyethylene glycol Polymers 0.000 description 1
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 235000013930 proline Nutrition 0.000 description 1
- 235000018102 proteins Nutrition 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 238000005215 recombination Methods 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 230000000717 retained effect Effects 0.000 description 1
- 239000012266 salt solution Substances 0.000 description 1
- 235000004400 serine Nutrition 0.000 description 1
- 238000007086 side reaction Methods 0.000 description 1
- 235000017557 sodium bicarbonate Nutrition 0.000 description 1
- 229910000030 sodium bicarbonate Inorganic materials 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 239000003549 soybean oil Substances 0.000 description 1
- 235000012424 soybean oil Nutrition 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 239000008117 stearic acid Substances 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- PJTTXANTBQDXME-UGDNZRGBSA-N sucrose 6(F)-phosphate Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@]1(CO)[C@@H](O)[C@H](O)[C@@H](COP(O)(O)=O)O1 PJTTXANTBQDXME-UGDNZRGBSA-N 0.000 description 1
- 150000008163 sugars Chemical class 0.000 description 1
- TUNFSRHWOTWDNC-HKGQFRNVSA-N tetradecanoic acid Chemical compound CCCCCCCCCCCCC[14C](O)=O TUNFSRHWOTWDNC-HKGQFRNVSA-N 0.000 description 1
- 229920001169 thermoplastic Polymers 0.000 description 1
- 229960003495 thiamine Drugs 0.000 description 1
- DPJRMOMPQZCRJU-UHFFFAOYSA-M thiamine hydrochloride Chemical compound Cl.[Cl-].CC1=C(CCO)SC=[N+]1CC1=CN=C(C)N=C1N DPJRMOMPQZCRJU-UHFFFAOYSA-M 0.000 description 1
- 235000008521 threonine Nutrition 0.000 description 1
- 229910021654 trace metal Inorganic materials 0.000 description 1
- 238000013519 translation Methods 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
- 229930003231 vitamin Natural products 0.000 description 1
- 229940088594 vitamin Drugs 0.000 description 1
- 235000010374 vitamin B1 Nutrition 0.000 description 1
- 239000011691 vitamin B1 Substances 0.000 description 1
- 235000019163 vitamin B12 Nutrition 0.000 description 1
- 239000011715 vitamin B12 Substances 0.000 description 1
- 235000019154 vitamin C Nutrition 0.000 description 1
- 239000011718 vitamin C Substances 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P7/00—Preparation of oxygen-containing organic compounds
- C12P7/62—Carboxylic acid esters
- C12P7/625—Polyesters of hydroxy carboxylic acids
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/195—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria
- C07K14/24—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria from Enterobacteriaceae (F), e.g. Citrobacter, Serratia, Proteus, Providencia, Morganella, Yersinia
- C07K14/245—Escherichia (G)
-
- 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
- C08G63/00—Macromolecular compounds obtained by reactions forming a carboxylic ester link in the main chain of the macromolecule
- C08G63/02—Polyesters derived from hydroxycarboxylic acids or from polycarboxylic acids and polyhydroxy compounds
- C08G63/06—Polyesters derived from hydroxycarboxylic acids or from polycarboxylic acids and polyhydroxy compounds derived from hydroxycarboxylic acids
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
- C12N15/52—Genes encoding for enzymes or proenzymes
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/10—Transferases (2.)
- C12N9/1025—Acyltransferases (2.3)
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/10—Transferases (2.)
- C12N9/1025—Acyltransferases (2.3)
- C12N9/1029—Acyltransferases (2.3) transferring groups other than amino-acyl groups (2.3.1)
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/10—Transferases (2.)
- C12N9/1048—Glycosyltransferases (2.4)
- C12N9/1051—Hexosyltransferases (2.4.1)
- C12N9/1062—Sucrose synthase (2.4.1.13)
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/14—Hydrolases (3)
- C12N9/24—Hydrolases (3) acting on glycosyl compounds (3.2)
- C12N9/2402—Hydrolases (3) acting on glycosyl compounds (3.2) hydrolysing O- and S- glycosyl compounds (3.2.1)
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y204/00—Glycosyltransferases (2.4)
- C12Y204/01—Hexosyltransferases (2.4.1)
- C12Y204/01013—Sucrose synthase (2.4.1.13)
Landscapes
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Genetics & Genomics (AREA)
- Engineering & Computer Science (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biotechnology (AREA)
- General Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Molecular Biology (AREA)
- Microbiology (AREA)
- Medicinal Chemistry (AREA)
- Biophysics (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Physics & Mathematics (AREA)
- Plant Pathology (AREA)
- General Chemical & Material Sciences (AREA)
- Gastroenterology & Hepatology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Virology (AREA)
- Polymers & Plastics (AREA)
- Tropical Medicine & Parasitology (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
Description
(1)配列番号1に記載のアミノ酸配列をコードするスクロース加水分解酵素遺伝子、又は該アミノ酸配列に対して90%以上の配列同一性を有し、スクロース加水分解酵素活性を有するポリペプチドをコードする遺伝子
(2)配列番号2に記載のアミノ酸配列をコードするスクロース透過酵素遺伝子、又は該アミノ酸配列に対して90%以上の配列同一性を有し、スクロース透過酵素活性を有するポリペプチドをコードする遺伝子
好ましくは、前記微生物が、カプリアビダス属に属する微生物を宿主とする形質転換体であり、より好ましくは、前記カプリアビダス属に属する微生物が、カプリアビダス・ネカトールである。好ましくは、前記微生物はグルコース資化性が付与又は強化されている。好ましくは、前記PHA合成酵素遺伝子が、P(3HB−co−3HH)を合成可能なPHA合成酵素遺伝子である。好ましくは、前記微生物は、さらにクロトニル−CoA還元酵素遺伝子およびエチルマロニル−CoA脱炭酸酵素遺伝子を有する。好ましくは、前記微生物は、アセトアセチルCoA還元酵素遺伝子が欠失した、またはその発現量が抑制されている。
本発明では、PHA合成酵素遺伝子を有する微生物に対し、異種生物由来のスクロース加水分解酵素遺伝子と異種生物由来のスクロース透過酵素遺伝子の両方を導入することによって、スクロース資化性が付与または強化され、スクロースを炭素源としてPHAを生産する微生物を提供する。
本発明の微生物を、炭素源としてスクロースを含む培地で培養することで、PHAを生産させ、得られたPHAを回収することでPHAを製造することができる。
まず、染色体置換用プラスミドの作製を行った。作製は以下のように行った。
まず、cscAおよびcscB遺伝子発現用プラスミドの作製を行った。作製は以下のように行った。
製造例2で作製したプラスミドベクターpCUP2−lacUV5−cscABを、製造例2と同様の方法で、製造例1で作製したKNK005ΔphaZ1,2,6/nagEG793C,dR株へ導入し、形質転換体pCUP2−lacUV5−cscAB in KNK005ΔphaZ1,2,6/nagEG793C,dR株を得た。
まず、cscA遺伝子発現用プラスミドの作製を行った。作製は以下のように行った。
まず、cscB遺伝子発現用プラスミドの作製を行った。作製は以下のように行った。
まず、特開2013−9627号公報に記載のプラスミドbAO/pBlu/SacB−Kmを用いて、以下のようにしてプロモーターおよびリボソーム結合配列挿入株ACP−bktB/ΔphaZ1,2,6/nagEG793C,dR株の作製を行った。
まず、プロモーター、リボソーム結合配列及び遺伝子挿入用プラスミドの作製を行った。作製は以下のように行った。
まず、遺伝子破壊用プラスミドの作製を行った。作製は以下のように行った。
製造例8に記載の遺伝子破壊用プラスミドベクターpNS2X−sacB+phaAB1UDを用いて、製造例7に記載のKNK−143S株を親株として、製造例1に記載の染色体置換株の作製と同様に、接合伝達、シモンズ寒天培地での選択、及び、15%のシュークロースを含むNutrient Agar培地での選択を行い、KNK142S株を作製した。KNK142S株はC.necator H16株の染色体上のphaZ6遺伝子及びphaZ1遺伝子の開始コドンから終止コドンまでを欠失し、さらにphaZ2遺伝子の16番目のコドンから終止コドンまでを欠失し、染色体上に配列番号4に記載のアミノ酸配列を有するPHA合成酵素をコードする遺伝子が導入され、nagE構造遺伝子の793番目の塩基であるGがCに置換され、nagR遺伝子の開始コドンから終止コドンまでを欠失し、bktB遺伝子の開始コドン直前にA. caviaeのphaC遺伝子のプロモーターおよびリボソーム結合配列を含む塩基配列からなるDNAが挿入され、phaJ4b遺伝子の開始コドン直前にtrcプロモーターおよびリボソーム結合配列を含む塩基配列からなるDNAが挿入され、元々はphaZ2遺伝子があった位置にtrcプロモーター、リボソーム結合配列、ccr遺伝子及びemd遺伝子が挿入され、さらにphaA遺伝子の開始コドンからphaB1遺伝子の終止コドンまでを欠失した株である。
種母培地の組成は1w/v% Meat−extract、1w/v% Bacto−Trypton、0.2w/v% Yeast−extract、0.9w/v% Na2HPO4・12H2O、0.15w/v% KH2PO4とした。種母培地でプラスミドベクター導入株を培養する場合には、カナマイシンを最終濃度100μg/mlとなるように種母培地に添加した。
種母培地の組成は比較例1〜4に記載のものと同様とした。種母培地でプラスミドベクター導入株を培養する場合には、カナマイシンを最終濃度100μg/mlとなるように種母培地に添加した。
種母培地の組成は比較例1〜4に記載のものと同様とした。種母培地でプラスミドベクター導入株を培養する場合には、カナマイシンを最終濃度100μg/mlとなるように種母培地に添加した。
種母培地の組成は比較例1〜4に記載のものと同様とした。種母培地でプラスミドベクター導入株を培養する場合には、カナマイシンを最終濃度100μg/mlとなるように種母培地に添加した。
Claims (9)
- PHA合成酵素遺伝子と、下記(1)及び(2)の異種生物由来遺伝子とを有するPHA生産微生物。
(1)配列番号1に記載のアミノ酸配列をコードするスクロース加水分解酵素遺伝子、又は該アミノ酸配列に対して90%以上の配列同一性を有し、スクロース加水分解酵素活性を有するポリペプチドをコードする遺伝子
(2)配列番号2に記載のアミノ酸配列をコードするスクロース透過酵素遺伝子、又は該アミノ酸配列に対して90%以上の配列同一性を有し、スクロース透過酵素活性を有するポリペプチドをコードする遺伝子 - 前記微生物が、カプリアビダス属に属する微生物を宿主とする形質転換体である、請求項1に記載の微生物。
- 前記カプリアビダス属に属する微生物が、カプリアビダス・ネカトールである、請求項2に記載の微生物。
- グルコース資化性が付与又は強化されている、請求項1〜3いずれか1項に記載の微生物。
- 前記PHA合成酵素遺伝子が、P(3HB−co−3HH)を合成可能なPHA合成酵素遺伝子である、請求項1〜4いずれか1項に記載の微生物。
- さらにクロトニル−CoA還元酵素遺伝子およびエチルマロニル−CoA脱炭酸酵素遺伝子を有する、請求項3〜5いずれか1項に記載の微生物。
- アセトアセチルCoA還元酵素遺伝子が欠失した、またはその発現量が抑制されている、請求項6に記載の微生物。
- 請求項1〜7いずれか1項に記載の微生物を、スクロースを炭素源として含む培地で培養する工程を含む、PHAの製造方法。
- PHAがP(3HB−co−3HH)である請求項8に記載の製造方法。
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP2015245214 | 2015-12-16 | ||
JP2015245214 | 2015-12-16 | ||
PCT/JP2016/087292 WO2017104722A1 (ja) | 2015-12-16 | 2016-12-14 | スクロース資化性を有するpha生産微生物、及び該微生物を用いたphaの製造方法 |
Publications (2)
Publication Number | Publication Date |
---|---|
JPWO2017104722A1 true JPWO2017104722A1 (ja) | 2018-10-04 |
JP6853787B2 JP6853787B2 (ja) | 2021-03-31 |
Family
ID=59056918
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP2017556109A Active JP6853787B2 (ja) | 2015-12-16 | 2016-12-14 | スクロース資化性を有するpha生産微生物、及び該微生物を用いたphaの製造方法 |
Country Status (3)
Country | Link |
---|---|
US (1) | US20180371509A1 (ja) |
JP (1) | JP6853787B2 (ja) |
WO (1) | WO2017104722A1 (ja) |
Families Citing this family (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US11186831B2 (en) | 2018-01-16 | 2021-11-30 | Kaneka Corporation | Mutant polyhydroxyalkanoate synthase, gene thereof and transformant, and method for producing polyhydroxyalkanoate |
CN111979163B (zh) * | 2019-05-24 | 2022-05-06 | 深圳蓝晶生物科技有限公司 | 一种重组罗氏真氧菌及其制备方法和应用 |
CN114480317B (zh) * | 2022-04-06 | 2022-07-29 | 深圳蓝晶生物科技有限公司 | 表达乙酰乙酰辅酶a还原酶变体的工程化微生物及提高pha产量的方法 |
Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2013025286A1 (en) * | 2011-08-16 | 2013-02-21 | E. I. Du Pont De Nemours And Company | Recombinant bacteria having improved sucrose utilization |
WO2014052923A2 (en) * | 2012-09-28 | 2014-04-03 | The Regents Of The University Of California | Trophic conversion of photoautotrophic bacteria for improved diurnal properties |
WO2014065253A1 (ja) * | 2012-10-22 | 2014-05-01 | 株式会社カネカ | 高分子量pha生産微生物とそれを用いた高分子量phaの製造方法 |
WO2016021604A1 (ja) * | 2014-08-04 | 2016-02-11 | 国立大学法人東京工業大学 | 糖質原料からの共重合ポリヒドロキシアルカン酸の製造法 |
-
2016
- 2016-12-14 WO PCT/JP2016/087292 patent/WO2017104722A1/ja active Application Filing
- 2016-12-14 US US16/061,923 patent/US20180371509A1/en not_active Abandoned
- 2016-12-14 JP JP2017556109A patent/JP6853787B2/ja active Active
Patent Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2013025286A1 (en) * | 2011-08-16 | 2013-02-21 | E. I. Du Pont De Nemours And Company | Recombinant bacteria having improved sucrose utilization |
WO2014052923A2 (en) * | 2012-09-28 | 2014-04-03 | The Regents Of The University Of California | Trophic conversion of photoautotrophic bacteria for improved diurnal properties |
WO2014065253A1 (ja) * | 2012-10-22 | 2014-05-01 | 株式会社カネカ | 高分子量pha生産微生物とそれを用いた高分子量phaの製造方法 |
WO2016021604A1 (ja) * | 2014-08-04 | 2016-02-11 | 国立大学法人東京工業大学 | 糖質原料からの共重合ポリヒドロキシアルカン酸の製造法 |
Non-Patent Citations (6)
Title |
---|
ARIFIN Y. ET AL.: "Metabolic engineering of sucrose utilizing Escherichia coli for polyhydroxybutyrate production.", JOURNAL OF BIOTECHNOLOGY, vol. 150S (2010), JPN6017006371, pages 72 - 73, ISSN: 0004405780 * |
FRIEHS K. ET AL., JOURNAL OF BIOTECHNOLOGY, vol. 10 (1989), JPN6017006373, 1989, pages 285 - 292, ISSN: 0004405781 * |
INSOMPHUN C. ET AL., METABOLIC ENGINEERING, vol. 27 (2014.10.30), JPN6017006375, pages 38 - 45, ISSN: 0004405783 * |
ORITA I. ET AL., JOURNAL OF BIOSCIENCE AND BIOENGINEERING, vol. 113(1) (2012), JPN6017006376, pages 63 - 69, ISSN: 0004405784 * |
PARK S.J. ET AL, BIOTECHNOLOGY AND BIOENGINEERING, vol. Vol.112, No.3 (2015.03), JPN6017006370, pages 638 - 643, ISSN: 0004405779 * |
SABRI S. ET AL., APPLIED AND ENVIRONMENTAL MICROBIOLOGY, vol. 79(2) (2013), JPN6017006374, 2013, pages 478 - 487, ISSN: 0004405782 * |
Also Published As
Publication number | Publication date |
---|---|
WO2017104722A1 (ja) | 2017-06-22 |
US20180371509A1 (en) | 2018-12-27 |
JP6853787B2 (ja) | 2021-03-31 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
JP7001596B2 (ja) | 3hh単位含有共重合phaを生産する形質転換体、及び当該phaの製造方法 | |
US11453896B2 (en) | Transformed microorganism for producing PHA copolymer comprising 3HH monomer unit at high composition rate and method for producing PHA using same | |
EP2295536B1 (en) | Microorganism capable of producing improved polyhydroxyalkanoate and method of producing polyhydroxyalkanoate by using the same | |
JP7256740B2 (ja) | グリセロールキナーゼ活性を強化したpha産生微生物とそれを用いたphaの製造方法 | |
JPWO2015115619A1 (ja) | R体特異的エノイル−CoAヒドラターゼ遺伝子の発現が調節された微生物及びそれを用いたポリヒドロキシアルカノエート共重合体の製造方法 | |
JPWO2005098001A1 (ja) | 新規形質転換体 | |
JP7360329B2 (ja) | 変異型ポリヒドロキシアルカン酸合成酵素、その遺伝子および形質転換体、並びに、ポリヒドロキシアルカン酸の製造方法 | |
JP6313708B2 (ja) | 高分子量pha生産微生物とそれを用いた高分子量phaの製造方法 | |
JP6853787B2 (ja) | スクロース資化性を有するpha生産微生物、及び該微生物を用いたphaの製造方法 | |
JP4960033B2 (ja) | 酵素活性を低下させた微生物を用いる共重合ポリエステルの製造方法 | |
JP2008086238A (ja) | ポリヒドロキシアルカノエートの製造方法 | |
CN114616320A (zh) | 转化微生物、以及使用了该微生物的聚羟基烷酸酯的制造方法 | |
US20090130731A1 (en) | Microorganism capable of accumulating ultra high molecular weight polyester | |
JP5839854B2 (ja) | 微生物の培養方法 | |
JP2009225775A (ja) | ポリヒドロキシアルカン酸の製造方法 | |
EP3960867A1 (en) | Gene for synthesizing high molecular weight copolymer | |
JP7425783B2 (ja) | 形質転換微生物、及びポリヒドロキシアルカン酸の製造方法 | |
WO2024058182A1 (ja) | 形質転換微生物、及び、共重合ポリエステルの製造方法 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
A621 | Written request for application examination |
Free format text: JAPANESE INTERMEDIATE CODE: A621 Effective date: 20191107 |
|
A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20201215 |
|
A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20210128 |
|
TRDD | Decision of grant or rejection written | ||
A01 | Written decision to grant a patent or to grant a registration (utility model) |
Free format text: JAPANESE INTERMEDIATE CODE: A01 Effective date: 20210216 |
|
A61 | First payment of annual fees (during grant procedure) |
Free format text: JAPANESE INTERMEDIATE CODE: A61 Effective date: 20210312 |
|
R150 | Certificate of patent or registration of utility model |
Ref document number: 6853787 Country of ref document: JP Free format text: JAPANESE INTERMEDIATE CODE: R150 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |