CN114774451A - 一种重组大肠杆菌及其生产酪醇、羟基酪醇或红景天苷的方法 - Google Patents
一种重组大肠杆菌及其生产酪醇、羟基酪醇或红景天苷的方法 Download PDFInfo
- Publication number
- CN114774451A CN114774451A CN202210466075.3A CN202210466075A CN114774451A CN 114774451 A CN114774451 A CN 114774451A CN 202210466075 A CN202210466075 A CN 202210466075A CN 114774451 A CN114774451 A CN 114774451A
- Authority
- CN
- China
- Prior art keywords
- gene
- tyrosol
- salidroside
- hydroxytyrosol
- recombinant
- 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
- YCCILVSKPBXVIP-UHFFFAOYSA-N 2-(4-hydroxyphenyl)ethanol Chemical compound OCCC1=CC=C(O)C=C1 YCCILVSKPBXVIP-UHFFFAOYSA-N 0.000 title claims abstract description 95
- JUUBCHWRXWPFFH-UHFFFAOYSA-N Hydroxytyrosol Chemical compound OCCC1=CC=C(O)C(O)=C1 JUUBCHWRXWPFFH-UHFFFAOYSA-N 0.000 title claims abstract description 58
- DBLDQZASZZMNSL-QMMMGPOBSA-N L-tyrosinol Natural products OC[C@@H](N)CC1=CC=C(O)C=C1 DBLDQZASZZMNSL-QMMMGPOBSA-N 0.000 title claims abstract description 47
- 235000004330 tyrosol Nutrition 0.000 title claims abstract description 47
- 235000003248 hydroxytyrosol Nutrition 0.000 title claims abstract description 28
- 229940095066 hydroxytyrosol Drugs 0.000 title claims abstract description 28
- ILRCGYURZSFMEG-UHFFFAOYSA-N Salidroside Natural products OC1C(O)C(O)C(CO)OC1OCCC1=CC=C(O)C=C1 ILRCGYURZSFMEG-UHFFFAOYSA-N 0.000 title claims abstract description 27
- ILRCGYURZSFMEG-RQICVUQASA-N salidroside Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)OC1OCCC1=CC=C(O)C=C1 ILRCGYURZSFMEG-RQICVUQASA-N 0.000 title claims abstract description 27
- 238000004519 manufacturing process Methods 0.000 title claims abstract description 10
- 241000588724 Escherichia coli Species 0.000 title abstract description 19
- 241000894006 Bacteria Species 0.000 claims abstract description 65
- 238000003786 synthesis reaction Methods 0.000 claims abstract description 23
- 230000015572 biosynthetic process Effects 0.000 claims abstract description 22
- 230000014509 gene expression Effects 0.000 claims abstract description 17
- 230000037361 pathway Effects 0.000 claims abstract description 12
- XQXPVVBIMDBYFF-UHFFFAOYSA-N 4-hydroxyphenylacetic acid Chemical compound OC(=O)CC1=CC=C(O)C=C1 XQXPVVBIMDBYFF-UHFFFAOYSA-N 0.000 claims abstract description 10
- 102000004190 Enzymes Human genes 0.000 claims abstract description 10
- 108090000790 Enzymes Proteins 0.000 claims abstract description 10
- 108700023372 Glycosyltransferases Proteins 0.000 claims abstract description 9
- 230000004927 fusion Effects 0.000 claims abstract description 9
- 238000003501 co-culture Methods 0.000 claims abstract description 4
- 102000051366 Glycosyltransferases Human genes 0.000 claims abstract description 3
- 108090000623 proteins and genes Proteins 0.000 claims description 11
- 239000013598 vector Substances 0.000 claims description 11
- 238000000034 method Methods 0.000 claims description 10
- 238000000855 fermentation Methods 0.000 claims description 9
- 230000004151 fermentation Effects 0.000 claims description 9
- 108010021809 Alcohol dehydrogenase Proteins 0.000 claims description 7
- 238000006555 catalytic reaction Methods 0.000 claims description 6
- 230000008569 process Effects 0.000 claims description 6
- 101710118140 Alpha-keto-acid decarboxylase Proteins 0.000 claims description 4
- 101000585554 Arabidopsis thaliana Phenylacetaldehyde synthase Proteins 0.000 claims description 3
- 101000758783 Bacillus subtilis (strain 168) Probable 4-hydroxyphenylacetate 3-monooxygenase Proteins 0.000 claims description 3
- 101000866605 Geobacillus sp. (strain PA-9) 4-hydroxyphenylacetate 3-monooxygenase oxygenase component Proteins 0.000 claims description 3
- 102100031794 Alcohol dehydrogenase 6 Human genes 0.000 claims description 2
- 101100319874 Escherichia coli (strain K12) yahK gene Proteins 0.000 claims description 2
- 101000775460 Homo sapiens Alcohol dehydrogenase 6 Proteins 0.000 claims description 2
- 101100216804 Saccharomyces cerevisiae (strain ATCC 204508 / S288c) ARO10 gene Proteins 0.000 claims description 2
- 101150032117 ipdC gene Proteins 0.000 claims description 2
- 238000002360 preparation method Methods 0.000 claims description 2
- 230000002503 metabolic effect Effects 0.000 abstract description 4
- 239000000543 intermediate Substances 0.000 abstract description 2
- 230000037353 metabolic pathway Effects 0.000 abstract description 2
- 230000035699 permeability Effects 0.000 abstract description 2
- 238000007363 ring formation reaction Methods 0.000 abstract description 2
- 231100000331 toxic Toxicity 0.000 abstract description 2
- 230000002588 toxic effect Effects 0.000 abstract description 2
- OUYCCCASQSFEME-QMMMGPOBSA-N L-tyrosine Chemical compound OC(=O)[C@@H](N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-QMMMGPOBSA-N 0.000 description 24
- OUYCCCASQSFEME-UHFFFAOYSA-N tyrosine Natural products OC(=O)C(N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-UHFFFAOYSA-N 0.000 description 24
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 18
- 210000004027 cell Anatomy 0.000 description 15
- 230000001580 bacterial effect Effects 0.000 description 13
- 239000000725 suspension Substances 0.000 description 11
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 description 6
- 241001013691 Escherichia coli BW25113 Species 0.000 description 6
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 6
- 239000002504 physiological saline solution Substances 0.000 description 6
- 239000003814 drug Substances 0.000 description 5
- 230000000694 effects Effects 0.000 description 5
- 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 4
- DTQVDTLACAAQTR-UHFFFAOYSA-N Trifluoroacetic acid Chemical compound OC(=O)C(F)(F)F DTQVDTLACAAQTR-UHFFFAOYSA-N 0.000 description 4
- 239000008103 glucose Substances 0.000 description 4
- 239000008363 phosphate buffer Substances 0.000 description 4
- ZZZCUOFIHGPKAK-UHFFFAOYSA-N D-erythro-ascorbic acid Natural products OCC1OC(=O)C(O)=C1O ZZZCUOFIHGPKAK-UHFFFAOYSA-N 0.000 description 3
- 229930003268 Vitamin C Natural products 0.000 description 3
- 230000003078 antioxidant effect Effects 0.000 description 3
- 238000005119 centrifugation Methods 0.000 description 3
- 239000012634 fragment Substances 0.000 description 3
- 238000010438 heat treatment Methods 0.000 description 3
- 238000004128 high performance liquid chromatography Methods 0.000 description 3
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 3
- 235000019341 magnesium sulphate Nutrition 0.000 description 3
- 239000002609 medium Substances 0.000 description 3
- 235000019154 vitamin C Nutrition 0.000 description 3
- 239000011718 vitamin C Substances 0.000 description 3
- 101710095468 Cyclase Proteins 0.000 description 2
- 239000012880 LB liquid culture medium Substances 0.000 description 2
- 206010028980 Neoplasm Diseases 0.000 description 2
- ILRCGYURZSFMEG-RKQHYHRCSA-N Salidroside Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1OCCC1=CC=C(O)C=C1 ILRCGYURZSFMEG-RKQHYHRCSA-N 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 150000001413 amino acids Chemical group 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 238000007036 catalytic synthesis reaction Methods 0.000 description 2
- 210000000349 chromosome Anatomy 0.000 description 2
- 238000012258 culturing Methods 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 238000010828 elution Methods 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 239000013604 expression vector Substances 0.000 description 2
- XBGGUPMXALFZOT-UHFFFAOYSA-N glycyl-L-tyrosine hemihydrate Natural products NCC(=O)NC(C(O)=O)CC1=CC=C(O)C=C1 XBGGUPMXALFZOT-UHFFFAOYSA-N 0.000 description 2
- 230000036541 health Effects 0.000 description 2
- 244000005700 microbiome Species 0.000 description 2
- 239000004006 olive oil Substances 0.000 description 2
- 235000008390 olive oil Nutrition 0.000 description 2
- 239000008055 phosphate buffer solution Substances 0.000 description 2
- 239000002243 precursor Substances 0.000 description 2
- ZVNPWFOVUDMGRP-UHFFFAOYSA-N 4-methylaminophenol sulfate Chemical compound OS(O)(=O)=O.CNC1=CC=C(O)C=C1.CNC1=CC=C(O)C=C1 ZVNPWFOVUDMGRP-UHFFFAOYSA-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
- HQJKCXHQNUCKMY-GHCJXIJMSA-N Ala-Ile-Asp Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)O)NC(=O)[C@H](C)N HQJKCXHQNUCKMY-GHCJXIJMSA-N 0.000 description 1
- KYDYGANDJHFBCW-DRZSPHRISA-N Ala-Phe-Gln Chemical compound C[C@@H](C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CCC(=O)N)C(=O)O)N KYDYGANDJHFBCW-DRZSPHRISA-N 0.000 description 1
- AAWLEICNDUHIJM-MBLNEYKQSA-N Ala-Thr-His Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC1=CN=CN1)C(=O)O)NC(=O)[C@H](C)N)O AAWLEICNDUHIJM-MBLNEYKQSA-N 0.000 description 1
- CREYEAPXISDKSB-FQPOAREZSA-N Ala-Thr-Tyr Chemical compound [H]N[C@@H](C)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O CREYEAPXISDKSB-FQPOAREZSA-N 0.000 description 1
- 102000007698 Alcohol dehydrogenase Human genes 0.000 description 1
- 206010002660 Anoxia Diseases 0.000 description 1
- 241000976983 Anoxia Species 0.000 description 1
- HKRXJBBCQBAGIM-FXQIFTODSA-N Arg-Asp-Ser Chemical compound C(C[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)N[C@@H](CO)C(=O)O)N)CN=C(N)N HKRXJBBCQBAGIM-FXQIFTODSA-N 0.000 description 1
- VXXHDZKEQNGXNU-QXEWZRGKSA-N Arg-Asp-Val Chemical compound CC(C)[C@@H](C(O)=O)NC(=O)[C@H](CC(O)=O)NC(=O)[C@@H](N)CCCN=C(N)N VXXHDZKEQNGXNU-QXEWZRGKSA-N 0.000 description 1
- AQPVUEJJARLJHB-BQBZGAKWSA-N Arg-Gly-Ala Chemical compound OC(=O)[C@H](C)NC(=O)CNC(=O)[C@@H](N)CCCN=C(N)N AQPVUEJJARLJHB-BQBZGAKWSA-N 0.000 description 1
- BKDDABUWNKGZCK-XHNCKOQMSA-N Asn-Glu-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CCC(=O)O)NC(=O)[C@H](CC(=O)N)N)C(=O)O BKDDABUWNKGZCK-XHNCKOQMSA-N 0.000 description 1
- DJIMLSXHXKWADV-CIUDSAMLSA-N Asn-Leu-Ser Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](N)CC(N)=O DJIMLSXHXKWADV-CIUDSAMLSA-N 0.000 description 1
- GZXOUBTUAUAVHD-ACZMJKKPSA-N Asn-Ser-Glu Chemical compound NC(=O)C[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@H](C(O)=O)CCC(O)=O GZXOUBTUAUAVHD-ACZMJKKPSA-N 0.000 description 1
- XAJRHVUUVUPFQL-ACZMJKKPSA-N Asp-Glu-Asp Chemical compound OC(=O)C[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(O)=O)C(O)=O XAJRHVUUVUPFQL-ACZMJKKPSA-N 0.000 description 1
- KQBVNNAPIURMPD-PEFMBERDSA-N Asp-Ile-Glu Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CCC(O)=O)C(O)=O KQBVNNAPIURMPD-PEFMBERDSA-N 0.000 description 1
- SPKCGKRUYKMDHP-GUDRVLHUSA-N Asp-Ile-Pro Chemical compound CC[C@H](C)[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CC(=O)O)N SPKCGKRUYKMDHP-GUDRVLHUSA-N 0.000 description 1
- NONWUQAWAANERO-BZSNNMDCSA-N Asp-Phe-Tyr Chemical compound C([C@H](NC(=O)[C@H](CC(O)=O)N)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(O)=O)C1=CC=CC=C1 NONWUQAWAANERO-BZSNNMDCSA-N 0.000 description 1
- 108090000489 Carboxy-Lyases Proteins 0.000 description 1
- 102000004031 Carboxy-Lyases Human genes 0.000 description 1
- 208000024172 Cardiovascular disease Diseases 0.000 description 1
- ZPDVKYLJTOFQJV-WDSKDSINSA-N Gln-Asn-Gly Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(N)=O)C(=O)NCC(O)=O ZPDVKYLJTOFQJV-WDSKDSINSA-N 0.000 description 1
- CLPQUWHBWXFJOX-BQBZGAKWSA-N Gln-Gly-Gln Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)NCC(=O)N[C@@H](CCC(N)=O)C(O)=O CLPQUWHBWXFJOX-BQBZGAKWSA-N 0.000 description 1
- SBHVGKBYOQKAEA-SDDRHHMPSA-N Gln-His-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CC2=CN=CN2)NC(=O)[C@H](CCC(=O)N)N)C(=O)O SBHVGKBYOQKAEA-SDDRHHMPSA-N 0.000 description 1
- MKRDNSWGJWTBKZ-GVXVVHGQSA-N Gln-Val-Lys Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CCCCN)C(=O)O)NC(=O)[C@H](CCC(=O)N)N MKRDNSWGJWTBKZ-GVXVVHGQSA-N 0.000 description 1
- KIEICAOUSNYOLM-NRPADANISA-N Glu-Val-Ala Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](C)C(O)=O KIEICAOUSNYOLM-NRPADANISA-N 0.000 description 1
- LCNXZQROPKFGQK-WHFBIAKZSA-N Gly-Asp-Ser Chemical compound NCC(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(O)=O LCNXZQROPKFGQK-WHFBIAKZSA-N 0.000 description 1
- FIQQRCFQXGLOSZ-WDSKDSINSA-N Gly-Glu-Asp Chemical compound [H]NCC(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(O)=O)C(O)=O FIQQRCFQXGLOSZ-WDSKDSINSA-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
- IUKIDFVOUHZRAK-QWRGUYRKSA-N Gly-Lys-His Chemical compound NCCCC[C@H](NC(=O)CN)C(=O)N[C@H](C(O)=O)CC1=CN=CN1 IUKIDFVOUHZRAK-QWRGUYRKSA-N 0.000 description 1
- MHXKHKWHPNETGG-QWRGUYRKSA-N Gly-Lys-Leu Chemical compound [H]NCC(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(O)=O MHXKHKWHPNETGG-QWRGUYRKSA-N 0.000 description 1
- CVFOYJJOZYYEPE-KBPBESRZSA-N Gly-Lys-Tyr Chemical compound [H]NCC(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O CVFOYJJOZYYEPE-KBPBESRZSA-N 0.000 description 1
- 206010020772 Hypertension Diseases 0.000 description 1
- 206010021143 Hypoxia Diseases 0.000 description 1
- IDMNOFVUXYYZPF-DKIMLUQUSA-N Ile-Lys-Phe Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)O)N IDMNOFVUXYYZPF-DKIMLUQUSA-N 0.000 description 1
- FQYQMFCIJNWDQZ-CYDGBPFRSA-N Ile-Pro-Pro Chemical compound CC[C@H](C)[C@H](N)C(=O)N1CCC[C@H]1C(=O)N1[C@H](C(O)=O)CCC1 FQYQMFCIJNWDQZ-CYDGBPFRSA-N 0.000 description 1
- PRTZQMBYUZFSFA-XEGUGMAKSA-N Ile-Tyr-Gly Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC1=CC=C(C=C1)O)C(=O)NCC(=O)O)N PRTZQMBYUZFSFA-XEGUGMAKSA-N 0.000 description 1
- ZYVTXBXHIKGZMD-QSFUFRPTSA-N Ile-Val-Asn Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(=O)N)C(=O)O)N ZYVTXBXHIKGZMD-QSFUFRPTSA-N 0.000 description 1
- FQZPTCNSNPWHLJ-AVGNSLFASA-N Leu-Gln-Lys Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCCCN)C(O)=O FQZPTCNSNPWHLJ-AVGNSLFASA-N 0.000 description 1
- AIRUUHAOKGVJAD-JYJNAYRXSA-N Leu-Phe-Glu Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CCC(O)=O)C(O)=O AIRUUHAOKGVJAD-JYJNAYRXSA-N 0.000 description 1
- DCRWPTBMWMGADO-AVGNSLFASA-N Lys-Glu-Leu Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(O)=O DCRWPTBMWMGADO-AVGNSLFASA-N 0.000 description 1
- VMTYLUGCXIEDMV-QWRGUYRKSA-N Lys-Leu-Gly Chemical compound OC(=O)CNC(=O)[C@H](CC(C)C)NC(=O)[C@@H](N)CCCCN VMTYLUGCXIEDMV-QWRGUYRKSA-N 0.000 description 1
- QBHGXFQJFPWJIH-XUXIUFHCSA-N Lys-Pro-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)[C@@H]1CCCN1C(=O)[C@@H](N)CCCCN QBHGXFQJFPWJIH-XUXIUFHCSA-N 0.000 description 1
- HYSVGEAWTGPMOA-IHRRRGAJSA-N Lys-Pro-Leu Chemical compound [H]N[C@@H](CCCCN)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CC(C)C)C(O)=O HYSVGEAWTGPMOA-IHRRRGAJSA-N 0.000 description 1
- PPNCMJARTHYNEC-MEYUZBJRSA-N Lys-Tyr-Thr Chemical compound NCCCC[C@H](N)C(=O)N[C@H](C(=O)N[C@@H]([C@H](O)C)C(O)=O)CC1=CC=C(O)C=C1 PPNCMJARTHYNEC-MEYUZBJRSA-N 0.000 description 1
- 208000029725 Metabolic bone disease Diseases 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
- 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
- 206010049088 Osteopenia Diseases 0.000 description 1
- WEMYTDDMDBLPMI-DKIMLUQUSA-N Phe-Ile-Lys Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCCCN)C(=O)O)NC(=O)[C@H](CC1=CC=CC=C1)N WEMYTDDMDBLPMI-DKIMLUQUSA-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
- YUPRIZTWANWWHK-DZKIICNBSA-N Phe-Val-Glu Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)O)NC(=O)[C@H](CC1=CC=CC=C1)N YUPRIZTWANWWHK-DZKIICNBSA-N 0.000 description 1
- KIZQGKLMXKGDIV-BQBZGAKWSA-N Pro-Ala-Gly Chemical compound OC(=O)CNC(=O)[C@H](C)NC(=O)[C@@H]1CCCN1 KIZQGKLMXKGDIV-BQBZGAKWSA-N 0.000 description 1
- SGCZFWSQERRKBD-BQBZGAKWSA-N Pro-Asp-Gly Chemical compound OC(=O)CNC(=O)[C@H](CC(O)=O)NC(=O)[C@@H]1CCCN1 SGCZFWSQERRKBD-BQBZGAKWSA-N 0.000 description 1
- 108010079005 RDV peptide Proteins 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- 235000014680 Saccharomyces cerevisiae Nutrition 0.000 description 1
- BNFVPSRLHHPQKS-WHFBIAKZSA-N Ser-Asp-Gly Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC(O)=O)C(=O)NCC(O)=O BNFVPSRLHHPQKS-WHFBIAKZSA-N 0.000 description 1
- WSTIOCFMWXNOCX-YUMQZZPRSA-N Ser-Gly-Lys Chemical compound C(CCN)C[C@@H](C(=O)O)NC(=O)CNC(=O)[C@H](CO)N WSTIOCFMWXNOCX-YUMQZZPRSA-N 0.000 description 1
- NNFMANHDYSVNIO-DCAQKATOSA-N Ser-Lys-Arg Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O NNFMANHDYSVNIO-DCAQKATOSA-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
- PXQUBKWZENPDGE-CIQUZCHMSA-N Thr-Ala-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)[C@H](C)NC(=O)[C@H]([C@@H](C)O)N PXQUBKWZENPDGE-CIQUZCHMSA-N 0.000 description 1
- FQPDRTDDEZXCEC-SVSWQMSJSA-N Thr-Ile-Ser Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CO)C(O)=O FQPDRTDDEZXCEC-SVSWQMSJSA-N 0.000 description 1
- MCDVZTRGHNXTGK-HJGDQZAQSA-N Thr-Met-Glu Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CCC(O)=O)C(O)=O MCDVZTRGHNXTGK-HJGDQZAQSA-N 0.000 description 1
- VGYVVSQFSSKZRJ-OEAJRASXSA-N Thr-Phe-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)[C@@H](N)[C@H](O)C)CC1=CC=CC=C1 VGYVVSQFSSKZRJ-OEAJRASXSA-N 0.000 description 1
- MXNAOGFNFNKUPD-JHYOHUSXSA-N Thr-Phe-Thr Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H]([C@@H](C)O)C(O)=O MXNAOGFNFNKUPD-JHYOHUSXSA-N 0.000 description 1
- NQQMWWVVGIXUOX-SVSWQMSJSA-N Thr-Ser-Ile Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O NQQMWWVVGIXUOX-SVSWQMSJSA-N 0.000 description 1
- GJOBRAHDRIDAPT-NGTWOADLSA-N Thr-Trp-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)[C@H](CC1=CNC2=CC=CC=C21)NC(=O)[C@H]([C@@H](C)O)N GJOBRAHDRIDAPT-NGTWOADLSA-N 0.000 description 1
- KAJRRNHOVMZYBL-IRIUXVKKSA-N Thr-Tyr-Gln Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CCC(N)=O)C(O)=O KAJRRNHOVMZYBL-IRIUXVKKSA-N 0.000 description 1
- BKIOKSLLAAZYTC-KKHAAJSZSA-N Thr-Val-Asn Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(N)=O)C(O)=O BKIOKSLLAAZYTC-KKHAAJSZSA-N 0.000 description 1
- BXPOOVDVGWEXDU-WZLNRYEVSA-N Tyr-Ile-Thr Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H]([C@@H](C)O)C(O)=O BXPOOVDVGWEXDU-WZLNRYEVSA-N 0.000 description 1
- KGSDLCMCDFETHU-YESZJQIVSA-N Tyr-Lys-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CCCCN)NC(=O)[C@H](CC2=CC=C(C=C2)O)N)C(=O)O KGSDLCMCDFETHU-YESZJQIVSA-N 0.000 description 1
- XJPXTYLVMUZGNW-IHRRRGAJSA-N Tyr-Pro-Asp Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CC(O)=O)C(O)=O XJPXTYLVMUZGNW-IHRRRGAJSA-N 0.000 description 1
- SZEIFUXUTBBQFQ-STQMWFEESA-N Tyr-Pro-Gly Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N1CCC[C@H]1C(=O)NCC(O)=O SZEIFUXUTBBQFQ-STQMWFEESA-N 0.000 description 1
- DNOOLPROHJWCSQ-RCWTZXSCSA-N Val-Arg-Thr Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H]([C@@H](C)O)C(O)=O DNOOLPROHJWCSQ-RCWTZXSCSA-N 0.000 description 1
- ZMDCGGKHRKNWKD-LAEOZQHASA-N Val-Asn-Glu Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N[C@@H](CCC(=O)O)C(=O)O)N ZMDCGGKHRKNWKD-LAEOZQHASA-N 0.000 description 1
- OGNMURQZFMHFFD-NHCYSSNCSA-N Val-Asn-Lys Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N[C@@H](CCCCN)C(=O)O)N OGNMURQZFMHFFD-NHCYSSNCSA-N 0.000 description 1
- OUUBKKIJQIAPRI-LAEOZQHASA-N Val-Gln-Asn Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(N)=O)C(O)=O OUUBKKIJQIAPRI-LAEOZQHASA-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
- GQMNEJMFMCJJTD-NHCYSSNCSA-N Val-Pro-Gln Chemical compound CC(C)[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CCC(N)=O)C(O)=O GQMNEJMFMCJJTD-NHCYSSNCSA-N 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- 239000013543 active substance Substances 0.000 description 1
- 230000032683 aging Effects 0.000 description 1
- 108010087924 alanylproline Proteins 0.000 description 1
- 230000007953 anoxia Effects 0.000 description 1
- 230000003266 anti-allergic effect Effects 0.000 description 1
- 230000000879 anti-atherosclerotic effect Effects 0.000 description 1
- 230000002929 anti-fatigue Effects 0.000 description 1
- 230000000141 anti-hypoxic effect Effects 0.000 description 1
- 230000003110 anti-inflammatory effect Effects 0.000 description 1
- 230000000845 anti-microbial effect Effects 0.000 description 1
- 230000002180 anti-stress Effects 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- 235000006708 antioxidants Nutrition 0.000 description 1
- 108010047857 aspartylglycine Proteins 0.000 description 1
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 1
- NWIUTZDMDHAVTP-UHFFFAOYSA-N betaxolol Chemical compound C1=CC(OCC(O)CNC(C)C)=CC=C1CCOCC1CC1 NWIUTZDMDHAVTP-UHFFFAOYSA-N 0.000 description 1
- 229960004324 betaxolol Drugs 0.000 description 1
- 230000000975 bioactive effect Effects 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 210000004899 c-terminal region Anatomy 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- 229940041514 candida albicans extract Drugs 0.000 description 1
- 239000002327 cardiovascular agent Substances 0.000 description 1
- 229940125692 cardiovascular agent Drugs 0.000 description 1
- 230000002113 chemopreventative effect Effects 0.000 description 1
- 238000003776 cleavage reaction Methods 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 238000003912 environmental pollution Methods 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 239000012847 fine chemical Substances 0.000 description 1
- 230000008014 freezing Effects 0.000 description 1
- 238000007710 freezing Methods 0.000 description 1
- 108010042598 glutamyl-aspartyl-glycine Proteins 0.000 description 1
- 230000013595 glycosylation Effects 0.000 description 1
- 238000006206 glycosylation reaction Methods 0.000 description 1
- 108010090037 glycyl-alanyl-isoleucine Proteins 0.000 description 1
- 108010027668 glycyl-alanyl-valine Proteins 0.000 description 1
- 108010089804 glycyl-threonine Proteins 0.000 description 1
- 108010015792 glycyllysine Proteins 0.000 description 1
- 230000033444 hydroxylation Effects 0.000 description 1
- 238000005805 hydroxylation reaction Methods 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 239000002054 inoculum Substances 0.000 description 1
- 108010078274 isoleucylvaline Proteins 0.000 description 1
- 150000004715 keto acids Chemical class 0.000 description 1
- 108010053037 kyotorphin Proteins 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 108010017391 lysylvaline Proteins 0.000 description 1
- 230000004792 oxidative damage Effects 0.000 description 1
- 239000002530 phenolic antioxidant Substances 0.000 description 1
- 230000008832 photodamage Effects 0.000 description 1
- 230000019612 pigmentation Effects 0.000 description 1
- 108010015796 prolylisoleucine Proteins 0.000 description 1
- 230000005855 radiation Effects 0.000 description 1
- 230000007017 scission Effects 0.000 description 1
- 238000011218 seed culture Methods 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 230000002194 synthesizing effect Effects 0.000 description 1
- 108010061238 threonyl-glycine Proteins 0.000 description 1
- 230000002936 tranquilizing effect Effects 0.000 description 1
- 239000012137 tryptone Substances 0.000 description 1
- OUYCCCASQSFEME-QMMMGPOBSA-M tyrosinate group Chemical group N[C@@H](CC1=CC=C(C=C1)O)C(=O)[O-] OUYCCCASQSFEME-QMMMGPOBSA-M 0.000 description 1
- 108700026220 vif Genes Proteins 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
- 101150061013 yahK gene Proteins 0.000 description 1
- 239000012138 yeast extract Substances 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
- C12N15/70—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
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/0004—Oxidoreductases (1.)
- C12N9/0006—Oxidoreductases (1.) acting on CH-OH groups as donors (1.1)
-
- 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
- C12N9/0004—Oxidoreductases (1.)
- C12N9/0071—Oxidoreductases (1.) acting on paired donors with incorporation of molecular oxygen (1.14)
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/10—Transferases (2.)
- C12N9/1048—Glycosyltransferases (2.4)
- C12N9/1051—Hexosyltransferases (2.4.1)
-
- 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
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/44—Preparation of O-glycosides, e.g. glucosides
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P7/00—Preparation of oxygen-containing organic compounds
- C12P7/02—Preparation of oxygen-containing organic compounds containing a hydroxy group
- C12P7/22—Preparation of oxygen-containing organic compounds containing a hydroxy group aromatic
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y101/00—Oxidoreductases acting on the CH-OH group of donors (1.1)
- C12Y101/01—Oxidoreductases acting on the CH-OH group of donors (1.1) with NAD+ or NADP+ as acceptor (1.1.1)
- C12Y101/01002—Alcohol dehydrogenase (NADP+) (1.1.1.2), i.e. aldehyde reductase
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y114/00—Oxidoreductases acting on paired donors, with incorporation or reduction of molecular oxygen (1.14)
- C12Y114/14—Oxidoreductases acting on paired donors, with incorporation or reduction of molecular oxygen (1.14) with reduced flavin or flavoprotein as one donor, and incorporation of one atom of oxygen (1.14.14)
- C12Y114/14009—4-Hydroxyphenylacetate 3-monooxygenase (1.14.14.9)
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y204/00—Glycosyltransferases (2.4)
- C12Y204/01—Hexosyltransferases (2.4.1)
- C12Y204/01203—Trans-zeatin O-beta-D-glucosyltransferase (2.4.1.203)
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y401/00—Carbon-carbon lyases (4.1)
- C12Y401/01—Carboxy-lyases (4.1.1)
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Genetics & Genomics (AREA)
- Wood Science & Technology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- Medicinal Chemistry (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Physics & Mathematics (AREA)
- Biophysics (AREA)
- Plant Pathology (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
本发明公开了一种重组大肠杆菌及其生产酪醇、羟基酪醇或红景天苷的方法,所述重组大肠杆菌为在宿主菌中表达了酪醇合成途径酶融合体来生产酪醇,再引入4‑羟基苯乙酸3‑羟化酶生产羟基酪醇,引入糖基转移酶生产红景天苷;Spy(或Snoop)环化还可通过热处理以增加细胞的通透性,避免细胞的传质限制。通过模块共培养因将长代谢途径分割成多个途径、并可利用不同宿主细胞进行表达,减轻了对宿主细胞的代谢负担;而且不同途径可选用各自最合适的宿主细胞进行表达;可通过调整模块菌的比例来控制模块的适配性;即插即用,容易组装成不同途径以合成不同产物。还能避免代谢中间物和目标产物毒性的影响。
Description
技术领域
本发明属于生物工程技术领域,具体涉及一种重组大肠杆菌及其生产酪醇、羟基酪醇或红景天苷的方法。
背景技术
酪醇,对羟基苯乙醇,是天然酚类抗氧化剂。它具有抗氧化、抗疲劳、抗缺氧、抗应激、抗寒冷、抗敏、镇静、预防心血管疾病、预防高血压、预防骨质减少、预防色素沉积等生物活性。它又是合成心血管药物倍他洛尔和美托尔的前体。酪醇还可作为合成羟基酪醇盒红景天苷的前体。羟基酪醇具有防治、红景天苷具有。所以,酪醇作为精细化学品和制药工业中的生物活性物,备受研究者的关注。酪醇最初来源于橄榄油,但在橄榄油中浓度低(25mg/kg)等问题,提取难以实现工业化。目前化学合成是其主要的生产方法。但化学合成过程繁琐、环境污染重,生物合成却是其最有效的替代方法。
羟基酪醇,3,4-二羟基苯乙醇,是酪醇的羟化产物。具有比酪醇更强的抗氧化活性。因其高抗氧化活性对人体健康大有益处,具有癌症癌症化学预防、抗动脉粥样硬化、抑制DNA氧化损伤、保护皮肤光损伤、抗炎和抗微生物等活性。
红景天苷是红景天的主要活性物质、酪醇的糖基化产物。,红景天苷不但有抗缺氧、抗寒冷、抗疲劳、抗微波辐射、抗病毒、抗肿瘤等明显功能,而且还具有增强注意力、提高工作效率、延缓机体衰老、防止老年疾病等功效,尤其在军事医学、航天医学、运动医学和保健医学等方面具有十分重要的应用价值,是一种极具开发前景的环境适应药物,近年来备受关注。
多项发明专利构建了产酪醇的重组大肠杆菌(ZL201310133238.7、ZL20140115011.4、ZL201710091999.9、ZL201711054680.5、ZL201811234765.6、CN202110292484.1和ZL201910754497.9)和重组酿酒酵母(ZL201810601213.8、ZL201711479443.3和CN201910911304.6。上述重组微生物通过表达-酮酸脱羧酶(或芳香醛合酶)和醇脱氢酶,实现了酪醇的从头合成。一些发明专利(ZLZL201910882020.9、CN202010582557.6、CN202110292118.6、CN202110546281.0、CN202110176003.0)公开了一些生产羟基酪醇的重组微生物及其构建方法。一些发明专利(ZL201410115011.4、ZL201610395330.4、ZL201610408741.2、ZL201610361309.2、ZL201711479443.3、CN201811116170.0、CN201910911304.6、CN202011466963.2、CN202111307645.6)公开了一些生产红景天苷的重组微生物及其构建方法。
上述重组微生物的生产水平还难以达到产业化水平,尚需提高。
发明内容
本发明的目的在于提供一种高效合成酪醇、羟基酪醇或红景天苷的新菌种,为酪醇、羟基酪醇或红景天苷的生产提供一种新方法。
本发明所采取的技术方案是:
本发明的第一方面,提供一种基因表达模块,所述基因为:酪醇合成途径酶融合体基因、4-羟基苯乙酸3-羟化酶基因、糖基转移酶基因中的至少一种,所述酪醇合成途径酶融合体、4-羟基苯乙酸3-羟化酶和糖基转移酶均为Spy/Snoop环化。
在本发明的一些实施方式中,所述酪醇合成途径酶融合体基因包括:芳香醛合成酶AAS(Uniprot Q06086)或α-酮酸脱羧酶基因,以及醇脱氢酶基因。
在本发明的一些实施方式中,所述α-酮酸脱羧酶基因为ipdC(Uniprot O07043)、ARO10(Uniprot Q06408)、KDC4(Uniprot C4R7I0)或PDC(Uniprot C5MDS4)。所述醇脱氢酶基因为yahK(Uniprot P75691)、ADH6(Uniprot Q04894)、ahr(Uniprot A0A024L8S1)或adh(Uniprot A0A7M3GDV1)。
在本发明的一些实施方式中,所述4-羟基苯乙酸3-羟化酶基因为EchpaBC(GenBank ACT46003.1、ACT46002.1)、hpaB-hpaC杂合体或sam5(GenBank ABC88666.1);所述hpaB-hpaC杂合体不限定物种来源。
在本发明的一些实施方式中,所述糖基转移酶基因为RrUGT33(UniprotA0A2I6B3R9)、AtUGT85A1(Uniprot W8Q3R5)、RsUGT73B6(Uniprot Q6QDB6)、RsAS(UniProtKB:Q9AR73)、GeGT1(GeneBank MW015078)或VvGT2(Uniprot G9FKG7)。
在本发明的一些实施方式中,所述Spy为SpyTag/SpyCatcher;所述Snoop为SnoopTag/SnoopCatcher。
SpyTag、SnoopTag、SpyCatcher、SnoopCatcher的氨基酸序列如下:
SpyTag:AHIVMVDAYKPTK;
SnoopTag:KLGDIEFIKVNK;
SpyCatcher:GDSATHIKFSKRDEDGKELAGATMELRDSSGKTISTWISDGQVKDFYLYPGKYTFVETAAPDGYEVATAITFTVNEQGQVTVNG;
SnoopCatcher:KPLRGAVFSLQKQHPDYPDIYGAIDQNGTYQNVRTGEDGKLTFKNLSDGKYRLFENSEPAGYKPVQNKPIVAFQIVNGEVRDVTSIVPQDIPATYEFTNGKHYITNEPIPPK。
本发明的第二方面,提供一种重组载体,包含本发明第一方面所述基因表达模块。
在本发明的一些实施方式中,所述重组载体的出发载体为pZBKBP、pZEABP、pZACBP、pZSABP或任何大肠杆菌表达载体。
本发明的第三方面,提供一种重组菌,包含本发明第一方面所述的基因表达模块或本发明第二方面所述的重组载体。
在本发明的一些实施方式中,所述重组菌为以下任一种:
(a)将本发明第一方面所述的酪醇合成途径酶融合体基因导入宿主菌中得重组菌1;
(b)将本发明第一方面所述的4-羟基苯乙酸3-羟化酶导入宿主菌中得重组菌2;
(c)将本发明第一方面所述的糖基转移酶基因导入宿主菌中得重组菌3。
在本发明的一些实施方式中,所述宿主菌是普通大肠杆菌,如K12衍生菌(BW25113、MG1665、W3110、DH10B、BW2952、MDS42及其衍生菌)、BL21(DE3)及其衍生菌、BREL606、W、DH1等,也可以是产酪氨酸的大肠杆菌,优选是产酪氨酸的大肠杆菌。
在本发明的一些实施方式,本发明第一方面所述的基因表达模块通过重组载体导入宿主菌中。
本发明的第四方面,提供本发明第一方面所述的基因表达模块或本发明第二方面所述的重组载体或本发明第三方面所述的重组菌在制备酪醇、羟基酪醇或红景天苷中的应用。
本发明的第五方面,提供一种制备酪醇、羟基酪醇或红景天苷的方法,应用本发明第三方面任一项所述的重组菌发酵,得到酪醇、羟基酪醇或红景天苷。
在本发明的一些实施方式中,具体为在含有葡萄糖、酪氨酸的培养基中应用本发明第三方面任一项所述的重组菌发酵,得到酪醇、羟基酪醇或红景天苷。
在本发明的一些实施方式中,所述酪氨酸可以是外源添加,也可以是添加酪氨酸产生菌。
在本发明的一些实施方式中,所述酪氨酸产生菌可以为任何产酪氨酸的大肠杆菌,如本实验之前公开的酪氨酸产生菌大肠杆菌TYR-14B1(Frontiers in Microbiology,12(2021)710405)。
在本发明的一些实施方式中,所述发酵可以采用单菌发酵法、模块共培养法和模块共催化法。
在本发明的一些优选实施方式中,优选为模块共催化法。
在本发明的一些实施方式中,对酪醇合成而言,采用酪氨酸产生菌和重组菌1进行模块共催化;对羟基酪醇合成而言,采用酪氨酸产生菌、重组菌1和重组菌2进行模块共催化;对红景天苷合成而言,采用酪氨酸产生菌、重组菌1和重组菌3进行模块共催化。
在本发明的一些实施方式中,对酪醇合成而言,酪氨酸产生菌和重组菌1的重量比为(1-5):(1-5)。
在本发明的一些实施方式中,对羟基酪醇合成而言,酪氨酸产生菌、重组菌1和重组菌2的重量比为(1-5):(1-5):(1-5)。
在本发明的一些实施方式中,对红景天苷合成而言,酪氨酸产生菌、重组菌1和重组菌3的重量比为(1-5):(1-5):(1-5)。
在本发明的一些实施方式中,所述菌的接种量为:起始OD600为20~50。
在本发明的一些实施方式中,所述催化条件为30~42℃、150~250rpm下生物催化20~40h。
本发明的有益效果是:
本发明在宿主菌中表达了酪醇合成途径酶融合体来生产酪醇,再引入4-羟基苯乙酸3-羟化酶HpaB生产羟基酪醇,引入糖基转移酶生产红景天苷;Spy(或Snoop)环化还可通过热处理以增加细胞的通透性,避免细胞的传质限制。通过模块共培养因将长代谢途径分割成多个途径、并可利用不同宿主细胞进行表达,减轻了对宿主细胞的代谢负担;而且不同途径可选用各自最合适的宿主细胞进行表达;可通过调整模块菌的比例来控制模块的适配性;即插即用,容易组装成不同途径以合成不同产物。还能避免代谢中间物和目标产物毒性的影响。
具体实施方式
以下将结合实施例对本发明的构思及产生的技术效果进行清楚、完整地描述,以充分地理解本发明的目的、特征和效果。显然,所描述的实施例只是本发明的一部分实施例,而不是全部实施例,基于本发明的实施例,本领域的技术人员在不付出创造性劳动的前提下所获得的其他实施例,均属于本发明保护的范围。
SpyTag、SnoopTag、SpyCatcher、SnoopCatcher的氨基酸序列如下:
SpyTag:AHIVMVDAYKPTK(SEQ ID NO.1);
SnoopTag:KLGDIEFIKVNK(SEQ ID NO.2);
SpyCatcher:GDSATHIKFSKRDEDGKELAGATMELRDSSGKTISTWISDGQVKDFYLYPGKYTFVETAAPDGYEVATAITFTVNEQGQVTVNG(SEQ ID NO.3);
SnoopCatcher:KPLRGAVFSLQKQHPDYPDIYGAIDQNGTYQNVRTGEDGKLTFKNLSDGKYRLFENSEPAGYKPVQNKPIVAFQIVNGEVRDVTSIVPQDIPATYEFTNGKHYITNEPIPPK(SEQ ID NO.4)。
实施例1模块菌的构建
提供酪氨酸的模块菌为产酪氨酸大肠杆菌,如本实验之前公开的酪氨酸产生菌大肠杆菌TYR-14B1为酪氨酸模块菌(Frontiers in Microbiology,12(2021)710405)。
AAS(Uniprot Q06086)、yahK基因(Uniprot P75691)(它们各自的序列可在Uniprot数据库中可以查到),用(GSG)2进行融合表达,而且在N端加上SpyTag序列、在C端加上SpyCatcher序列,委托商业公司进行全基因合成SpyTag-(GSG)2-aas-(GSG)2-yahK-(GSG)2-SpyCatcher片段,NheI/KpnI酶切连接到实验室之前构建的表达载体pZBKBP(Journal of Industrial Microbiology Biotechnology 42(2015)627-636),得到pZBK-ST-AAS-yahK-SC;转化到大肠杆菌BW25113化学感受态细胞中,得到酪醇模块菌大肠杆菌BW25113(pZBK-ST-AAS-yahK-SC),即前述重组菌1。
EchpaBC(GenBank ACT46003.1、ACT46002.1),在N端加上SpyTag序列、在C端加上SpyCatcher序列,委托商业公司进行全基因合成SpyTag-(GSG)2-hpaBC-(GSG)2-SpyCatcher片段,NheI/KpnI酶切连接到pZBKBP载体,得到pZBK-ST-hpaBC-SC;转化到大肠杆菌BW25113化学感受态细胞中,得到羟基酪醇模块菌大肠杆菌BW25113(pZBK-ST-hpaBC-SC)。
AtUGT85A1(Uniprot W8Q3R5),在N端加上SpyTag序列、在C端加上SpyCatcher序列,委托商业公司进行全基因合成SpyTag-(GSG)2-AtUGT85A1-(GSG)2-SpyCatcher片段,NheI/KpnI酶切连接到pZBKBP载体得到pZBK-ST-AtUGT85A1-SC;转化到大肠杆菌BW25113化学感受态细胞中,得到红景天苷模块菌大肠杆菌BW25113(pZBK-ST-AtUGT85A1-SC)。
实施例2模块共催化合成酪醇
挑取LB(1%胰蛋白胨、0.5%酵母抽提物、1%NaCl)平板上的酪氨酸模块菌和酪醇模块菌单菌落,接种至LB液体培养基中,于37℃、200rpm下培养约14h。于4℃、2800g离心15min冷冻离心、并用冷的生理盐水洗涤、收集菌体。将酪醇模块菌重悬在0.1M pH7.0磷酸盐缓冲液中至OD600为100制备菌悬液。菌悬液置于55℃热处理20min。菌悬液再次冷冻离心并用冷生理盐水洗涤收集菌体。最后将酪氨酸模块菌和酪醇模块菌按4:1(g/g)比例重悬至10mL含20g/L葡萄糖、10mM硫酸镁和1.5g/L维生素C的0.1M pH7.0磷酸盐缓冲液中至OD600为25,于37℃、200rpm下生物催化24h。HPLC(LC-20A,岛津)分析酪醇产量。
色谱条件:色谱柱Inertsil ODS-SP柱(5μm,4.6×150mm,GL Sciences Inc.);流动相:A为0.2%三氟乙酸、B相为100%甲醇;梯度洗脱:0-20min、甲醇浓度由14%升至45%;20-30min、甲醇浓度由45%降至14%;流速0.5mL/min;柱温30℃;检测波长280nm。
结果可产1.3g/L酪醇。
实施例3模块共催化合成羟基酪醇
挑取LB平板上的酪氨酸模块菌、酪醇模块菌和羟基酪醇模块菌单菌落,接种至LB液体培养基中,于37℃、200rpm下培养约14h。冷冻离心并用冷的生理盐水洗涤、收集菌体。将酪醇模块菌和羟基酪醇重悬在0.1M pH7.0磷酸盐缓冲液中至OD600为100制备菌悬液。菌悬液置于55℃热处理20min。菌悬液再次冷冻离心并用冷生理盐水洗涤收集菌体。最后将酪氨酸模块菌:酪醇模块菌:羟基酪醇按4:1:1(g/g)比例重悬至10mL含20g/L葡萄糖、10mM硫酸镁和1.5g/L维生素C的0.1M pH7.0磷酸盐缓冲液中至OD600为25,于37℃、200rpm下生物催化24h。HPLC(LC-20A,岛津)分析羟基酪醇产量。
色谱条件:色谱柱Inertsil ODS-SP柱(5μm,4.6×150mm,GL Sciences Inc.);流动相:A为0.2%三氟乙酸、B相为100%甲醇;梯度洗脱:0-20min、甲醇浓度由14%升至45%;20-30min、甲醇浓度由45%降至14%;流速0.5mL/min;柱温30℃;检测波长280nm。
结果可产193.2mg/L羟基酪醇。
实施例4模块共催化合成红景天苷
挑取LB平板上的酪氨酸模块菌、酪醇模块菌和红景天苷模块菌单菌落,接种至LB液体培养基中,于37℃、200rpm下培养约14h。冷冻离心并用冷的生理盐水洗涤、收集菌体。将酪醇模块菌和红景天苷模块菌重悬在0.1M pH7.0磷酸盐缓冲液中至OD600为100制备菌悬液。菌悬液置于55℃热处理20min。菌悬液再次冷冻离心并用冷生理盐水洗涤收集菌体。最后将酪氨酸模块菌:酪醇模块菌:红景天苷模块菌按4:1:1(g/g)比例重悬至10mL含20g/L葡萄糖、10mM硫酸镁和1.5g/L维生素C的0.1M pH7.0磷酸盐缓冲液中至OD600为25,于37℃、200rpm下生物催化24h。
按实施例2方法HPLC(LC-20A,岛津)分析红景天苷产量,可产146.1mg/L红景天苷。
除了本实施例提及的方式外,还可采用模块共培养的方法生产酪醇、羟基酪醇或红景天苷。即不同模块菌分别进行种子培养,按一定比例接种到发酵培养基中进行发酵,也可以单菌发酵法。环化酶也可以直接转化到酪氨酸产生菌中进行单菌发酵。环化酶基因也可以整合到大肠杆菌染色体上,构建相应的重组大肠杆菌。
除了本实施例提及的方式外,相应酶还可以用其它表达载体进行表达,也可以整合到大肠杆菌染色体上进行表达。
上述具体实施方式对本发明作了详细说明,但是本发明不限于上述实施例,在所属技术领域普通技术人员所具备的知识范围内,还可以在不脱离本发明宗旨的前提下作出各种变化。此外,在不冲突的情况下,本发明的实施例及实施例中的特征可以相互组合。
SEQUENCE LISTING
<110> 中山大学
<120> 一种重组大肠杆菌及其生产酪醇、羟基酪醇或红景天苷的方法
<130>
<160> 4
<170> PatentIn version 3.5
<210> 1
<211> 13
<212> PRT
<213> SpyTag
<400> 1
Ala His Ile Val Met Val Asp Ala Tyr Lys Pro Thr Lys
1 5 10
<210> 2
<211> 12
<212> PRT
<213> SnoopTag
<400> 2
Lys Leu Gly Asp Ile Glu Phe Ile Lys Val Asn Lys
1 5 10
<210> 3
<211> 84
<212> PRT
<213> SpyCatcher
<400> 3
Gly Asp Ser Ala Thr His Ile Lys Phe Ser Lys Arg Asp Glu Asp Gly
1 5 10 15
Lys Glu Leu Ala Gly Ala Thr Met Glu Leu Arg Asp Ser Ser Gly Lys
20 25 30
Thr Ile Ser Thr Trp Ile Ser Asp Gly Gln Val Lys Asp Phe Tyr Leu
35 40 45
Tyr Pro Gly Lys Tyr Thr Phe Val Glu Thr Ala Ala Pro Asp Gly Tyr
50 55 60
Glu Val Ala Thr Ala Ile Thr Phe Thr Val Asn Glu Gln Gly Gln Val
65 70 75 80
Thr Val Asn Gly
<210> 4
<211> 112
<212> PRT
<213> SnoopCatcher
<400> 4
Lys Pro Leu Arg Gly Ala Val Phe Ser Leu Gln Lys Gln His Pro Asp
1 5 10 15
Tyr Pro Asp Ile Tyr Gly Ala Ile Asp Gln Asn Gly Thr Tyr Gln Asn
20 25 30
Val Arg Thr Gly Glu Asp Gly Lys Leu Thr Phe Lys Asn Leu Ser Asp
35 40 45
Gly Lys Tyr Arg Leu Phe Glu Asn Ser Glu Pro Ala Gly Tyr Lys Pro
50 55 60
Val Gln Asn Lys Pro Ile Val Ala Phe Gln Ile Val Asn Gly Glu Val
65 70 75 80
Arg Asp Val Thr Ser Ile Val Pro Gln Asp Ile Pro Ala Thr Tyr Glu
85 90 95
Phe Thr Asn Gly Lys His Tyr Ile Thr Asn Glu Pro Ile Pro Pro Lys
100 105 110
Claims (10)
1.一种基因表达模块,所述基因表达模块包含:酪醇合成途径酶融合体基因、4-羟基苯乙酸3-羟化酶基因、糖基转移酶基因中的至少一种;所述酪醇合成途径酶融合体、4-羟基苯乙酸3-羟化酶和糖基转移酶均为Spy/Snoop环化。
2.根据权利要求1所述的基因表达模块,其特征在于,所述酪醇合成途径酶融合体基因包含:芳香醛合成酶AAS/α-酮酸脱羧酶基因和醇脱氢酶基因;所述α-酮酸脱羧酶基因为ipdC、ARO10、KDC4或PDC;所述醇脱氢酶基因为yahK、ADH6、ahr或adh。
3.根据权利要求1所述的基因表达模块,其特征在于,所述4-羟基苯乙酸3-羟化酶基因为EchpaBC、hpaB-hpaC杂合体或sam5。
4.根据权利要求1所述的基因表达模块,其特征在于,所述糖基转移酶基因为RrUGT33、AtUGT85A1、RsUGT73B6、RsAS、GeGT1或VvGT2。
5.一种重组载体,包含权利要求1~4任一项所述的基因表达模块。
6.一种重组菌,包含权利要求1~4任一项所述的基因表达模块或权利要求5所述的重组载体。
7.根据权利要求6所述的重组菌,其特征在于,所述重组菌为以下任一种:
(a)将权利要求1~4任一项所述的酪醇合成途径酶融合体基因导入受体菌中;
(b)将权利要求1~4任一项所述的4-羟基苯乙酸3-羟化酶基因导入受体菌中;
(c)将权利要求1~4任一项所述的糖基转移酶基因导入受体菌中。
8.权利要求1~4任一项所述的基因表达模块或权利要求5所述的重组载体或权利要求6~7任一项所述的重组菌在制备酪醇、羟基酪醇或红景天苷中的应用。
9.一种制备酪醇、羟基酪醇或红景天苷的方法,其特征在于,通过权利要求7~8任一项所述的重组菌发酵。
10.根据权利要求9所述的方法,其特征在于,所述发酵方法为单菌发酵法、模块共培养或模块共催化法;优选为模块共催化法。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202210466075.3A CN114774451A (zh) | 2022-04-29 | 2022-04-29 | 一种重组大肠杆菌及其生产酪醇、羟基酪醇或红景天苷的方法 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202210466075.3A CN114774451A (zh) | 2022-04-29 | 2022-04-29 | 一种重组大肠杆菌及其生产酪醇、羟基酪醇或红景天苷的方法 |
Publications (1)
Publication Number | Publication Date |
---|---|
CN114774451A true CN114774451A (zh) | 2022-07-22 |
Family
ID=82434326
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202210466075.3A Pending CN114774451A (zh) | 2022-04-29 | 2022-04-29 | 一种重组大肠杆菌及其生产酪醇、羟基酪醇或红景天苷的方法 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN114774451A (zh) |
Citations (11)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN103352031A (zh) * | 2013-04-26 | 2013-10-16 | 中山大学 | 一种糖基转移酶基因及应用 |
KR101632697B1 (ko) * | 2015-04-28 | 2016-06-22 | 건국대학교 산학협력단 | 대사 조절된 대장균을 이용한 살리드로사이드의 제조방법 |
CN107201331A (zh) * | 2016-03-18 | 2017-09-26 | 中国科学院天津工业生物技术研究所 | 表达羟基酪醇和羟基酪醇葡萄糖苷的大肠杆菌及构建方法及应用 |
CN107586794A (zh) * | 2017-11-01 | 2018-01-16 | 北京化工大学 | 异源代谢途径生产酪醇及羟基酪醇的方法 |
CN109295113A (zh) * | 2018-10-23 | 2019-02-01 | 江南大学 | 一种生产羟基酪醇的方法 |
CN109988722A (zh) * | 2017-12-29 | 2019-07-09 | 中国科学院天津工业生物技术研究所 | 一种重组酿酒酵母菌株及其应用和生产酪醇和/或红景天苷的方法 |
CN111411128A (zh) * | 2020-03-03 | 2020-07-14 | 湖北大学 | 一种生产α,ω-二元羧酸的整细胞生物催化方法及其应用 |
CN111748508A (zh) * | 2020-06-23 | 2020-10-09 | 厦门大学 | 一种高产羟基酪醇的大肠杆菌构建方法及应用 |
US20210254081A1 (en) * | 2018-06-12 | 2021-08-19 | Shandong Henglu Biotech. Co., Ltd | Yeast producing tyrosol or hydroxytyrosol, and construction methods thereof |
CN113897325A (zh) * | 2021-11-05 | 2022-01-07 | 江南大学 | 一种生产红景天苷的重组大肠杆菌及其构建方法和应用 |
CN114381484A (zh) * | 2021-12-09 | 2022-04-22 | 山东大学 | UGT85A1或RrUGT3在催化多种底物生成糖苷化合物中的应用 |
-
2022
- 2022-04-29 CN CN202210466075.3A patent/CN114774451A/zh active Pending
Patent Citations (11)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN103352031A (zh) * | 2013-04-26 | 2013-10-16 | 中山大学 | 一种糖基转移酶基因及应用 |
KR101632697B1 (ko) * | 2015-04-28 | 2016-06-22 | 건국대학교 산학협력단 | 대사 조절된 대장균을 이용한 살리드로사이드의 제조방법 |
CN107201331A (zh) * | 2016-03-18 | 2017-09-26 | 中国科学院天津工业生物技术研究所 | 表达羟基酪醇和羟基酪醇葡萄糖苷的大肠杆菌及构建方法及应用 |
CN107586794A (zh) * | 2017-11-01 | 2018-01-16 | 北京化工大学 | 异源代谢途径生产酪醇及羟基酪醇的方法 |
CN109988722A (zh) * | 2017-12-29 | 2019-07-09 | 中国科学院天津工业生物技术研究所 | 一种重组酿酒酵母菌株及其应用和生产酪醇和/或红景天苷的方法 |
US20210254081A1 (en) * | 2018-06-12 | 2021-08-19 | Shandong Henglu Biotech. Co., Ltd | Yeast producing tyrosol or hydroxytyrosol, and construction methods thereof |
CN109295113A (zh) * | 2018-10-23 | 2019-02-01 | 江南大学 | 一种生产羟基酪醇的方法 |
CN111411128A (zh) * | 2020-03-03 | 2020-07-14 | 湖北大学 | 一种生产α,ω-二元羧酸的整细胞生物催化方法及其应用 |
CN111748508A (zh) * | 2020-06-23 | 2020-10-09 | 厦门大学 | 一种高产羟基酪醇的大肠杆菌构建方法及应用 |
CN113897325A (zh) * | 2021-11-05 | 2022-01-07 | 江南大学 | 一种生产红景天苷的重组大肠杆菌及其构建方法和应用 |
CN114381484A (zh) * | 2021-12-09 | 2022-04-22 | 山东大学 | UGT85A1或RrUGT3在催化多种底物生成糖苷化合物中的应用 |
Non-Patent Citations (6)
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN107435049B (zh) | 一种生产红景天苷的重组大肠杆菌及构建方法及应用 | |
CN107099516A (zh) | 7β‑羟基甾醇脱氢酶突变体及其在熊脱氧胆酸合成中的应用 | |
CN110791493B (zh) | 一种天冬氨酸氨裂合酶突变体及其应用 | |
CN103131721B (zh) | 瘤胃菌d-塔格糖3-差向异构酶的核苷酸序列及其应用 | |
CN101824404B (zh) | 白藜芦醇合酶及其编码基因与应用 | |
CN109988722B (zh) | 一种重组酿酒酵母菌株及其应用和生产酪醇和/或红景天苷的方法 | |
KR101511361B1 (ko) | 헴 생산성이 증가된 재조합 대장균 및 이를 이용한 헴 생산 방법 | |
CN112813013B (zh) | 一种生产羟基酪醇的重组大肠杆菌及其应用 | |
CN110616180B (zh) | 一种生产羟基酪醇的工程菌及其应用 | |
CN108949852B (zh) | 一种利用全细胞催化制备木糖醇的方法 | |
KR20110070977A (ko) | 생물학적 헴철 생산 방법 및 그에 의해 생산된 헴철 추출물을 포함하는 철분보충 조성물 | |
WO2020244031A1 (zh) | 一种石莼多糖裂解酶及其编码基因与应用 | |
KR20220158770A (ko) | 칸나비노이드 및 칸나비노이드 전구체의 생합성 | |
CN109652481A (zh) | 一种环糊精糖基转移酶在生产α-糖基橙皮苷中的应用 | |
CN106947772B (zh) | 一种羰基还原酶突变体及其在手性醇制备中的应用 | |
Zhang et al. | Biocatalytic conversion of a lactose-rich dairy waste into D-tagatose, D-arabitol and galactitol using sequential whole cell and fermentation technologies | |
CN114621883B (zh) | 一种具有强化防晒效果的酿酒酵母及其应用 | |
CN117467627B (zh) | 一种烯醛还原酶突变体及其编码基因和应用 | |
CN110616205B (zh) | 一种用于黄酮糖苷合成制备的黄酮合酶 | |
CN114774451A (zh) | 一种重组大肠杆菌及其生产酪醇、羟基酪醇或红景天苷的方法 | |
CN109628476B (zh) | 一种利用全细胞转化生产4-羟基异亮氨酸的方法 | |
CN110669713A (zh) | 一种合成d-柠檬烯的基因工程菌及其构建方法与应用 | |
CN116731886A (zh) | 产糖基化虾青素的工程菌及其构建方法与应用 | |
CN109234216B (zh) | 一种生产鲨烯的基因工程菌及其方法 | |
CN110004099A (zh) | 一种红景天苷的发酵生产方法 |
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 |