CN116555133A - 一种用于生产虾青素的转基因蓝藻及其应用 - Google Patents
一种用于生产虾青素的转基因蓝藻及其应用 Download PDFInfo
- Publication number
- CN116555133A CN116555133A CN202210110355.0A CN202210110355A CN116555133A CN 116555133 A CN116555133 A CN 116555133A CN 202210110355 A CN202210110355 A CN 202210110355A CN 116555133 A CN116555133 A CN 116555133A
- Authority
- CN
- China
- Prior art keywords
- plasmid
- blue algae
- transgenic
- astaxanthin
- crtw
- 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.)
- Pending
Links
- JEBFVOLFMLUKLF-IFPLVEIFSA-N Astaxanthin Natural products CC(=C/C=C/C(=C/C=C/C1=C(C)C(=O)C(O)CC1(C)C)/C)C=CC=C(/C)C=CC=C(/C)C=CC2=C(C)C(=O)C(O)CC2(C)C JEBFVOLFMLUKLF-IFPLVEIFSA-N 0.000 title claims abstract description 54
- 235000013793 astaxanthin Nutrition 0.000 title claims abstract description 54
- 239000001168 astaxanthin Substances 0.000 title claims abstract description 54
- MQZIGYBFDRPAKN-ZWAPEEGVSA-N astaxanthin Chemical compound C([C@H](O)C(=O)C=1C)C(C)(C)C=1/C=C/C(/C)=C/C=C/C(/C)=C/C=C/C=C(C)C=CC=C(C)C=CC1=C(C)C(=O)[C@@H](O)CC1(C)C MQZIGYBFDRPAKN-ZWAPEEGVSA-N 0.000 title claims abstract description 54
- 229940022405 astaxanthin Drugs 0.000 title claims abstract description 54
- 241000195493 Cryptophyta Species 0.000 title claims abstract description 46
- 230000009261 transgenic effect Effects 0.000 title claims abstract description 37
- 238000004519 manufacturing process Methods 0.000 claims abstract description 23
- 235000021466 carotenoid Nutrition 0.000 claims abstract description 21
- 150000001747 carotenoids Chemical class 0.000 claims abstract description 21
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 21
- 239000013612 plasmid Substances 0.000 claims description 62
- 241000192700 Cyanobacteria Species 0.000 claims description 24
- 241000192581 Synechocystis sp. Species 0.000 claims description 11
- 238000000034 method Methods 0.000 claims description 10
- 238000010276 construction Methods 0.000 claims description 9
- 239000002773 nucleotide Substances 0.000 claims description 8
- 125000003729 nucleotide group Chemical group 0.000 claims description 8
- 241000192656 Nostoc Species 0.000 claims description 7
- 238000010367 cloning Methods 0.000 claims description 7
- 238000012258 culturing Methods 0.000 claims description 6
- 101150093858 psbA1 gene Proteins 0.000 claims description 6
- 229960005091 chloramphenicol Drugs 0.000 claims description 5
- WIIZWVCIJKGZOK-RKDXNWHRSA-N chloramphenicol Chemical compound ClC(Cl)C(=O)N[C@H](CO)[C@H](O)C1=CC=C([N+]([O-])=O)C=C1 WIIZWVCIJKGZOK-RKDXNWHRSA-N 0.000 claims description 5
- 108700039691 Genetic Promoter Regions Proteins 0.000 claims description 4
- 238000011144 upstream manufacturing Methods 0.000 claims description 4
- 238000000855 fermentation Methods 0.000 claims description 3
- 230000004151 fermentation Effects 0.000 claims description 3
- 230000001131 transforming effect Effects 0.000 claims description 3
- 241001464430 Cyanobacterium Species 0.000 claims 2
- 244000005700 microbiome Species 0.000 abstract description 8
- 241000192673 Nostoc sp. Species 0.000 abstract description 7
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 abstract description 6
- 239000001569 carbon dioxide Substances 0.000 abstract description 3
- 229910002092 carbon dioxide Inorganic materials 0.000 abstract description 3
- 241000192593 Synechocystis sp. PCC 6803 Species 0.000 description 24
- 108020004414 DNA Proteins 0.000 description 20
- 230000015572 biosynthetic process Effects 0.000 description 10
- 230000008569 process Effects 0.000 description 8
- JKQXZKUSFCKOGQ-JLGXGRJMSA-N (3R,3'R)-beta,beta-carotene-3,3'-diol Chemical compound C([C@H](O)CC=1C)C(C)(C)C=1/C=C/C(/C)=C/C=C/C(/C)=C/C=C/C=C(C)C=CC=C(C)C=CC1=C(C)C[C@@H](O)CC1(C)C JKQXZKUSFCKOGQ-JLGXGRJMSA-N 0.000 description 7
- JKQXZKUSFCKOGQ-LQFQNGICSA-N Z-zeaxanthin Natural products C([C@H](O)CC=1C)C(C)(C)C=1C=CC(C)=CC=CC(C)=CC=CC=C(C)C=CC=C(C)C=CC1=C(C)C[C@@H](O)CC1(C)C JKQXZKUSFCKOGQ-LQFQNGICSA-N 0.000 description 7
- QOPRSMDTRDMBNK-RNUUUQFGSA-N Zeaxanthin Natural products CC(=C/C=C/C=C(C)/C=C/C=C(C)/C=C/C1=C(C)CCC(O)C1(C)C)C=CC=C(/C)C=CC2=C(C)CC(O)CC2(C)C QOPRSMDTRDMBNK-RNUUUQFGSA-N 0.000 description 7
- JKQXZKUSFCKOGQ-LOFNIBRQSA-N all-trans-Zeaxanthin Natural products CC(=C/C=C/C=C(C)/C=C/C=C(C)/C=C/C1=C(C)CC(O)CC1(C)C)C=CC=C(/C)C=CC2=C(C)CC(O)CC2(C)C JKQXZKUSFCKOGQ-LOFNIBRQSA-N 0.000 description 7
- KBPHJBAIARWVSC-XQIHNALSSA-N trans-lutein Natural products CC(=C/C=C/C=C(C)/C=C/C=C(C)/C=C/C1=C(C)CC(O)CC1(C)C)C=CC=C(/C)C=CC2C(=CC(O)CC2(C)C)C KBPHJBAIARWVSC-XQIHNALSSA-N 0.000 description 7
- 235000010930 zeaxanthin Nutrition 0.000 description 7
- 239000001775 zeaxanthin Substances 0.000 description 7
- 229940043269 zeaxanthin Drugs 0.000 description 7
- 210000004027 cell Anatomy 0.000 description 6
- NUHSROFQTUXZQQ-UHFFFAOYSA-N isopentenyl diphosphate Chemical compound CC(=C)CCO[P@](O)(=O)OP(O)(O)=O NUHSROFQTUXZQQ-UHFFFAOYSA-N 0.000 description 6
- CBIDRCWHNCKSTO-UHFFFAOYSA-N prenyl diphosphate Chemical compound CC(C)=CCO[P@](O)(=O)OP(O)(O)=O CBIDRCWHNCKSTO-UHFFFAOYSA-N 0.000 description 6
- 241000168517 Haematococcus lacustris Species 0.000 description 5
- 241000192584 Synechocystis Species 0.000 description 5
- 230000002068 genetic effect Effects 0.000 description 5
- 230000009466 transformation Effects 0.000 description 5
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 4
- 238000012408 PCR amplification Methods 0.000 description 4
- 238000004458 analytical method Methods 0.000 description 4
- 229910052799 carbon Inorganic materials 0.000 description 4
- 239000012634 fragment Substances 0.000 description 4
- 230000010354 integration Effects 0.000 description 4
- 230000000243 photosynthetic effect Effects 0.000 description 4
- 108090000790 Enzymes Proteins 0.000 description 3
- 102000004190 Enzymes Human genes 0.000 description 3
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- OENHQHLEOONYIE-UKMVMLAPSA-N all-trans beta-carotene Natural products CC=1CCCC(C)(C)C=1/C=C/C(/C)=C/C=C/C(/C)=C/C=C/C=C(C)C=CC=C(C)C=CC1=C(C)CCCC1(C)C OENHQHLEOONYIE-UKMVMLAPSA-N 0.000 description 3
- 230000008901 benefit Effects 0.000 description 3
- 235000013734 beta-carotene Nutrition 0.000 description 3
- 239000011648 beta-carotene Substances 0.000 description 3
- TUPZEYHYWIEDIH-WAIFQNFQSA-N beta-carotene Natural products CC(=C/C=C/C=C(C)/C=C/C=C(C)/C=C/C1=C(C)CCCC1(C)C)C=CC=C(/C)C=CC2=CCCCC2(C)C TUPZEYHYWIEDIH-WAIFQNFQSA-N 0.000 description 3
- 229960002747 betacarotene Drugs 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- 238000004128 high performance liquid chromatography Methods 0.000 description 3
- 239000012528 membrane Substances 0.000 description 3
- 230000037353 metabolic pathway Effects 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- 230000037361 pathway Effects 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- OENHQHLEOONYIE-JLTXGRSLSA-N β-Carotene Chemical compound CC=1CCCC(C)(C)C=1\C=C\C(\C)=C\C=C\C(\C)=C\C=C\C=C(/C)\C=C\C=C(/C)\C=C\C1=C(C)CCCC1(C)C OENHQHLEOONYIE-JLTXGRSLSA-N 0.000 description 3
- YVLPJIGOMTXXLP-UHFFFAOYSA-N 15-cis-phytoene Chemical compound CC(C)=CCCC(C)=CCCC(C)=CCCC(C)=CC=CC=C(C)CCC=C(C)CCC=C(C)CCC=C(C)C YVLPJIGOMTXXLP-UHFFFAOYSA-N 0.000 description 2
- OINNEUNVOZHBOX-QIRCYJPOSA-K 2-trans,6-trans,10-trans-geranylgeranyl diphosphate(3-) Chemical compound CC(C)=CCC\C(C)=C\CC\C(C)=C\CC\C(C)=C\COP([O-])(=O)OP([O-])([O-])=O OINNEUNVOZHBOX-QIRCYJPOSA-K 0.000 description 2
- OSJPPGNTCRNQQC-UWTATZPHSA-N 3-phospho-D-glyceric acid Chemical compound OC(=O)[C@H](O)COP(O)(O)=O OSJPPGNTCRNQQC-UWTATZPHSA-N 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 2
- -1 C10) Chemical compound 0.000 description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- VWFJDQUYCIWHTN-UHFFFAOYSA-N Farnesyl pyrophosphate Natural products CC(C)=CCCC(C)=CCCC(C)=CCOP(O)(=O)OP(O)(O)=O VWFJDQUYCIWHTN-UHFFFAOYSA-N 0.000 description 2
- OINNEUNVOZHBOX-XBQSVVNOSA-N Geranylgeranyl diphosphate Natural products [P@](=O)(OP(=O)(O)O)(OC/C=C(\CC/C=C(\CC/C=C(\CC/C=C(\C)/C)/C)/C)/C)O OINNEUNVOZHBOX-XBQSVVNOSA-N 0.000 description 2
- SWQALSGKVLYKDT-UHFFFAOYSA-N Gly-Ile-Ala Natural products NCC(=O)NC(C(C)CC)C(=O)NC(C)C(O)=O SWQALSGKVLYKDT-UHFFFAOYSA-N 0.000 description 2
- BRTVHXHCUSXYRI-CIUDSAMLSA-N Leu-Ser-Ser Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@@H](CO)C(O)=O BRTVHXHCUSXYRI-CIUDSAMLSA-N 0.000 description 2
- LCTONWCANYUPML-UHFFFAOYSA-N Pyruvic acid Chemical compound CC(=O)C(O)=O LCTONWCANYUPML-UHFFFAOYSA-N 0.000 description 2
- MOINZPRHJGTCHZ-MMWGEVLESA-N Ser-Ile-Pro Chemical compound CC[C@H](C)[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CO)N MOINZPRHJGTCHZ-MMWGEVLESA-N 0.000 description 2
- PMKQKNBISAOSRI-XHSDSOJGSA-N Val-Tyr-Pro Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC1=CC=C(C=C1)O)C(=O)N2CCC[C@@H]2C(=O)O)N PMKQKNBISAOSRI-XHSDSOJGSA-N 0.000 description 2
- XMWHRVNVKDKBRG-CRCLSJGQSA-N [(2s,3r)-2,3,4-trihydroxy-3-methylbutyl] dihydrogen phosphate Chemical compound OC[C@](O)(C)[C@@H](O)COP(O)(O)=O XMWHRVNVKDKBRG-CRCLSJGQSA-N 0.000 description 2
- 150000001413 amino acids Chemical group 0.000 description 2
- 108010080924 beta-carotene ketolase Proteins 0.000 description 2
- 230000003115 biocidal effect Effects 0.000 description 2
- 230000010261 cell growth Effects 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 238000009833 condensation Methods 0.000 description 2
- 230000005494 condensation Effects 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 238000010353 genetic engineering Methods 0.000 description 2
- GVVPGTZRZFNKDS-JXMROGBWSA-N geranyl diphosphate Chemical compound CC(C)=CCC\C(C)=C\CO[P@](O)(=O)OP(O)(O)=O GVVPGTZRZFNKDS-JXMROGBWSA-N 0.000 description 2
- 230000034659 glycolysis Effects 0.000 description 2
- 108010089804 glycyl-threonine Proteins 0.000 description 2
- 108010015792 glycyllysine Proteins 0.000 description 2
- 230000012010 growth Effects 0.000 description 2
- 108010028295 histidylhistidine Proteins 0.000 description 2
- 108010083476 phenylalanyltryptophan Proteins 0.000 description 2
- AWUCVROLDVIAJX-GSVOUGTGSA-N sn-glycerol 3-phosphate Chemical compound OC[C@@H](O)COP(O)(O)=O AWUCVROLDVIAJX-GSVOUGTGSA-N 0.000 description 2
- HEMHJVSKTPXQMS-UHFFFAOYSA-M sodium hydroxide Inorganic materials [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 2
- 238000003786 synthesis reaction Methods 0.000 description 2
- YVLPJIGOMTXXLP-UUKUAVTLSA-N 15,15'-cis-Phytoene Natural products C(=C\C=C/C=C(\CC/C=C(\CC/C=C(\CC/C=C(\C)/C)/C)/C)/C)(\CC/C=C(\CC/C=C(\CC/C=C(\C)/C)/C)/C)/C YVLPJIGOMTXXLP-UUKUAVTLSA-N 0.000 description 1
- YVLPJIGOMTXXLP-BAHRDPFUSA-N 15Z-phytoene Natural products CC(=CCCC(=CCCC(=CCCC(=CC=C/C=C(C)/CCC=C(/C)CCC=C(/C)CCC=C(C)C)C)C)C)C YVLPJIGOMTXXLP-BAHRDPFUSA-N 0.000 description 1
- XMWHRVNVKDKBRG-UHNVWZDZSA-N 2-C-Methyl-D-erythritol 4-phosphate Natural products OC[C@@](O)(C)[C@H](O)COP(O)(O)=O XMWHRVNVKDKBRG-UHNVWZDZSA-N 0.000 description 1
- VWFJDQUYCIWHTN-YFVJMOTDSA-N 2-trans,6-trans-farnesyl diphosphate Chemical compound CC(C)=CCC\C(C)=C\CC\C(C)=C\CO[P@](O)(=O)OP(O)(O)=O VWFJDQUYCIWHTN-YFVJMOTDSA-N 0.000 description 1
- 229920001817 Agar Polymers 0.000 description 1
- YWWATNIVMOCSAV-UBHSHLNASA-N Ala-Arg-Phe Chemical compound NC(=N)NCCC[C@H](NC(=O)[C@@H](N)C)C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 YWWATNIVMOCSAV-UBHSHLNASA-N 0.000 description 1
- ZIBWKCRKNFYTPT-ZKWXMUAHSA-N Ala-Asn-Val Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](C(C)C)C(O)=O ZIBWKCRKNFYTPT-ZKWXMUAHSA-N 0.000 description 1
- KIUYPHAMDKDICO-WHFBIAKZSA-N Ala-Asp-Gly Chemical compound C[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)NCC(O)=O KIUYPHAMDKDICO-WHFBIAKZSA-N 0.000 description 1
- ZPXCNXMJEZKRLU-LSJOCFKGSA-N Ala-His-Arg Chemical compound NC(N)=NCCC[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)[C@@H](N)C)CC1=CN=CN1 ZPXCNXMJEZKRLU-LSJOCFKGSA-N 0.000 description 1
- SHKGHIFSEAGTNL-DLOVCJGASA-N Ala-His-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)[C@@H](N)C)CC1=CN=CN1 SHKGHIFSEAGTNL-DLOVCJGASA-N 0.000 description 1
- MNZHHDPWDWQJCQ-YUMQZZPRSA-N Ala-Leu-Gly Chemical compound C[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)NCC(O)=O MNZHHDPWDWQJCQ-YUMQZZPRSA-N 0.000 description 1
- DPNZTBKGAUAZQU-DLOVCJGASA-N Ala-Leu-His Chemical compound C[C@@H](C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC1=CN=CN1)C(=O)O)N DPNZTBKGAUAZQU-DLOVCJGASA-N 0.000 description 1
- WUHJHHGYVVJMQE-BJDJZHNGSA-N Ala-Leu-Ile Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O WUHJHHGYVVJMQE-BJDJZHNGSA-N 0.000 description 1
- 102000007698 Alcohol dehydrogenase Human genes 0.000 description 1
- 108010021809 Alcohol dehydrogenase Proteins 0.000 description 1
- 241000192542 Anabaena Species 0.000 description 1
- DPXDVGDLWJYZBH-GUBZILKMSA-N Arg-Asn-Arg Chemical compound NC(N)=NCCC[C@H](N)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CCCN=C(N)N)C(O)=O DPXDVGDLWJYZBH-GUBZILKMSA-N 0.000 description 1
- FRMQITGHXMUNDF-GMOBBJLQSA-N Arg-Ile-Asn Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)O)NC(=O)[C@H](CCCN=C(N)N)N FRMQITGHXMUNDF-GMOBBJLQSA-N 0.000 description 1
- OGSQONVYSTZIJB-WDSOQIARSA-N Arg-Leu-Trp Chemical compound CC(C)C[C@H](NC(=O)[C@@H](N)CCCN=C(N)N)C(=O)N[C@@H](Cc1c[nH]c2ccccc12)C(O)=O OGSQONVYSTZIJB-WDSOQIARSA-N 0.000 description 1
- AZHXYLJRGVMQKW-UMPQAUOISA-N Arg-Trp-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CC1=CNC2=CC=CC=C21)NC(=O)[C@H](CCCN=C(N)N)N)O AZHXYLJRGVMQKW-UMPQAUOISA-N 0.000 description 1
- DAPLJWATMAXPPZ-CIUDSAMLSA-N Asn-Asn-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](CC(N)=O)NC(=O)[C@@H](N)CC(N)=O DAPLJWATMAXPPZ-CIUDSAMLSA-N 0.000 description 1
- GWNMUVANAWDZTI-YUMQZZPRSA-N Asn-Gly-His Chemical compound C1=C(NC=N1)C[C@@H](C(=O)O)NC(=O)CNC(=O)[C@H](CC(=O)N)N GWNMUVANAWDZTI-YUMQZZPRSA-N 0.000 description 1
- PTSDPWIHOYMRGR-UGYAYLCHSA-N Asn-Ile-Asn Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC(N)=O)C(O)=O PTSDPWIHOYMRGR-UGYAYLCHSA-N 0.000 description 1
- NCFJQJRLQJEECD-NHCYSSNCSA-N Asn-Leu-Val Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C(C)C)C(O)=O NCFJQJRLQJEECD-NHCYSSNCSA-N 0.000 description 1
- GFGUPLIETCNQGF-DCAQKATOSA-N Asn-Pro-His Chemical compound C1C[C@H](N(C1)C(=O)[C@H](CC(=O)N)N)C(=O)N[C@@H](CC2=CN=CN2)C(=O)O GFGUPLIETCNQGF-DCAQKATOSA-N 0.000 description 1
- JWQWPRCDYWNVNM-ACZMJKKPSA-N Asn-Ser-Gln Chemical compound C(CC(=O)N)[C@@H](C(=O)O)NC(=O)[C@H](CO)NC(=O)[C@H](CC(=O)N)N JWQWPRCDYWNVNM-ACZMJKKPSA-N 0.000 description 1
- HCZQKHSRYHCPSD-IUKAMOBKSA-N Asn-Thr-Ile Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O HCZQKHSRYHCPSD-IUKAMOBKSA-N 0.000 description 1
- CGYKCTPUGXFPMG-IHPCNDPISA-N Asn-Tyr-Trp Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC1=CNC2=C1C=CC=C2)C(O)=O CGYKCTPUGXFPMG-IHPCNDPISA-N 0.000 description 1
- MJIJBEYEHBKTIM-BYULHYEWSA-N Asn-Val-Asn Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)O)NC(=O)[C@H](CC(=O)N)N MJIJBEYEHBKTIM-BYULHYEWSA-N 0.000 description 1
- CXBOKJPLEYUPGB-FXQIFTODSA-N Asp-Ala-Met Chemical compound C[C@@H](C(=O)N[C@@H](CCSC)C(=O)O)NC(=O)[C@H](CC(=O)O)N CXBOKJPLEYUPGB-FXQIFTODSA-N 0.000 description 1
- NJIKKGUVGUBICV-ZLUOBGJFSA-N Asp-Ala-Ser Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)[C@@H](N)CC(O)=O NJIKKGUVGUBICV-ZLUOBGJFSA-N 0.000 description 1
- CELPEWWLSXMVPH-CIUDSAMLSA-N Asp-Asp-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@H](CC(O)=O)NC(=O)[C@@H](N)CC(O)=O CELPEWWLSXMVPH-CIUDSAMLSA-N 0.000 description 1
- DXQOQMCLWWADMU-ACZMJKKPSA-N Asp-Gln-Ser Chemical compound OC(=O)C[C@H](N)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CO)C(O)=O DXQOQMCLWWADMU-ACZMJKKPSA-N 0.000 description 1
- UJGRZQYSNYTCAX-SRVKXCTJSA-N Asp-Leu-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](N)CC(O)=O UJGRZQYSNYTCAX-SRVKXCTJSA-N 0.000 description 1
- KESWRFKUZRUTAH-FXQIFTODSA-N Asp-Pro-Asp Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CC(O)=O)C(O)=O KESWRFKUZRUTAH-FXQIFTODSA-N 0.000 description 1
- AHWRSSLYSGLBGD-CIUDSAMLSA-N Asp-Pro-Glu Chemical compound OC(=O)C[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CCC(O)=O)C(O)=O AHWRSSLYSGLBGD-CIUDSAMLSA-N 0.000 description 1
- GCACQYDBDHRVGE-LKXGYXEUSA-N Asp-Thr-Ser Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H]([C@H](O)C)NC(=O)[C@@H](N)CC(O)=O GCACQYDBDHRVGE-LKXGYXEUSA-N 0.000 description 1
- ALMIMUZAWTUNIO-BZSNNMDCSA-N Asp-Tyr-Tyr Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O ALMIMUZAWTUNIO-BZSNNMDCSA-N 0.000 description 1
- AMRLSQGGERHDHJ-FXQIFTODSA-N Cys-Ala-Arg Chemical compound [H]N[C@@H](CS)C(=O)N[C@@H](C)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O AMRLSQGGERHDHJ-FXQIFTODSA-N 0.000 description 1
- PORWNQWEEIOIRH-XHNCKOQMSA-N Cys-Gln-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CCC(=O)N)NC(=O)[C@H](CS)N)C(=O)O PORWNQWEEIOIRH-XHNCKOQMSA-N 0.000 description 1
- 241000588724 Escherichia coli Species 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- 108700007698 Genetic Terminator Regions Proteins 0.000 description 1
- GVVPGTZRZFNKDS-YFHOEESVSA-N Geranyl diphosphate Natural products CC(C)=CCC\C(C)=C/COP(O)(=O)OP(O)(O)=O GVVPGTZRZFNKDS-YFHOEESVSA-N 0.000 description 1
- XXLBHPPXDUWYAG-XQXXSGGOSA-N Gln-Ala-Thr Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O XXLBHPPXDUWYAG-XQXXSGGOSA-N 0.000 description 1
- JSYULGSPLTZDHM-NRPADANISA-N Gln-Ala-Val Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](C(C)C)C(O)=O JSYULGSPLTZDHM-NRPADANISA-N 0.000 description 1
- GMGKDVVBSVVKCT-NUMRIWBASA-N Gln-Asn-Thr Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O GMGKDVVBSVVKCT-NUMRIWBASA-N 0.000 description 1
- MWERYIXRDZDXOA-QEWYBTABSA-N Gln-Ile-Phe Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O MWERYIXRDZDXOA-QEWYBTABSA-N 0.000 description 1
- SHAUZYVSXAMYAZ-JYJNAYRXSA-N Gln-Leu-Phe Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)O)NC(=O)[C@H](CCC(=O)N)N SHAUZYVSXAMYAZ-JYJNAYRXSA-N 0.000 description 1
- ZBKUIQNCRIYVGH-SDDRHHMPSA-N Gln-Leu-Pro Chemical compound CC(C)C[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CCC(=O)N)N ZBKUIQNCRIYVGH-SDDRHHMPSA-N 0.000 description 1
- XQDGOJPVMSWZSO-SRVKXCTJSA-N Gln-Pro-Leu Chemical compound CC(C)C[C@@H](C(=O)O)NC(=O)[C@@H]1CCCN1C(=O)[C@H](CCC(=O)N)N XQDGOJPVMSWZSO-SRVKXCTJSA-N 0.000 description 1
- RONJIBWTGKVKFY-HTUGSXCWSA-N Gln-Thr-Phe Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)O)NC(=O)[C@H](CCC(=O)N)N)O RONJIBWTGKVKFY-HTUGSXCWSA-N 0.000 description 1
- WTMZXOPHTIVFCP-QEWYBTABSA-N Glu-Ile-Phe Chemical compound OC(=O)CC[C@H](N)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 WTMZXOPHTIVFCP-QEWYBTABSA-N 0.000 description 1
- DMYACXMQUABZIQ-NRPADANISA-N Glu-Ser-Val Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](C(C)C)C(O)=O DMYACXMQUABZIQ-NRPADANISA-N 0.000 description 1
- KXRORHJIRAOQPG-SOUVJXGZSA-N Glu-Tyr-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CC2=CC=C(C=C2)O)NC(=O)[C@H](CCC(=O)O)N)C(=O)O KXRORHJIRAOQPG-SOUVJXGZSA-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
- BULIVUZUDBHKKZ-WDSKDSINSA-N Gly-Gln-Asn Chemical compound NCC(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(N)=O)C(O)=O BULIVUZUDBHKKZ-WDSKDSINSA-N 0.000 description 1
- LLZXNUUIBOALNY-QWRGUYRKSA-N Gly-Leu-Lys Chemical compound NCC(=O)N[C@@H](CC(C)C)C(=O)N[C@H](C(O)=O)CCCCN LLZXNUUIBOALNY-QWRGUYRKSA-N 0.000 description 1
- TVUWMSBGMVAHSJ-KBPBESRZSA-N Gly-Leu-Phe Chemical compound NCC(=O)N[C@@H](CC(C)C)C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 TVUWMSBGMVAHSJ-KBPBESRZSA-N 0.000 description 1
- UUYBFNKHOCJCHT-VHSXEESVSA-N Gly-Leu-Pro Chemical compound CC(C)C[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)CN UUYBFNKHOCJCHT-VHSXEESVSA-N 0.000 description 1
- WMGHDYWNHNLGBV-ONGXEEELSA-N Gly-Phe-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@@H](NC(=O)CN)CC1=CC=CC=C1 WMGHDYWNHNLGBV-ONGXEEELSA-N 0.000 description 1
- IBYOLNARKHMLBG-WHOFXGATSA-N Gly-Phe-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)CN)CC1=CC=CC=C1 IBYOLNARKHMLBG-WHOFXGATSA-N 0.000 description 1
- VDCRBJACQKOSMS-JSGCOSHPSA-N Gly-Phe-Val Chemical compound [H]NCC(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](C(C)C)C(O)=O VDCRBJACQKOSMS-JSGCOSHPSA-N 0.000 description 1
- HFPVRZWORNJRRC-UWVGGRQHSA-N Gly-Pro-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@@H]1CCCN1C(=O)CN HFPVRZWORNJRRC-UWVGGRQHSA-N 0.000 description 1
- IRJWAYCXIYUHQE-WHFBIAKZSA-N Gly-Ser-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@H](CO)NC(=O)CN IRJWAYCXIYUHQE-WHFBIAKZSA-N 0.000 description 1
- KBBFOULZCHWGJX-KBPBESRZSA-N Gly-Tyr-His Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)N[C@@H](CC2=CN=CN2)C(=O)O)NC(=O)CN)O KBBFOULZCHWGJX-KBPBESRZSA-N 0.000 description 1
- GBYYQVBXFVDJPJ-WLTAIBSBSA-N Gly-Tyr-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CC1=CC=C(C=C1)O)NC(=O)CN)O GBYYQVBXFVDJPJ-WLTAIBSBSA-N 0.000 description 1
- PROLDOGUBQJNPG-RWMBFGLXSA-N His-Arg-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CC2=CN=CN2)N)C(=O)O PROLDOGUBQJNPG-RWMBFGLXSA-N 0.000 description 1
- MAABHGXCIBEYQR-XVYDVKMFSA-N His-Asn-Ala Chemical compound C[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)N)NC(=O)[C@H](CC1=CN=CN1)N MAABHGXCIBEYQR-XVYDVKMFSA-N 0.000 description 1
- VTMLJMNQHKBPON-QWRGUYRKSA-N His-Gly-His Chemical compound C([C@H](N)C(=O)NCC(=O)N[C@@H](CC=1NC=NC=1)C(O)=O)C1=CN=CN1 VTMLJMNQHKBPON-QWRGUYRKSA-N 0.000 description 1
- NTXIJPDAHXSHNL-ONGXEEELSA-N His-Gly-Val Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)NCC(=O)N[C@@H](C(C)C)C(O)=O NTXIJPDAHXSHNL-ONGXEEELSA-N 0.000 description 1
- KWBISLAEQZUYIC-UWJYBYFXSA-N His-His-Ala Chemical compound C[C@@H](C(=O)O)NC(=O)[C@H](CC1=CN=CN1)NC(=O)[C@H](CC2=CN=CN2)N KWBISLAEQZUYIC-UWJYBYFXSA-N 0.000 description 1
- DYKZGTLPSNOFHU-DEQVHRJGSA-N His-Ile-Pro Chemical compound CC[C@H](C)[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CC2=CN=CN2)N DYKZGTLPSNOFHU-DEQVHRJGSA-N 0.000 description 1
- UROVZOUMHNXPLZ-AVGNSLFASA-N His-Leu-Gln Chemical compound NC(=O)CC[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](N)CC1=CN=CN1 UROVZOUMHNXPLZ-AVGNSLFASA-N 0.000 description 1
- VDHOMPFVSABJKU-ULQDDVLXSA-N His-Phe-Val Chemical compound CC(C)[C@@H](C(=O)O)NC(=O)[C@H](CC1=CC=CC=C1)NC(=O)[C@H](CC2=CN=CN2)N VDHOMPFVSABJKU-ULQDDVLXSA-N 0.000 description 1
- JMSONHOUHFDOJH-GUBZILKMSA-N His-Ser-Glu Chemical compound OC(=O)CC[C@@H](C(O)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](N)CC1=CN=CN1 JMSONHOUHFDOJH-GUBZILKMSA-N 0.000 description 1
- LNVILFYCPVOHPV-IHPCNDPISA-N His-Trp-Leu Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CC1=CNC2=C1C=CC=C2)C(=O)N[C@@H](CC(C)C)C(O)=O LNVILFYCPVOHPV-IHPCNDPISA-N 0.000 description 1
- QADCTXFNLZBZAB-GHCJXIJMSA-N Ile-Asn-Ala Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N[C@@H](C)C(=O)O)N QADCTXFNLZBZAB-GHCJXIJMSA-N 0.000 description 1
- AMSYMDIIIRJRKZ-HJPIBITLSA-N Ile-His-His Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC1=CN=CN1)C(=O)N[C@@H](CC2=CN=CN2)C(=O)O)N AMSYMDIIIRJRKZ-HJPIBITLSA-N 0.000 description 1
- VNDQNDYEPSXHLU-JUKXBJQTSA-N Ile-His-Tyr Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC1=CN=CN1)C(=O)N[C@@H](CC2=CC=C(C=C2)O)C(=O)O)N VNDQNDYEPSXHLU-JUKXBJQTSA-N 0.000 description 1
- HUWYGQOISIJNMK-SIGLWIIPSA-N Ile-Ile-His Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC1=CN=CN1)C(=O)O)N HUWYGQOISIJNMK-SIGLWIIPSA-N 0.000 description 1
- DMSVBUWGDLYNLC-IAVJCBSLSA-N Ile-Ile-Phe Chemical compound CC[C@H](C)[C@H](N)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 DMSVBUWGDLYNLC-IAVJCBSLSA-N 0.000 description 1
- OUUCIIJSBIBCHB-ZPFDUUQYSA-N Ile-Leu-Asp Chemical compound CC[C@H](C)[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(O)=O)C(O)=O OUUCIIJSBIBCHB-ZPFDUUQYSA-N 0.000 description 1
- USXAYNCLFSUSBA-MGHWNKPDSA-N Ile-Phe-His Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC2=CN=CN2)C(=O)O)N USXAYNCLFSUSBA-MGHWNKPDSA-N 0.000 description 1
- CNMOKANDJMLAIF-CIQUZCHMSA-N Ile-Thr-Ala Chemical compound CC[C@H](C)[C@H](N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C)C(O)=O CNMOKANDJMLAIF-CIQUZCHMSA-N 0.000 description 1
- ZSESFIFAYQEKRD-CYDGBPFRSA-N Ile-Val-Met Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCSC)C(=O)O)N ZSESFIFAYQEKRD-CYDGBPFRSA-N 0.000 description 1
- RRHGJUQNOFWUDK-UHFFFAOYSA-N Isoprene Chemical group CC(=C)C=C RRHGJUQNOFWUDK-UHFFFAOYSA-N 0.000 description 1
- KFKWRHQBZQICHA-STQMWFEESA-N L-leucyl-L-phenylalanine Natural products CC(C)C[C@H](N)C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 KFKWRHQBZQICHA-STQMWFEESA-N 0.000 description 1
- 241000880493 Leptailurus serval Species 0.000 description 1
- USTCFDAQCLDPBD-XIRDDKMYSA-N Leu-Asn-Trp Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N[C@@H](CC1=CNC2=CC=CC=C21)C(=O)O)N USTCFDAQCLDPBD-XIRDDKMYSA-N 0.000 description 1
- MMEDVBWCMGRKKC-GARJFASQSA-N Leu-Asp-Pro Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)N1CCC[C@@H]1C(=O)O)N MMEDVBWCMGRKKC-GARJFASQSA-N 0.000 description 1
- NEEOBPIXKWSBRF-IUCAKERBSA-N Leu-Glu-Gly Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(=O)NCC(O)=O NEEOBPIXKWSBRF-IUCAKERBSA-N 0.000 description 1
- VBZOAGIPCULURB-QWRGUYRKSA-N Leu-Gly-His Chemical compound CC(C)C[C@@H](C(=O)NCC(=O)N[C@@H](CC1=CN=CN1)C(=O)O)N VBZOAGIPCULURB-QWRGUYRKSA-N 0.000 description 1
- CCQLQKZTXZBXTN-NHCYSSNCSA-N Leu-Gly-Ile Chemical compound [H]N[C@@H](CC(C)C)C(=O)NCC(=O)N[C@@H]([C@@H](C)CC)C(O)=O CCQLQKZTXZBXTN-NHCYSSNCSA-N 0.000 description 1
- HYIFFZAQXPUEAU-QWRGUYRKSA-N Leu-Gly-Leu Chemical compound CC(C)C[C@H](N)C(=O)NCC(=O)N[C@H](C(O)=O)CC(C)C HYIFFZAQXPUEAU-QWRGUYRKSA-N 0.000 description 1
- KUIDCYNIEJBZBU-AJNGGQMLSA-N Leu-Ile-Leu Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC(C)C)C(O)=O KUIDCYNIEJBZBU-AJNGGQMLSA-N 0.000 description 1
- QLDHBYRUNQZIJQ-DKIMLUQUSA-N Leu-Ile-Phe Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O QLDHBYRUNQZIJQ-DKIMLUQUSA-N 0.000 description 1
- XVZCXCTYGHPNEM-UHFFFAOYSA-N Leu-Leu-Pro Natural products CC(C)CC(N)C(=O)NC(CC(C)C)C(=O)N1CCCC1C(O)=O XVZCXCTYGHPNEM-UHFFFAOYSA-N 0.000 description 1
- DDVHDMSBLRAKNV-IHRRRGAJSA-N Leu-Met-Leu Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CC(C)C)C(O)=O DDVHDMSBLRAKNV-IHRRRGAJSA-N 0.000 description 1
- VULJUQZPSOASBZ-SRVKXCTJSA-N Leu-Pro-Glu Chemical compound [H]N[C@@H](CC(C)C)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CCC(O)=O)C(O)=O VULJUQZPSOASBZ-SRVKXCTJSA-N 0.000 description 1
- KWLWZYMNUZJKMZ-IHRRRGAJSA-N Leu-Pro-Leu Chemical compound CC(C)C[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CC(C)C)C(O)=O KWLWZYMNUZJKMZ-IHRRRGAJSA-N 0.000 description 1
- UCXQIIIFOOGYEM-ULQDDVLXSA-N Leu-Pro-Tyr Chemical compound CC(C)C[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 UCXQIIIFOOGYEM-ULQDDVLXSA-N 0.000 description 1
- RGUXWMDNCPMQFB-YUMQZZPRSA-N Leu-Ser-Gly Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CO)C(=O)NCC(O)=O RGUXWMDNCPMQFB-YUMQZZPRSA-N 0.000 description 1
- MVHXGBZUJLWZOH-BJDJZHNGSA-N Leu-Ser-Ile Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O MVHXGBZUJLWZOH-BJDJZHNGSA-N 0.000 description 1
- ONHCDMBHPQIPAI-YTQUADARSA-N Leu-Trp-Pro Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CC1=CNC2=CC=CC=C21)C(=O)N3CCC[C@@H]3C(=O)O)N ONHCDMBHPQIPAI-YTQUADARSA-N 0.000 description 1
- BGGTYDNTOYRTTR-MEYUZBJRSA-N Leu-Tyr-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CC1=CC=C(C=C1)O)NC(=O)[C@H](CC(C)C)N)O BGGTYDNTOYRTTR-MEYUZBJRSA-N 0.000 description 1
- QQXJROOJCMIHIV-AVGNSLFASA-N Leu-Val-Met Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCSC)C(O)=O QQXJROOJCMIHIV-AVGNSLFASA-N 0.000 description 1
- DGAAQRAUOFHBFJ-CIUDSAMLSA-N Lys-Asn-Ala Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](C)C(O)=O DGAAQRAUOFHBFJ-CIUDSAMLSA-N 0.000 description 1
- YVSHZSUKQHNDHD-KKUMJFAQSA-N Lys-Asn-Phe Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)N)NC(=O)[C@H](CCCCN)N YVSHZSUKQHNDHD-KKUMJFAQSA-N 0.000 description 1
- NCTDKZKNBDZDOL-GARJFASQSA-N Lys-Asn-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CC(=O)N)NC(=O)[C@H](CCCCN)N)C(=O)O NCTDKZKNBDZDOL-GARJFASQSA-N 0.000 description 1
- KZOHPCYVORJBLG-AVGNSLFASA-N Lys-Glu-His Chemical compound C1=C(NC=N1)C[C@@H](C(=O)O)NC(=O)[C@H](CCC(=O)O)NC(=O)[C@H](CCCCN)N KZOHPCYVORJBLG-AVGNSLFASA-N 0.000 description 1
- DKTNGXVSCZULPO-YUMQZZPRSA-N Lys-Gly-Cys Chemical compound NCCCC[C@H](N)C(=O)NCC(=O)N[C@@H](CS)C(O)=O DKTNGXVSCZULPO-YUMQZZPRSA-N 0.000 description 1
- DTUZCYRNEJDKSR-NHCYSSNCSA-N Lys-Gly-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)CNC(=O)[C@@H](N)CCCCN DTUZCYRNEJDKSR-NHCYSSNCSA-N 0.000 description 1
- SKRGVGLIRUGANF-AVGNSLFASA-N Lys-Leu-Glu Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(O)=O SKRGVGLIRUGANF-AVGNSLFASA-N 0.000 description 1
- OIQSIMFSVLLWBX-VOAKCMCISA-N Lys-Leu-Thr Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O OIQSIMFSVLLWBX-VOAKCMCISA-N 0.000 description 1
- LJADEBULDNKJNK-IHRRRGAJSA-N Lys-Leu-Val Chemical compound CC(C)C[C@H](NC(=O)[C@@H](N)CCCCN)C(=O)N[C@@H](C(C)C)C(O)=O LJADEBULDNKJNK-IHRRRGAJSA-N 0.000 description 1
- JQSIGLHQNSZZRL-KKUMJFAQSA-N Lys-Lys-His Chemical compound C1=C(NC=N1)C[C@@H](C(=O)O)NC(=O)[C@H](CCCCN)NC(=O)[C@H](CCCCN)N JQSIGLHQNSZZRL-KKUMJFAQSA-N 0.000 description 1
- IOQWIOPSKJOEKI-SRVKXCTJSA-N Lys-Ser-Leu Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(O)=O IOQWIOPSKJOEKI-SRVKXCTJSA-N 0.000 description 1
- OCRSGGIJBDUXHU-WDSOQIARSA-N Met-Leu-Trp Chemical compound C1=CC=C2C(C[C@H](NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](N)CCSC)C(O)=O)=CNC2=C1 OCRSGGIJBDUXHU-WDSOQIARSA-N 0.000 description 1
- CNFMPVYIVQUJOO-NHCYSSNCSA-N Met-Val-Gln Chemical compound CSCC[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)N[C@H](C(O)=O)CCC(N)=O CNFMPVYIVQUJOO-NHCYSSNCSA-N 0.000 description 1
- 108010074633 Mixed Function Oxygenases Proteins 0.000 description 1
- 102000008109 Mixed Function Oxygenases Human genes 0.000 description 1
- XMBSYZWANAQXEV-UHFFFAOYSA-N N-alpha-L-glutamyl-L-phenylalanine Natural products OC(=O)CCC(N)C(=O)NC(C(O)=O)CC1=CC=CC=C1 XMBSYZWANAQXEV-UHFFFAOYSA-N 0.000 description 1
- BQVUABVGYYSDCJ-UHFFFAOYSA-N Nalpha-L-Leucyl-L-tryptophan Natural products C1=CC=C2C(CC(NC(=O)C(N)CC(C)C)C(O)=O)=CNC2=C1 BQVUABVGYYSDCJ-UHFFFAOYSA-N 0.000 description 1
- 108091028043 Nucleic acid sequence Proteins 0.000 description 1
- 101100245405 Parasynechococcus marenigrum (strain WH8102) psbA2 gene Proteins 0.000 description 1
- UNLYPPYNDXHGDG-IHRRRGAJSA-N Phe-Gln-Glu Chemical compound OC(=O)CC[C@@H](C(O)=O)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@@H](N)CC1=CC=CC=C1 UNLYPPYNDXHGDG-IHRRRGAJSA-N 0.000 description 1
- WFHRXJOZEXUKLV-IRXDYDNUSA-N Phe-Gly-Tyr Chemical compound C([C@H](N)C(=O)NCC(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(O)=O)C1=CC=CC=C1 WFHRXJOZEXUKLV-IRXDYDNUSA-N 0.000 description 1
- MJQFZGOIVBDIMZ-WHOFXGATSA-N Phe-Ile-Gly Chemical compound N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H]([C@@H](C)CC)C(=O)NCC(=O)O MJQFZGOIVBDIMZ-WHOFXGATSA-N 0.000 description 1
- RSPUIENXSJYZQO-JYJNAYRXSA-N Phe-Leu-Gln Chemical compound NC(=O)CC[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](N)CC1=CC=CC=C1 RSPUIENXSJYZQO-JYJNAYRXSA-N 0.000 description 1
- SMFGCTXUBWEPKM-KBPBESRZSA-N Phe-Leu-Gly Chemical compound OC(=O)CNC(=O)[C@H](CC(C)C)NC(=O)[C@@H](N)CC1=CC=CC=C1 SMFGCTXUBWEPKM-KBPBESRZSA-N 0.000 description 1
- KZRQONDKKJCAOL-DKIMLUQUSA-N Phe-Leu-Ile Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O KZRQONDKKJCAOL-DKIMLUQUSA-N 0.000 description 1
- FUAIIFPQELBNJF-ULQDDVLXSA-N Phe-Met-Lys Chemical compound CSCC[C@@H](C(=O)N[C@@H](CCCCN)C(=O)O)NC(=O)[C@H](CC1=CC=CC=C1)N FUAIIFPQELBNJF-ULQDDVLXSA-N 0.000 description 1
- KAJLHCWRWDSROH-BZSNNMDCSA-N Phe-Phe-Asp Chemical compound C([C@H](N)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H](CC(O)=O)C(O)=O)C1=CC=CC=C1 KAJLHCWRWDSROH-BZSNNMDCSA-N 0.000 description 1
- AXIOGMQCDYVTNY-ACRUOGEOSA-N Phe-Phe-Leu Chemical compound C([C@@H](C(=O)N[C@@H](CC(C)C)C(O)=O)NC(=O)[C@@H](N)CC=1C=CC=CC=1)C1=CC=CC=C1 AXIOGMQCDYVTNY-ACRUOGEOSA-N 0.000 description 1
- RBRNEFJTEHPDSL-ACRUOGEOSA-N Phe-Phe-Lys Chemical compound C([C@@H](C(=O)N[C@@H](CCCCN)C(O)=O)NC(=O)[C@@H](N)CC=1C=CC=CC=1)C1=CC=CC=C1 RBRNEFJTEHPDSL-ACRUOGEOSA-N 0.000 description 1
- ZJPGOXWRFNKIQL-JYJNAYRXSA-N Phe-Pro-Pro Chemical compound C([C@H](N)C(=O)N1[C@@H](CCC1)C(=O)N1[C@@H](CCC1)C(O)=O)C1=CC=CC=C1 ZJPGOXWRFNKIQL-JYJNAYRXSA-N 0.000 description 1
- ZLAKUZDMKVKFAI-JYJNAYRXSA-N Phe-Pro-Val Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N1CCC[C@H]1C(=O)N[C@@H](C(C)C)C(O)=O ZLAKUZDMKVKFAI-JYJNAYRXSA-N 0.000 description 1
- IIEOLPMQYRBZCN-SRVKXCTJSA-N Phe-Ser-Cys Chemical compound N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CO)C(=O)N[C@@H](CS)C(=O)O IIEOLPMQYRBZCN-SRVKXCTJSA-N 0.000 description 1
- NHHZWPNMYQUNEH-ACRUOGEOSA-N Phe-Tyr-His Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)N[C@@H](CC2=CC=C(C=C2)O)C(=O)N[C@@H](CC3=CN=CN3)C(=O)O)N NHHZWPNMYQUNEH-ACRUOGEOSA-N 0.000 description 1
- VIIRRNQMMIHYHQ-XHSDSOJGSA-N Phe-Val-Pro Chemical compound CC(C)[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CC2=CC=CC=C2)N VIIRRNQMMIHYHQ-XHSDSOJGSA-N 0.000 description 1
- ODPIUQVTULPQEP-CIUDSAMLSA-N Pro-Gln-Asn Chemical compound NC(=O)C[C@@H](C(O)=O)NC(=O)[C@H](CCC(=O)N)NC(=O)[C@@H]1CCCN1 ODPIUQVTULPQEP-CIUDSAMLSA-N 0.000 description 1
- AFXCXDQNRXTSBD-FJXKBIBVSA-N Pro-Gly-Thr Chemical compound [H]N1CCC[C@H]1C(=O)NCC(=O)N[C@@H]([C@@H](C)O)C(O)=O AFXCXDQNRXTSBD-FJXKBIBVSA-N 0.000 description 1
- BAKAHWWRCCUDAF-IHRRRGAJSA-N Pro-His-Lys Chemical compound C([C@@H](C(=O)N[C@@H](CCCCN)C(O)=O)NC(=O)[C@H]1NCCC1)C1=CN=CN1 BAKAHWWRCCUDAF-IHRRRGAJSA-N 0.000 description 1
- XQPHBAKJJJZOBX-SRVKXCTJSA-N Pro-Lys-Glu Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCC(O)=O)C(O)=O XQPHBAKJJJZOBX-SRVKXCTJSA-N 0.000 description 1
- ITUDDXVFGFEKPD-NAKRPEOUSA-N Pro-Ser-Ile Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O ITUDDXVFGFEKPD-NAKRPEOUSA-N 0.000 description 1
- 108010011939 Pyruvate Decarboxylase Proteins 0.000 description 1
- NFDYGNFETJVMSE-BQBZGAKWSA-N Ser-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@@H](N)CO NFDYGNFETJVMSE-BQBZGAKWSA-N 0.000 description 1
- GZGFSPWOMUKKCV-NAKRPEOUSA-N Ser-Pro-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)[C@@H]1CCCN1C(=O)[C@@H](N)CO GZGFSPWOMUKKCV-NAKRPEOUSA-N 0.000 description 1
- PPCZVWHJWJFTFN-ZLUOBGJFSA-N Ser-Ser-Asp Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(O)=O)C(O)=O PPCZVWHJWJFTFN-ZLUOBGJFSA-N 0.000 description 1
- BMKNXTJLHFIAAH-CIUDSAMLSA-N Ser-Ser-Leu Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(O)=O BMKNXTJLHFIAAH-CIUDSAMLSA-N 0.000 description 1
- PURRNJBBXDDWLX-ZDLURKLDSA-N Ser-Thr-Gly Chemical compound C[C@H]([C@@H](C(=O)NCC(=O)O)NC(=O)[C@H](CO)N)O PURRNJBBXDDWLX-ZDLURKLDSA-N 0.000 description 1
- BIWBTRRBHIEVAH-IHPCNDPISA-N Ser-Tyr-Trp Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC1=CNC2=C1C=CC=C2)C(O)=O BIWBTRRBHIEVAH-IHPCNDPISA-N 0.000 description 1
- YEDSOSIKVUMIJE-DCAQKATOSA-N Ser-Val-Leu Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(C)C)C(O)=O YEDSOSIKVUMIJE-DCAQKATOSA-N 0.000 description 1
- BQCADISMDOOEFD-UHFFFAOYSA-N Silver Chemical compound [Ag] BQCADISMDOOEFD-UHFFFAOYSA-N 0.000 description 1
- 241000192707 Synechococcus Species 0.000 description 1
- 241000192589 Synechococcus elongatus PCC 7942 Species 0.000 description 1
- JNQZPAWOPBZGIX-RCWTZXSCSA-N Thr-Arg-Val Chemical compound CC(C)[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)[C@@H](N)[C@@H](C)O)CCCN=C(N)N JNQZPAWOPBZGIX-RCWTZXSCSA-N 0.000 description 1
- WPAKPLPGQNUXGN-OSUNSFLBSA-N Thr-Ile-Arg Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O WPAKPLPGQNUXGN-OSUNSFLBSA-N 0.000 description 1
- XYFISNXATOERFZ-OSUNSFLBSA-N Thr-Ile-Val Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](C(C)C)C(=O)O)NC(=O)[C@H]([C@@H](C)O)N XYFISNXATOERFZ-OSUNSFLBSA-N 0.000 description 1
- WFAUDCSNCWJJAA-KXNHARMFSA-N Thr-Lys-Pro Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@@H](CCCCN)C(=O)N1CCC[C@@H]1C(O)=O WFAUDCSNCWJJAA-KXNHARMFSA-N 0.000 description 1
- WNQJTLATMXYSEL-OEAJRASXSA-N Thr-Phe-Leu Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC(C)C)C(O)=O WNQJTLATMXYSEL-OEAJRASXSA-N 0.000 description 1
- ILUOMMDDGREELW-OSUNSFLBSA-N Thr-Val-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)[C@H](C(C)C)NC(=O)[C@@H](N)[C@@H](C)O ILUOMMDDGREELW-OSUNSFLBSA-N 0.000 description 1
- MNYNCKZAEIAONY-XGEHTFHBSA-N Thr-Val-Ser Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CO)C(O)=O MNYNCKZAEIAONY-XGEHTFHBSA-N 0.000 description 1
- LJCLHMPCYYXVPR-VJBMBRPKSA-N Trp-Gln-Trp Chemical compound C1=CC=C2C(=C1)C(=CN2)C[C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)N[C@@H](CC3=CNC4=CC=CC=C43)C(=O)O)N LJCLHMPCYYXVPR-VJBMBRPKSA-N 0.000 description 1
- IQXWAJUIAQLZNX-IHPCNDPISA-N Trp-Leu-His Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CC1=CN=CN1)C(=O)O)NC(=O)[C@H](CC2=CNC3=CC=CC=C32)N IQXWAJUIAQLZNX-IHPCNDPISA-N 0.000 description 1
- RIKLKPANMFNREP-FDARSICLSA-N Trp-Met-Ile Chemical compound C1=CC=C2C(C[C@H](N)C(=O)N[C@@H](CCSC)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O)=CNC2=C1 RIKLKPANMFNREP-FDARSICLSA-N 0.000 description 1
- GBEAUNVBIMLWIB-IHPCNDPISA-N Trp-Ser-Phe Chemical compound C([C@H](NC(=O)[C@H](CO)NC(=O)[C@H](CC=1C2=CC=CC=C2NC=1)N)C(O)=O)C1=CC=CC=C1 GBEAUNVBIMLWIB-IHPCNDPISA-N 0.000 description 1
- XXJDYWYVZBHELV-TUSQITKMSA-N Trp-Trp-Lys Chemical compound C1=CC=C2C(=C1)C(=CN2)C[C@@H](C(=O)N[C@@H](CC3=CNC4=CC=CC=C43)C(=O)N[C@@H](CCCCN)C(=O)O)N XXJDYWYVZBHELV-TUSQITKMSA-N 0.000 description 1
- FJKXUIJOMUWCDD-FHWLQOOXSA-N Tyr-Gln-Tyr Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)N[C@@H](CC2=CC=C(C=C2)O)C(=O)O)N)O FJKXUIJOMUWCDD-FHWLQOOXSA-N 0.000 description 1
- KCPFDGNYAMKZQP-KBPBESRZSA-N Tyr-Gly-Leu Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)NCC(=O)N[C@@H](CC(C)C)C(O)=O KCPFDGNYAMKZQP-KBPBESRZSA-N 0.000 description 1
- WPXKRJVHBXYLDT-JUKXBJQTSA-N Tyr-His-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)[C@H](CC1=CN=CN1)NC(=O)[C@H](CC2=CC=C(C=C2)O)N WPXKRJVHBXYLDT-JUKXBJQTSA-N 0.000 description 1
- FBHBVXUBTYVCRU-BZSNNMDCSA-N Tyr-His-Leu Chemical compound C([C@@H](C(=O)N[C@@H](CC(C)C)C(O)=O)NC(=O)[C@@H](N)CC=1C=CC(O)=CC=1)C1=CN=CN1 FBHBVXUBTYVCRU-BZSNNMDCSA-N 0.000 description 1
- ARSHSYUZHSIYKR-ACRUOGEOSA-N Tyr-His-Phe Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O ARSHSYUZHSIYKR-ACRUOGEOSA-N 0.000 description 1
- YKCXQOBTISTQJD-BZSNNMDCSA-N Tyr-Leu-His Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CC1=CN=CN1)C(=O)O)NC(=O)[C@H](CC2=CC=C(C=C2)O)N YKCXQOBTISTQJD-BZSNNMDCSA-N 0.000 description 1
- ARJASMXQBRNAGI-YESZJQIVSA-N Tyr-Leu-Pro Chemical compound CC(C)C[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CC2=CC=C(C=C2)O)N ARJASMXQBRNAGI-YESZJQIVSA-N 0.000 description 1
- OKDNSNWJEXAMSU-IRXDYDNUSA-N Tyr-Phe-Gly Chemical compound C([C@H](N)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)NCC(O)=O)C1=CC=C(O)C=C1 OKDNSNWJEXAMSU-IRXDYDNUSA-N 0.000 description 1
- VXCAZHCVDBQMTP-NRPADANISA-N Val-Cys-Gln Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CS)C(=O)N[C@@H](CCC(=O)N)C(=O)O)N VXCAZHCVDBQMTP-NRPADANISA-N 0.000 description 1
- PMDOQZFYGWZSTK-LSJOCFKGSA-N Val-Gly-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)CNC(=O)[C@@H](N)C(C)C PMDOQZFYGWZSTK-LSJOCFKGSA-N 0.000 description 1
- BZDGLJPROOOUOZ-XGEHTFHBSA-N Val-Thr-Cys Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CS)C(=O)O)NC(=O)[C@H](C(C)C)N)O BZDGLJPROOOUOZ-XGEHTFHBSA-N 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- 108010070944 alanylhistidine Proteins 0.000 description 1
- 229960000723 ampicillin Drugs 0.000 description 1
- AVKUERGKIZMTKX-NJBDSQKTSA-N ampicillin Chemical compound C1([C@@H](N)C(=O)N[C@H]2[C@H]3SC([C@@H](N3C2=O)C(O)=O)(C)C)=CC=CC=C1 AVKUERGKIZMTKX-NJBDSQKTSA-N 0.000 description 1
- 239000003674 animal food additive Substances 0.000 description 1
- 239000003242 anti bacterial agent Substances 0.000 description 1
- 229940088710 antibiotic agent Drugs 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- 230000003078 antioxidant effect Effects 0.000 description 1
- 235000006708 antioxidants Nutrition 0.000 description 1
- 238000009360 aquaculture Methods 0.000 description 1
- 244000144974 aquaculture Species 0.000 description 1
- 108010038633 aspartylglutamate Proteins 0.000 description 1
- 108010092854 aspartyllysine Proteins 0.000 description 1
- 108010068265 aspartyltyrosine Proteins 0.000 description 1
- MQZIGYBFDRPAKN-UWFIBFSHSA-N astaxanthin Chemical compound C([C@H](O)C(=O)C=1C)C(C)(C)C=1\C=C\C(\C)=C\C=C\C(\C)=C\C=C\C=C(/C)\C=C\C=C(/C)\C=C\C1=C(C)C(=O)[C@@H](O)CC1(C)C MQZIGYBFDRPAKN-UWFIBFSHSA-N 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 108010091656 beta-carotene hydroxylase Proteins 0.000 description 1
- 230000036978 cell physiology Effects 0.000 description 1
- 239000006285 cell suspension Substances 0.000 description 1
- 210000003850 cellular structure Anatomy 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- 239000003086 colorant Substances 0.000 description 1
- 230000002860 competitive effect Effects 0.000 description 1
- 238000011109 contamination Methods 0.000 description 1
- 239000013256 coordination polymer Substances 0.000 description 1
- 108010069495 cysteinyltyrosine Proteins 0.000 description 1
- 235000015872 dietary supplement Nutrition 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 150000002191 fatty alcohols Chemical class 0.000 description 1
- 239000000446 fuel Substances 0.000 description 1
- 238000012239 gene modification Methods 0.000 description 1
- 230000005017 genetic modification Effects 0.000 description 1
- 235000013617 genetically modified food Nutrition 0.000 description 1
- 230000004110 gluconeogenesis Effects 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 108010049041 glutamylalanine Proteins 0.000 description 1
- 238000003306 harvesting Methods 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 108010036413 histidylglycine Proteins 0.000 description 1
- 108010025306 histidylleucine Proteins 0.000 description 1
- 108010092114 histidylphenylalanine Proteins 0.000 description 1
- 210000000987 immune system Anatomy 0.000 description 1
- 239000000411 inducer Substances 0.000 description 1
- 238000003780 insertion Methods 0.000 description 1
- 230000037431 insertion Effects 0.000 description 1
- 230000003834 intracellular effect Effects 0.000 description 1
- 101150044508 key gene Proteins 0.000 description 1
- 108010051673 leucyl-glycyl-phenylalanine Proteins 0.000 description 1
- 108010034529 leucyl-lysine Proteins 0.000 description 1
- 108010044056 leucyl-phenylalanine Proteins 0.000 description 1
- 108010057821 leucylproline Proteins 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 108010054155 lysyllysine Proteins 0.000 description 1
- 230000001404 mediated effect Effects 0.000 description 1
- 230000002503 metabolic effect Effects 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 238000005502 peroxidation Methods 0.000 description 1
- 108010065135 phenylalanyl-phenylalanyl-phenylalanine Proteins 0.000 description 1
- 108010073101 phenylalanylleucine Proteins 0.000 description 1
- 108010073025 phenylalanylphenylalanine Proteins 0.000 description 1
- 230000029553 photosynthesis Effects 0.000 description 1
- 238000010672 photosynthesis Methods 0.000 description 1
- 210000001916 photosynthetic cell Anatomy 0.000 description 1
- 235000011765 phytoene Nutrition 0.000 description 1
- 230000037039 plant physiology Effects 0.000 description 1
- 239000013600 plasmid vector Substances 0.000 description 1
- 239000002243 precursor Substances 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 108010031719 prolyl-serine Proteins 0.000 description 1
- 108010070643 prolylglutamic acid Proteins 0.000 description 1
- 108010090894 prolylleucine Proteins 0.000 description 1
- 229940107700 pyruvic acid Drugs 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000004007 reversed phase HPLC Methods 0.000 description 1
- 238000007363 ring formation reaction Methods 0.000 description 1
- 108010048818 seryl-histidine Proteins 0.000 description 1
- 239000004332 silver Substances 0.000 description 1
- 229910052709 silver Inorganic materials 0.000 description 1
- AKHNMLFCWUSKQB-UHFFFAOYSA-L sodium thiosulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=S AKHNMLFCWUSKQB-UHFFFAOYSA-L 0.000 description 1
- 235000019345 sodium thiosulphate Nutrition 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 150000003505 terpenes Chemical class 0.000 description 1
- 108010061238 threonyl-glycine Proteins 0.000 description 1
- 108010071097 threonyl-lysyl-proline Proteins 0.000 description 1
- 150000003626 triacylglycerols Chemical class 0.000 description 1
- 108010080629 tryptophan-leucine Proteins 0.000 description 1
- 108010038745 tryptophylglycine Proteins 0.000 description 1
- 108010051110 tyrosyl-lysine Proteins 0.000 description 1
- 238000010792 warming Methods 0.000 description 1
- 108010027345 wheylin-1 peptide Proteins 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- 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
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/74—Vectors or expression systems specially adapted for prokaryotic hosts other than E. coli, e.g. Lactobacillus, Micromonospora
-
- 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
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/65—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression using markers
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/0004—Oxidoreductases (1.)
-
- 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
- C12P23/00—Preparation of compounds containing a cyclohexene ring having an unsaturated side chain containing at least ten carbon atoms bound by conjugated double bonds, e.g. carotenes
Landscapes
- Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biotechnology (AREA)
- General Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Microbiology (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Molecular Biology (AREA)
- Plant Pathology (AREA)
- Biophysics (AREA)
- Physics & Mathematics (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Medicinal Chemistry (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
本发明公开了一种用于生产虾青素的转基因蓝藻及其应用，涉及微生物学和分子生物学领域。一种用于生产虾青素的转基因蓝藻，以蓝藻作为出发菌株，通过导入来自念珠藻Nostoc sp.ATCC 27893的类胡萝卜素酮醇酶CrtW的编码基因获得所述藻类工程菌。本发明的转基因蓝藻能够有效地将二氧化碳转化为虾青素，其虾青素产量最高可达2.6mg/L，为虾青素的异源微生物生产提供更加经济、效率、简便的方式。
Description
技术领域
本发明涉及微生物学和分子生物学领域，具体涉及一种用于生产虾青素的转基因蓝藻及其应用。
背景技术
随着全球变暖的影响日益加剧，开发环保生物技术备受关注。使用光合蓝藻整合CO2捕获与生物合成的技术为生产碳中和且高附加值的产品提供了一种可能性。虾青素的生物合成基于类胡萝卜素的合成代谢途径。由于蓝藻天然具有广泛的类胡萝卜素合成途径，因此在蓝藻中进行虾青素的生物合成具有显着的遗传优势。此外，蓝藻，特别是蓝藻中的集胞藻Synechocystis sp.PCC 6803天然可积累虾青素的前提物玉米黄素(Makino T.,Harada H.,Ikenaga H.,Matsuda S.,Takaichi S.,Shindo K.,Sandmann G.,Ogata T.,Misawa N..2008.Characterization of cyanobacterial carotenoid ketolase CrtWand hydroxylase CrtR by complementation analysis in Escherichia coli.Plantand Cell Physiology,49:1867-1878.)，作为虾青素的生产宿主，蓝藻中的集胞藻Synechocystis sp.PCC 6803更具竞争力。
虾青素(3,3'-dihydroxy-β,β-carotene-4,4'-dione，astaxanthin)是一种酮类胡萝卜素，被认为是一种强抗氧化剂。在水产养殖中主要用作营养增补剂、饲料添加剂与着色剂。由于虾青素具有防止脂质膜过氧化与增强免疫系统等对人体健康的益处，因此近年来虾青素的生物合成已引起广泛关注。
天然虾青素是由某些微藻、细菌与真菌合成的。目前，雨生红球藻(Haematococcuspluvialis)是工业上用于虾青素生物合成的主要微生物之一。然而，雨生红球藻在虾青素的产量与成本上仍存在诸多挑战。比如在大规模的培养过程中，关于细胞生长和培养过程中的污染等问题。因此，开发用于能高效生产虾青素的替代菌株有着显著意义。
近年来，开发蓝藻作为光合细胞工厂受到科研工作者的广泛关注。作为最简单的光合模式原核生物之一，蓝藻呈现出兼具原核微生物和光自养微生物两种特性的表型，其相对充分的遗传信息及简单的细胞结构为研究光合作用与碳固定提供了理想平台(DucatD.C.,Way J.C.,Silver P.A..2011.Engineering cyanobacteria to generate high-value products.Trends in Biotechnology,29:95-103.)。
利用蓝藻固定CO2的能力与代谢途径工程，蓝藻已被用于燃料和多种化学品的光合生物制造。例如，授权号为US06699696B2的专利申请公开了一种通过基因工程改造蓝藻Synechococcus PCC 7942异源表达丙酮酸脱羧酶和醇脱氢酶生产乙醇的方法。授权号为US2011/0256599A1的专利申请公开了通过基因工程改造蓝藻生产脂肪醇的过程。授权号为US2010/0081178A1的专利申请公开了一种用于生产三酰基甘油酯的转基因蓝藻的过程。
作为光合微生物，蓝藻的快速生长、简单的碳源需求与相对容易的遗传改造过程为进一步提升当前基于藻类的类胡萝卜素制造提供了新的机遇。相比细菌或真菌等利用有机碳源生产虾青素的模式(如CN107129995A，CN105861538B等)，利用光能与CO2的生产模式更具可持续性。
发明内容
本发明提供了一种转基因蓝藻用于生产虾青素。经遗传改造的蓝藻经由包含编码类胡萝卜素酮醇酶CrtW的核苷酸序列的重组质粒pNX31-Crt W所转化。重组质粒pNX31-CrtW用于整合至蓝藻中的集胞藻Synechocy stis sp.PCC 6803的基因组psbA1基因座。类胡萝卜素酮醇酶CrtW在改造过的蓝藻细胞内表达，能够将其天然合成的玉米黄素转化为虾青素，因而拓展了类胡萝卜素的代谢途径。
本发明提供了一种用于生产虾青素的转基因蓝藻，以蓝藻作为出发菌株，通过导入来自念珠藻(Nostoc sp.)的类胡萝卜素酮醇酶CrtW的编码基因获得所述转基因蓝藻。
所述念珠藻(Nostoc sp.)可以为念珠藻Nostoc sp.ATCC 27893。
优选的，所述类胡萝卜素酮醇酶CrtW的编码基因的核苷酸序列如SEQ ID NO.15所示。
优选的，还导入了来自集胞藻(Synechocystis sp.)的CrtR基因，CrtR基因的核苷酸序列如SEQ ID NO.17所示。
所述集胞藻(Synechocystis sp.)可以为集胞藻Synechocystis sp.PCC 6803。
优选的，作为出发菌株的蓝藻为集胞藻(Synechocystis sp.)。
本发明还提供了上述转基因蓝藻的构建方法，包括以下步骤：
(1)构建重组质粒pNX31-CrtW：
将氯霉素抗性基因连接到质粒pGEM T-Easy上，得到质粒pGEM-Cm；将psbA1基因的上游序列以及下游序列连接到质粒pGEM-Cm上，获得质粒pGEM-Arm；再将从质粒pTAC-MAT-TAG-1克隆的操纵子序列，包括tac启动子区域、多克隆位点与T1终止子整合到质粒pGEM-Arm上，获得质粒pNX31；将来自蓝藻中集胞藻(Synechocystis sp.)的启动子rbc插入到质粒pNX31中，取代tac启动子，得到质粒pNX31-rbc；将来自念珠藻(Nostoc sp.)的类胡萝卜素酮醇酶CrtW的编码基因连接到质粒pNX31-rbc上，获得重组质粒pNX31-CrtW；
(2)将步骤(1)中得到的质粒pNX31-CrtW转化到蓝藻中的集胞藻(Synechocystissp.)中，培养获得转基因蓝藻。
本发明还提供了上述转基因蓝藻在生产虾青素中的应用。
本发明还提供了一种生产虾青素的方法，发酵培养上述转基因蓝藻，结束后收集转基因蓝藻并提取虾青素。
发酵培养条件为：通入含体积比浓度为1％的CO2的空气，25℃，30-50μmol/s/m2光子下培养。
本发明的有益效果：
本发明的转基因蓝藻能够有效地将二氧化碳转化为虾青素，其虾青素产量最高可达2.6mg/L。异源基因被整合至基因组上且为组成型表达，培养过程中无需添加抗生素或者诱导剂。此外，用蓝藻进行的虾青素生产与当下普遍使用的雨生红球藻的光自养生产过程完全兼容，这可直接为基于藻类的类胡萝卜素生产技术提供替代菌株与升级。本发明为虾青素的异源微生物生产提供更加经济、效率、简便的方式。
附图说明
图1为蓝藻中的集胞藻Synechocystis sp.PCC 6803中虾青素生物合成的代谢途径图。
图2为质粒pNX31-crtW的构建过程图。
图3为比较转基因与野生型Synechocystis sp.PCC 6803中虾青素的HPLC分析图。
图4为转基因蓝藻Synechocystis sp.PCC 6803生产虾青素的产量图。
具体实施方式
本发明提供了一种用于生产虾青素的替代微生物。如本文所述，蓝藻中的集胞藻Synechocystis sp.PCC 6803经过基因改造可用以生产虾青素。遗传修饰的蓝藻经由包含编码类胡萝卜素酮醇酶CrtW的DNA序列的重组质粒所转化。类胡萝卜素酮醇酶CrtW在改造后的蓝藻体内进行异源表达，能够催化玉米黄素向虾青素的转化。
在本发明中，使用了多个术语。在此提供以下定义。
“虾青素生产”是指通过微生物在胞内生物合成虾青素。
“蓝藻”是指能够进行光合自养微生物，例如Synechocystis，Nostoc(Anabaena)，Synechococcus。
“类胡萝卜素酮醇酶”是指能够催化玉米黄素转化为虾青素的酶，例如β-胡萝卜素酮醇酶(CrtW/CrtO)。
“转化”是指利用质粒载体介导的核苷酸分子对微生物进行遗传转化，包括整合型质粒与非整合型质粒。
“pGEM-Cm”是指克隆了pHSG398(TaKaRa，大连，中国)中氯霉素抗性基因到pGEM T-Easy(Promega，Madison，WI)的质粒。
“pNX31-Arm”是指一个蓝藻转化质粒，其包含了来自蓝藻中的集胞Synechocystissp.的psbA1基因片段上游和下游的同源序列并整合至pGEM-Cm质粒中。
“pNX31”是指一个蓝藻转化质粒。将来自pTAC-MAT-TAG-1(Sigma，St.Louis，MO)的tac启动子区域、多克隆位点和T1终止子区域克隆到pNX31-Arm中构建而成。
“pNX31-rbc”是指一个包含来自Synechocystis sp.PCC 6803的rbc启动子的蓝藻转化质粒，rbc启动子整合进入pNX31以取代tac启动子。
“pNX31-crtW”是指将来自念珠藻Nostoc sp.ATCC 27893的CrtW基因克隆到pNX31-rbc质粒中的虾青素生产质粒。
在蓝藻中，起始碳源CO2经过开尔文循环转化为3-磷酸甘油酸(3-PG)，用于连接糖酵解和糖异生途径。类胡萝卜素的生物合成始于糖酵解中两个代谢节点的缩合，即甘油-3-磷酸(G3P)与丙酮酸。在Synechocystis sp.PCC 6803中，2-C-甲基-D-赤藓糖醇-4-磷酸(MEP)途径被用于合成异戊二烯单元异戊烯基二磷酸(IPP)和二甲基烯丙基二磷酸(DMAPP)。从IPP和DMAPP的缩合开始，合成了一组类异戊二烯，包括焦磷酸香叶酯(GPP，C10)、焦磷酸法呢酯(FPP，C15)、焦磷酸香叶基香叶酯(GGPP，C20)和八氢番茄红素(C40)。经过两步去饱和与两步环化，生成β-胡萝卜素。从β-胡萝卜素合成虾青素依赖于两个关键酶，即β-胡萝卜素羟化酶(CrtR/CrtZ)和β-胡萝卜素酮醇酶(CrtW/CrtO)(图1)。由于玉米黄素在蓝藻中的集胞藻Synechocystis sp.PCC 6803中天然积累，因此通过异源表达类胡萝卜素酮醇酶(CrtW)的转基因蓝藻能够将天然合成的玉米黄素转化为虾青素。
本发明的一个优点在于用蓝藻进行的虾青素生产与当下普遍使用的雨生红球藻的光自养生产过程完全兼容。这可直接为基于藻类的类胡萝卜素生产技术提供替代菌株与升级。转基因蓝藻能够有效地将二氧化碳转化为虾青素。此外，Synechocystis sp.PCC6803独特的玉米黄素积累和相对完整的遗传学研究可有效减少遗传操作的步骤，从而显著提高重组菌株的遗传稳定性和稳健性。与野生型Synechocystis sp.相比，重组蓝藻的生长没有受到显著影响。
质粒pNX31与pNX31-rbc被用于整合至Synechocystis sp.基因组psbA1的基因座。所述质粒中的tac/rbc启动子、多克隆位点与终止子构成了质粒的操纵子，可应用于单个或多个基因的构建与表达，以实现对Synechocystis sp.的遗传改造。
以下示例旨在说明本发明的实施例，但并不受限于如下应用。
实施例1
质粒pNX31-CrtW的构建。
(1)质粒pNX31-CrtW来自于质粒pGEM-T-Easy(Promega，Madison，WI)。为了构建该质粒，首先使用引物(如SEQ ID NO.1和SEQ ID NO.2所示)从质粒pHSG398(TaKaRa，大连，中国)克隆氯霉素抗性基因。然后，通过使用T-A克隆将克隆的DNA片段插入到质粒pGEM-T-Easy中以生成质粒pGEM-Cm，图谱如图2所示。
(2)由于psbA1基因(GenBank ID:CP028094.1)被设计为Synechocystis基因组中的插入位点，psbA1基因的上游(PCR扩增引物：如SEQ ID NO.3和SEQ ID NO.4所示)和下游序列(PCR扩增引物：如SEQ ID NO.5和SEQ ID NO.6所示)从Synechocystis sp.PCC 6803的基因组中克隆出来分别插入到质粒pGEM-Cm，获得质粒pNX31-Arm，图谱如图2所示。
(3)在进一步整合了从质粒pTAC-MAT-TAG-1(Sigma，St.Louis，MO)克隆的操纵子序列，包括tac启动子区域、多克隆位点与T1终止子(PCR扩增引物：如SEQ ID NO.7和SEQ IDNO.8所示)，获得质粒pNX31，图谱如图2所示。
(4)为了构建替代启动子的质粒，rbc启动子(核苷酸序列如SEQ ID NO.19所示)PCR扩增引物：如SEQ ID NO.9和SEQ ID NO.10所示)从Synechocystis sp.PCC 6803中克隆并插入质粒pNX31取代tac启动子获得质粒pNX31-rbc，图谱如图2所示。
(5)通过使用引物(引物序列如SEQ ID NO.11和SEQ ID NO.12所示)克隆来自Nostoc sp.ATCC 27893的CrtW基因(基因序列如SEQ ID NO.15所示，氨基酸序列如SEQ IDNO.16所示)的DNA片段至质粒pNX31-rbc以构建成质粒pNX31-CrtW，图谱如图2所示。pNX31-CrtW的构建过程见图2。
大肠杆菌NEB 5α感受态细胞用于所有质粒的构建。大肠杆菌使用Luria-Bertani(LB)培养基于37℃生长，并根据需要将100μg/mL氨苄青霉素添加到LB培养基中。
实施例2
重组质粒pNX31-CrtW转化蓝藻Synechocystis sp.PCC 6803的构建过程。
Synechocystis sp.PCC 6803菌株在BG-11培养基中于25℃下在30-50μmol/s/m2光子的光强度下生长。BG-11培养基平板包含15g/L的琼脂，10mM TES-NaOH(pH 8.2)与3g/L硫代硫酸钠(Eaton-Rye，2004)。
收获指数生长期的Synechocystis sp.PCC 6803细胞并使用新鲜的BG-11培养基以1×109细胞/mL的最终密度进行重悬浮。在0.5mL细胞悬液中加入2-10μg质粒pNX31-CrtW，光照培养6小时。将细胞-DNA混合物铺在附有Nuclepore膜(Whatman，Florham Park，NJ，USA)的不含抗生素的BG-11平板上过夜生长。然后将Nuclepore膜转移到含有5mM葡萄糖与25μg/mL氯霉素的BG-11平板上。经1-2周的培养后，转基因蓝藻Synechocystis sp.PCC6803菌落可出现在含抗生素的平板上。挑取单菌落在含抗生素的BG-11平板上重新划线以获得纯合转化子。
实施例3
转基因蓝藻Synechocystis sp.PCC 6803生产虾青素的过程及产品分析。
转基因蓝藻Synechocystis sp.PCC 6803菌株在装有150mL BG-11培养基的250mL摇瓶中生长。摇瓶中通入含1％(体积比)CO2浓度的空气，25℃，在30-50μmol/s/m2光子下生长。通过测量730nm处的光密度(OD)来监测细胞生长，在培养后，收集细胞用于虾青素的提取和分析。使用甲醇从细胞中提取类胡萝卜素，并利用反相高效液相色谱(HPLC)进行分析。虾青素的分析可通过与标准品的比较进行，并通过积分峰面积进行定量。HPLC结果表明，与野生型菌株相比，转基因蓝藻可合成虾青素(图3)。
由于自β-胡萝卜素合成虾青素依赖于两个关键酶(CrtR与CrtW)，我们在转基因蓝藻Synechocystis sp.PCC 6803中进一步过量表达了另外一个关键基因CrtR。通过使用引物(引物序列如SEQ ID NO.13和SEQ ID NO.14所示)克隆来自蓝藻中的集胞藻Synechocystis sp.PCC 6803的CrtR基因(核苷酸序列如SEQ ID NO.17所示，氨基酸序列如SEQ ID NO.18所示)的DNA片段至质粒pNX31-CrtW的XbaI与KpnI位点以构建成质粒pNX31-CrtWR，并用于转化蓝藻Synechocystis sp.PCC 6803获得重组菌株。图4展示了转基因蓝藻Synechocystis sp.PCC 6803虾青素的生产量，最高可达2.6mg/L(1.5mg/g DCW)。
序列表
<110> 浙江大学杭州国际科创中心
<120> 一种用于生产虾青素的转基因蓝藻及其应用
<160> 19
<170> SIPOSequenceListing 1.0
<210> 1
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 1
tcaaaaacac catcatacac t 21
<210> 2
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 2
catccgctta ttatcactta t 21
<210> 3
<211> 32
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 3
acatgcatgc ctgaaattca agggaagtta at 32
<210> 4
<211> 30
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 4
taatgcatgc cggcgacggg ggcgctatag 30
<210> 5
<211> 32
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 5
ccgcctgcag ctgccactgc tactttgtta at 32
<210> 6
<211> 30
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 6
ttccgagctc aatggcgggg gcgttcaggg 30
<210> 7
<211> 35
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 7
gcggccgcta ctagtgggct tatcgactgc acggt 35
<210> 8
<211> 32
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 8
aaaactgcag cgttgcttcg caacgttcaa at 32
<210> 9
<211> 36
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 9
gcggccgcta ctagtatcac catttggaca aaacat 36
<210> 10
<211> 35
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 10
gggtatcata tgctaggtca gtcctccata aacat 35
<210> 11
<211> 34
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 11
gggaattcca tatggttcag tgtcaaccat catc 34
<210> 12
<211> 38
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 12
gcttctcgag ttctagagtt ataaagatat tttgtgag 38
<210> 13
<211> 48
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 13
ccgctactag tatatggagg tttaaatgtg ccaggagtcc gtcatagt 48
<210> 14
<211> 44
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 14
ccggggtacc ttctagagct agggcttgtc agatgattga gaat 44
<210> 15
<211> 777
<212> DNA
<213> 念珠藻(Nostoc sp. ATCC 27893)
<400> 15
atggttcagt gtcaaccatc atctctgcat tcagaaaaac tggtgttatt gtcatcgaca 60
atcagagatg ataaaaatat taataagggt atatttattg cctgctttat cttattttta 120
tgggcaatta gtttaatctt attactctca atagatacat ccataattca taagagctta 180
ttaggtatag ccatgctttg gcagaccttc ttatatacag gtttatttat tactgctcat 240
gatgccatgc acggcgtagt ttatcccaaa aatcccagaa taaataattt tataggtaag 300
ctcactctaa tcttgtatgg actactccct tataaagatt tattgaaaaa acattggtta 360
caccacggac atcctggtac tgatttagac cctgattatt acaatggtca tccccaaaac 420
ttctttcttt ggtatctaca ttttatgaag tcttattggc gatggacgca aattttcgga 480
ttagtgatga tttttcatgg acttaaaaat ctggtgcata taccagaaaa taatttaatt 540
atattttgga tgataccttc tattttaagt tcagtacaac tattttattt tggtacattt 600
ttgcctcata aaaagctaga aggtggttat actaaccccc attgtgcgcg cagtatccca 660
ttacctcttt tttggtcttt tgttacttgt tatcacttcg gctaccacaa ggaacatcac 720
gaataccctc aacttccttg gtggaaatta cctgaagctc acaaaatatc tttataa 777
<210> 16
<211> 258
<212> PRT
<213> 念珠藻(Nostoc sp. ATCC 27893)
<400> 16
Met Val Gln Cys Gln Pro Ser Ser Leu His Ser Glu Lys Leu Val Leu
1 5 10 15
Leu Ser Ser Thr Ile Arg Asp Asp Lys Asn Ile Asn Lys Gly Ile Phe
20 25 30
Ile Ala Cys Phe Ile Leu Phe Leu Trp Ala Ile Ser Leu Ile Leu Leu
35 40 45
Leu Ser Ile Asp Thr Ser Ile Ile His Lys Ser Leu Leu Gly Ile Ala
50 55 60
Met Leu Trp Gln Thr Phe Leu Tyr Thr Gly Leu Phe Ile Thr Ala His
65 70 75 80
Asp Ala Met His Gly Val Val Tyr Pro Lys Asn Pro Arg Ile Asn Asn
85 90 95
Phe Ile Gly Lys Leu Thr Leu Ile Leu Tyr Gly Leu Leu Pro Tyr Lys
100 105 110
Asp Leu Leu Lys Lys His Trp Leu His His Gly His Pro Gly Thr Asp
115 120 125
Leu Asp Pro Asp Tyr Tyr Asn Gly His Pro Gln Asn Phe Phe Leu Trp
130 135 140
Tyr Leu His Phe Met Lys Ser Tyr Trp Arg Trp Thr Gln Ile Phe Gly
145 150 155 160
Leu Val Met Ile Phe His Gly Leu Lys Asn Leu Val His Ile Pro Glu
165 170 175
Asn Asn Leu Ile Ile Phe Trp Met Ile Pro Ser Ile Leu Ser Ser Val
180 185 190
Gln Leu Phe Tyr Phe Gly Thr Phe Leu Pro His Lys Lys Leu Glu Gly
195 200 205
Gly Tyr Thr Asn Pro His Cys Ala Arg Ser Ile Pro Leu Pro Leu Phe
210 215 220
Trp Ser Phe Val Thr Cys Tyr His Phe Gly Tyr His Lys Glu His His
225 230 235 240
Glu Tyr Pro Gln Leu Pro Trp Trp Lys Leu Pro Glu Ala His Lys Ile
245 250 255
Ser Leu
<210> 17
<211> 939
<212> DNA
<213> 集胞藻(Synechocystis PCC6803)
<400> 17
gtgtgccagg agtccgtcat agtaatgcag gcgacccaac cgctgcaaac cgtttcccaa 60
gctgtcccaa aagagttttt acaggcggac ggcggcttca atcccaacgt ggccatgttc 120
gggatagcta ttctcttaat gctcgctaac gtttttggct actggcaatg ggggctgccc 180
cactggcttt gttttagttg ttcggtgctg gcgctgcacc tgtcaggcac agtgatccat 240
gatgcatccc acaatgcggc ccatcggaac accattatta atgcagtgct tggccacggt 300
agtgccttaa tgttgggctt tgcttttccc gtctttaccc gggttcatct ccaacaccac 360
gccaacgtca atgaccctga aaatgaccca gaccattttg tttccaccgg cggtcccctc 420
ttcctcattg ccgcccggtt cttctaccat gagatctttt tctttaaacg gcggttatgg 480
cgcaaatatg agctactaga gtggttctta agtcggcttg tgttgttcac gatcgttttt 540
ctcggcattc attacggctt tatcggcttt gtgatgaatt actggtttgt gcctgcttta 600
attgttggca ttgccctggg actgtttttt gattacctgc cccatcgacc tttccaagaa 660
cgcaaccgtt ggaaaaatgc cagggtttat cccagcccca ttttaaattg gctcattttc 720
gggcaaaatt accacctgat ccaccacctt tggccttcta ttccttggta tcagtaccaa 780
aacacctatc acatcaccaa gcccattttg gatgagaagg gttgtgatca atccctggga 840
ttactggaag ggaaaaattt ctggagcttc ctctatgatg ttttccttgg tattcgtttt 900
cacggccata ataattctca atcatctgac aagccctag 939
<210> 18
<211> 312
<212> PRT
<213> 集胞藻(Synechocystis PCC6803)
<400> 18
Val Cys Gln Glu Ser Val Ile Val Met Gln Ala Thr Gln Pro Leu Gln
1 5 10 15
Thr Val Ser Gln Ala Val Pro Lys Glu Phe Leu Gln Ala Asp Gly Gly
20 25 30
Phe Asn Pro Asn Val Ala Met Phe Gly Ile Ala Ile Leu Leu Met Leu
35 40 45
Ala Asn Val Phe Gly Tyr Trp Gln Trp Gly Leu Pro His Trp Leu Cys
50 55 60
Phe Ser Cys Ser Val Leu Ala Leu His Leu Ser Gly Thr Val Ile His
65 70 75 80
Asp Ala Ser His Asn Ala Ala His Arg Asn Thr Ile Ile Asn Ala Val
85 90 95
Leu Gly His Gly Ser Ala Leu Met Leu Gly Phe Ala Phe Pro Val Phe
100 105 110
Thr Arg Val His Leu Gln His His Ala Asn Val Asn Asp Pro Glu Asn
115 120 125
Asp Pro Asp His Phe Val Ser Thr Gly Gly Pro Leu Phe Leu Ile Ala
130 135 140
Ala Arg Phe Phe Tyr His Glu Ile Phe Phe Phe Lys Arg Arg Leu Trp
145 150 155 160
Arg Lys Tyr Glu Leu Leu Glu Trp Phe Leu Ser Arg Leu Val Leu Phe
165 170 175
Thr Ile Val Phe Leu Gly Ile His Tyr Gly Phe Ile Gly Phe Val Met
180 185 190
Asn Tyr Trp Phe Val Pro Ala Leu Ile Val Gly Ile Ala Leu Gly Leu
195 200 205
Phe Phe Asp Tyr Leu Pro His Arg Pro Phe Gln Glu Arg Asn Arg Trp
210 215 220
Lys Asn Ala Arg Val Tyr Pro Ser Pro Ile Leu Asn Trp Leu Ile Phe
225 230 235 240
Gly Gln Asn Tyr His Leu Ile His His Leu Trp Pro Ser Ile Pro Trp
245 250 255
Tyr Gln Tyr Gln Asn Thr Tyr His Ile Thr Lys Pro Ile Leu Asp Glu
260 265 270
Lys Gly Cys Asp Gln Ser Leu Gly Leu Leu Glu Gly Lys Asn Phe Trp
275 280 285
Ser Phe Leu Tyr Asp Val Phe Leu Gly Ile Arg Phe His Gly His Asn
290 295 300
Asn Ser Gln Ser Ser Asp Lys Pro
305 310
<210> 19
<211> 277
<212> DNA
<213> 集胞藻(Synechocystis PCC6803)
<400> 19
tcaccatttg gacaaaacat caggaattct aattagaaag tccaaaaatt gtaatttaaa 60
aaacagtcaa tggagagcat tgccataagt aaaggcatcc cctgcgtgat aagattacct 120
tcagaaaaca gatagttgct gggttatcgc agatttttct cgcaaccaaa taactgtaaa 180
taataactgt ctctggggcg acggtaggct ttatattgcc aaatttcgcc cgtgggagaa 240
agctaggcta ttcaatgttt atggaggact gacctag 277
Claims (9)
1.一种用于生产虾青素的转基因蓝藻，其特征在于，以蓝藻作为出发菌株，通过导入来自念珠藻(Nostoc sp.)的类胡萝卜素酮醇酶CrtW的编码基因获得所述转基因蓝藻。
2.如权利要求1所述的转基因蓝藻，其特征在于，所述类胡萝卜素酮醇酶CrtW的编码基因的核苷酸序列如SEQ ID NO.15所示。
3.如权利要求1所述的转基因蓝藻，其特征在于，还导入了来自集胞藻(Synechocystissp.)的CrtR基因。
4.如权利要求3所述的转基因蓝藻，其特征在于，CrtR基因的核苷酸序列如SEQ IDNO.17所示。
5.如权利要求1所述的转基因蓝藻，其特征在于，作为出发菌株的蓝藻为集胞藻(Synechocystis sp.)。
6.如权利要求1-5任一项所述转基因蓝藻的构建方法，其特征在于，包括以下步骤：
(1)构建重组质粒pNX31-CrtW：
将氯霉素抗性基因连接到质粒pGEM T-Easy上，得到质粒pGEM-Cm；将psbA1基因的上游序列以及下游序列连接到质粒pGEM-Cm上，获得质粒pGEM-Arm；再将从质粒pTAC-MAT-TAG-1克隆的操纵子序列，包括tac启动子区域、多克隆位点与T1终止子整合到质粒pGEM-Arm上，获得质粒pNX31；将来自蓝藻中集胞藻(Synechocystis sp.)的启动子rbc插入到质粒pNX31中，取代tac启动子，得到质粒pNX31-rbc；将来自念珠藻(Nostoc sp.)的类胡萝卜素酮醇酶CrtW的编码基因连接到质粒pNX31-rbc上，获得重组质粒pNX31-CrtW；
(2)将步骤(1)中得到的质粒pNX31-CrtW转化到集胞藻(Synechocystis sp.)中，培养获得转基因蓝藻。
7.如权利要求1-5任一项所述转基因蓝藻在生产虾青素中的应用。
8.一种生产虾青素的方法，其特征在于，发酵培养权利要求1-5任一项所述转基因蓝藻，结束后收集转基因蓝藻并提取虾青素。
9.如权利要求8所述的方法，其特征在于，发酵培养条件为：通入含体积比浓度为1％的CO2的空气，25℃，30-50μmol/s/m2光子下培养。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210110355.0A CN116555133A (zh) | 2022-01-29 | 2022-01-29 | 一种用于生产虾青素的转基因蓝藻及其应用 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210110355.0A CN116555133A (zh) | 2022-01-29 | 2022-01-29 | 一种用于生产虾青素的转基因蓝藻及其应用 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN116555133A true CN116555133A (zh) | 2023-08-08 |
Family
ID=87484907
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202210110355.0A Pending CN116555133A (zh) | 2022-01-29 | 2022-01-29 | 一种用于生产虾青素的转基因蓝藻及其应用 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN116555133A (zh) |
-
2022
- 2022-01-29 CN CN202210110355.0A patent/CN116555133A/zh active Pending
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Desai et al. | Photosynthetic approaches to chemical biotechnology | |
| Scaife et al. | Characterization of cyanobacterial β‐carotene ketolase and hydroxylase genes in Escherichia coli, and their application for astaxanthin biosynthesis | |
| US20110039323A1 (en) | Isoprene Production | |
| Chien et al. | Solar-to-bioH2 production enhanced by homologous overexpression of hydrogenase in green alga Chlorella sp. DT | |
| US10435720B2 (en) | Recombinant microorganism having enhanced D(-) 2,3-butanediol producing ability and method for producing D(-) 2,3-butanediol using the same | |
| CN108728471A (zh) | 产3-羟基丙酸的重组菌及其制备方法与应用 | |
| CN109689857A (zh) | 重组细胞、以及异戊二烯或萜的生产方法 | |
| CN111088175A (zh) | 一种产红没药烯的解脂耶氏酵母及其构建方法与用途 | |
| Yuzbasheva et al. | Large-scale bioproduction of natural astaxanthin in Yarrowia lipolytica | |
| Sun et al. | Conversion of carbon dioxide into valencene and other sesquiterpenes with metabolic engineered Synechocystis sp. PCC 6803 cell factories | |
| AU2012245238B2 (en) | Diffusion-based method for obtaining volatile hydrocarbons produced by photosynthetic microorganisms in two-phase bioreactors | |
| CA3026569A1 (en) | Methods of producing astaxanthin or precursors thereof | |
| CN116555133A (zh) | 一种用于生产虾青素的转基因蓝藻及其应用 | |
| US10138489B2 (en) | Cyanobacterial strains capable of utilizing phosphite | |
| CN114806909B (zh) | 一种生产β-胡萝卜素的菌株及其应用 | |
| CN105087627B (zh) | 一种sll0659基因在合成集胞藻类胡萝卜素中的应用 | |
| US20220220512A1 (en) | Carbon compound production in photosynthetic microorganisms | |
| AU2022257089A9 (en) | Methods of isoprenoid synthesis using a genetically engineered hydrocarbonoclastic organism in a biofilm bioreactor | |
| Tan et al. | Production of photosynthetic biofuels by genetically engineering cyanobacteria | |
| CN103571785B (zh) | 一种高效利用脂肪酸生产甲羟戊酸的方法及构建得到的基因工程菌 | |
| KR20160138897A (ko) | 글리세롤을 탄소원으로 이용하는 고생장성 대장균 | |
| KR20210047992A (ko) | 알파-휴물렌 생산용 형질전환 메탄자화균 및 이의 용도 | |
| Yoo et al. | Metabolic engineering for redirecting carbon to enhance the fatty acid content of synechocystis sp. PCC6803 | |
| RU2794980C1 (ru) | Модифицированная нуклеотидная последовательность, кодирующая полипептид, обладающий активностью фитоен синтазы и геранилгеранилпирофосфат синтазы (варианты), ее использование при конструировании трансформантов дрожжей Yarrowia lipolytica, продуцирующих каротиноиды | |
| US20170283832A1 (en) | Transformed synechococcus elongatus strain having capability of producing squalene from carbon dioxide and method for producing squalene using the same |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination |