CN112961875B - 一种生物法生产四氢嘧啶的工程菌株的构建方法 - Google Patents
一种生物法生产四氢嘧啶的工程菌株的构建方法 Download PDFInfo
- Publication number
- CN112961875B CN112961875B CN202110243895.1A CN202110243895A CN112961875B CN 112961875 B CN112961875 B CN 112961875B CN 202110243895 A CN202110243895 A CN 202110243895A CN 112961875 B CN112961875 B CN 112961875B
- Authority
- CN
- China
- Prior art keywords
- tetrahydropyrimidine
- gly
- gene cluster
- ala
- nde
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- OTPDWCMLUKMQNO-UHFFFAOYSA-N 1,2,3,4-tetrahydropyrimidine Chemical compound C1NCC=CN1 OTPDWCMLUKMQNO-UHFFFAOYSA-N 0.000 title claims abstract description 107
- 238000010276 construction Methods 0.000 title claims abstract description 11
- 238000010170 biological method Methods 0.000 title claims abstract description 6
- 238000006243 chemical reaction Methods 0.000 claims abstract description 38
- 230000015572 biosynthetic process Effects 0.000 claims abstract description 28
- 108091008053 gene clusters Proteins 0.000 claims abstract description 28
- 238000003786 synthesis reaction Methods 0.000 claims abstract description 28
- 238000004519 manufacturing process Methods 0.000 claims abstract description 27
- 238000000034 method Methods 0.000 claims abstract description 27
- WTWSHHITWMVLBX-DKWTVANSSA-M sodium;(2s)-2-aminobutanedioate;hydron Chemical compound [Na+].[O-]C(=O)[C@@H](N)CC(O)=O WTWSHHITWMVLBX-DKWTVANSSA-M 0.000 claims abstract description 24
- 241000588724 Escherichia coli Species 0.000 claims abstract description 21
- 108090000364 Ligases Proteins 0.000 claims abstract description 21
- 102000003960 Ligases Human genes 0.000 claims abstract description 20
- 102000003929 Transaminases Human genes 0.000 claims abstract description 15
- 108090000340 Transaminases Proteins 0.000 claims abstract description 15
- 102000005421 acetyltransferase Human genes 0.000 claims abstract description 15
- 108020002494 acetyltransferase Proteins 0.000 claims abstract description 15
- 230000008569 process Effects 0.000 claims abstract description 7
- 241001466077 Salina Species 0.000 claims abstract description 4
- 238000000855 fermentation Methods 0.000 claims description 33
- 230000004151 fermentation Effects 0.000 claims description 33
- WCUXLLCKKVVCTQ-UHFFFAOYSA-M Potassium chloride Chemical compound [Cl-].[K+] WCUXLLCKKVVCTQ-UHFFFAOYSA-M 0.000 claims description 28
- 239000013598 vector Substances 0.000 claims description 24
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 20
- 101000702488 Rattus norvegicus High affinity cationic amino acid transporter 1 Proteins 0.000 claims description 19
- 102000004190 Enzymes Human genes 0.000 claims description 17
- 108090000790 Enzymes Proteins 0.000 claims description 17
- 239000012634 fragment Substances 0.000 claims description 16
- 241000894006 Bacteria Species 0.000 claims description 15
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 claims description 15
- 238000012258 culturing Methods 0.000 claims description 15
- 238000004128 high performance liquid chromatography Methods 0.000 claims description 15
- 239000001888 Peptone Substances 0.000 claims description 14
- 108010080698 Peptones Proteins 0.000 claims description 14
- 229940041514 candida albicans extract Drugs 0.000 claims description 14
- 239000007788 liquid Substances 0.000 claims description 14
- 235000019319 peptone Nutrition 0.000 claims description 14
- 239000001103 potassium chloride Substances 0.000 claims description 14
- 235000011164 potassium chloride Nutrition 0.000 claims description 14
- 239000012138 yeast extract Substances 0.000 claims description 14
- 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 13
- 229940124277 aminobutyric acid Drugs 0.000 claims description 13
- BTCSSZJGUNDROE-UHFFFAOYSA-N gamma-aminobutyric acid Chemical compound NCCCC(O)=O BTCSSZJGUNDROE-UHFFFAOYSA-N 0.000 claims description 13
- 239000008103 glucose Substances 0.000 claims description 13
- 238000005119 centrifugation Methods 0.000 claims description 12
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 11
- BLCJBICVQSYOIF-UHFFFAOYSA-N 2,2-diaminobutanoic acid Chemical compound CCC(N)(N)C(O)=O BLCJBICVQSYOIF-UHFFFAOYSA-N 0.000 claims description 10
- 241001052560 Thallis Species 0.000 claims description 10
- 238000001976 enzyme digestion Methods 0.000 claims description 10
- 239000002609 medium Substances 0.000 claims description 10
- 239000011780 sodium chloride Substances 0.000 claims description 10
- 239000000872 buffer Substances 0.000 claims description 9
- 239000013612 plasmid Substances 0.000 claims description 8
- 239000008213 purified water Substances 0.000 claims description 8
- 108700005078 Synthetic Genes Proteins 0.000 claims description 7
- 239000000843 powder Substances 0.000 claims description 7
- 239000000243 solution Substances 0.000 claims description 7
- 230000002194 synthesizing effect Effects 0.000 claims description 7
- 229920001817 Agar Polymers 0.000 claims description 6
- 239000008272 agar Substances 0.000 claims description 6
- 239000008363 phosphate buffer Substances 0.000 claims description 6
- 230000035939 shock Effects 0.000 claims description 6
- 239000007787 solid Substances 0.000 claims description 6
- 230000009466 transformation Effects 0.000 claims description 6
- 238000001816 cooling Methods 0.000 claims description 5
- 238000000605 extraction Methods 0.000 claims description 5
- BPHPUYQFMNQIOC-NXRLNHOXSA-N isopropyl beta-D-thiogalactopyranoside Chemical compound CC(C)S[C@@H]1O[C@H](CO)[C@H](O)[C@H](O)[C@H]1O BPHPUYQFMNQIOC-NXRLNHOXSA-N 0.000 claims description 5
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 claims description 4
- 230000001939 inductive effect Effects 0.000 claims description 4
- 229910000402 monopotassium phosphate Inorganic materials 0.000 claims description 4
- 235000019796 monopotassium phosphate Nutrition 0.000 claims description 4
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 claims description 4
- 230000001954 sterilising effect Effects 0.000 claims description 4
- 239000000758 substrate Substances 0.000 claims description 4
- 230000001131 transforming effect Effects 0.000 claims description 4
- 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 description 3
- 238000001962 electrophoresis Methods 0.000 claims description 3
- 239000001963 growth medium Substances 0.000 claims description 3
- VHUUQVKOLVNVRT-UHFFFAOYSA-N Ammonium hydroxide Chemical compound [NH4+].[OH-] VHUUQVKOLVNVRT-UHFFFAOYSA-N 0.000 claims description 2
- 239000012880 LB liquid culture medium Substances 0.000 claims description 2
- 235000011114 ammonium hydroxide Nutrition 0.000 claims description 2
- 239000000499 gel Substances 0.000 claims description 2
- 125000003275 alpha amino acid group Chemical group 0.000 claims 3
- OGNSCSPNOLGXSM-VKHMYHEASA-O L-2,4-diazaniumylbutyrate Chemical compound [NH3+]CC[C@H]([NH3+])C([O-])=O OGNSCSPNOLGXSM-VKHMYHEASA-O 0.000 claims 1
- 230000029087 digestion Effects 0.000 claims 1
- 238000004321 preservation Methods 0.000 claims 1
- 241000545263 Salacia <hydroid> Species 0.000 abstract description 6
- 241000623228 Salinicola Species 0.000 abstract description 6
- 244000005700 microbiome Species 0.000 abstract description 3
- 238000006911 enzymatic reaction Methods 0.000 abstract description 2
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 abstract description 2
- 108020004414 DNA Proteins 0.000 description 20
- 229940088598 enzyme Drugs 0.000 description 15
- 241000206595 Halomonas elongata Species 0.000 description 9
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 9
- 150000001413 amino acids Chemical group 0.000 description 7
- 239000000523 sample Substances 0.000 description 7
- 238000006555 catalytic reaction Methods 0.000 description 6
- 239000008055 phosphate buffer solution Substances 0.000 description 6
- 239000000126 substance Substances 0.000 description 6
- WQXNXVUDBPYKBA-UHFFFAOYSA-N Ectoine Natural products CC1=NCCC(C(O)=O)N1 WQXNXVUDBPYKBA-UHFFFAOYSA-N 0.000 description 5
- 230000001580 bacterial effect Effects 0.000 description 5
- 230000036983 biotransformation Effects 0.000 description 5
- 239000000047 product Substances 0.000 description 5
- WQXNXVUDBPYKBA-YFKPBYRVSA-N ectoine Chemical compound CC1=[NH+][C@H](C([O-])=O)CCN1 WQXNXVUDBPYKBA-YFKPBYRVSA-N 0.000 description 4
- 238000009776 industrial production Methods 0.000 description 4
- 108010061238 threonyl-glycine Proteins 0.000 description 4
- YTSVAIMKVLZUDU-YUMQZZPRSA-N Gly-Leu-Asp Chemical compound [H]NCC(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(O)=O)C(O)=O YTSVAIMKVLZUDU-YUMQZZPRSA-N 0.000 description 3
- 241000206596 Halomonas Species 0.000 description 3
- 108010047495 alanylglycine Proteins 0.000 description 3
- 108010013835 arginine glutamate Proteins 0.000 description 3
- 238000001514 detection method Methods 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- 230000002503 metabolic effect Effects 0.000 description 3
- 239000002773 nucleotide Substances 0.000 description 3
- 125000003729 nucleotide group Chemical group 0.000 description 3
- 230000003204 osmotic effect Effects 0.000 description 3
- 108090000623 proteins and genes Proteins 0.000 description 3
- 239000013595 supernatant sample Substances 0.000 description 3
- DSPQRJXOIXHOHK-WDSKDSINSA-N Glu-Asp-Gly Chemical compound OC(=O)CC[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)NCC(O)=O DSPQRJXOIXHOHK-WDSKDSINSA-N 0.000 description 2
- APQYGMBHIVXFML-OSUNSFLBSA-N Ile-Val-Thr Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](C(C)C)C(=O)N[C@@H]([C@@H](C)O)C(=O)O)N APQYGMBHIVXFML-OSUNSFLBSA-N 0.000 description 2
- CQQGCWPXDHTTNF-GUBZILKMSA-N Leu-Ala-Glu Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@H](C(O)=O)CCC(O)=O CQQGCWPXDHTTNF-GUBZILKMSA-N 0.000 description 2
- RVVBWTWPNFDYBE-SRVKXCTJSA-N Leu-Glu-Arg Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O RVVBWTWPNFDYBE-SRVKXCTJSA-N 0.000 description 2
- 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 2
- 108010087924 alanylproline Proteins 0.000 description 2
- 108010010430 asparagine-proline-alanine Proteins 0.000 description 2
- 108010068265 aspartyltyrosine Proteins 0.000 description 2
- 239000006227 byproduct Substances 0.000 description 2
- 230000006378 damage Effects 0.000 description 2
- 239000000706 filtrate Substances 0.000 description 2
- 238000001914 filtration Methods 0.000 description 2
- 108010089804 glycyl-threonine Proteins 0.000 description 2
- 108010050848 glycylleucine Proteins 0.000 description 2
- 108010015792 glycyllysine Proteins 0.000 description 2
- 238000010438 heat treatment Methods 0.000 description 2
- 239000003906 humectant Substances 0.000 description 2
- 238000002347 injection Methods 0.000 description 2
- 239000007924 injection Substances 0.000 description 2
- 229930027917 kanamycin Natural products 0.000 description 2
- 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 2
- 229960000318 kanamycin Drugs 0.000 description 2
- 229930182823 kanamycin A Natural products 0.000 description 2
- 239000012528 membrane Substances 0.000 description 2
- 238000012269 metabolic engineering Methods 0.000 description 2
- 238000000926 separation method Methods 0.000 description 2
- 239000012086 standard solution Substances 0.000 description 2
- 238000003756 stirring Methods 0.000 description 2
- 230000001502 supplementing effect Effects 0.000 description 2
- UKAUYVFTDYCKQA-UHFFFAOYSA-N -2-Amino-4-hydroxybutanoic acid Natural products OC(=O)C(N)CCO UKAUYVFTDYCKQA-UHFFFAOYSA-N 0.000 description 1
- VBDMWOKJZDCFJM-FXQIFTODSA-N Ala-Ala-Met Chemical compound CSCC[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)[C@H](C)N VBDMWOKJZDCFJM-FXQIFTODSA-N 0.000 description 1
- SDMAQFGBPOJFOM-GUBZILKMSA-N Ala-Arg-Arg Chemical compound NC(=N)NCCC[C@H](NC(=O)[C@@H](N)C)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O SDMAQFGBPOJFOM-GUBZILKMSA-N 0.000 description 1
- SKHCUBQVZJHOFM-NAKRPEOUSA-N Ala-Arg-Ile Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O SKHCUBQVZJHOFM-NAKRPEOUSA-N 0.000 description 1
- NHCPCLJZRSIDHS-ZLUOBGJFSA-N Ala-Asp-Ala Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](C)C(O)=O NHCPCLJZRSIDHS-ZLUOBGJFSA-N 0.000 description 1
- PBAMJJXWDQXOJA-FXQIFTODSA-N Ala-Asp-Arg Chemical compound C[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@H](C(O)=O)CCCN=C(N)N PBAMJJXWDQXOJA-FXQIFTODSA-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
- LSLIRHLIUDVNBN-CIUDSAMLSA-N Ala-Asp-Lys Chemical compound C[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@H](C(O)=O)CCCCN LSLIRHLIUDVNBN-CIUDSAMLSA-N 0.000 description 1
- ZVFVBBGVOILKPO-WHFBIAKZSA-N Ala-Gly-Ala Chemical compound C[C@H](N)C(=O)NCC(=O)N[C@@H](C)C(O)=O ZVFVBBGVOILKPO-WHFBIAKZSA-N 0.000 description 1
- LMFXXZPPZDCPTA-ZKWXMUAHSA-N Ala-Gly-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)CNC(=O)[C@H](C)N LMFXXZPPZDCPTA-ZKWXMUAHSA-N 0.000 description 1
- FDAZDMAFZYTHGS-XVYDVKMFSA-N Ala-His-Asp Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CC(O)=O)C(O)=O FDAZDMAFZYTHGS-XVYDVKMFSA-N 0.000 description 1
- YHKANGMVQWRMAP-DCAQKATOSA-N Ala-Leu-Arg Chemical compound C[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@H](C(O)=O)CCCN=C(N)N YHKANGMVQWRMAP-DCAQKATOSA-N 0.000 description 1
- MEFILNJXAVSUTO-JXUBOQSCSA-N Ala-Leu-Thr Chemical compound C[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O MEFILNJXAVSUTO-JXUBOQSCSA-N 0.000 description 1
- MFMDKJIPHSWSBM-GUBZILKMSA-N Ala-Lys-Glu Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCC(O)=O)C(O)=O MFMDKJIPHSWSBM-GUBZILKMSA-N 0.000 description 1
- 108010011667 Ala-Phe-Ala Proteins 0.000 description 1
- XRUJOVRWNMBAAA-NHCYSSNCSA-N Ala-Phe-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@@H](NC(=O)[C@@H](N)C)CC1=CC=CC=C1 XRUJOVRWNMBAAA-NHCYSSNCSA-N 0.000 description 1
- CYBJZLQSUJEMAS-LFSVMHDDSA-N Ala-Phe-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CC1=CC=CC=C1)NC(=O)[C@H](C)N)O CYBJZLQSUJEMAS-LFSVMHDDSA-N 0.000 description 1
- PEEYDECOOVQKRZ-DLOVCJGASA-N Ala-Ser-Phe Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O PEEYDECOOVQKRZ-DLOVCJGASA-N 0.000 description 1
- UCDOXFBTMLKASE-HERUPUMHSA-N Ala-Ser-Trp Chemical compound C[C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CC1=CNC2=CC=CC=C21)C(=O)O)N UCDOXFBTMLKASE-HERUPUMHSA-N 0.000 description 1
- OEVCHROQUIVQFZ-YTLHQDLWSA-N Ala-Thr-Ala Chemical compound C[C@H](N)C(=O)N[C@@H]([C@H](O)C)C(=O)N[C@@H](C)C(O)=O OEVCHROQUIVQFZ-YTLHQDLWSA-N 0.000 description 1
- KUFVXLQLDHJVOG-SHGPDSBTSA-N Ala-Thr-Thr Chemical compound C[C@H]([C@@H](C(=O)N[C@@H]([C@@H](C)O)C(=O)O)NC(=O)[C@H](C)N)O KUFVXLQLDHJVOG-SHGPDSBTSA-N 0.000 description 1
- ZJLORAAXDAJLDC-CQDKDKBSSA-N Ala-Tyr-Leu Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC(C)C)C(O)=O ZJLORAAXDAJLDC-CQDKDKBSSA-N 0.000 description 1
- JAYIQMNQDMOBFY-KKUMJFAQSA-N Arg-Glu-Tyr Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O JAYIQMNQDMOBFY-KKUMJFAQSA-N 0.000 description 1
- HQIZDMIGUJOSNI-IUCAKERBSA-N Arg-Gly-Arg Chemical compound N[C@@H](CCCNC(N)=N)C(=O)NCC(=O)N[C@@H](CCCNC(N)=N)C(O)=O HQIZDMIGUJOSNI-IUCAKERBSA-N 0.000 description 1
- RFXXUWGNVRJTNQ-QXEWZRGKSA-N Arg-Gly-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)CNC(=O)[C@H](CCCN=C(N)N)N RFXXUWGNVRJTNQ-QXEWZRGKSA-N 0.000 description 1
- QEHMMRSQJMOYNO-DCAQKATOSA-N Arg-His-Asn Chemical compound C1=C(NC=N1)C[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)O)NC(=O)[C@H](CCCN=C(N)N)N QEHMMRSQJMOYNO-DCAQKATOSA-N 0.000 description 1
- UAOSDDXCTBIPCA-QXEWZRGKSA-N Arg-Ile-Gly Chemical compound CC[C@H](C)[C@@H](C(=O)NCC(=O)O)NC(=O)[C@H](CCCN=C(N)N)N UAOSDDXCTBIPCA-QXEWZRGKSA-N 0.000 description 1
- LLUGJARLJCGLAR-CYDGBPFRSA-N Arg-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](CCCN=C(N)N)N LLUGJARLJCGLAR-CYDGBPFRSA-N 0.000 description 1
- NOZYDJOPOGKUSR-AVGNSLFASA-N Arg-Leu-Met Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCSC)C(O)=O NOZYDJOPOGKUSR-AVGNSLFASA-N 0.000 description 1
- NGTYEHIRESTSRX-UWVGGRQHSA-N Arg-Lys-Gly Chemical compound NCCCC[C@@H](C(=O)NCC(O)=O)NC(=O)[C@@H](N)CCCN=C(N)N NGTYEHIRESTSRX-UWVGGRQHSA-N 0.000 description 1
- XSPKAHFVDKRGRL-DCAQKATOSA-N Arg-Pro-Glu Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CCC(O)=O)C(O)=O XSPKAHFVDKRGRL-DCAQKATOSA-N 0.000 description 1
- UZSQXCMNUPKLCC-FJXKBIBVSA-N Arg-Thr-Gly Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H]([C@@H](C)O)C(=O)NCC(O)=O UZSQXCMNUPKLCC-FJXKBIBVSA-N 0.000 description 1
- DXVMJJNAOVECBA-WHFBIAKZSA-N Asn-Gly-Asn Chemical compound NC(=O)C[C@H](N)C(=O)NCC(=O)N[C@@H](CC(N)=O)C(O)=O DXVMJJNAOVECBA-WHFBIAKZSA-N 0.000 description 1
- PBSQFBAJKPLRJY-BYULHYEWSA-N Asn-Gly-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)CNC(=O)[C@H](CC(=O)N)N PBSQFBAJKPLRJY-BYULHYEWSA-N 0.000 description 1
- RAQMSGVCGSJKCL-FOHZUACHSA-N Asn-Gly-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)CNC(=O)[C@@H](N)CC(N)=O RAQMSGVCGSJKCL-FOHZUACHSA-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
- PLTGTJAZQRGMPP-FXQIFTODSA-N Asn-Pro-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@@H]1CCCN1C(=O)[C@@H](N)CC(N)=O PLTGTJAZQRGMPP-FXQIFTODSA-N 0.000 description 1
- UGXYFDQFLVCDFC-CIUDSAMLSA-N Asn-Ser-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](N)CC(N)=O UGXYFDQFLVCDFC-CIUDSAMLSA-N 0.000 description 1
- YSYTWUMRHSFODC-QWRGUYRKSA-N Asn-Tyr-Gly Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)NCC(O)=O YSYTWUMRHSFODC-QWRGUYRKSA-N 0.000 description 1
- YBMUFUWSMIKJQA-GUBZILKMSA-N Asp-Gln-His Chemical compound C1=C(NC=N1)C[C@@H](C(=O)O)NC(=O)[C@H](CCC(=O)N)NC(=O)[C@H](CC(=O)O)N YBMUFUWSMIKJQA-GUBZILKMSA-N 0.000 description 1
- HRGGPWBIMIQANI-GUBZILKMSA-N Asp-Gln-Leu Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(C)C)C(O)=O HRGGPWBIMIQANI-GUBZILKMSA-N 0.000 description 1
- QCLHLXDWRKOHRR-GUBZILKMSA-N Asp-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](CC(=O)O)N QCLHLXDWRKOHRR-GUBZILKMSA-N 0.000 description 1
- YNCHFVRXEQFPBY-BQBZGAKWSA-N Asp-Gly-Arg Chemical compound OC(=O)C[C@H](N)C(=O)NCC(=O)N[C@H](C(O)=O)CCCN=C(N)N YNCHFVRXEQFPBY-BQBZGAKWSA-N 0.000 description 1
- WBDWQKRLTVCDSY-WHFBIAKZSA-N Asp-Gly-Asp Chemical compound OC(=O)C[C@H](N)C(=O)NCC(=O)N[C@@H](CC(O)=O)C(O)=O WBDWQKRLTVCDSY-WHFBIAKZSA-N 0.000 description 1
- BIVYLQMZPHDUIH-WHFBIAKZSA-N Asp-Gly-Cys Chemical compound C([C@@H](C(=O)NCC(=O)N[C@@H](CS)C(=O)O)N)C(=O)O BIVYLQMZPHDUIH-WHFBIAKZSA-N 0.000 description 1
- PGUYEUCYVNZGGV-QWRGUYRKSA-N Asp-Gly-Tyr Chemical compound OC(=O)C[C@H](N)C(=O)NCC(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 PGUYEUCYVNZGGV-QWRGUYRKSA-N 0.000 description 1
- RTXQQDVBACBSCW-CFMVVWHZSA-N Asp-Ile-Tyr Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O RTXQQDVBACBSCW-CFMVVWHZSA-N 0.000 description 1
- JNNVNVRBYUJYGS-CIUDSAMLSA-N Asp-Leu-Ala Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C)C(O)=O JNNVNVRBYUJYGS-CIUDSAMLSA-N 0.000 description 1
- LTCKTLYKRMCFOC-KKUMJFAQSA-N Asp-Phe-Leu Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC(C)C)C(O)=O LTCKTLYKRMCFOC-KKUMJFAQSA-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
- FIAKNCXQFFKSSI-ZLUOBGJFSA-N Asp-Ser-Cys Chemical compound OC(=O)C[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@@H](CS)C(O)=O FIAKNCXQFFKSSI-ZLUOBGJFSA-N 0.000 description 1
- YIDFBWRHIYOYAA-LKXGYXEUSA-N Asp-Ser-Thr Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)O)C(O)=O YIDFBWRHIYOYAA-LKXGYXEUSA-N 0.000 description 1
- GFYOIYJJMSHLSN-QXEWZRGKSA-N Asp-Val-Arg Chemical compound OC(=O)C[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCCN=C(N)N)C(O)=O GFYOIYJJMSHLSN-QXEWZRGKSA-N 0.000 description 1
- PLOKOIJSGCISHE-BYULHYEWSA-N Asp-Val-Asn Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(N)=O)C(O)=O PLOKOIJSGCISHE-BYULHYEWSA-N 0.000 description 1
- MFDPBZAFCRKYEY-LAEOZQHASA-N Asp-Val-Gln Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCC(N)=O)C(O)=O MFDPBZAFCRKYEY-LAEOZQHASA-N 0.000 description 1
- DQUWSUWXPWGTQT-DCAQKATOSA-N Cys-Pro-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@@H]1CCCN1C(=O)[C@@H](N)CS DQUWSUWXPWGTQT-DCAQKATOSA-N 0.000 description 1
- 230000005778 DNA damage Effects 0.000 description 1
- 231100000277 DNA damage Toxicity 0.000 description 1
- 108030003594 Diaminopimelate decarboxylases Proteins 0.000 description 1
- 241000660147 Escherichia coli str. K-12 substr. MG1655 Species 0.000 description 1
- 241001302584 Escherichia coli str. K-12 substr. W3110 Species 0.000 description 1
- WQWMZOIPXWSZNE-WDSKDSINSA-N Gln-Asp-Gly Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(O)=O)C(=O)NCC(O)=O WQWMZOIPXWSZNE-WDSKDSINSA-N 0.000 description 1
- MCAVASRGVBVPMX-FXQIFTODSA-N Gln-Glu-Ala Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C)C(O)=O MCAVASRGVBVPMX-FXQIFTODSA-N 0.000 description 1
- JEFZIKRIDLHOIF-BYPYZUCNSA-N Gln-Gly Chemical compound NC(=O)CC[C@H](N)C(=O)NCC(O)=O JEFZIKRIDLHOIF-BYPYZUCNSA-N 0.000 description 1
- VSXBYIJUAXPAAL-WDSKDSINSA-N Gln-Gly-Ala Chemical compound OC(=O)[C@H](C)NC(=O)CNC(=O)[C@@H](N)CCC(N)=O VSXBYIJUAXPAAL-WDSKDSINSA-N 0.000 description 1
- MFJAPSYJQJCQDN-BQBZGAKWSA-N Gln-Gly-Glu Chemical compound NC(=O)CC[C@H](N)C(=O)NCC(=O)N[C@@H](CCC(O)=O)C(O)=O MFJAPSYJQJCQDN-BQBZGAKWSA-N 0.000 description 1
- SFAFZYYMAWOCIC-KKUMJFAQSA-N Gln-Phe-Arg Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)N[C@@H](CCCN=C(N)N)C(=O)O)NC(=O)[C@H](CCC(=O)N)N SFAFZYYMAWOCIC-KKUMJFAQSA-N 0.000 description 1
- ZZLDMBMFKZFQMU-NRPADANISA-N Gln-Val-Ala Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](C)C(O)=O ZZLDMBMFKZFQMU-NRPADANISA-N 0.000 description 1
- SOEXCCGNHQBFPV-DLOVCJGASA-N Gln-Val-Val Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](C(C)C)C(O)=O SOEXCCGNHQBFPV-DLOVCJGASA-N 0.000 description 1
- LKDIBBOKUAASNP-FXQIFTODSA-N Glu-Ala-Glu Chemical compound OC(=O)CC[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@@H](CCC(O)=O)C(O)=O LKDIBBOKUAASNP-FXQIFTODSA-N 0.000 description 1
- FYBSCGZLICNOBA-XQXXSGGOSA-N Glu-Ala-Thr Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O FYBSCGZLICNOBA-XQXXSGGOSA-N 0.000 description 1
- CKRUHITYRFNUKW-WDSKDSINSA-N Glu-Asn-Gly Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(N)=O)C(=O)NCC(O)=O CKRUHITYRFNUKW-WDSKDSINSA-N 0.000 description 1
- JVSBYEDSSRZQGV-GUBZILKMSA-N Glu-Asp-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](CC(O)=O)NC(=O)[C@@H](N)CCC(O)=O JVSBYEDSSRZQGV-GUBZILKMSA-N 0.000 description 1
- ILGFBUGLBSAQQB-GUBZILKMSA-N Glu-Glu-Arg Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O ILGFBUGLBSAQQB-GUBZILKMSA-N 0.000 description 1
- OGNJZUXUTPQVBR-BQBZGAKWSA-N Glu-Gly-Glu Chemical compound OC(=O)CC[C@H](N)C(=O)NCC(=O)N[C@@H](CCC(O)=O)C(O)=O OGNJZUXUTPQVBR-BQBZGAKWSA-N 0.000 description 1
- PJBVXVBTTFZPHJ-GUBZILKMSA-N Glu-Leu-Asp Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)O)NC(=O)[C@H](CCC(=O)O)N PJBVXVBTTFZPHJ-GUBZILKMSA-N 0.000 description 1
- DDXZHOHEABQXSE-NKIYYHGXSA-N Glu-Thr-His Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC1=CN=CN1)C(=O)O)NC(=O)[C@H](CCC(=O)O)N)O DDXZHOHEABQXSE-NKIYYHGXSA-N 0.000 description 1
- RGJKYNUINKGPJN-RWRJDSDZSA-N Glu-Thr-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)[C@H]([C@@H](C)O)NC(=O)[C@H](CCC(=O)O)N RGJKYNUINKGPJN-RWRJDSDZSA-N 0.000 description 1
- HJTSRYLPAYGEEC-SIUGBPQLSA-N Glu-Tyr-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)[C@H](CC1=CC=C(C=C1)O)NC(=O)[C@H](CCC(=O)O)N HJTSRYLPAYGEEC-SIUGBPQLSA-N 0.000 description 1
- MLILEEIVMRUYBX-NHCYSSNCSA-N Glu-Val-Arg Chemical compound OC(=O)CC[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCCN=C(N)N)C(O)=O MLILEEIVMRUYBX-NHCYSSNCSA-N 0.000 description 1
- FGGKGJHCVMYGCD-UKJIMTQDSA-N Glu-Val-Ile Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O FGGKGJHCVMYGCD-UKJIMTQDSA-N 0.000 description 1
- VSVZIEVNUYDAFR-YUMQZZPRSA-N Gly-Ala-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)CN VSVZIEVNUYDAFR-YUMQZZPRSA-N 0.000 description 1
- OGCIHJPYKVSMTE-YUMQZZPRSA-N Gly-Arg-Glu Chemical compound [H]NCC(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(O)=O)C(O)=O OGCIHJPYKVSMTE-YUMQZZPRSA-N 0.000 description 1
- DWUKOTKSTDWGAE-BQBZGAKWSA-N Gly-Asn-Arg Chemical compound NCC(=O)N[C@@H](CC(N)=O)C(=O)N[C@H](C(O)=O)CCCN=C(N)N DWUKOTKSTDWGAE-BQBZGAKWSA-N 0.000 description 1
- ZRZILYKEJBMFHY-BQBZGAKWSA-N Gly-Asp-Met Chemical compound CSCC[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)O)NC(=O)CN ZRZILYKEJBMFHY-BQBZGAKWSA-N 0.000 description 1
- CEXINUGNTZFNRY-BYPYZUCNSA-N Gly-Cys-Gly Chemical compound [NH3+]CC(=O)N[C@@H](CS)C(=O)NCC([O-])=O CEXINUGNTZFNRY-BYPYZUCNSA-N 0.000 description 1
- JLJLBWDKDRYOPA-RYUDHWBXSA-N Gly-Gln-Tyr Chemical compound NCC(=O)N[C@@H](CCC(N)=O)C(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 JLJLBWDKDRYOPA-RYUDHWBXSA-N 0.000 description 1
- XTQFHTHIAKKCTM-YFKPBYRVSA-N Gly-Glu-Gly Chemical compound NCC(=O)N[C@@H](CCC(O)=O)C(=O)NCC(O)=O XTQFHTHIAKKCTM-YFKPBYRVSA-N 0.000 description 1
- JSNNHGHYGYMVCK-XVKPBYJWSA-N Gly-Glu-Val Chemical compound [H]NCC(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C(C)C)C(O)=O JSNNHGHYGYMVCK-XVKPBYJWSA-N 0.000 description 1
- GDOZQTNZPCUARW-YFKPBYRVSA-N Gly-Gly-Glu Chemical compound NCC(=O)NCC(=O)N[C@H](C(O)=O)CCC(O)=O GDOZQTNZPCUARW-YFKPBYRVSA-N 0.000 description 1
- XMPXVJIDADUOQB-RCOVLWMOSA-N Gly-Gly-Ile Chemical compound CC[C@H](C)[C@@H](C([O-])=O)NC(=O)CNC(=O)C[NH3+] XMPXVJIDADUOQB-RCOVLWMOSA-N 0.000 description 1
- BHPQOIPBLYJNAW-NGZCFLSTSA-N Gly-Ile-Pro Chemical compound CC[C@H](C)[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)CN BHPQOIPBLYJNAW-NGZCFLSTSA-N 0.000 description 1
- YSDLIYZLOTZZNP-UWVGGRQHSA-N Gly-Leu-Met Chemical compound CSCC[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)CN YSDLIYZLOTZZNP-UWVGGRQHSA-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
- UWQDKRIZSROAKS-FJXKBIBVSA-N Gly-Met-Thr Chemical compound [H]NCC(=O)N[C@@H](CCSC)C(=O)N[C@@H]([C@@H](C)O)C(O)=O UWQDKRIZSROAKS-FJXKBIBVSA-N 0.000 description 1
- DHNXGWVNLFPOMQ-KBPBESRZSA-N Gly-Phe-His Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)N[C@@H](CC2=CN=CN2)C(=O)O)NC(=O)CN DHNXGWVNLFPOMQ-KBPBESRZSA-N 0.000 description 1
- GGAPHLIUUTVYMX-QWRGUYRKSA-N Gly-Phe-Ser Chemical compound OC[C@@H](C([O-])=O)NC(=O)[C@@H](NC(=O)C[NH3+])CC1=CC=CC=C1 GGAPHLIUUTVYMX-QWRGUYRKSA-N 0.000 description 1
- GLACUWHUYFBSPJ-FJXKBIBVSA-N Gly-Pro-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)[C@@H]1CCCN1C(=O)CN GLACUWHUYFBSPJ-FJXKBIBVSA-N 0.000 description 1
- ZZWUYQXMIFTIIY-WEDXCCLWSA-N Gly-Thr-Leu Chemical compound [H]NCC(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(O)=O ZZWUYQXMIFTIIY-WEDXCCLWSA-N 0.000 description 1
- ZVXMEWXHFBYJPI-LSJOCFKGSA-N Gly-Val-Ile Chemical compound [H]NCC(=O)N[C@@H](C(C)C)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O ZVXMEWXHFBYJPI-LSJOCFKGSA-N 0.000 description 1
- AFMOTCMSEBITOE-YEPSODPASA-N Gly-Val-Thr Chemical compound NCC(=O)N[C@@H](C(C)C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O AFMOTCMSEBITOE-YEPSODPASA-N 0.000 description 1
- JBCLFWXMTIKCCB-UHFFFAOYSA-N H-Gly-Phe-OH Natural products NCC(=O)NC(C(O)=O)CC1=CC=CC=C1 JBCLFWXMTIKCCB-UHFFFAOYSA-N 0.000 description 1
- WMKXFMUJRCEGRP-SRVKXCTJSA-N His-Asn-His Chemical compound C1=C(NC=N1)C[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N[C@@H](CC2=CN=CN2)C(=O)O)N WMKXFMUJRCEGRP-SRVKXCTJSA-N 0.000 description 1
- SKOKHBGDXGTDDP-MELADBBJSA-N His-Leu-Pro Chemical compound CC(C)C[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CC2=CN=CN2)N SKOKHBGDXGTDDP-MELADBBJSA-N 0.000 description 1
- ZUELLZFHJUPFEC-PMVMPFDFSA-N His-Phe-Trp Chemical compound C([C@H](N)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(O)=O)C1=CN=CN1 ZUELLZFHJUPFEC-PMVMPFDFSA-N 0.000 description 1
- QCBYAHHNOHBXIH-UWVGGRQHSA-N His-Pro-Gly Chemical compound C([C@H](N)C(=O)N1[C@@H](CCC1)C(=O)NCC(O)=O)C1=CN=CN1 QCBYAHHNOHBXIH-UWVGGRQHSA-N 0.000 description 1
- FFYYUUWROYYKFY-IHRRRGAJSA-N His-Val-Leu Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(C)C)C(O)=O FFYYUUWROYYKFY-IHRRRGAJSA-N 0.000 description 1
- 241000282414 Homo sapiens Species 0.000 description 1
- 101150017040 I gene Proteins 0.000 description 1
- YOTNPRLPIPHQSB-XUXIUFHCSA-N Ile-Arg-Lys Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CCCCN)C(=O)O)N YOTNPRLPIPHQSB-XUXIUFHCSA-N 0.000 description 1
- SJIGTGZVQGLMGG-NAKRPEOUSA-N Ile-Cys-Arg Chemical compound N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CS)C(=O)N[C@@H](CCCNC(N)=N)C(=O)O SJIGTGZVQGLMGG-NAKRPEOUSA-N 0.000 description 1
- ZDNORQNHCJUVOV-KBIXCLLPSA-N Ile-Gln-Ala Chemical compound CC[C@H](C)[C@H](N)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](C)C(O)=O ZDNORQNHCJUVOV-KBIXCLLPSA-N 0.000 description 1
- WZDCVAWMBUNDDY-KBIXCLLPSA-N Ile-Glu-Ala Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)N[C@@H](C)C(=O)O)N WZDCVAWMBUNDDY-KBIXCLLPSA-N 0.000 description 1
- AREBLHSMLMRICD-PYJNHQTQSA-N Ile-His-Arg Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC1=CN=CN1)C(=O)N[C@@H](CCCN=C(N)N)C(=O)O)N AREBLHSMLMRICD-PYJNHQTQSA-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
- UIEZQYNXCYHMQS-BJDJZHNGSA-N Ile-Lys-Ala Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C)C(=O)O)N UIEZQYNXCYHMQS-BJDJZHNGSA-N 0.000 description 1
- JZNVOBUNTWNZPW-GHCJXIJMSA-N Ile-Ser-Asp Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(=O)O)C(=O)O)N JZNVOBUNTWNZPW-GHCJXIJMSA-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
- WCNWGAUZWWSYDG-SVSWQMSJSA-N Ile-Thr-Ser Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CO)C(=O)O)N WCNWGAUZWWSYDG-SVSWQMSJSA-N 0.000 description 1
- WIYDLTIBHZSPKY-HJWJTTGWSA-N Ile-Val-Phe Chemical compound CC[C@H](C)[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 WIYDLTIBHZSPKY-HJWJTTGWSA-N 0.000 description 1
- FADYJNXDPBKVCA-UHFFFAOYSA-N L-Phenylalanyl-L-lysin Natural products NCCCCC(C(O)=O)NC(=O)C(N)CC1=CC=CC=C1 FADYJNXDPBKVCA-UHFFFAOYSA-N 0.000 description 1
- SITWEMZOJNKJCH-UHFFFAOYSA-N L-alanine-L-arginine Natural products CC(N)C(=O)NC(C(O)=O)CCCNC(N)=N SITWEMZOJNKJCH-UHFFFAOYSA-N 0.000 description 1
- UKAUYVFTDYCKQA-VKHMYHEASA-N L-homoserine Chemical compound OC(=O)[C@@H](N)CCO UKAUYVFTDYCKQA-VKHMYHEASA-N 0.000 description 1
- RCFDOSNHHZGBOY-UHFFFAOYSA-N L-isoleucyl-L-alanine Natural products CCC(C)C(N)C(=O)NC(C)C(O)=O RCFDOSNHHZGBOY-UHFFFAOYSA-N 0.000 description 1
- GUBGYTABKSRVRQ-QKKXKWKRSA-N Lactose Natural products OC[C@H]1O[C@@H](O[C@H]2[C@H](O)[C@@H](O)C(O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@H]1O GUBGYTABKSRVRQ-QKKXKWKRSA-N 0.000 description 1
- XBBKIIGCUMBKCO-JXUBOQSCSA-N Leu-Ala-Thr Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O XBBKIIGCUMBKCO-JXUBOQSCSA-N 0.000 description 1
- KSZCCRIGNVSHFH-UWVGGRQHSA-N Leu-Arg-Gly Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)NCC(O)=O KSZCCRIGNVSHFH-UWVGGRQHSA-N 0.000 description 1
- VIWUBXKCYJGNCL-SRVKXCTJSA-N Leu-Asn-His Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@H](C(O)=O)CC1=CN=CN1 VIWUBXKCYJGNCL-SRVKXCTJSA-N 0.000 description 1
- KTFHTMHHKXUYPW-ZPFDUUQYSA-N Leu-Asp-Ile Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O KTFHTMHHKXUYPW-ZPFDUUQYSA-N 0.000 description 1
- XVSJMWYYLHPDKY-DCAQKATOSA-N Leu-Asp-Met Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCSC)C(O)=O XVSJMWYYLHPDKY-DCAQKATOSA-N 0.000 description 1
- QLQHWWCSCLZUMA-KKUMJFAQSA-N Leu-Asp-Tyr Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 QLQHWWCSCLZUMA-KKUMJFAQSA-N 0.000 description 1
- ZFNLIDNJUWNIJL-WDCWCFNPSA-N Leu-Glu-Thr Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O ZFNLIDNJUWNIJL-WDCWCFNPSA-N 0.000 description 1
- SEMUSFOBZGKBGW-YTFOTSKYSA-N Leu-Ile-Ile Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O SEMUSFOBZGKBGW-YTFOTSKYSA-N 0.000 description 1
- HRTRLSRYZZKPCO-BJDJZHNGSA-N Leu-Ile-Ser Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CO)C(O)=O HRTRLSRYZZKPCO-BJDJZHNGSA-N 0.000 description 1
- FAELBUXXFQLUAX-AJNGGQMLSA-N Leu-Leu-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](N)CC(C)C FAELBUXXFQLUAX-AJNGGQMLSA-N 0.000 description 1
- RXGLHDWAZQECBI-SRVKXCTJSA-N Leu-Leu-Ser Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(O)=O RXGLHDWAZQECBI-SRVKXCTJSA-N 0.000 description 1
- XXXXOVFBXRERQL-ULQDDVLXSA-N Leu-Pro-Phe Chemical compound CC(C)C[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 XXXXOVFBXRERQL-ULQDDVLXSA-N 0.000 description 1
- ZJZNLRVCZWUONM-JXUBOQSCSA-N Leu-Thr-Ala Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C)C(O)=O ZJZNLRVCZWUONM-JXUBOQSCSA-N 0.000 description 1
- LFSQWRSVPNKJGP-WDCWCFNPSA-N Leu-Thr-Glu Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@H](C(O)=O)CCC(O)=O LFSQWRSVPNKJGP-WDCWCFNPSA-N 0.000 description 1
- FBNPMTNBFFAMMH-UHFFFAOYSA-N Leu-Val-Arg Natural products CC(C)CC(N)C(=O)NC(C(C)C)C(=O)NC(C(O)=O)CCCN=C(N)N FBNPMTNBFFAMMH-UHFFFAOYSA-N 0.000 description 1
- AIMGJYMCTAABEN-GVXVVHGQSA-N Leu-Val-Glu Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCC(O)=O)C(O)=O AIMGJYMCTAABEN-GVXVVHGQSA-N 0.000 description 1
- RVOMPSJXSRPFJT-DCAQKATOSA-N Lys-Ala-Arg Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](C)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O RVOMPSJXSRPFJT-DCAQKATOSA-N 0.000 description 1
- VHFFQUSNFFIZBT-CIUDSAMLSA-N Lys-Ala-Cys Chemical compound C[C@@H](C(=O)N[C@@H](CS)C(=O)O)NC(=O)[C@H](CCCCN)N VHFFQUSNFFIZBT-CIUDSAMLSA-N 0.000 description 1
- YIBOAHAOAWACDK-QEJZJMRPSA-N Lys-Ala-Phe Chemical compound NCCCC[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 YIBOAHAOAWACDK-QEJZJMRPSA-N 0.000 description 1
- GQUDMNDPQTXZRV-DCAQKATOSA-N Lys-Arg-Asp Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(O)=O)C(O)=O GQUDMNDPQTXZRV-DCAQKATOSA-N 0.000 description 1
- YNNPKXBBRZVIRX-IHRRRGAJSA-N Lys-Arg-Leu Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(C)C)C(O)=O YNNPKXBBRZVIRX-IHRRRGAJSA-N 0.000 description 1
- MKBIVWXCFINCLE-SRVKXCTJSA-N Lys-Asn-His Chemical compound C1=C(NC=N1)C[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)N)NC(=O)[C@H](CCCCN)N MKBIVWXCFINCLE-SRVKXCTJSA-N 0.000 description 1
- SSYOBDBNBQBSQE-SRVKXCTJSA-N Lys-Cys-Leu Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CS)C(=O)N[C@@H](CC(C)C)C(O)=O SSYOBDBNBQBSQE-SRVKXCTJSA-N 0.000 description 1
- IMAKMJCBYCSMHM-AVGNSLFASA-N Lys-Glu-Lys Chemical compound NCCCC[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@H](C(O)=O)CCCCN IMAKMJCBYCSMHM-AVGNSLFASA-N 0.000 description 1
- YWJQHDDBFAXNIR-MXAVVETBSA-N Lys-Ile-His Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC1=CN=CN1)C(=O)O)NC(=O)[C@H](CCCCN)N YWJQHDDBFAXNIR-MXAVVETBSA-N 0.000 description 1
- XFOAWKDQMRMCDN-ULQDDVLXSA-N Lys-Phe-Arg Chemical compound NC(N)=NCCC[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)[C@@H](N)CCCCN)CC1=CC=CC=C1 XFOAWKDQMRMCDN-ULQDDVLXSA-N 0.000 description 1
- IPTUBUUIFRZMJK-ACRUOGEOSA-N Lys-Phe-Phe Chemical compound C([C@H](NC(=O)[C@@H](N)CCCCN)C(=O)N[C@@H](CC=1C=CC=CC=1)C(O)=O)C1=CC=CC=C1 IPTUBUUIFRZMJK-ACRUOGEOSA-N 0.000 description 1
- AFLBTVGQCQLOFJ-AVGNSLFASA-N Lys-Pro-Arg Chemical compound NCCCC[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CCCN=C(N)N)C(O)=O AFLBTVGQCQLOFJ-AVGNSLFASA-N 0.000 description 1
- WGILOYIKJVQUPT-DCAQKATOSA-N Lys-Pro-Asp Chemical compound [H]N[C@@H](CCCCN)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CC(O)=O)C(O)=O WGILOYIKJVQUPT-DCAQKATOSA-N 0.000 description 1
- SBQDRNOLGSYHQA-YUMQZZPRSA-N Lys-Ser-Gly Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CO)C(=O)NCC(O)=O SBQDRNOLGSYHQA-YUMQZZPRSA-N 0.000 description 1
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 description 1
- 239000004472 Lysine Substances 0.000 description 1
- ZAJNRWKGHWGPDQ-SDDRHHMPSA-N Met-Arg-Pro Chemical compound CSCC[C@@H](C(=O)N[C@@H](CCCN=C(N)N)C(=O)N1CCC[C@@H]1C(=O)O)N ZAJNRWKGHWGPDQ-SDDRHHMPSA-N 0.000 description 1
- HLQWFLJOJRFXHO-CIUDSAMLSA-N Met-Glu-Ser Chemical compound [H]N[C@@H](CCSC)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CO)C(O)=O HLQWFLJOJRFXHO-CIUDSAMLSA-N 0.000 description 1
- BCRQJDMZQUHQSV-STQMWFEESA-N Met-Gly-Tyr Chemical compound [H]N[C@@H](CCSC)C(=O)NCC(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O BCRQJDMZQUHQSV-STQMWFEESA-N 0.000 description 1
- WXUUEPIDLLQBLJ-DCAQKATOSA-N Met-Met-Gln Chemical compound CSCC[C@@H](C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CCC(=O)N)C(=O)O)N WXUUEPIDLLQBLJ-DCAQKATOSA-N 0.000 description 1
- YLDSJJOGQNEQJK-AVGNSLFASA-N Met-Pro-Leu Chemical compound CSCC[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CC(C)C)C(O)=O YLDSJJOGQNEQJK-AVGNSLFASA-N 0.000 description 1
- RIIFMEBFDDXGCV-VEVYYDQMSA-N Met-Thr-Asn Chemical compound CSCC[C@H](N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@H](C(O)=O)CC(N)=O RIIFMEBFDDXGCV-VEVYYDQMSA-N 0.000 description 1
- YBAFDPFAUTYYRW-UHFFFAOYSA-N N-L-alpha-glutamyl-L-leucine Natural products CC(C)CC(C(O)=O)NC(=O)C(N)CCC(O)=O YBAFDPFAUTYYRW-UHFFFAOYSA-N 0.000 description 1
- PESQCPHRXOFIPX-UHFFFAOYSA-N N-L-methionyl-L-tyrosine Natural products CSCCC(N)C(=O)NC(C(O)=O)CC1=CC=C(O)C=C1 PESQCPHRXOFIPX-UHFFFAOYSA-N 0.000 description 1
- KZNQNBZMBZJQJO-UHFFFAOYSA-N N-glycyl-L-proline Natural products NCC(=O)N1CCCC1C(O)=O KZNQNBZMBZJQJO-UHFFFAOYSA-N 0.000 description 1
- AJHCSUXXECOXOY-UHFFFAOYSA-N N-glycyl-L-tryptophan Natural products C1=CC=C2C(CC(NC(=O)CN)C(O)=O)=CNC2=C1 AJHCSUXXECOXOY-UHFFFAOYSA-N 0.000 description 1
- CGOMLCQJEMWMCE-STQMWFEESA-N Phe-Arg-Gly Chemical compound NC(N)=NCCC[C@@H](C(=O)NCC(O)=O)NC(=O)[C@@H](N)CC1=CC=CC=C1 CGOMLCQJEMWMCE-STQMWFEESA-N 0.000 description 1
- KIEPQOIQHFKQLK-PCBIJLKTSA-N Phe-Asn-Ile Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O KIEPQOIQHFKQLK-PCBIJLKTSA-N 0.000 description 1
- LNIIRLODKOWQIY-IHRRRGAJSA-N Phe-Asn-Met Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CCSC)C(O)=O LNIIRLODKOWQIY-IHRRRGAJSA-N 0.000 description 1
- HOYQLNNGMHXZDW-KKUMJFAQSA-N Phe-Glu-Arg Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O HOYQLNNGMHXZDW-KKUMJFAQSA-N 0.000 description 1
- PSKRILMFHNIUAO-JYJNAYRXSA-N Phe-Glu-Lys Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)N[C@@H](CCCCN)C(=O)O)N PSKRILMFHNIUAO-JYJNAYRXSA-N 0.000 description 1
- HQPWNHXERZCIHP-PMVMPFDFSA-N Phe-Leu-Trp Chemical compound C([C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(O)=O)C1=CC=CC=C1 HQPWNHXERZCIHP-PMVMPFDFSA-N 0.000 description 1
- IAOZOFPONWDXNT-IXOXFDKPSA-N Phe-Ser-Thr Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)O)C(O)=O IAOZOFPONWDXNT-IXOXFDKPSA-N 0.000 description 1
- JHSRGEODDALISP-XVSYOHENSA-N Phe-Thr-Asn Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(N)=O)C(O)=O JHSRGEODDALISP-XVSYOHENSA-N 0.000 description 1
- MWQXFDIQXIXPMS-UNQGMJICSA-N Phe-Val-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](C(C)C)NC(=O)[C@H](CC1=CC=CC=C1)N)O MWQXFDIQXIXPMS-UNQGMJICSA-N 0.000 description 1
- XQLBWXHVZVBNJM-FXQIFTODSA-N Pro-Ala-Ser Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)[C@@H]1CCCN1 XQLBWXHVZVBNJM-FXQIFTODSA-N 0.000 description 1
- TXPUNZXZDVJUJQ-LPEHRKFASA-N Pro-Asn-Pro Chemical compound C1C[C@H](NC1)C(=O)N[C@@H](CC(=O)N)C(=O)N2CCC[C@@H]2C(=O)O TXPUNZXZDVJUJQ-LPEHRKFASA-N 0.000 description 1
- CJZTUKSFZUSNCC-FXQIFTODSA-N Pro-Asp-Asn Chemical compound NC(=O)C[C@@H](C(O)=O)NC(=O)[C@H](CC(O)=O)NC(=O)[C@@H]1CCCN1 CJZTUKSFZUSNCC-FXQIFTODSA-N 0.000 description 1
- LSIWVWRUTKPXDS-DCAQKATOSA-N Pro-Gln-Arg Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O LSIWVWRUTKPXDS-DCAQKATOSA-N 0.000 description 1
- RMODQFBNDDENCP-IHRRRGAJSA-N Pro-Lys-Leu Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(O)=O RMODQFBNDDENCP-IHRRRGAJSA-N 0.000 description 1
- ZUZINZIJHJFJRN-UBHSHLNASA-N Pro-Phe-Ala Chemical compound C([C@@H](C(=O)N[C@@H](C)C(O)=O)NC(=O)[C@H]1NCCC1)C1=CC=CC=C1 ZUZINZIJHJFJRN-UBHSHLNASA-N 0.000 description 1
- AWQGDZBKQTYNMN-IHRRRGAJSA-N Pro-Phe-Asp Chemical compound C1C[C@H](NC1)C(=O)N[C@@H](CC2=CC=CC=C2)C(=O)N[C@@H](CC(=O)O)C(=O)O AWQGDZBKQTYNMN-IHRRRGAJSA-N 0.000 description 1
- CGSOWZUPLOKYOR-AVGNSLFASA-N Pro-Pro-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@@H]1CCCN1C(=O)[C@H]1NCCC1 CGSOWZUPLOKYOR-AVGNSLFASA-N 0.000 description 1
- POQFNPILEQEODH-FXQIFTODSA-N Pro-Ser-Ala Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CO)C(=O)N[C@@H](C)C(O)=O POQFNPILEQEODH-FXQIFTODSA-N 0.000 description 1
- GZNYIXWOIUFLGO-ZJDVBMNYSA-N Pro-Thr-Thr Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O GZNYIXWOIUFLGO-ZJDVBMNYSA-N 0.000 description 1
- CXGLFEOYCJFKPR-RCWTZXSCSA-N Pro-Thr-Val Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C(C)C)C(O)=O CXGLFEOYCJFKPR-RCWTZXSCSA-N 0.000 description 1
- 108010079005 RDV peptide Proteins 0.000 description 1
- MESDJCNHLZBMEP-ZLUOBGJFSA-N Ser-Asp-Asp Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(O)=O)C(O)=O MESDJCNHLZBMEP-ZLUOBGJFSA-N 0.000 description 1
- OQPNSDWGAMFJNU-QWRGUYRKSA-N Ser-Gly-Tyr Chemical compound OC[C@H](N)C(=O)NCC(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 OQPNSDWGAMFJNU-QWRGUYRKSA-N 0.000 description 1
- YUJLIIRMIAGMCQ-CIUDSAMLSA-N Ser-Leu-Ser Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(O)=O YUJLIIRMIAGMCQ-CIUDSAMLSA-N 0.000 description 1
- BUYHXYIUQUBEQP-AVGNSLFASA-N Ser-Phe-Glu Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)O)NC(=O)[C@H](CO)N BUYHXYIUQUBEQP-AVGNSLFASA-N 0.000 description 1
- RWDVVSKYZBNDCO-MELADBBJSA-N Ser-Phe-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CC2=CC=CC=C2)NC(=O)[C@H](CO)N)C(=O)O RWDVVSKYZBNDCO-MELADBBJSA-N 0.000 description 1
- PIQRHJQWEPWFJG-UWJYBYFXSA-N Ser-Tyr-Ala Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](C)C(O)=O PIQRHJQWEPWFJG-UWJYBYFXSA-N 0.000 description 1
- IGROJMCBGRFRGI-YTLHQDLWSA-N Thr-Ala-Ala Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@@H](C)C(O)=O IGROJMCBGRFRGI-YTLHQDLWSA-N 0.000 description 1
- TYVAWPFQYFPSBR-BFHQHQDPSA-N Thr-Ala-Gly Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C)C(=O)NCC(O)=O TYVAWPFQYFPSBR-BFHQHQDPSA-N 0.000 description 1
- MQBTXMPQNCGSSZ-OSUNSFLBSA-N Thr-Arg-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)[C@@H](N)[C@@H](C)O)CCCN=C(N)N MQBTXMPQNCGSSZ-OSUNSFLBSA-N 0.000 description 1
- XDARBNMYXKUFOJ-GSSVUCPTSA-N Thr-Asp-Thr Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O XDARBNMYXKUFOJ-GSSVUCPTSA-N 0.000 description 1
- UHBPFYOQQPFKQR-JHEQGTHGSA-N Thr-Gln-Gly Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCC(N)=O)C(=O)NCC(O)=O UHBPFYOQQPFKQR-JHEQGTHGSA-N 0.000 description 1
- GARULAKWZGFIKC-RWRJDSDZSA-N Thr-Gln-Ile Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O GARULAKWZGFIKC-RWRJDSDZSA-N 0.000 description 1
- LGNBRHZANHMZHK-NUMRIWBASA-N Thr-Glu-Asp Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)N[C@@H](CC(=O)O)C(=O)O)N)O LGNBRHZANHMZHK-NUMRIWBASA-N 0.000 description 1
- QQWNRERCGGZOKG-WEDXCCLWSA-N Thr-Gly-Leu Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)NCC(=O)N[C@@H](CC(C)C)C(O)=O QQWNRERCGGZOKG-WEDXCCLWSA-N 0.000 description 1
- GXUWHVZYDAHFSV-FLBSBUHZSA-N Thr-Ile-Thr Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H]([C@@H](C)O)C(O)=O GXUWHVZYDAHFSV-FLBSBUHZSA-N 0.000 description 1
- VTVVYQOXJCZVEB-WDCWCFNPSA-N Thr-Leu-Glu Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(O)=O VTVVYQOXJCZVEB-WDCWCFNPSA-N 0.000 description 1
- KRDSCBLRHORMRK-JXUBOQSCSA-N Thr-Lys-Ala Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C)C(O)=O KRDSCBLRHORMRK-JXUBOQSCSA-N 0.000 description 1
- CZSMNLQMRWPGQF-XEGUGMAKSA-N Trp-Gln-Ala Chemical compound [H]N[C@@H](CC1=CNC2=C1C=CC=C2)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](C)C(O)=O CZSMNLQMRWPGQF-XEGUGMAKSA-N 0.000 description 1
- DNUJCLUFRGGSDJ-YLVFBTJISA-N Trp-Gly-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)CNC(=O)[C@H](CC1=CNC2=CC=CC=C21)N DNUJCLUFRGGSDJ-YLVFBTJISA-N 0.000 description 1
- RRVUOLRWIZXBRQ-IHPCNDPISA-N Trp-Leu-Lys Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CCCCN)C(=O)O)NC(=O)[C@H](CC1=CNC2=CC=CC=C21)N RRVUOLRWIZXBRQ-IHPCNDPISA-N 0.000 description 1
- VDUJEEQMRQCLHB-YTQUADARSA-N Trp-Lys-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CCCCN)NC(=O)[C@H](CC2=CNC3=CC=CC=C32)N)C(=O)O VDUJEEQMRQCLHB-YTQUADARSA-N 0.000 description 1
- NZFCWALTLNFHHC-JYJNAYRXSA-N Tyr-Glu-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@@H](N)CC1=CC=C(O)C=C1 NZFCWALTLNFHHC-JYJNAYRXSA-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
- KSCVLGXNQXKUAR-JYJNAYRXSA-N Tyr-Leu-Glu Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(O)=O KSCVLGXNQXKUAR-JYJNAYRXSA-N 0.000 description 1
- KHCSOLAHNLOXJR-BZSNNMDCSA-N Tyr-Leu-Leu Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(C)C)C(O)=O KHCSOLAHNLOXJR-BZSNNMDCSA-N 0.000 description 1
- PMHLLBKTDHQMCY-ULQDDVLXSA-N Tyr-Lys-Val Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C(C)C)C(O)=O PMHLLBKTDHQMCY-ULQDDVLXSA-N 0.000 description 1
- RWOKVQUCENPXGE-IHRRRGAJSA-N Tyr-Ser-Arg Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O RWOKVQUCENPXGE-IHRRRGAJSA-N 0.000 description 1
- WOCYUGQDXPTQPY-FXQIFTODSA-N Val-Ala-Cys Chemical compound C[C@@H](C(=O)N[C@@H](CS)C(=O)O)NC(=O)[C@H](C(C)C)N WOCYUGQDXPTQPY-FXQIFTODSA-N 0.000 description 1
- SLLKXDSRVAOREO-KZVJFYERSA-N Val-Ala-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](C)NC(=O)[C@H](C(C)C)N)O SLLKXDSRVAOREO-KZVJFYERSA-N 0.000 description 1
- HZYOWMGWKKRMBZ-BYULHYEWSA-N Val-Asp-Asp Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)N[C@@H](CC(=O)O)C(=O)O)N HZYOWMGWKKRMBZ-BYULHYEWSA-N 0.000 description 1
- XWYUBUYQMOUFRQ-IFFSRLJSSA-N Val-Glu-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CCC(=O)O)NC(=O)[C@H](C(C)C)N)O XWYUBUYQMOUFRQ-IFFSRLJSSA-N 0.000 description 1
- PMXBARDFIAPBGK-DZKIICNBSA-N Val-Glu-Tyr Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 PMXBARDFIAPBGK-DZKIICNBSA-N 0.000 description 1
- WFENBJPLZMPVAX-XVKPBYJWSA-N Val-Gly-Glu Chemical compound CC(C)[C@H](N)C(=O)NCC(=O)N[C@H](C(O)=O)CCC(O)=O WFENBJPLZMPVAX-XVKPBYJWSA-N 0.000 description 1
- PTFPUAXGIKTVNN-ONGXEEELSA-N Val-His-Gly Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC1=CN=CN1)C(=O)NCC(=O)O)N PTFPUAXGIKTVNN-ONGXEEELSA-N 0.000 description 1
- SYSWVVCYSXBVJG-RHYQMDGZSA-N Val-Leu-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](C(C)C)N)O SYSWVVCYSXBVJG-RHYQMDGZSA-N 0.000 description 1
- YQMILNREHKTFBS-IHRRRGAJSA-N Val-Phe-Cys Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CS)C(=O)O)N YQMILNREHKTFBS-IHRRRGAJSA-N 0.000 description 1
- KISFXYYRKKNLOP-IHRRRGAJSA-N Val-Phe-Ser Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CO)C(=O)O)N KISFXYYRKKNLOP-IHRRRGAJSA-N 0.000 description 1
- XBJKAZATRJBDCU-GUBZILKMSA-N Val-Pro-Ala Chemical compound CC(C)[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](C)C(O)=O XBJKAZATRJBDCU-GUBZILKMSA-N 0.000 description 1
- ZXYPHBKIZLAQTL-QXEWZRGKSA-N Val-Pro-Asp Chemical compound CC(C)[C@@H](C(=O)N1CCC[C@H]1C(=O)N[C@@H](CC(=O)O)C(=O)O)N ZXYPHBKIZLAQTL-QXEWZRGKSA-N 0.000 description 1
- AEFJNECXZCODJM-UWVGGRQHSA-N Val-Val-Gly Chemical compound CC(C)[C@H]([NH3+])C(=O)N[C@@H](C(C)C)C(=O)NCC([O-])=O AEFJNECXZCODJM-UWVGGRQHSA-N 0.000 description 1
- 108010084217 alanyl-glutamyl-aspartyl-glycine Proteins 0.000 description 1
- 108010041407 alanylaspartic acid Proteins 0.000 description 1
- 108010005233 alanylglutamic acid Proteins 0.000 description 1
- 108010044940 alanylglutamine Proteins 0.000 description 1
- 108010070944 alanylhistidine Proteins 0.000 description 1
- KOSRFJWDECSPRO-UHFFFAOYSA-N alpha-L-glutamyl-L-glutamic acid Natural products OC(=O)CCC(N)C(=O)NC(CCC(O)=O)C(O)=O KOSRFJWDECSPRO-UHFFFAOYSA-N 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- PYMYPHUHKUWMLA-WDCZJNDASA-N arabinose Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)C=O PYMYPHUHKUWMLA-WDCZJNDASA-N 0.000 description 1
- PYMYPHUHKUWMLA-UHFFFAOYSA-N arabinose Natural products OCC(O)C(O)C(O)C=O PYMYPHUHKUWMLA-UHFFFAOYSA-N 0.000 description 1
- 108010080488 arginyl-arginyl-leucine Proteins 0.000 description 1
- 108010062796 arginyllysine Proteins 0.000 description 1
- 108010077245 asparaginyl-proline Proteins 0.000 description 1
- 108010093581 aspartyl-proline Proteins 0.000 description 1
- 108010038633 aspartylglutamate Proteins 0.000 description 1
- 108010092854 aspartyllysine Proteins 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- SRBFZHDQGSBBOR-UHFFFAOYSA-N beta-D-Pyranose-Lyxose Natural products OC1COC(O)C(O)C1O SRBFZHDQGSBBOR-UHFFFAOYSA-N 0.000 description 1
- 239000003124 biologic agent Substances 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- 238000007036 catalytic synthesis reaction Methods 0.000 description 1
- 108091092356 cellular DNA Proteins 0.000 description 1
- 238000012824 chemical production Methods 0.000 description 1
- 239000012295 chemical reaction liquid Substances 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 238000010367 cloning Methods 0.000 description 1
- 230000001447 compensatory effect Effects 0.000 description 1
- 230000000536 complexating effect Effects 0.000 description 1
- 239000002537 cosmetic Substances 0.000 description 1
- 239000013078 crystal Substances 0.000 description 1
- 108010016616 cysteinylglycine Proteins 0.000 description 1
- 108010060199 cysteinylproline Proteins 0.000 description 1
- 108010069495 cysteinyltyrosine Proteins 0.000 description 1
- 229940079919 digestives enzyme preparation Drugs 0.000 description 1
- 238000005265 energy consumption Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 239000013604 expression vector Substances 0.000 description 1
- 238000007710 freezing Methods 0.000 description 1
- 230000008014 freezing Effects 0.000 description 1
- 108010006664 gamma-glutamyl-glycyl-glycine Proteins 0.000 description 1
- 239000003292 glue Substances 0.000 description 1
- 108010078144 glutaminyl-glycine Proteins 0.000 description 1
- 108010042598 glutamyl-aspartyl-glycine Proteins 0.000 description 1
- 108010055341 glutamyl-glutamic acid Proteins 0.000 description 1
- 108010049041 glutamylalanine Proteins 0.000 description 1
- 108010079547 glutamylmethionine Proteins 0.000 description 1
- VPZXBVLAVMBEQI-UHFFFAOYSA-N glycyl-DL-alpha-alanine Natural products OC(=O)C(C)NC(=O)CN VPZXBVLAVMBEQI-UHFFFAOYSA-N 0.000 description 1
- 108010000434 glycyl-alanyl-leucine Proteins 0.000 description 1
- 108010026364 glycyl-glycyl-leucine Proteins 0.000 description 1
- 108010048994 glycyl-tyrosyl-alanine Proteins 0.000 description 1
- 108010020688 glycylhistidine Proteins 0.000 description 1
- 108010081551 glycylphenylalanine Proteins 0.000 description 1
- 108010077515 glycylproline Proteins 0.000 description 1
- -1 heterocyclic amino acid derivative Chemical class 0.000 description 1
- 108010036413 histidylglycine Proteins 0.000 description 1
- 108010025306 histidylleucine Proteins 0.000 description 1
- 108010085325 histidylproline Proteins 0.000 description 1
- 230000002779 inactivation Effects 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 230000006698 induction Effects 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 239000000543 intermediate Substances 0.000 description 1
- 230000002427 irreversible effect Effects 0.000 description 1
- 108010044374 isoleucyl-tyrosine Proteins 0.000 description 1
- 239000008101 lactose Substances 0.000 description 1
- 108010044311 leucyl-glycyl-glycine Proteins 0.000 description 1
- 101150033534 lysA gene Proteins 0.000 description 1
- 108010064235 lysylglycine Proteins 0.000 description 1
- 238000002844 melting Methods 0.000 description 1
- 230000008018 melting Effects 0.000 description 1
- 230000037353 metabolic pathway Effects 0.000 description 1
- GBMDVOWEEQVZKZ-UHFFFAOYSA-N methanol;hydrate Chemical compound O.OC GBMDVOWEEQVZKZ-UHFFFAOYSA-N 0.000 description 1
- 108010034507 methionyltryptophan Proteins 0.000 description 1
- 230000002906 microbiologic effect Effects 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 230000003020 moisturizing effect Effects 0.000 description 1
- 238000010369 molecular cloning Methods 0.000 description 1
- 108020004707 nucleic acids Proteins 0.000 description 1
- 102000039446 nucleic acids Human genes 0.000 description 1
- 150000007523 nucleic acids Chemical class 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 108010012581 phenylalanylglutamate Proteins 0.000 description 1
- 108010051242 phenylalanylserine Proteins 0.000 description 1
- 230000000243 photosynthetic effect Effects 0.000 description 1
- 230000035790 physiological processes and functions Effects 0.000 description 1
- 238000011020 pilot scale process Methods 0.000 description 1
- 239000002244 precipitate Substances 0.000 description 1
- 230000001376 precipitating effect Effects 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 239000002243 precursor Substances 0.000 description 1
- 108010014614 prolyl-glycyl-proline Proteins 0.000 description 1
- 108010015796 prolylisoleucine Proteins 0.000 description 1
- 239000012474 protein marker Substances 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 230000005855 radiation Effects 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 230000008439 repair process Effects 0.000 description 1
- 108091008146 restriction endonucleases Proteins 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 230000003248 secreting effect Effects 0.000 description 1
- 239000013049 sediment Substances 0.000 description 1
- 238000011218 seed culture Methods 0.000 description 1
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 108010038745 tryptophylglycine Proteins 0.000 description 1
- 108010051110 tyrosyl-lysine Proteins 0.000 description 1
- 238000002525 ultrasonication Methods 0.000 description 1
- 108010073969 valyllysine Proteins 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
Images
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
- C12N15/70—Vectors or expression systems specially adapted for E. coli
-
- 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)
- 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.)
- C12N9/1096—Transferases (2.) transferring nitrogenous groups (2.6)
-
- 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
- C12N9/88—Lyases (4.)
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P17/00—Preparation of heterocyclic carbon compounds with only O, N, S, Se or Te as ring hetero atoms
- C12P17/10—Nitrogen as only ring hetero atom
- C12P17/12—Nitrogen as only ring hetero atom containing a six-membered hetero ring
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y203/00—Acyltransferases (2.3)
- C12Y203/01—Acyltransferases (2.3) transferring groups other than amino-acyl groups (2.3.1)
- C12Y203/01178—Diaminobutyrate acetyltransferase (2.3.1.178)
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y206/00—Transferases transferring nitrogenous groups (2.6)
- C12Y206/01—Transaminases (2.6.1)
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y402/00—Carbon-oxygen lyases (4.2)
- C12Y402/01—Hydro-lyases (4.2.1)
- C12Y402/01108—Ectoine synthase (4.2.1.108)
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)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Biotechnology (AREA)
- Biomedical Technology (AREA)
- Microbiology (AREA)
- Molecular Biology (AREA)
- Medicinal Chemistry (AREA)
- Physics & Mathematics (AREA)
- Biophysics (AREA)
- Plant Pathology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
本发明公开了一种生物法生产四氢嘧啶工程菌株的构建方法及其在四氢嘧啶生产中的应用。本发明提供的工程菌株利用海洋微生物盐栖盐田菌Salinicola salaria的四氢嘧啶合成基因簇的异源表达，使氨基丁酸乙酰基转移酶、二氨基丁酸氨基转移酶和四氢嘧啶合成酶在大肠杆菌中串联高表达，再经过酶促反应催化天冬氨酸钠生成四氢嘧啶。该四氢嘧啶的生物转化方法具有反应温度温和、工艺简单、转化率高的特点。
Description
技术领域
本发明属于医药原料生产技术领域，具体涉及一种生产四氢嘧啶的工程菌株构建方法，以及该菌株生产应用。
背景技术
四氢嘧啶(Ectoine)的化学名称为1，4，5，6-四氢-2-甲基4-嘧啶羧酸、依克多因，是一种杂环氨基酸衍生物。它化学性质稳定，为白色结晶或结晶性粉末，熔点280℃，具极性、易溶于水，溶于甲醇，在生理酸碱度范围内不带电荷。
1985年科学家Galinski等人在研究生存于海洋中高pH值、极端盐环境里的光合细菌时，首次发现嗜盐绿外硫红螺菌Ectothiorhodospira halochloris体内中存在一种相容性溶质——四氢嘧啶。作为一种重要的渗透压补偿性、相容性溶质，四氢嘧啶能够维持内的渗透压平衡，帮助嗜盐微生物在高盐、高渗环境下生存；同时其独特分子结构具有很强的水分子络合能力，能使细胞内的游离水结构化，是非常优秀的天然保湿剂；近年来发现，四氢嘧啶还可以对处于高温、冷冻、射线等极端环境中细菌细胞的蛋白质、酶、核酸等起到保护作用，对抗紫外线对皮肤的伤害，修复紫外线导致的细胞DNA损伤。因此，目前四氢嘧啶主要应用于生物制剂、酶制剂等工业领域，以及具有保湿修复功能的护肤品和化妆品行业，这对人类的生产生活有着极其重要的意义。
由于四氢嘧啶的化学生产工艺复杂，且容易出现结构相似的副产物，导致后续分离成本过高，因此目前四氢嘧啶多通过生物发酵转化法进行生产。德国默克公司利用延长盐单胞菌Halomonas elongate发酵合成目的产物，再通过“细菌挤奶”的渗透压法提取纯化获得四氢嘧啶；近年来，国内外学者通过代谢工程手段，构建了一系列的大肠杆菌工程菌株进行全细胞催化合成生产，具有一定的市场优势。
文献He et al.Microbial Cell Factories(2015)14:55DOI 10.1186/s12934-015-0238-0描述了一种异源生物合成四氢嘧啶的方法，将来源于延长盐单胞菌Halomonaselongata的四氢嘧啶合成基因簇插入pBAD载体，在大肠杆菌中表达出氨基丁酸乙酰基转移酶、二氨基丁酸氨基转移酶和四氢嘧啶合成酶；摇瓶反应体系中添加OD600＝5的发酵菌体、100mM天冬氨酸钠、100mM甘油、100mM氯化钾以及100mM pH 7.0磷酸缓冲溶液，转化条件为30℃反应，摇床转数为200rpm，反应24小后，获得四氢嘧啶的产量为2.67g/L。通过发酵罐进行高密度菌体生产转化，30℃反应24小时后，获得的四氢嘧啶的产量为25.1g/L。
文献Y.Ning et al.Metabolic Engineering 36(2016)10-18DOI 10.1016/j.ymben.2016.02.013报道了多株合成四氢嘧啶的大肠杆菌W3110工程菌株，将来源于延长盐单胞菌Halomonas elongata的四氢嘧啶合成基因簇插入pTrc99a载体，在工程菌株中表达四氢嘧啶合成所需要的三种酶，同时对代谢通路进行了改造，提高了代谢流通路，并敲除了高丝氨酸和赖氨酸的代谢分支以积累代谢中间产物；在摇瓶反应体系中添加60％的葡萄糖，转化条件为36℃反应，摇床转数为200rpm，反应24小后，获得四氢嘧啶的产量为4.88～13.6g/L。通过发酵罐进行高密度菌体生产转化，36℃反应24小时后，获得的四氢嘧啶的产量为25.1g/L。
公布号CN104560844A的专利公布了一种四氢嘧啶高产大肠杆菌工程菌及其应用，将含有延长盐单胞菌Halomonas elongata的四氢嘧啶合成基因簇EctABC的重组质粒导入大肠杆菌后获得重组菌株，实现了四氢嘧啶合成的三个关键酶在阿拉伯糖启动子的调控下可溶性表达，诱导表达后的菌体以天冬氨酸钠为前体通过30℃条件下的生物转化，实现了四氢嘧啶的高效分泌型合成。每克菌体可以合成1.1克四氢嘧啶，其中超过90％的四氢嘧啶分泌至胞外。通过发酵罐进行高密度菌体生产转化，30℃反应36小时后，获得的四氢嘧啶的产量为21g/L。
公布号CN109182236A的专利公布了一种重组大肠杆菌及合成四氢嘧啶的应用，将大肠杆菌MG1655的二氨基庚二酸脱羧酶lysA基因敲除，并导入延长盐单胞菌Halomonaselongata的四氢嘧啶合成基因簇ectABC获得重组菌株，诱导表达后的菌体以L-天冬氨酸钠为底物，40℃条件下的生物转化24h后，四氢嘧啶产量达到3.24g/L。
目前现有的生产菌株均依赖于延长盐单胞菌Halomonas elongata的四氢嘧啶合成基因簇，其合成反应的最佳温度为30-40℃，较高的反应温度对冷热设备的要求较高，增加加热及降温过程的生产成本；再者中高温度会大大降低酶促反应中关键因素--酶的稳定性，造成生产的效率和产量下降。
目前现有的生产菌株及方法制约了四氢嘧啶的工业化生产和广泛领域的应用，因此开发一株新型、高效的低温生产菌株从而提高合成效率、降低生产成本，对四氢嘧啶的生产和应用具有重大意义。
发明内容
本发明的目的在于提供一株串联表达新型氨基丁酸乙酰基转移酶、二氨基丁酸氨基转移酶和四氢嘧啶合成酶的重组大肠杆菌工程菌株。本发明提供的新型氨基丁酸乙酰基转移酶、二氨基丁酸氨基转移酶和四氢嘧啶合成酶基因来源于海洋微生物盐栖盐田菌Salinicola salaria的四氢嘧啶合成基因簇，通过将该基因簇与表达载体pET24连接，并导入大肠杆菌宿主菌BL21(DE3)，从而获得工程菌株。该工程菌株通过诱导成功表达新型氨基丁酸乙酰基转移酶、二氨基丁酸氨基转移酶和四氢嘧啶合成酶。
为解决本发明的技术问题，提供的技术方案为：一种生物法生产四氢嘧啶的工程菌株的构建方法，包括如下步骤：
(1)用EcoR I和Nde I分别双酶切来自于盐栖盐田菌Salinicola salaria的四氢嘧啶合成基因簇，克隆至用EcoR I和Nde I酶切过的pET24载体上，得重组载体pET-ECT；
(2)将重组载体pET-ECT转入大肠杆菌宿主菌BL21(DE3)，得工程菌BL21-ECT；
所构建的重组菌株通过聚合酶启动转录四氢嘧啶合成基因簇，串联高表达其中的氨基丁酸乙酰基转移酶、二氨基丁酸氨基转移酶和四氢嘧啶合成酶；
所述的四氢嘧啶合成基因簇的序列如SEQ ID NO：1所示；
所述的氨基丁酸乙酰基转移酶的氨基酸序列如SEQ ID NO：2所示；
所述的二氨基丁酸氨基转移酶的氨基酸序列如SEQ ID NO：3所示；
所述的四氢嘧啶合成酶的氨基酸序列如SEQ ID NO：4所示。
优选的，步骤(1)包括如下步骤：
用EcoR I和Nde I双酶切合成的长度为2416bp的四氢嘧啶合成基因簇DNA片段；用EcoR I和Nde I双酶切载体pET24，酶切体系：DNA 43μL，Buffer R 5μL，Nde I 1μL，EcoR I1μL，37℃保温3小时；酶切的DNA片段胶回收，用T4连接酶连接四氢嘧啶合成基因簇DNA和载体片段，连接体系如下：DNA 7.5μL，pET24载体1.5μL，Buffer 1μL，T4连接酶1μL，保温16℃过夜，连接产物采用热激法转化至大肠杆菌宿主菌DH5α中；涂布到含有1％蛋白胨，0.5％酵母膏，1％氯化钠及1.5％琼脂粉的LB固体培养基上；LB平板在37℃培养至生长出转化子，挑取单菌落，用LB液体培养基37℃培养过夜，12000rpm离心1分钟，提取质粒，提取方法按照试剂盒说明书操作；用EcoR I和Nde I双酶切重组载体，酶切体系：DNA 43μL，Buffer R 5μL，Nde I 1μL，EcoR I 1μL，37℃保温3小时；电泳确定含有四氢嘧啶合成基因簇的DNA片段。
优选的，步骤(2)包括如下步骤：采用热激法将重组载体转化至大肠杆菌宿主菌BL21(DE3)中；涂布到含有1％蛋白胨，0.5％酵母膏，1％氯化钠及1.5％琼脂粉的LB固体培养基上；LB平板在37℃培养至生长出转化子，挑取单菌落，获得工程菌BL21-ECT。
优选的，包括如下步骤：
a.诱导发酵获得大量表达的氨基丁酸乙酰基转移酶、二氨基丁酸氨基转移酶和四氢嘧啶合成酶；
b.收集菌体、重悬浮作为酶液，并添加天冬氨酸钠底物，转化生成产物四氢嘧啶。
优选的，步骤a中包括：配种子液100mL，种子液含有蛋白胨1％，酵母提取物0.5％，氯化钠1％，余量为纯净水，氨水调pH 7.0～7.2；装在250mL三角瓶中灭菌后，接种平板培养基上的单菌落，摇床转数为200rpm；37℃培养16小时后，接种入含有100mL发酵液的500mL三角瓶中，发酵液含有蛋白胨1.2％，酵母提取物2.4％，甘油0.4％，磷酸二氢钾0.23％，磷酸氢二钾1.25％，余量为纯净水；发酵培养条件：37℃培养，摇床转数为200rpm，接种后2小时后，降温至25～28℃，加入终浓度0.2mM IPTG，培养12小时。
优选的，步骤b中包括：发酵培养结束后，4500rpm 4℃低温离心收集菌体；在100mLpH 6.5的磷酸缓冲液体系中，分别加入OD600＝5-20菌体，100-300mM天冬氨酸钠，50-150mM葡萄糖，0-200mM氯化钾，转化条件：20-35℃反应，摇床转数为180rpm；24小时后，通过高效液相色谱(HPLC)检测四氢嘧啶产量。
优选的，步骤b中包括：发酵培养结束后，4500rpm 4℃低温离心收集菌体；在100mLpH 6.5的磷酸缓冲液体系中，分别加入OD600＝10，200mM天冬氨酸钠，100mM葡萄糖，50mM氯化钾；转化条件：25℃反应，摇床转数为180rpm；24小时后，通过高效液相色谱(HPLC)检测四氢嘧啶产量。
本发明的另一个目的是提供一种在较低温度条件下生产四氢嘧啶的大肠杆菌工程菌株及生物转化方法。由于较高的反应温度对冷热设备的要求较高，同时工业生产过程中升温，以及后续分离提取降温过程都将消耗大量的能源成本；再者，大部分的生化酶类对温度较为敏感，长时间中高温度处理，会对酶的三级结构和生理功能造成不可逆破坏，从而导致造成生产的效率和产量下降。本发明提供的大肠杆菌工程菌株能够在20-25℃条件下高效地催化四氢嘧啶合成生产，从而避免酶的热失活并有效地减低工业生产产本。
为了实现上述目的，本发明提供了一种生产四氢嘧啶的大肠杆菌工程菌株，所述工程菌株含有重组质粒pET-ECT，所述重组质粒包含有人工合成自于盐栖盐田菌Salinicola salaria的四氢嘧啶合成基因簇的DNA片段，核苷酸序列如序列表中的SEQIDNO：1。所述四氢嘧啶合成基因簇编码的三个四氢嘧啶合成相关酶类，分别为氨基丁酸乙酰基转移酶、二氨基丁酸氨基转移酶和四氢嘧啶合成酶。所述氨基丁酸乙酰基转移酶与来源于延长盐单胞菌Halomonas elongata的氨基丁酸乙酰基转移酶同源性为82.9％，氨基酸序列如序列表中的SEQID NO：2。所述二氨基丁酸氨基转移酶与来源于延长盐单胞菌Halomonas elongata的二氨基丁酸氨基转移酶同源性为85.5％，氨基酸序列如序列表中的SEQUENCE LISTING NO：3。四氢嘧啶合成酶与来源于延长盐单胞菌Halomonas elongata的四氢嘧啶合成酶同源性为72.1％，氨基酸序列如序列表中的SEQID NO：4。
本发明还提供一种在较低温度条件下生产四氢嘧啶的生物转化方法，所述是生物转化方法包括以下步骤：(1)所述的工程菌株接种于种子培养基中培养；(2)将培养所得菌液转接至发酵培养基中进行诱导发酵；(3)将发酵后的菌液进行离心收集菌体；在100mL pH6.5的磷酸缓冲液体系中，分别加入OD600＝5-20菌体，优选OD600＝10，100-300mM天冬氨酸钠，优选200mM天冬氨酸钠，50-150mM葡萄糖，优选100mM葡萄糖，0-200mM氯化钾，优选50mM氯化钾。转化条件：20-25℃反应，优选25℃反应，摇床转数为180rpm；24小时后，通过高效液相色谱(HPLC)检测四氢嘧啶产量。
与目前现有技术相比，本发明构建的工程菌株，导入了来源于盐栖盐田菌Salinicola salaria的四氢嘧啶合成基因簇的DNA片段。该基因簇的功能为首次报道。其编码表达的三个四氢嘧啶合成相关酶类，分别为氨基丁酸乙酰基转移酶、二氨基丁酸氨基转移酶和四氢嘧啶合成酶，与来源于延长盐单胞菌Halomonas elongata的相类似酶的氨基酸序列同源性分别为82.9％、85.5％和72.1％。这三个酶赋予了本发明所构建工程菌株能够在25℃的反应温度下合成四氢嘧啶，其发酵罐产量为24.8g/L；现有的大肠杆菌菌株需在30-36℃的反应温度下合成，其最高产量为25.1g/L，因此本发明具有在更低温度条产生四氢嘧啶，且产量与目前技术的效果相似、。
本发明提供的生产四氢嘧啶的生物转化方法，可以有效地降低对工业生产的设备要求，20-25℃的工业反应温度可以节约可观的工业能耗，同时也对生化酶类起到一定的低温保护作用。反应结束后，底物转化率高，副产物少，纯化简单易行，生产效率高，具有较大的工业化潜力。
附图说明
图1为工程菌株中四氢嘧啶基因簇的表达情况；M：蛋白质Marker，1：空载菌株破碎上清液样品，2：空载菌株破碎沉淀样品，3：未诱导工程菌株破碎上清液样品，4：未诱导工程菌株破碎沉淀样品，5：诱导工程菌株破碎上清液样品，6：诱导工程菌株破碎沉淀样品。
图2为HPLC检测鉴定标准品和反应液四氢嘧啶含量；A：四氢嘧啶标准品，B：反应液。
图3为不同反应条件对四氢嘧啶合成的影响；A：天冬氨酸钠，B：氯化钾，C：温度，D：pH值。
具体实施例
下面结合实施例对本发明作进一步的详细描述。实施例中的实验方法，按常规条件进行，参考如萨姆布鲁克等，分子克隆实验指南(第四版，2017年，科学出版社)中所述实验方法。
实施例中所用的材料、试剂等，均可从商业途径得到。
实施例中Salinicola salaria的四氢嘧啶合成基因簇DNA片段，其核苷酸序列公布于美国国家生物技术信息中心(National Center for Biotechnology Information，NCBI)建立GenBank序列数据库中，登录编号为NZ_NHOU01000014.1。DNA实物从商业生物技术公司，如金斯瑞生物科技股份有限公司合成得到。
实施例1：重组菌株的构建及鉴定
1.含重组质粒pET-ECT的菌株构建
1)质粒构建
质粒pET24大小为4.97kb，含有卡那霉素抗性基因，乳糖抑制子lac I基因，启动子是tac，并具有多个限制性内切酶位点。
商业人工合成自于盐栖盐田菌Salinicola salaria的四氢嘧啶合成基因簇的DNA片段，核苷酸序列如序列表中的SEQ ID NO：1。
用EcoR I和Nde I双酶切合成的四氢嘧啶合成基因簇DNA片段，用EcoR I和Nde I双酶切载体pET24。酶切体系：DNA 43μL，Buffer R 5μL，Nde I 1μL，EcoR I 1μL，37℃保温3小时。
酶切的DNA片段胶回收，用T4连接酶连接四氢嘧啶合成基因簇和pET24载体DNA片段，连接体系如下：四氢嘧啶合成基因簇DNA 7.5μL，pET24载体1.5μL，Buffer 1μL，T4连接酶1μL，保温16℃过夜，连接产物采用热激法转化至大肠杆菌宿主菌DH5α中，购于美国赛默飞世尔公司(Thermo Fisher Scientific)，货号为18265017。涂布到含有1％蛋白胨，0.5％酵母膏，1％氯化钠及1.5％琼脂粉卡那霉素的LB固体培养基上。
2)重组菌株鉴定
LB平板在37℃培养至生长出转化子，挑取单菌落，用LB液体培养基37℃培养过夜，12000rpm离心1分钟，提取质粒，提取方法按照试剂盒说明书操作。
用EcoR I和Nde I双酶切重组载体，酶切体系：DNA 43μL，buffer R 5μL，Nde I 1μL，EcoR I 1μL，37℃保温3小时。电泳确定含有目的四氢嘧啶合成基因簇的DNA片段。
2.构建含pET-ECT重组载体的生产菌株
采用热激法将重组载体转化至大肠杆菌宿主菌BL21(DE3)中，购于美国赛默飞世尔公司(Thermo Fisher Scientific)，，货号为C601003。涂布到含有1％蛋白胨，0.5％酵母膏，1％氯化钠及1.5％琼脂粉的LB固体培养基上。LB平板在37℃培养至生长出转化子，挑取单菌落，获得预期工程菌。
实施例2：重组菌株的发酵
配种子液100mL，种子液含有蛋白胨1％，酵母提取物0.5％，氯化钠1％，余量为纯净水。装在250mL三角瓶中灭菌后，接种平板培养基上的单菌落，摇床转数为200rpm。37℃培养16小时后，接种入含有100mL发酵液的500mL三角瓶中，发酵液含有蛋白胨1.2％，酵母提取物2.4％，甘油0.4％，磷酸二氢钾0.23％，磷酸氢二钾1.25％，余量为纯净水。发酵培养条件：按发酵体积1％量接种，37℃培养，摇床转数为200rpm，接种后2小时后，降温至25～28℃，加入终浓度0.2mM IPTG，培养12小时。发酵培养结束后，取1mL菌体4000rpm 4℃低温离心收集，超声破碎后12000rpm离心，收集上清液和沉淀，通过SDS-PAGE检测酶的表达情况，结果见图1。
实施例3：催化天冬氨酸钠生成四氢嘧啶
发酵培养结束后，4000rpm 4℃低温离心收集菌体，用20mL pH 6.5的磷酸缓冲液悬浮菌体，以及200mM天冬氨酸钠，100mM葡萄糖，50mM氯化钾，转入100mL三角瓶中，转化条件为20℃反应，摇床转数为180rpm。24小时后，通过高效液相色谱(HPLC)检测得四氢嘧啶产量为2.15g/L。
实施例4：催化天冬氨酸钠生成四氢嘧啶
发酵培养结束后，4000rpm 4℃低温离心收集菌体，用20mL pH 6.5的磷酸缓冲液悬浮菌体，以及200mM天冬氨酸钠，100mM葡萄糖，50mM氯化钾，转入100mL三角瓶中，转化条件为25℃反应，摇床转数为180rpm。24小时后，通过高效液相色谱(HPLC)检测得四氢嘧啶产量为3.41g/L。检测结果见图3。该实施例为目前大肠杆菌在25℃温度条件下摇瓶催化合成四氢嘧啶的最高产量。
实施例5：催化天冬氨酸钠生成四氢嘧啶
发酵培养结束后，4000rpm 4℃低温离心收集菌体，用20mL pH 6.5的磷酸缓冲液悬浮菌体，以及200mM天冬氨酸钠，100mM葡萄糖，50mM氯化钾，转入100mL三角瓶中，转化条件为30℃反应，摇床转数为180rpm。24小时后，通过高效液相色谱(HPLC)检测得四氢嘧啶产量为2.75g/L。检测结果见图3。
实施例6：催化天冬氨酸钠生成四氢嘧啶
发酵培养结束后，4000rpm 4℃低温离心收集菌体，用20mL pH 6.5的磷酸缓冲液悬浮菌体，以及200mM天冬氨酸钠，100mM葡萄糖，50mM氯化钾，转入100mL三角瓶中，转化条件为35℃反应，摇床转数为180rpm。24小时后，通过高效液相色谱(HPLC)检测得四氢嘧啶产量为2.58g/L。
实施例7：催化天冬氨酸钠生成四氢嘧啶
发酵培养结束后，4000rpm 4℃低温离心收集菌体，用20mL pH 6.5的磷酸缓冲液悬浮菌体，以及200mM天冬氨酸钠，100mM葡萄糖，50mM氯化钾，转入100mL三角瓶中，转化条件为40℃反应，摇床转数为180rpm。24小时后，通过高效液相色谱(HPLC)检测得四氢嘧啶产量为1.74g/L。
实施例8：发酵罐小试生产四氢嘧啶
配种子液200mL，种子液含有蛋白胨1％，酵母提取物0.5％，氯化钠1％，余量为纯净水。装在500mL三角瓶中灭菌后，接种平板培养基上的单菌落，摇床转数为200rpm。37℃培养16小时后，接种入含有3L发酵液的5L发酵罐中，发酵液含有蛋白胨1.2％，酵母提取物2.4％，甘油0.4％，磷酸二氢钾0.23％，磷酸氢二钾1.25％，余量为纯净水。发酵培养条件：按发酵体积2％量接种，37℃培养，控制pH为7，溶氧30％，搅拌桨转数为500rpm。菌液浓度数值OD600达到20后，降温至25℃，加入终浓度0.2mM IPTG，培养至菌体浓度不再增加。
发酵培养结束后，4500rpm 4℃低温离心收集菌体，1L反应体系中，含pH 6.5磷酸缓冲液悬浮、浓度OD600为20的菌体，200mM天冬氨酸钠，100mM葡萄糖，50mM氯化钾；转化条件为25℃反应，搅拌桨转数为500rpm，补料速度为20mL/h，补料液中含有2M天冬氨酸钠，2M葡萄糖。24小时后，通过高效液相色谱(HPLC)检测得四氢嘧啶产量为24.8g/L。
目前已报道的大肠杆菌工程菌株，在发酵罐反应器中合成的最高产量为25.1g/L，其包含了延长盐单胞菌Halomonas elongata的四氢嘧啶合成基因簇，且反应温度需在30-36℃条件下进行。本发明在25℃的反应条件下，应用新颖的基因族，达到了相似的效果，具有较大的工业应用潜力。
实施例9：高效液相色谱法检测四氢嘧啶含量
标准品处理：称取标准品四氢嘧啶0.1克，定溶于100mL 90％甲醇水溶液，配制成浓度为1g/L的四氢嘧啶标准溶液，取标准溶液1mL，0.45μm滤膜过滤，取滤液转入进样瓶待测。
样品处理：取转化液100μL，加入900μL甲醇充分混匀，10000rpm离心2min，0.45μm滤膜过滤，取滤液转入进样瓶待测。
色谱条件：色谱柱为日本岛津NH2氨基色谱柱(250mm)，流动相为90％甲醇水溶液，流速为0.8mL/min，柱温35℃，检测波长为210nm；进样量5μL。检测结果见图2。
SEQUENCE LISTING
<110> 安徽师范大学
<120> 一种生物法生产四氢嘧啶的工程菌株构建方法
<130> asd1
<160> 4
<170> PatentIn version 3.5
<210> 1
<211> 2416
<212> DNA
<213> 人工序列
<400> 1
atgaccaacc ccaatccggc atttacgccg tcggcagact tggcaaggcc taccgtggcc 60
gacgccgtcg ttggcaagcc cgatatgcca ctgttcattc gcaagcccac cacggaagac 120
gggtggggga tctatgaact ggtcaaggcc tgcccgccgc ttgatgtcaa ttccggttat 180
gcctatctgt tgctggcaac gcagttccgc gatagctgcg ccgtcgcgac gacagaagat 240
ggggaaatcg tcggattcgt gtccggctat gtcaaagcgg ccgcacccga tatctatttc 300
ctgtggcaag tcgccgtcgg ggaaaaggcg cggggcaccg gcctggccag gcgcttggtg 360
gaagccgtac tgactcggcc ggagttggca gagctgaacc atctcgagac cacaatcacg 420
ccggacaatc aggcctcatg gggcttgttc aagcgtctag cggatcgctg gcaggccccc 480
ctcatcagtc gcgaatactt ttcgaccgat caactgggcg gagagcatga cccggaaaat 540
ctcgtccgga tcggcccgtt cgacccccaa cgtatccagg gcgggtaagg tctgcgttgt 600
cacgcatgtc ttatgcaagc acgccttatt cacgctcccc acgactatcc acactcccat 660
tacggaggtt gtatgcagac tcaaattctc gaacgcatgg agtccgaggt ccggacctat 720
tcccgctcct ttccgattgt tttcaccaag gcccagggcg cgcgtctcac tgccgaggac 780
ggccgcgagt acatcgattt cctcgctggc gccggcacgc tcaattacgg tcataaccat 840
ccgaagcttc aggaagcgct ggtcgaatac atcacttcca acggcatcgt ccatggtctg 900
gacatgtgga ccgccgccaa gcgtgactac ctcgagacgc tggaagaagt gatcttcaaa 960
ccgcgcggac tggactacaa ggttcatctg ccgggaccga cgggcaccaa cgctaccgaa 1020
gccgcgattc gcctggctcg cgtggcgaag gggcgtcata acatcgtcac cttcaccaat 1080
ggcttccatg gcgtcaccat gggcgcactg gcgacgaccg gcaaccgcaa gttccgcgaa 1140
gcgaccggtg gtatcccgac ccagggtgcc agcttcctgc cgttcgatgg ctatctcggg 1200
gaaggcactg acacactcga ctatttcgag aagctgctga gcgacaagtc tggcgggctc 1260
gacgtgccgg ccggcgtgat cgtcgagacg gtgcagggcg agggcggtat caacccggca 1320
ggcctggatt ggctgaagcg tctggaaggc atctgccgcg cgcatgacat cctgctgatc 1380
gtcgatgaca tccaggcagg ttgtggccgt accggcaagt tcttcagctt cgagcatgcc 1440
ggcatcgtgc cggacatcgt caccaactcg aagtcgctat ccgcttacgg gctgccgttc 1500
gcccacgtgc tgatgcgccc tgaactggat aagtggaagc ctggccagta caacggcacg 1560
ttccgcggct tcaacatggc tttcgtgacg gcgaccgccg cgctcagaca tttctggagc 1620
gatgacacct tcgagcgcga cgtccagcgt aaagggcgta tcgtcgaaga gcgcttcgcc 1680
agaattgccg cgatgctgac cgaaatgggt tatcccgcga gcgaacgcgg ccgcggtctg 1740
atgcgcggta tcgatgtacg cgacggtgac gtggcggaca agatcacggc aaaagccttc 1800
gaaaacgggc tcatcatcga aaccgcgggg caggatggtc aagtcgtgaa gtgcctgtgc 1860
ccgctggtga tcagcgatga agatctcctc gaaggtctgg atattctgga gcgcagtgcc 1920
aaggaagcct ttgcctgagg cggaaatgac gctgaaaggc ggccttcgat ggccgtcttt 1980
cataccgcct tttcacgtag tcacccacca ggagttgtaa tcgatgatcg tacgtaatct 2040
tgcagagtgt atgaaaaccg atcgttttgt cgaagccgag aacggtaact gggacagcac 2100
tcgcttgatg ctggctgaag acggctgtgg gttttcattc aacatcacgc gtattcatcc 2160
gggtacggaa acccacattc actacaagaa ccacatcgaa gcggtgttct gctacgaagg 2220
tgagggtgaa gtggaaacca tcgccgatgg caagatccat accatcaagg ccggcgacat 2280
gtatttgctc gaccagcacg atgagcattg gctgcgtggc aaggaaaagg gcatgaccgt 2340
ggcctgtgtc ttcagccctg cgctaactgg ccgtgaaatt caccgcgaag acggctccta 2400
cgcaccggcc gagtaa 2416
<210> 2
<211> 195
<212> PRT
<213> 人工序列
<400> 2
Met Thr Asn Pro Asn Pro Ala Phe Thr Pro Ser Ala Asp Leu Ala Arg
1 5 10 15
Pro Thr Val Ala Asp Ala Val Val Gly Lys Pro Asp Met Pro Leu Phe
20 25 30
Ile Arg Lys Pro Thr Thr Glu Asp Gly Trp Gly Ile Tyr Glu Leu Val
35 40 45
Lys Ala Cys Pro Pro Leu Asp Val Asn Ser Gly Tyr Ala Tyr Leu Leu
50 55 60
Leu Ala Thr Gln Phe Arg Asp Ser Cys Ala Val Ala Thr Thr Glu Asp
65 70 75 80
Gly Glu Ile Val Gly Phe Val Ser Gly Tyr Val Lys Ala Ala Ala Pro
85 90 95
Asp Ile Tyr Phe Leu Trp Gln Val Ala Val Gly Glu Lys Ala Arg Gly
100 105 110
Thr Gly Leu Ala Arg Arg Leu Val Glu Ala Val Leu Thr Arg Pro Glu
115 120 125
Leu Ala Glu Leu Asn His Leu Glu Thr Thr Ile Thr Pro Asp Asn Gln
130 135 140
Ala Ser Trp Gly Leu Phe Lys Arg Leu Ala Asp Arg Trp Gln Ala Pro
145 150 155 160
Leu Ile Ser Arg Glu Tyr Phe Ser Thr Asp Gln Leu Gly Gly Glu His
165 170 175
Asp Pro Glu Asn Leu Val Arg Ile Gly Pro Phe Asp Pro Gln Arg Ile
180 185 190
Gln Gly Gly
195
<210> 3
<211> 422
<212> PRT
<213> 人工序列
<400> 3
Met Met Gln Thr Gln Ile Leu Glu Arg Met Glu Ser Glu Val Arg Thr
1 5 10 15
Tyr Ser Arg Ser Phe Pro Ile Val Phe Thr Lys Ala Gln Gly Ala Arg
20 25 30
Leu Thr Ala Glu Asp Gly Arg Glu Tyr Ile Asp Phe Leu Ala Gly Ala
35 40 45
Gly Thr Leu Asn Tyr Gly His Asn His Pro Lys Leu Gln Glu Ala Leu
50 55 60
Val Glu Tyr Ile Thr Ser Asn Gly Ile Val His Gly Leu Asp Met Trp
65 70 75 80
Thr Ala Ala Lys Arg Asp Tyr Leu Glu Thr Leu Glu Glu Val Ile Phe
85 90 95
Lys Pro Arg Gly Leu Asp Tyr Lys Val His Leu Pro Gly Pro Thr Gly
100 105 110
Thr Asn Ala Thr Glu Ala Ala Ile Arg Leu Ala Arg Val Ala Lys Gly
115 120 125
Arg His Asn Ile Val Thr Phe Thr Asn Gly Phe His Gly Val Thr Met
130 135 140
Gly Ala Leu Ala Thr Thr Gly Asn Arg Lys Phe Arg Glu Ala Thr Gly
145 150 155 160
Gly Ile Pro Thr Gln Gly Ala Ser Phe Leu Pro Phe Asp Gly Tyr Leu
165 170 175
Gly Glu Gly Thr Asp Thr Leu Asp Tyr Phe Glu Lys Leu Leu Ser Asp
180 185 190
Lys Ser Gly Gly Leu Asp Val Pro Ala Gly Val Ile Val Glu Thr Val
195 200 205
Gln Gly Glu Gly Gly Ile Asn Pro Ala Gly Leu Asp Trp Leu Lys Arg
210 215 220
Leu Glu Gly Ile Cys Arg Ala His Asp Ile Leu Leu Ile Val Asp Asp
225 230 235 240
Ile Gln Ala Gly Cys Gly Arg Thr Gly Lys Phe Phe Ser Phe Glu His
245 250 255
Ala Gly Ile Val Pro Asp Ile Val Thr Asn Ser Lys Ser Leu Ser Ala
260 265 270
Tyr Gly Leu Pro Phe Ala His Val Leu Met Arg Pro Glu Leu Asp Lys
275 280 285
Trp Lys Pro Gly Gln Tyr Asn Gly Thr Phe Arg Gly Phe Asn Met Ala
290 295 300
Phe Val Thr Ala Thr Ala Ala Leu Arg His Phe Trp Ser Asp Asp Thr
305 310 315 320
Phe Glu Arg Asp Val Gln Arg Lys Gly Arg Ile Val Glu Glu Arg Phe
325 330 335
Ala Arg Ile Ala Ala Met Leu Thr Glu Met Gly Tyr Pro Ala Ser Glu
340 345 350
Arg Gly Arg Gly Leu Met Arg Gly Ile Asp Val Arg Asp Gly Asp Val
355 360 365
Ala Asp Lys Ile Thr Ala Lys Ala Phe Glu Asn Gly Leu Ile Ile Glu
370 375 380
Thr Ala Gly Gln Asp Gly Gln Val Val Lys Cys Leu Cys Pro Leu Val
385 390 395 400
Ile Ser Asp Glu Asp Leu Leu Glu Gly Leu Asp Ile Leu Glu Arg Ser
405 410 415
Ala Lys Glu Ala Phe Ala
420
<210> 4
<211> 130
<212> PRT
<213> 人工序列
<400> 4
Met Ile Val Arg Asn Leu Ala Glu Cys Met Lys Thr Asp Arg Phe Val
1 5 10 15
Glu Ala Glu Asn Gly Asn Trp Asp Ser Thr Arg Leu Met Leu Ala Glu
20 25 30
Asp Gly Cys Gly Phe Ser Phe Asn Ile Thr Arg Ile His Pro Gly Thr
35 40 45
Glu Thr His Ile His Tyr Lys Asn His Ile Glu Ala Val Phe Cys Tyr
50 55 60
Glu Gly Glu Gly Glu Val Glu Thr Ile Ala Asp Gly Lys Ile His Thr
65 70 75 80
Ile Lys Ala Gly Asp Met Tyr Leu Leu Asp Gln His Asp Glu His Trp
85 90 95
Leu Arg Gly Lys Glu Lys Gly Met Thr Val Ala Cys Val Phe Ser Pro
100 105 110
Ala Leu Thr Gly Arg Glu Ile His Arg Glu Asp Gly Ser Tyr Ala Pro
115 120 125
Ala Glu
130
Claims (7)
1.一种生物法生产四氢嘧啶的工程菌株的构建方法，其特征在于，包括如下步骤：
(1)用EcoR I和Nde I分别双酶切来自于盐栖盐田菌Salinicola salaria的四氢嘧啶合成基因簇，克隆至用EcoR I和Nde I酶切过的pET24载体上，得重组载体pET-ECT；
(2)将重组载体pET-ECT转入大肠杆菌宿主菌BL21(DE3)，得工程菌BL21-ECT；
所构建的重组菌株通过聚合酶启动转录四氢嘧啶合成基因簇，串联高表达其中的氨基丁酸乙酰基转移酶、二氨基丁酸氨基转移酶和四氢嘧啶合成酶；
所述的四氢嘧啶合成基因簇的序列如SEQ ID NO：1所示；
所述的氨基丁酸乙酰基转移酶的氨基酸序列如SEQ ID NO：2所示；
所述的二氨基丁酸氨基转移酶的氨基酸序列如SEQ ID NO：3所示；
所述的四氢嘧啶合成酶的氨基酸序列如SEQ ID NO：4所示。
2.根据权利要求1所述生物法生产四氢嘧啶的工程菌株的构建方法，其特征在于，步骤(1)包括如下步骤：
用EcoR I和Nde I双酶切合成的长度为2416bp的四氢嘧啶合成基因簇DNA片段；用EcoRI和Nde I双酶切载体pET24，酶切体系：DNA 43μL，Buffer R 5μL，Nde I 1μL，EcoR I 1μL，37℃保温3小时；酶切的DNA片段胶回收，用T4连接酶连接四氢嘧啶合成基因簇DNA和载体片段，连接体系如下：DNA 7.5μL，pET24载体1.5μL，Buffer 1μL，T4连接酶1μL，保温16℃过夜，连接产物采用热激法转化至大肠杆菌宿主菌DH5α中；涂布到含有1％蛋白胨，0.5％酵母膏，1％氯化钠及1.5％琼脂粉的LB固体培养基上；LB平板在37℃培养至生长出转化子，挑取单菌落，用LB液体培养基37℃培养过夜，12000rpm离心1分钟，提取质粒，提取方法按照试剂盒说明书操作；用EcoR I和Nde I双酶切重组载体，酶切体系：DNA 43μL，Buffer R 5μL，Nde I1μL，EcoR I 1μL，37℃保温3小时；
电泳确定含有四氢嘧啶合成基因簇的DNA片段。
3.根据权利要求1或2所述的生物法生产四氢嘧啶的工程菌株的构建方法，其特征在于，步骤(2)包括如下步骤：采用热激法将重组载体转化至大肠杆菌宿主菌BL21(DE3)中；涂布到含有1％蛋白胨，0.5％酵母膏，1％氯化钠及1.5％琼脂粉的LB固体培养基上；LB平板在37℃培养至生长出转化子，挑取单菌落，获得工程菌BL21-ECT。
4.一种权利要求1所述构建方法所构建的四氢嘧啶工程菌株的生物法转化方法，其特征在于，包括如下步骤：
a.诱导发酵获得大量表达的氨基丁酸乙酰基转移酶、二氨基丁酸氨基转移酶和四氢嘧啶合成酶；
b.收集菌体、重悬浮作为酶液，并添加天冬氨酸钠底物，转化生成产物四氢嘧啶。
5.根据权利要求4所述的生物法转化方法，其特征在于，步骤a中包括：配种子液100mL，种子液含有蛋白胨1％，酵母提取物0.5％，氯化钠1％，余量为纯净水，氨水调pH 7.0～7.2；装在250mL三角瓶中灭菌后，接种平板培养基上的单菌落，摇床转数为200rpm；37℃培养16小时后，接种入含有100mL发酵液的500mL三角瓶中，发酵液含有蛋白胨1.2％，酵母提取物2.4％，甘油0.4％，磷酸二氢钾0.23％，磷酸氢二钾1.25％，余量为纯净水；发酵培养条件：37℃培养，摇床转数为200rpm，接种后2小时后，降温至25～28℃，加入终浓度0.2mM IPTG，培养12小时。
6.根据权利要求4或5所述的生物法转化方法，其特征在于，步骤b中包括：发酵培养结束后，4500rpm 4℃低温离心收集菌体；在100mL pH 6.5的磷酸缓冲液体系中，分别加入OD600＝5-20菌体，100-300mM天冬氨酸钠，50-150mM葡萄糖，0-200mM氯化钾，转化条件：20-35℃反应，摇床转数为180rpm；24小时后，通过高效液相色谱HPLC检测四氢嘧啶产量。
7.根据权利要求6所述的生物法转化方法，其特征在于，步骤b中包括：发酵培养结束后，4500rpm 4℃低温离心收集菌体；在100mL pH 6.5的磷酸缓冲液体系中，分别加入OD600＝10，200mM天冬氨酸钠，100mM葡萄糖，50mM氯化钾；转化条件：25℃反应，摇床转数为180rpm；24小时后，通过高效液相色谱HPLC检测四氢嘧啶产量。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110243895.1A CN112961875B (zh) | 2021-03-05 | 2021-03-05 | 一种生物法生产四氢嘧啶的工程菌株的构建方法 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110243895.1A CN112961875B (zh) | 2021-03-05 | 2021-03-05 | 一种生物法生产四氢嘧啶的工程菌株的构建方法 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN112961875A CN112961875A (zh) | 2021-06-15 |
| CN112961875B true CN112961875B (zh) | 2022-07-19 |
Family
ID=76276753
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202110243895.1A Active CN112961875B (zh) | 2021-03-05 | 2021-03-05 | 一种生物法生产四氢嘧啶的工程菌株的构建方法 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN112961875B (zh) |
Families Citing this family (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113564090B (zh) * | 2021-06-28 | 2023-07-25 | 福建师范大学 | 一种用于产四氢嘧啶重组菌的构建方法及其应用 |
| CN114410664A (zh) * | 2021-12-20 | 2022-04-29 | 自然资源部第三海洋研究所 | 四氢嘧啶生物合成基因及其应用 |
| CN114621968B (zh) * | 2022-05-17 | 2022-07-15 | 深圳中科翎碳生物科技有限公司 | 四氢嘧啶生物合成基因簇、突变体及制备四氢嘧啶的方法 |
| CN114806995B (zh) * | 2022-05-30 | 2024-03-12 | 深圳中科欣扬生物科技有限公司 | 一种基于乙酰辅酶a代谢改造后高效合成四氢嘧啶的基因工程菌的构建和应用 |
| CN114806974B (zh) * | 2022-06-10 | 2023-08-08 | 中国科学院微生物研究所 | 一株盐单胞菌菌株及其应用 |
| CN116410950B (zh) * | 2023-06-06 | 2023-08-15 | 云合(天津)生物技术有限公司 | 四氢嘧啶生物合成基因簇及发酵生产四氢嘧啶的方法 |
| CN116426584A (zh) * | 2023-06-14 | 2023-07-14 | 山东福瑞达生物科技有限公司 | 一种提高四氢嘧啶发酵产量的方法 |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103820375A (zh) * | 2013-12-13 | 2014-05-28 | 安徽师范大学 | 一种生物法生产阿魏酸工程菌株及其构建方法 |
| CN104593442A (zh) * | 2013-11-01 | 2015-05-06 | 南京众惠生物材料科技有限公司 | 一种重组大肠杆菌高密度培养生产四氢嘧啶的方法 |
| CN109182236A (zh) * | 2018-08-29 | 2019-01-11 | 浙江工业大学 | 一种重组大肠杆菌及合成四氢嘧啶的应用 |
| JP2019162080A (ja) * | 2018-03-20 | 2019-09-26 | 学校法人 東洋大学 | 醤油諸味粕を分解する方法および醤油諸味粕分解用組成物 |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US7630836B2 (en) * | 2001-05-30 | 2009-12-08 | The Kitasato Institute | Polynucleotides |
-
2021
- 2021-03-05 CN CN202110243895.1A patent/CN112961875B/zh active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104593442A (zh) * | 2013-11-01 | 2015-05-06 | 南京众惠生物材料科技有限公司 | 一种重组大肠杆菌高密度培养生产四氢嘧啶的方法 |
| CN103820375A (zh) * | 2013-12-13 | 2014-05-28 | 安徽师范大学 | 一种生物法生产阿魏酸工程菌株及其构建方法 |
| JP2019162080A (ja) * | 2018-03-20 | 2019-09-26 | 学校法人 東洋大学 | 醤油諸味粕を分解する方法および醤油諸味粕分解用組成物 |
| CN109182236A (zh) * | 2018-08-29 | 2019-01-11 | 浙江工业大学 | 一种重组大肠杆菌及合成四氢嘧啶的应用 |
Non-Patent Citations (3)
| Title |
|---|
| Ectoine Production Using Novel Heterologous EctABCS. Salaries from Marine Bacterium Salinicola salarius;Yue Su et al.;《Applied Sciences》;20210726;第11卷;第1-18页 * |
| High production of ectoine from aspartate and glycerol by use of whole-cell biocatalysis in recombinant Escherichia coli;Yong-Zhi He et al.;《Microbial Cell Factories》;20151231;第14卷;第1-10页 * |
| 密旋链霉菌Act12中四氢嘧啶合成基因簇的鉴定及其功能分析;高波等;《西北农林科技大学学报(自然科学版)》;20170510;第45卷(第6期);第213-220页 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN112961875A (zh) | 2021-06-15 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN112961875B (zh) | 一种生物法生产四氢嘧啶的工程菌株的构建方法 | |
| CN105296456B (zh) | 一种pH稳定性提高的谷氨酸脱羧酶突变体及其应用 | |
| CN112391372B (zh) | 一种谷氨酸脱羧酶突变体、基因工程菌及其应用 | |
| CN104152505A (zh) | 一种利用重组菌株转化制备4-羟基-l-异亮氨酸的方法 | |
| CN104152498A (zh) | 一种酶法生产α-酮戊二酸的方法 | |
| CN109593702B (zh) | 一种基因工程菌株实现全细胞转化合成l-苯乳酸的方法 | |
| CN112831488B (zh) | 一种谷氨酸脱羧酶及γ-氨基丁酸高产菌株 | |
| RU2546239C1 (ru) | РЕКОМБИНАНТНЫЙ ШТАММ Escherichia coli, ОБЛАДАЮЩИЙ КОНСТИТУТИВНОЙ АСПАРТАЗНОЙ АКТИВНОСТЬЮ И СПОСОБ СИНТЕЗА L-АСПАРАГИНОВОЙ КИСЛОТЫ С ИСПОЛЬЗОВАНИЕМ ЭТОГО ШТАММА В КАЧЕСТВЕ БИОКАТАЛИЗАТОРА | |
| CN113549633A (zh) | 一种l-半胱氨酸转运蛋白突变体及其在生产l半胱氨酸中的应用 | |
| CN112708569B (zh) | 发酵生产硫酸软骨素的酵母工程菌及其应用 | |
| CN113502305B (zh) | 一种利用重组酰亚胺酶合成(r)-异丁基戊二酸单酰胺的方法 | |
| US11760988B2 (en) | L-aspartate alpha-decarboxylase mutant and application thereof | |
| CN110923223B (zh) | 一种新型腈水解酶及其应用 | |
| Lee et al. | Mass production of thermostable D‐hydantoinase by batch culture of recombinant Escherichia coli with a constitutive expression system | |
| CN113930376A (zh) | 一种用于催化生产d-对羟基苯甘氨酸的工程菌、高密度培养方法及催化生产方法 | |
| CN113528495A (zh) | 一种稳定表达几丁二糖脱乙酰酶的枯草芽孢杆菌及其构建方法与应用 | |
| CN107916271B (zh) | 一种重组腈水合酶的高效表达方法 | |
| CN108004275B (zh) | 一种产己二酸的大肠杆菌重组菌及其应用 | |
| CN115247144B (zh) | 生产l-苏式-3-羟基天冬氨酸的基因工程菌及其应用 | |
| CN114606212B (zh) | 一种圆锥铁线莲来源的香豆素合酶、基因、载体及其应用 | |
| CN114317476B (zh) | 葡萄糖基甘油的生物催化生产工艺及其蔗糖磷酸化酶 | |
| CN114381412B (zh) | 一种合成3-羟基丙酸的重组菌及其构建方法与应用 | |
| CN114480315B (zh) | 一种Baeyer-Villiger单加氧酶及其在布立西坦合成中的应用 | |
| CN112522171A (zh) | 一种工程菌、含鸟氨酸溶液的处理方法及试剂盒 | |
| CN112175929A (zh) | 一种重组海因消旋酶工程菌及其构建方法和用途 |
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 | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |