CN105247067A - 融合蛋白酶 - Google Patents
融合蛋白酶 Download PDFInfo
- Publication number
- CN105247067A CN105247067A CN201480029964.7A CN201480029964A CN105247067A CN 105247067 A CN105247067 A CN 105247067A CN 201480029964 A CN201480029964 A CN 201480029964A CN 105247067 A CN105247067 A CN 105247067A
- Authority
- CN
- China
- Prior art keywords
- protease
- fusion
- protein
- bifunctional fusion
- fusion protease
- 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.)
- Withdrawn
Links
- 108091005804 Peptidases Proteins 0.000 title claims abstract description 409
- 239000004365 Protease Substances 0.000 title claims abstract description 406
- 230000004927 fusion Effects 0.000 title claims abstract description 302
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 title claims abstract 23
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 187
- 230000001588 bifunctional effect Effects 0.000 claims abstract description 185
- 102000004169 proteins and genes Human genes 0.000 claims abstract description 179
- 102000004400 Aminopeptidases Human genes 0.000 claims abstract description 7
- 108090000915 Aminopeptidases Proteins 0.000 claims abstract description 7
- 102000004190 Enzymes Human genes 0.000 claims description 76
- 108090000790 Enzymes Proteins 0.000 claims description 76
- 238000003776 cleavage reaction Methods 0.000 claims description 54
- 241000709664 Picornaviridae Species 0.000 claims description 49
- 230000000694 effects Effects 0.000 claims description 41
- 108090000765 processed proteins & peptides Proteins 0.000 claims description 40
- 238000000034 method Methods 0.000 claims description 30
- 230000002797 proteolythic effect Effects 0.000 claims description 20
- 238000006467 substitution reaction Methods 0.000 claims description 15
- 102000004196 processed proteins & peptides Human genes 0.000 claims description 14
- 238000012217 deletion Methods 0.000 claims description 12
- 230000037430 deletion Effects 0.000 claims description 12
- 125000003275 alpha amino acid group Chemical group 0.000 claims description 11
- 108010091324 3C proteases Proteins 0.000 claims description 7
- 230000003197 catalytic effect Effects 0.000 claims description 6
- 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 5
- 241000894006 Bacteria Species 0.000 claims description 5
- 241000430519 Human rhinovirus sp. Species 0.000 claims description 5
- 102400000108 N-terminal peptide Human genes 0.000 claims description 4
- 101800000597 N-terminal peptide Proteins 0.000 claims description 4
- 101800000135 N-terminal protein Proteins 0.000 claims description 4
- 101800001452 P1 proteinase Proteins 0.000 claims description 4
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 claims description 4
- 241000194017 Streptococcus Species 0.000 claims description 3
- 239000004310 lactic acid Substances 0.000 claims description 2
- 235000014655 lactic acid Nutrition 0.000 claims description 2
- 102000035195 Peptidases Human genes 0.000 abstract description 386
- 108020001507 fusion proteins Proteins 0.000 abstract description 61
- 102000037865 fusion proteins Human genes 0.000 abstract description 61
- 238000004519 manufacturing process Methods 0.000 abstract description 16
- 241001144416 Picornavirales Species 0.000 abstract 1
- 235000019419 proteases Nutrition 0.000 description 337
- 235000018102 proteins Nutrition 0.000 description 165
- 229940088598 enzyme Drugs 0.000 description 72
- 238000006243 chemical reaction Methods 0.000 description 48
- 230000014509 gene expression Effects 0.000 description 46
- 239000000758 substrate Substances 0.000 description 43
- 210000004027 cell Anatomy 0.000 description 40
- 230000007017 scission Effects 0.000 description 35
- 125000000539 amino acid group Chemical group 0.000 description 30
- DTHNMHAUYICORS-KTKZVXAJSA-N Glucagon-like peptide 1 Chemical compound C([C@@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](C)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCCCN)C(=O)NCC(=O)N[C@@H](CCCNC(N)=N)C(N)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CCCCN)NC(=O)[C@H](C)NC(=O)[C@H](C)NC(=O)[C@H](CCC(N)=O)NC(=O)CNC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CO)NC(=O)[C@H](CO)NC(=O)[C@@H](NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](NC(=O)[C@H](CC=1C=CC=CC=1)NC(=O)[C@@H](NC(=O)CNC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](C)NC(=O)[C@@H](N)CC=1N=CNC=1)[C@@H](C)O)[C@@H](C)O)C(C)C)C1=CC=CC=C1 DTHNMHAUYICORS-KTKZVXAJSA-N 0.000 description 29
- 238000000746 purification Methods 0.000 description 29
- MASNOZXLGMXCHN-ZLPAWPGGSA-N glucagon Chemical compound C([C@@H](C(=O)N[C@H](C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O)C(C)C)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@H](C)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H](CO)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CCCCN)NC(=O)[C@H](CO)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](NC(=O)[C@H](CC=1C=CC=CC=1)NC(=O)[C@@H](NC(=O)CNC(=O)[C@H](CCC(N)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](N)CC=1NC=NC=1)[C@@H](C)O)[C@@H](C)O)C1=CC=CC=C1 MASNOZXLGMXCHN-ZLPAWPGGSA-N 0.000 description 24
- 229960004666 glucagon Drugs 0.000 description 24
- 102220281712 rs777988634 Human genes 0.000 description 24
- AUHJXHCVECGTKR-DQNUUZSMSA-N dnc007903 Chemical compound CC[C@H](C)[C@H](N)C(=O)N[C@@H](CCCCN)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C)C(=O)N1[C@H](C(=O)NCC(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](CO)C(=O)N2[C@@H](CCC2)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC=2C=CC(O)=CC=2)C(=O)N[C@@H](CC=2C=CC(O)=CC=2)C(=O)N[C@@H](C)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC=2NC=NC=2)C(=O)N[C@@H](CC=2C=CC(O)=CC=2)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC=2C=CC(O)=CC=2)C(N)=O)CCC1 AUHJXHCVECGTKR-DQNUUZSMSA-N 0.000 description 23
- 238000011534 incubation Methods 0.000 description 22
- 241000588724 Escherichia coli Species 0.000 description 20
- 235000001014 amino acid Nutrition 0.000 description 20
- 239000002953 phosphate buffered saline Substances 0.000 description 20
- 238000001819 mass spectrum Methods 0.000 description 19
- 239000000872 buffer Substances 0.000 description 18
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 17
- 244000057717 Streptococcus lactis Species 0.000 description 17
- 101800000224 Glucagon-like peptide 1 Proteins 0.000 description 16
- 102100040918 Pro-glucagon Human genes 0.000 description 16
- 238000006911 enzymatic reaction Methods 0.000 description 16
- 239000013612 plasmid Substances 0.000 description 16
- 108091028043 Nucleic acid sequence Proteins 0.000 description 15
- 239000013598 vector Substances 0.000 description 15
- 229940024606 amino acid Drugs 0.000 description 14
- 238000004458 analytical method Methods 0.000 description 13
- 238000004895 liquid chromatography mass spectrometry Methods 0.000 description 13
- 102000051325 Glucagon Human genes 0.000 description 12
- 108060003199 Glucagon Proteins 0.000 description 12
- RAXXELZNTBOGNW-UHFFFAOYSA-N imidazole Natural products C1=CNC=N1 RAXXELZNTBOGNW-UHFFFAOYSA-N 0.000 description 12
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 12
- 108010076504 Protein Sorting Signals Proteins 0.000 description 11
- 241000194021 Streptococcus suis Species 0.000 description 11
- 150000001413 amino acids Chemical class 0.000 description 11
- 210000004899 c-terminal region Anatomy 0.000 description 11
- 241000709675 Coxsackievirus B3 Species 0.000 description 10
- 235000014897 Streptococcus lactis Nutrition 0.000 description 10
- 238000007792 addition Methods 0.000 description 9
- 238000001597 immobilized metal affinity chromatography Methods 0.000 description 9
- 238000005119 centrifugation Methods 0.000 description 8
- 239000012634 fragment Substances 0.000 description 8
- 150000007523 nucleic acids Chemical group 0.000 description 8
- 239000011780 sodium chloride Substances 0.000 description 8
- YNXLOPYTAAFMTN-SBUIBGKBSA-N C([C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CCCCN)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C)C(=O)N1[C@@H](CCC1)C(=O)NCC(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](CO)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)N[C@@H](C)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC=1NC=NC=1)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(N)=O)C1=CC=C(O)C=C1 Chemical compound C([C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CCCCN)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C)C(=O)N1[C@@H](CCC1)C(=O)NCC(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](CO)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)N[C@@H](C)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC=1NC=NC=1)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(N)=O)C1=CC=C(O)C=C1 YNXLOPYTAAFMTN-SBUIBGKBSA-N 0.000 description 7
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 7
- 239000001488 sodium phosphate Substances 0.000 description 7
- 229910000162 sodium phosphate Inorganic materials 0.000 description 7
- 238000013518 transcription Methods 0.000 description 7
- 230000035897 transcription Effects 0.000 description 7
- RYFMWSXOAZQYPI-UHFFFAOYSA-K trisodium phosphate Chemical compound [Na+].[Na+].[Na+].[O-]P([O-])([O-])=O RYFMWSXOAZQYPI-UHFFFAOYSA-K 0.000 description 7
- 108010016626 Dipeptides Proteins 0.000 description 6
- 241000709661 Enterovirus Species 0.000 description 6
- 241000709721 Hepatovirus A Species 0.000 description 6
- 101710175625 Maltose/maltodextrin-binding periplasmic protein Proteins 0.000 description 6
- 108010088847 Peptide YY Proteins 0.000 description 6
- 102100029909 Peptide YY Human genes 0.000 description 6
- 102100036407 Thioredoxin Human genes 0.000 description 6
- 238000013461 design Methods 0.000 description 6
- 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 description 6
- 238000003259 recombinant expression Methods 0.000 description 6
- 108060008226 thioredoxin Proteins 0.000 description 6
- 241000193830 Bacillus <bacterium> Species 0.000 description 5
- 241000709687 Coxsackievirus Species 0.000 description 5
- 101000930822 Giardia intestinalis Dipeptidyl-peptidase 4 Proteins 0.000 description 5
- 102000007056 Recombinant Fusion Proteins Human genes 0.000 description 5
- 108010008281 Recombinant Fusion Proteins Proteins 0.000 description 5
- 230000008901 benefit Effects 0.000 description 5
- 229960003669 carbenicillin Drugs 0.000 description 5
- FPPNZSSZRUTDAP-UWFZAAFLSA-N carbenicillin Chemical compound N([C@H]1[C@H]2SC([C@@H](N2C1=O)C(O)=O)(C)C)C(=O)C(C(O)=O)C1=CC=CC=C1 FPPNZSSZRUTDAP-UWFZAAFLSA-N 0.000 description 5
- 230000002255 enzymatic effect Effects 0.000 description 5
- 238000002474 experimental method Methods 0.000 description 5
- 239000013604 expression vector Substances 0.000 description 5
- 230000000155 isotopic effect Effects 0.000 description 5
- 229920001184 polypeptide Polymers 0.000 description 5
- 239000011347 resin Substances 0.000 description 5
- 229920005989 resin Polymers 0.000 description 5
- 108090000907 ribosomal protein L9 Proteins 0.000 description 5
- 102000004346 ribosomal protein L9 Human genes 0.000 description 5
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 4
- 241001522878 Escherichia coli B Species 0.000 description 4
- 241000193385 Geobacillus stearothermophilus Species 0.000 description 4
- 125000003338 L-glutaminyl group Chemical group O=C([*])[C@](N([H])[H])([H])C([H])([H])C([H])([H])C(=O)N([H])[H] 0.000 description 4
- 241000700605 Viruses Species 0.000 description 4
- 230000001580 bacterial effect Effects 0.000 description 4
- 239000003814 drug Substances 0.000 description 4
- 239000013613 expression plasmid Substances 0.000 description 4
- 239000000499 gel Substances 0.000 description 4
- 238000004128 high performance liquid chromatography Methods 0.000 description 4
- 239000012535 impurity Substances 0.000 description 4
- 229930182817 methionine Natural products 0.000 description 4
- 108020004707 nucleic acids Proteins 0.000 description 4
- 102000039446 nucleic acids Human genes 0.000 description 4
- 239000008188 pellet Substances 0.000 description 4
- 239000002243 precursor Substances 0.000 description 4
- 230000008569 process Effects 0.000 description 4
- 238000012545 processing Methods 0.000 description 4
- 238000000926 separation method Methods 0.000 description 4
- 239000006228 supernatant Substances 0.000 description 4
- 244000063299 Bacillus subtilis Species 0.000 description 3
- 235000014469 Bacillus subtilis Nutrition 0.000 description 3
- 102100025012 Dipeptidyl peptidase 4 Human genes 0.000 description 3
- 241001198387 Escherichia coli BL21(DE3) Species 0.000 description 3
- 241001646716 Escherichia coli K-12 Species 0.000 description 3
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 3
- FFEARJCKVFRZRR-BYPYZUCNSA-N L-methionine Chemical compound CSCC[C@H](N)C(O)=O FFEARJCKVFRZRR-BYPYZUCNSA-N 0.000 description 3
- KZNQNBZMBZJQJO-UHFFFAOYSA-N N-glycyl-L-proline Natural products NCC(=O)N1CCCC1C(O)=O KZNQNBZMBZJQJO-UHFFFAOYSA-N 0.000 description 3
- 125000000729 N-terminal amino-acid group Chemical group 0.000 description 3
- 241000204666 Thermotoga maritima Species 0.000 description 3
- -1 amyL Chemical class 0.000 description 3
- 239000003153 chemical reaction reagent Substances 0.000 description 3
- 238000010367 cloning Methods 0.000 description 3
- LOKCTEFSRHRXRJ-UHFFFAOYSA-I dipotassium trisodium dihydrogen phosphate hydrogen phosphate dichloride Chemical compound P(=O)(O)(O)[O-].[K+].P(=O)(O)([O-])[O-].[Na+].[Na+].[Cl-].[K+].[Cl-].[Na+] LOKCTEFSRHRXRJ-UHFFFAOYSA-I 0.000 description 3
- 229940079593 drug Drugs 0.000 description 3
- 238000002523 gelfiltration Methods 0.000 description 3
- KZNQNBZMBZJQJO-YFKPBYRVSA-N glyclproline Chemical group NCC(=O)N1CCC[C@H]1C(O)=O KZNQNBZMBZJQJO-YFKPBYRVSA-N 0.000 description 3
- 108010077515 glycylproline Proteins 0.000 description 3
- 230000006698 induction Effects 0.000 description 3
- 150000002500 ions Chemical class 0.000 description 3
- 238000012531 mass spectrometric analysis of intact mass Methods 0.000 description 3
- 239000002609 medium Substances 0.000 description 3
- 230000000813 microbial effect Effects 0.000 description 3
- 239000011535 reaction buffer Substances 0.000 description 3
- 230000035484 reaction time Effects 0.000 description 3
- 238000011084 recovery Methods 0.000 description 3
- 230000010076 replication Effects 0.000 description 3
- 230000003248 secreting effect Effects 0.000 description 3
- 238000005063 solubilization Methods 0.000 description 3
- 230000007928 solubilization Effects 0.000 description 3
- 230000003381 solubilizing effect Effects 0.000 description 3
- 239000002904 solvent Substances 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- HNSDLXPSAYFUHK-UHFFFAOYSA-N 1,4-bis(2-ethylhexyl) sulfosuccinate Chemical compound CCCCC(CC)COC(=O)CC(S(O)(=O)=O)C(=O)OCC(CC)CCCC HNSDLXPSAYFUHK-UHFFFAOYSA-N 0.000 description 2
- QNAYBMKLOCPYGJ-UHFFFAOYSA-N Alanine Chemical compound CC([NH3+])C([O-])=O QNAYBMKLOCPYGJ-UHFFFAOYSA-N 0.000 description 2
- 241000193744 Bacillus amyloliquefaciens Species 0.000 description 2
- 101000695691 Bacillus licheniformis Beta-lactamase Proteins 0.000 description 2
- 108010029675 Bacillus licheniformis alpha-amylase Proteins 0.000 description 2
- 102000004860 Dipeptidases Human genes 0.000 description 2
- 108090001081 Dipeptidases Proteins 0.000 description 2
- 108010067722 Dipeptidyl Peptidase 4 Proteins 0.000 description 2
- 241001466953 Echovirus Species 0.000 description 2
- 241000991587 Enterovirus C Species 0.000 description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- 108700007698 Genetic Terminator Regions Proteins 0.000 description 2
- 101100369308 Geobacillus stearothermophilus nprS gene Proteins 0.000 description 2
- JEFZIKRIDLHOIF-BYPYZUCNSA-N Gln-Gly Chemical compound NC(=O)CC[C@H](N)C(=O)NCC(O)=O JEFZIKRIDLHOIF-BYPYZUCNSA-N 0.000 description 2
- 102400000324 Glucagon-like peptide 1(7-37) Human genes 0.000 description 2
- 101800004266 Glucagon-like peptide 1(7-37) Proteins 0.000 description 2
- 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 2
- 102000005720 Glutathione transferase Human genes 0.000 description 2
- 108010070675 Glutathione transferase Proteins 0.000 description 2
- 101000585528 Homo sapiens Peptide YY Proteins 0.000 description 2
- 239000007836 KH2PO4 Substances 0.000 description 2
- 239000006142 Luria-Bertani Agar Substances 0.000 description 2
- 241000124008 Mammalia Species 0.000 description 2
- 241001195348 Nusa Species 0.000 description 2
- WCUXLLCKKVVCTQ-UHFFFAOYSA-M Potassium chloride Chemical compound [Cl-].[K+] WCUXLLCKKVVCTQ-UHFFFAOYSA-M 0.000 description 2
- 241000219977 Vigna Species 0.000 description 2
- 238000002835 absorbance Methods 0.000 description 2
- 229960000723 ampicillin Drugs 0.000 description 2
- AVKUERGKIZMTKX-NJBDSQKTSA-N ampicillin Chemical compound C1([C@@H](N)C(=O)N[C@H]2[C@H]3SC([C@@H](N3C2=O)C(O)=O)(C)C)=CC=CC=C1 AVKUERGKIZMTKX-NJBDSQKTSA-N 0.000 description 2
- 125000000637 arginyl group Chemical group N[C@@H](CCCNC(N)=N)C(=O)* 0.000 description 2
- 230000004071 biological effect Effects 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 238000012411 cloning technique Methods 0.000 description 2
- 239000002299 complementary DNA Substances 0.000 description 2
- 239000000356 contaminant Substances 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 238000009826 distribution Methods 0.000 description 2
- NLEBIOOXCVAHBD-QKMCSOCLSA-N dodecyl beta-D-maltoside Chemical compound O[C@@H]1[C@@H](O)[C@H](OCCCCCCCCCCCC)O[C@H](CO)[C@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 NLEBIOOXCVAHBD-QKMCSOCLSA-N 0.000 description 2
- 230000009977 dual effect Effects 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 238000011156 evaluation Methods 0.000 description 2
- 108010061330 glucan 1,4-alpha-maltohydrolase Proteins 0.000 description 2
- 239000008103 glucose Substances 0.000 description 2
- 108010078144 glutaminyl-glycine Proteins 0.000 description 2
- HNDVDQJCIGZPNO-UHFFFAOYSA-N histidine Natural products OC(=O)C(N)CC1=CN=CN1 HNDVDQJCIGZPNO-UHFFFAOYSA-N 0.000 description 2
- 125000000487 histidyl group Chemical group [H]N([H])C(C(=O)O*)C([H])([H])C1=C([H])N([H])C([H])=N1 0.000 description 2
- 238000001727 in vivo Methods 0.000 description 2
- 230000003834 intracellular effect Effects 0.000 description 2
- 238000002955 isolation Methods 0.000 description 2
- 239000012139 lysis buffer Substances 0.000 description 2
- 238000004949 mass spectrometry Methods 0.000 description 2
- 238000005259 measurement Methods 0.000 description 2
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 2
- 235000019796 monopotassium phosphate Nutrition 0.000 description 2
- 230000037361 pathway Effects 0.000 description 2
- 239000011148 porous material Substances 0.000 description 2
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 description 2
- 230000000644 propagated effect Effects 0.000 description 2
- 235000019833 protease Nutrition 0.000 description 2
- 230000012846 protein folding Effects 0.000 description 2
- 230000017854 proteolysis Effects 0.000 description 2
- 239000012521 purified sample Substances 0.000 description 2
- 238000012552 review Methods 0.000 description 2
- 239000000523 sample Substances 0.000 description 2
- 230000028327 secretion Effects 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- 230000003595 spectral effect Effects 0.000 description 2
- 238000003786 synthesis reaction Methods 0.000 description 2
- 229940094937 thioredoxin Drugs 0.000 description 2
- 230000002103 transcriptional effect Effects 0.000 description 2
- LWIHDJKSTIGBAC-UHFFFAOYSA-K tripotassium phosphate Chemical compound [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 description 2
- JKMHFZQWWAIEOD-UHFFFAOYSA-N 2-[4-(2-hydroxyethyl)piperazin-1-yl]ethanesulfonic acid Chemical compound OCC[NH+]1CCN(CCS([O-])(=O)=O)CC1 JKMHFZQWWAIEOD-UHFFFAOYSA-N 0.000 description 1
- GOJUJUVQIVIZAV-UHFFFAOYSA-N 2-amino-4,6-dichloropyrimidine-5-carbaldehyde Chemical group NC1=NC(Cl)=C(C=O)C(Cl)=N1 GOJUJUVQIVIZAV-UHFFFAOYSA-N 0.000 description 1
- OSWFIVFLDKOXQC-UHFFFAOYSA-N 4-(3-methoxyphenyl)aniline Chemical compound COC1=CC=CC(C=2C=CC(N)=CC=2)=C1 OSWFIVFLDKOXQC-UHFFFAOYSA-N 0.000 description 1
- 102100025601 60S ribosomal protein L27 Human genes 0.000 description 1
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 1
- GUBGYTABKSRVRQ-XLOQQCSPSA-N Alpha-Lactose Chemical compound O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H](CO)O[C@H](O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-XLOQQCSPSA-N 0.000 description 1
- 108700016171 Aspartate ammonia-lyases Proteins 0.000 description 1
- 241000186000 Bifidobacterium Species 0.000 description 1
- 125000001433 C-terminal amino-acid group Chemical group 0.000 description 1
- 108091026890 Coding region Proteins 0.000 description 1
- 241000723655 Cowpea mosaic virus Species 0.000 description 1
- 241001429382 Coxsackievirus A16 Species 0.000 description 1
- 108010005843 Cysteine Proteases Proteins 0.000 description 1
- 102000005927 Cysteine Proteases Human genes 0.000 description 1
- QNAYBMKLOCPYGJ-UWTATZPHSA-N D-alanine Chemical compound C[C@@H](N)C(O)=O QNAYBMKLOCPYGJ-UWTATZPHSA-N 0.000 description 1
- 108020004414 DNA Proteins 0.000 description 1
- 108010059378 Endopeptidases Proteins 0.000 description 1
- 102000005593 Endopeptidases Human genes 0.000 description 1
- 241000701867 Enterobacteria phage T7 Species 0.000 description 1
- 108010013369 Enteropeptidase Proteins 0.000 description 1
- 102100029727 Enteropeptidase Human genes 0.000 description 1
- 241001529459 Enterovirus A71 Species 0.000 description 1
- 101000867232 Escherichia coli Heat-stable enterotoxin II Proteins 0.000 description 1
- 102000018389 Exopeptidases Human genes 0.000 description 1
- 108010091443 Exopeptidases Proteins 0.000 description 1
- 108010074860 Factor Xa Proteins 0.000 description 1
- 229920002670 Fructan Polymers 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- 101100001650 Geobacillus stearothermophilus amyM gene Proteins 0.000 description 1
- 101100080316 Geobacillus stearothermophilus nprT gene Proteins 0.000 description 1
- BCCRXDTUTZHDEU-VKHMYHEASA-N Gly-Ser Chemical compound NCC(=O)N[C@@H](CO)C(O)=O BCCRXDTUTZHDEU-VKHMYHEASA-N 0.000 description 1
- 241000238631 Hexapoda Species 0.000 description 1
- 101150062179 II gene Proteins 0.000 description 1
- 102000036770 Islet Amyloid Polypeptide Human genes 0.000 description 1
- 108010041872 Islet Amyloid Polypeptide Proteins 0.000 description 1
- QIVBCDIJIAJPQS-VIFPVBQESA-N L-tryptophane Chemical compound C1=CC=C2C(C[C@H](N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-VIFPVBQESA-N 0.000 description 1
- 241000186660 Lactobacillus Species 0.000 description 1
- 241000186673 Lactobacillus delbrueckii Species 0.000 description 1
- 241000194036 Lactococcus Species 0.000 description 1
- 241000194034 Lactococcus lactis subsp. cremoris Species 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
- 108010014251 Muramidase Proteins 0.000 description 1
- 102000016943 Muramidase Human genes 0.000 description 1
- 108010062010 N-Acetylmuramoyl-L-alanine Amidase Proteins 0.000 description 1
- 125000001429 N-terminal alpha-amino-acid group Chemical group 0.000 description 1
- 108010079246 OMPA outer membrane proteins Proteins 0.000 description 1
- 108700006385 OmpF Proteins 0.000 description 1
- 102100040444 P2X purinoceptor 1 Human genes 0.000 description 1
- 101710189973 P2X purinoceptor 1 Proteins 0.000 description 1
- 101800001672 Peptide YY(3-36) Proteins 0.000 description 1
- 108010076039 Polyproteins Proteins 0.000 description 1
- RWCOTTLHDJWHRS-YUMQZZPRSA-N Pro-Pro Chemical group OC(=O)[C@@H]1CCCN1C(=O)[C@H]1NCCC1 RWCOTTLHDJWHRS-YUMQZZPRSA-N 0.000 description 1
- 101710093543 Probable non-specific lipid-transfer protein Proteins 0.000 description 1
- 101710184309 Probable sucrose-6-phosphate hydrolase Proteins 0.000 description 1
- 108090001066 Racemases and epimerases Proteins 0.000 description 1
- 108020004511 Recombinant DNA Proteins 0.000 description 1
- 241000710122 Rhinovirus B14 Species 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- 229920002684 Sepharose Polymers 0.000 description 1
- WOUIMBGNEUWXQG-VKHMYHEASA-N Ser-Gly Chemical group OC[C@H](N)C(=O)NCC(O)=O WOUIMBGNEUWXQG-VKHMYHEASA-N 0.000 description 1
- MTCFGRXMJLQNBG-UHFFFAOYSA-N Serine Natural products OCC(N)C(O)=O MTCFGRXMJLQNBG-UHFFFAOYSA-N 0.000 description 1
- 108010034546 Serratia marcescens nuclease Proteins 0.000 description 1
- DBMJMQXJHONAFJ-UHFFFAOYSA-M Sodium laurylsulphate Chemical group [Na+].CCCCCCCCCCCCOS([O-])(=O)=O DBMJMQXJHONAFJ-UHFFFAOYSA-M 0.000 description 1
- 235000014962 Streptococcus cremoris Nutrition 0.000 description 1
- 101100309436 Streptococcus mutans serotype c (strain ATCC 700610 / UA159) ftf gene Proteins 0.000 description 1
- 241000194020 Streptococcus thermophilus Species 0.000 description 1
- 108090000787 Subtilisin Proteins 0.000 description 1
- 101710112652 Sucrose-6-phosphate hydrolase Proteins 0.000 description 1
- 108700005078 Synthetic Genes Proteins 0.000 description 1
- 101710137500 T7 RNA polymerase Proteins 0.000 description 1
- 108010076818 TEV protease Proteins 0.000 description 1
- 101100157012 Thermoanaerobacterium saccharolyticum (strain DSM 8691 / JW/SL-YS485) xynB gene Proteins 0.000 description 1
- 108090000190 Thrombin Proteins 0.000 description 1
- 239000007983 Tris buffer Substances 0.000 description 1
- 108090000631 Trypsin Proteins 0.000 description 1
- 102000004142 Trypsin Human genes 0.000 description 1
- QIVBCDIJIAJPQS-UHFFFAOYSA-N Tryptophan Natural products C1=CC=C2C(CC(N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-UHFFFAOYSA-N 0.000 description 1
- 235000010726 Vigna sinensis Nutrition 0.000 description 1
- 108700010756 Viral Polyproteins Proteins 0.000 description 1
- 108010067390 Viral Proteins Proteins 0.000 description 1
- 108030005913 Xaa-Pro dipeptidyl-peptidases Proteins 0.000 description 1
- 238000011481 absorbance measurement Methods 0.000 description 1
- 108090000637 alpha-Amylases Proteins 0.000 description 1
- 101150070828 alr gene Proteins 0.000 description 1
- 239000003242 anti bacterial agent Substances 0.000 description 1
- 229940088710 antibiotic agent Drugs 0.000 description 1
- 239000012736 aqueous medium Substances 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- 239000008366 buffered solution Substances 0.000 description 1
- 238000005277 cation exchange chromatography Methods 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 230000002759 chromosomal effect Effects 0.000 description 1
- 210000000349 chromosome Anatomy 0.000 description 1
- 108090001092 clostripain Proteins 0.000 description 1
- 230000002860 competitive effect Effects 0.000 description 1
- 230000000295 complement effect Effects 0.000 description 1
- 239000012141 concentrate Substances 0.000 description 1
- 239000000470 constituent Substances 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 210000000172 cytosol Anatomy 0.000 description 1
- 238000004090 dissolution Methods 0.000 description 1
- 238000011143 downstream manufacturing Methods 0.000 description 1
- 239000002158 endotoxin Substances 0.000 description 1
- 210000003527 eukaryotic cell Anatomy 0.000 description 1
- 230000002349 favourable effect Effects 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 235000019253 formic acid Nutrition 0.000 description 1
- HQVFCQRVQFYGRJ-UHFFFAOYSA-N formic acid;hydrate Chemical compound O.OC=O HQVFCQRVQFYGRJ-UHFFFAOYSA-N 0.000 description 1
- 238000004108 freeze drying Methods 0.000 description 1
- 108091008053 gene clusters Proteins 0.000 description 1
- 230000002068 genetic effect Effects 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 230000000977 initiatory effect Effects 0.000 description 1
- 238000007689 inspection Methods 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 238000004255 ion exchange chromatography Methods 0.000 description 1
- 229940039696 lactobacillus Drugs 0.000 description 1
- 239000008101 lactose Substances 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 238000004811 liquid chromatography Methods 0.000 description 1
- 239000006166 lysate Substances 0.000 description 1
- 239000004325 lysozyme Substances 0.000 description 1
- 229960000274 lysozyme Drugs 0.000 description 1
- 235000010335 lysozyme Nutrition 0.000 description 1
- 210000004962 mammalian cell Anatomy 0.000 description 1
- 239000003550 marker Substances 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 230000002906 microbiologic effect Effects 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 108091005573 modified proteins Proteins 0.000 description 1
- 102000035118 modified proteins Human genes 0.000 description 1
- 238000010369 molecular cloning Methods 0.000 description 1
- LWGJTAZLEJHCPA-UHFFFAOYSA-N n-(2-chloroethyl)-n-nitrosomorpholine-4-carboxamide Chemical compound ClCCN(N=O)C(=O)N1CCOCC1 LWGJTAZLEJHCPA-UHFFFAOYSA-N 0.000 description 1
- 239000013642 negative control Substances 0.000 description 1
- 101150105920 npr gene Proteins 0.000 description 1
- 101150017837 nprM gene Proteins 0.000 description 1
- 244000052769 pathogen Species 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
- 101150019841 penP gene Proteins 0.000 description 1
- 239000008194 pharmaceutical composition Substances 0.000 description 1
- 229920002401 polyacrylamide Polymers 0.000 description 1
- 230000004481 post-translational protein modification Effects 0.000 description 1
- 239000001103 potassium chloride Substances 0.000 description 1
- 235000011164 potassium chloride Nutrition 0.000 description 1
- 229910000160 potassium phosphate Inorganic materials 0.000 description 1
- 235000011009 potassium phosphates Nutrition 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 230000000135 prohibitive effect Effects 0.000 description 1
- 108010077112 prolyl-proline Proteins 0.000 description 1
- 229940024999 proteolytic enzymes for treatment of wounds and ulcers Drugs 0.000 description 1
- 101150108007 prs gene Proteins 0.000 description 1
- 101150086435 prs1 gene Proteins 0.000 description 1
- 101150070305 prsA gene Proteins 0.000 description 1
- 108700022487 rRNA Genes Proteins 0.000 description 1
- 239000012429 reaction media Substances 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 230000003362 replicative effect Effects 0.000 description 1
- 238000004007 reversed phase HPLC Methods 0.000 description 1
- 108010067528 ribosomal proteins L27 Proteins 0.000 description 1
- 210000003705 ribosome Anatomy 0.000 description 1
- 101150025220 sacB gene Proteins 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 230000035939 shock Effects 0.000 description 1
- 238000007086 side reaction Methods 0.000 description 1
- 239000002002 slurry Substances 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 150000003573 thiols Chemical class 0.000 description 1
- 229960004072 thrombin Drugs 0.000 description 1
- 230000005030 transcription termination Effects 0.000 description 1
- 108010020589 trehalose-6-phosphate synthase Proteins 0.000 description 1
- 108020003272 trehalose-phosphatase Proteins 0.000 description 1
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 description 1
- 239000012588 trypsin Substances 0.000 description 1
- 230000009452 underexpressoin Effects 0.000 description 1
- 241001515965 unidentified phage Species 0.000 description 1
- 239000011534 wash buffer Substances 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
- 239000003643 water by type Substances 0.000 description 1
- 101150110790 xylB gene Proteins 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P21/00—Preparation of peptides or proteins
- C12P21/06—Preparation of peptides or proteins produced by the hydrolysis of a peptide bond, e.g. hydrolysate products
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/14—Hydrolases (3)
- C12N9/48—Hydrolases (3) acting on peptide bonds (3.4)
- C12N9/485—Exopeptidases (3.4.11-3.4.19)
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/14—Hydrolases (3)
- C12N9/48—Hydrolases (3) acting on peptide bonds (3.4)
- C12N9/50—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25)
- C12N9/503—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from viruses
- C12N9/506—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from viruses derived from RNA viruses
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y304/00—Hydrolases acting on peptide bonds, i.e. peptidases (3.4)
- C12Y304/14—Dipeptidyl-peptidases and tripeptidyl-peptidases (3.4.14)
- C12Y304/14011—Xaa-Pro dipeptidyl-peptidase (3.4.14.11)
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y304/00—Hydrolases acting on peptide bonds, i.e. peptidases (3.4)
- C12Y304/22—Cysteine endopeptidases (3.4.22)
- C12Y304/22028—Picornain 3C (3.4.22.28)
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Genetics & Genomics (AREA)
- General Health & Medical Sciences (AREA)
- General Engineering & Computer Science (AREA)
- Biochemistry (AREA)
- Molecular Biology (AREA)
- Microbiology (AREA)
- Biotechnology (AREA)
- Medicinal Chemistry (AREA)
- Biomedical Technology (AREA)
- Virology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Enzymes And Modification Thereof (AREA)
- Peptides Or Proteins (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Applications Claiming Priority (11)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| EP13169173 | 2013-05-24 | ||
| EP13169173.5 | 2013-05-24 | ||
| EP13171191 | 2013-06-10 | ||
| EP13171191.3 | 2013-06-10 | ||
| US201361834100P | 2013-06-12 | 2013-06-12 | |
| US61/834100 | 2013-06-12 | ||
| EP13194053 | 2013-11-22 | ||
| EP13194053.8 | 2013-11-22 | ||
| US201361909412P | 2013-11-27 | 2013-11-27 | |
| US61/909412 | 2013-11-27 | ||
| PCT/EP2014/060696 WO2014187974A1 (en) | 2013-05-24 | 2014-05-23 | Fusion protease |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN105247067A true CN105247067A (zh) | 2016-01-13 |
Family
ID=51932984
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201480029964.7A Withdrawn CN105247067A (zh) | 2013-05-24 | 2014-05-23 | 融合蛋白酶 |
Country Status (5)
| Country | Link |
|---|---|
| US (1) | US20160122793A1 (enExample) |
| EP (1) | EP3004369A1 (enExample) |
| JP (1) | JP2016518855A (enExample) |
| CN (1) | CN105247067A (enExample) |
| WO (2) | WO2014187974A1 (enExample) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110128540A (zh) * | 2019-04-18 | 2019-08-16 | 江南大学 | 一种基于便携式血糖仪的广谱二抗 |
| CN112011497A (zh) * | 2020-08-28 | 2020-12-01 | 上海交通大学 | 分泌表达猪源性抗产肠毒素大肠杆菌k88菌毛单链抗体的重组乳酸乳球菌及其制备方法 |
Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20160060312A1 (en) | 2014-08-27 | 2016-03-03 | Cellivery Therapeutics, Inc. | Development of Protein-Based Biotherapeutics That Penetrates Cell-Membrane and Induces Anti-Pancreatic Cancer Effect - Improved Cell-Permeable Suppressor of Cytokine Signaling (iCP-SOCS3) Proteins, Polynucleotides Encoding the Same, and Anti-Pancreatic Cancer Compositions Comprising the Same |
| CN110592057B (zh) * | 2019-09-27 | 2022-01-28 | 昆明理工大学 | 嵌合裂解酶ILTphg和编码此酶的多核苷酸 |
| US20250122489A1 (en) * | 2023-10-16 | 2025-04-17 | Zhenyu Zhu | Protein Thermostability Reduction by Tag Fusion and Its Usage |
Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CA1200773A (en) * | 1980-02-29 | 1986-02-18 | William J. Rutter | Expression linkers |
| US4769326A (en) * | 1980-02-29 | 1988-09-06 | The Regents Of The University Of California | Expression linkers |
| NZ236819A (en) * | 1990-02-03 | 1993-07-27 | Max Planck Gesellschaft | Enzymatic cleavage of fusion proteins; fusion proteins; recombinant dna and pharmaceutical compositions |
-
2014
- 2014-05-23 WO PCT/EP2014/060696 patent/WO2014187974A1/en not_active Ceased
- 2014-05-23 US US14/889,993 patent/US20160122793A1/en not_active Abandoned
- 2014-05-23 EP EP14726158.0A patent/EP3004369A1/en not_active Withdrawn
- 2014-05-23 CN CN201480029964.7A patent/CN105247067A/zh not_active Withdrawn
- 2014-05-23 WO PCT/EP2014/060669 patent/WO2014187960A1/en not_active Ceased
- 2014-05-23 JP JP2016514440A patent/JP2016518855A/ja not_active Withdrawn
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110128540A (zh) * | 2019-04-18 | 2019-08-16 | 江南大学 | 一种基于便携式血糖仪的广谱二抗 |
| CN110128540B (zh) * | 2019-04-18 | 2021-12-14 | 江南大学 | 一种基于便携式血糖仪的二抗 |
| CN112011497A (zh) * | 2020-08-28 | 2020-12-01 | 上海交通大学 | 分泌表达猪源性抗产肠毒素大肠杆菌k88菌毛单链抗体的重组乳酸乳球菌及其制备方法 |
| CN112011497B (zh) * | 2020-08-28 | 2022-06-07 | 上海交通大学 | 分泌表达猪源性抗产肠毒素大肠杆菌k88菌毛单链抗体的重组乳酸乳球菌及其制备方法 |
Also Published As
| Publication number | Publication date |
|---|---|
| JP2016518855A (ja) | 2016-06-30 |
| US20160122793A1 (en) | 2016-05-05 |
| WO2014187960A1 (en) | 2014-11-27 |
| WO2014187974A1 (en) | 2014-11-27 |
| EP3004369A1 (en) | 2016-04-13 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US12054541B2 (en) | Split inteins, conjugates and uses thereof | |
| Guerrero et al. | Tandem SUMO fusion vectors for improving soluble protein expression and purification | |
| JP2019506163A (ja) | 例外的なスプライシング活性を有するスプリットインテイン | |
| JP2010505437A (ja) | 塩基性タンパク質タグに融合したプロセシング酵素 | |
| CN105247067A (zh) | 融合蛋白酶 | |
| CN109790205B (zh) | 酶促肽连接的方法 | |
| WO2010104114A1 (ja) | ポリペプチドライブラリーを調製する方法 | |
| JP2024513126A (ja) | ループまたは末端でペプチドタグと相互作用するポリペプチドおよびその使用 | |
| CN103797122A (zh) | 用于大肠杆菌中异源蛋白质生产的新的表达和分泌载体系统 | |
| EP1527181A1 (en) | Staphylococcal nuclease fusion proteins for the production of recombinant peptides | |
| JP2023508893A (ja) | Whepドメイン融合による目的タンパク質の水溶性増進方法 | |
| JP2010071744A (ja) | 化合物のスクリーニング方法、並びに、スクリーニング用キット | |
| CN1860226B (zh) | 使用OmpT蛋白酶突变体的多肽的切断方法 | |
| JP4891581B2 (ja) | ポリペプチドの製造方法およびキット | |
| Kuo et al. | Process intensification for the removal of poly‐histidine fusion tags from recombinant proteins by an exopeptidase | |
| CN101381739B (zh) | 一种周质分泌融合表达型前t载体及其制备与应用 | |
| JP5020487B2 (ja) | 融合タンパク質発現用新規dna及び該dnaを用いたタンパク質の製造方法 | |
| US20140065673A1 (en) | Membrane protein expression vector comprising major envelope protein p9 of systovirus phi12 as a fusion partner and method for producing membrane protein using the same | |
| EP2558482B1 (en) | Engineered plant cysteine proteases and their uses | |
| JP2018000174A (ja) | ヒトFcRnをコードするポリヌクレオチドおよび当該ポリヌクレオチドを利用したヒトFcRnの製造方法 | |
| Ramesh | Engineering Substrate Specificity of Mammalian Proteases |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| WW01 | Invention patent application withdrawn after publication |
Application publication date: 20160113 |
|
| WW01 | Invention patent application withdrawn after publication |