JP2021501603A - β−ラクタマーゼ変異体 - Google Patents
β−ラクタマーゼ変異体 Download PDFInfo
- Publication number
- JP2021501603A JP2021501603A JP2020543711A JP2020543711A JP2021501603A JP 2021501603 A JP2021501603 A JP 2021501603A JP 2020543711 A JP2020543711 A JP 2020543711A JP 2020543711 A JP2020543711 A JP 2020543711A JP 2021501603 A JP2021501603 A JP 2021501603A
- Authority
- JP
- Japan
- Prior art keywords
- polypeptide
- seq
- vim
- positions
- nucleic acid
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 108090000204 Dipeptidase 1 Proteins 0.000 title claims abstract description 10
- 102000006635 beta-lactamase Human genes 0.000 title claims abstract description 10
- 108090000765 processed proteins & peptides Proteins 0.000 claims abstract description 158
- 102000004196 processed proteins & peptides Human genes 0.000 claims abstract description 156
- 229920001184 polypeptide Polymers 0.000 claims abstract description 154
- 230000000694 effects Effects 0.000 claims abstract description 54
- 150000007523 nucleic acids Chemical group 0.000 claims abstract description 35
- 210000004027 cell Anatomy 0.000 claims description 58
- 238000000034 method Methods 0.000 claims description 38
- 238000006467 substitution reaction Methods 0.000 claims description 34
- 239000000203 mixture Substances 0.000 claims description 28
- 239000003782 beta lactam antibiotic agent Substances 0.000 claims description 27
- 239000002132 β-lactam antibiotic Substances 0.000 claims description 26
- 229940124586 β-lactam antibiotics Drugs 0.000 claims description 26
- 102220550120 Caveolae-associated protein 2_E130D_mutation Human genes 0.000 claims description 22
- 102220261130 rs148851677 Human genes 0.000 claims description 22
- 210000001072 colon Anatomy 0.000 claims description 21
- 108091028043 Nucleic acid sequence Proteins 0.000 claims description 20
- 125000003275 alpha amino acid group Chemical group 0.000 claims description 20
- 210000000936 intestine Anatomy 0.000 claims description 18
- 210000003405 ileum Anatomy 0.000 claims description 16
- 210000004534 cecum Anatomy 0.000 claims description 15
- 102220098555 rs878853237 Human genes 0.000 claims description 15
- 108010076504 Protein Sorting Signals Proteins 0.000 claims description 14
- 102000039446 nucleic acids Human genes 0.000 claims description 14
- 108020004707 nucleic acids Proteins 0.000 claims description 14
- 238000012986 modification Methods 0.000 claims description 11
- 230000004048 modification Effects 0.000 claims description 11
- 208000035143 Bacterial infection Diseases 0.000 claims description 10
- 208000022362 bacterial infectious disease Diseases 0.000 claims description 10
- 210000004899 c-terminal region Anatomy 0.000 claims description 8
- 239000012634 fragment Substances 0.000 claims description 6
- 210000001630 jejunum Anatomy 0.000 claims description 5
- 230000000415 inactivating effect Effects 0.000 claims description 3
- 125000001360 methionine group Chemical group N[C@@H](CCSC)C(=O)* 0.000 claims description 3
- 102220062675 rs786204152 Human genes 0.000 claims 1
- 230000000968 intestinal effect Effects 0.000 abstract description 19
- 239000003242 anti bacterial agent Substances 0.000 abstract description 16
- 229940088710 antibiotic agent Drugs 0.000 abstract description 14
- 108091005804 Peptidases Proteins 0.000 abstract description 7
- 239000004365 Protease Substances 0.000 abstract description 7
- 230000006872 improvement Effects 0.000 abstract description 2
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 abstract 1
- 102000004190 Enzymes Human genes 0.000 description 36
- 108090000790 Enzymes Proteins 0.000 description 36
- 229940088598 enzyme Drugs 0.000 description 36
- 108090000623 proteins and genes Proteins 0.000 description 31
- 102220625296 Dol-P-Man:Man(5)GlcNAc(2)-PP-Dol alpha-1,3-mannosyltransferase_Q22H_mutation Human genes 0.000 description 21
- 102000004169 proteins and genes Human genes 0.000 description 20
- 241000894006 Bacteria Species 0.000 description 19
- 238000004519 manufacturing process Methods 0.000 description 17
- -1 β-lactam compounds Chemical class 0.000 description 15
- 239000002609 medium Substances 0.000 description 14
- 150000001413 amino acids Chemical group 0.000 description 12
- 239000000284 extract Substances 0.000 description 12
- 150000003952 β-lactams Chemical class 0.000 description 11
- 230000003115 biocidal effect Effects 0.000 description 10
- 239000000758 substrate Substances 0.000 description 9
- 108091026890 Coding region Proteins 0.000 description 8
- 108010052650 beta-lactamase bla(vim-2) Proteins 0.000 description 8
- 230000007062 hydrolysis Effects 0.000 description 8
- 238000006460 hydrolysis reaction Methods 0.000 description 8
- 125000003729 nucleotide group Chemical group 0.000 description 8
- 239000013598 vector Substances 0.000 description 8
- 108020004414 DNA Proteins 0.000 description 7
- 241000206602 Eukaryota Species 0.000 description 7
- 241000282414 Homo sapiens Species 0.000 description 7
- 238000011534 incubation Methods 0.000 description 7
- 239000002773 nucleotide Substances 0.000 description 7
- 108091033319 polynucleotide Proteins 0.000 description 7
- 102000040430 polynucleotide Human genes 0.000 description 7
- 239000002157 polynucleotide Substances 0.000 description 7
- 102000035195 Peptidases Human genes 0.000 description 6
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 6
- 230000003197 catalytic effect Effects 0.000 description 6
- 230000014616 translation Effects 0.000 description 6
- 241000193830 Bacillus <bacterium> Species 0.000 description 5
- 108091034117 Oligonucleotide Proteins 0.000 description 5
- 230000001580 bacterial effect Effects 0.000 description 5
- 230000002538 fungal effect Effects 0.000 description 5
- 238000000338 in vitro Methods 0.000 description 5
- 230000035772 mutation Effects 0.000 description 5
- 230000009466 transformation Effects 0.000 description 5
- 241000124008 Mammalia Species 0.000 description 4
- 241001465754 Metazoa Species 0.000 description 4
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 4
- 235000014680 Saccharomyces cerevisiae Nutrition 0.000 description 4
- 125000000539 amino acid group Chemical group 0.000 description 4
- 230000002255 enzymatic effect Effects 0.000 description 4
- 238000001952 enzyme assay Methods 0.000 description 4
- 238000000855 fermentation Methods 0.000 description 4
- 230000004151 fermentation Effects 0.000 description 4
- 238000003780 insertion Methods 0.000 description 4
- 230000037431 insertion Effects 0.000 description 4
- 235000015097 nutrients Nutrition 0.000 description 4
- 230000008569 process Effects 0.000 description 4
- 238000002708 random mutagenesis Methods 0.000 description 4
- 238000013519 translation Methods 0.000 description 4
- 108020004705 Codon Proteins 0.000 description 3
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 3
- 108091081024 Start codon Proteins 0.000 description 3
- WKDDRNSBRWANNC-UHFFFAOYSA-N Thienamycin Natural products C1C(SCCN)=C(C(O)=O)N2C(=O)C(C(O)C)C21 WKDDRNSBRWANNC-UHFFFAOYSA-N 0.000 description 3
- JLCPHMBAVCMARE-UHFFFAOYSA-N [3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-[[3-[[3-[[3-[[3-[[3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-hydroxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methyl [5-(6-aminopurin-9-yl)-2-(hydroxymethyl)oxolan-3-yl] hydrogen phosphate Polymers Cc1cn(C2CC(OP(O)(=O)OCC3OC(CC3OP(O)(=O)OCC3OC(CC3O)n3cnc4c3nc(N)[nH]c4=O)n3cnc4c3nc(N)[nH]c4=O)C(COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3CO)n3cnc4c(N)ncnc34)n3ccc(N)nc3=O)n3cnc4c(N)ncnc34)n3ccc(N)nc3=O)n3ccc(N)nc3=O)n3ccc(N)nc3=O)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cc(C)c(=O)[nH]c3=O)n3cc(C)c(=O)[nH]c3=O)n3ccc(N)nc3=O)n3cc(C)c(=O)[nH]c3=O)n3cnc4c3nc(N)[nH]c4=O)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)O2)c(=O)[nH]c1=O JLCPHMBAVCMARE-UHFFFAOYSA-N 0.000 description 3
- 238000003556 assay Methods 0.000 description 3
- 238000005119 centrifugation Methods 0.000 description 3
- 239000003795 chemical substances by application Substances 0.000 description 3
- 239000013604 expression vector Substances 0.000 description 3
- ZSKVGTPCRGIANV-ZXFLCMHBSA-N imipenem Chemical compound C1C(SCC\N=C\N)=C(C(O)=O)N2C(=O)[C@H]([C@H](O)C)[C@H]21 ZSKVGTPCRGIANV-ZXFLCMHBSA-N 0.000 description 3
- 229960002182 imipenem Drugs 0.000 description 3
- 208000015181 infectious disease Diseases 0.000 description 3
- 108060004734 metallo-beta-lactamase Proteins 0.000 description 3
- 102000020235 metallo-beta-lactamase Human genes 0.000 description 3
- 238000012544 monitoring process Methods 0.000 description 3
- 239000013612 plasmid Substances 0.000 description 3
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- 210000001938 protoplast Anatomy 0.000 description 3
- 238000000746 purification Methods 0.000 description 3
- 239000011780 sodium chloride Substances 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 230000001225 therapeutic effect Effects 0.000 description 3
- 238000002560 therapeutic procedure Methods 0.000 description 3
- 238000013518 transcription Methods 0.000 description 3
- 230000035897 transcription Effects 0.000 description 3
- 230000014621 translational initiation Effects 0.000 description 3
- 125000002987 valine group Chemical group [H]N([H])C([H])(C(*)=O)C([H])(C([H])([H])[H])C([H])([H])[H] 0.000 description 3
- 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 2
- 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 description 2
- 101000695713 Aeromonas hydrophila Metallo-beta-lactamase type 2 Proteins 0.000 description 2
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 241000193422 Bacillus lentus Species 0.000 description 2
- 241000193764 Brevibacillus brevis Species 0.000 description 2
- 241000193163 Clostridioides difficile Species 0.000 description 2
- 102000053602 DNA Human genes 0.000 description 2
- 206010012735 Diarrhoea Diseases 0.000 description 2
- 241000588724 Escherichia coli Species 0.000 description 2
- 241000282326 Felis catus Species 0.000 description 2
- 239000007995 HEPES buffer Substances 0.000 description 2
- 206010020751 Hypersensitivity Diseases 0.000 description 2
- 241000276498 Pollachius virens Species 0.000 description 2
- 241000589517 Pseudomonas aeruginosa Species 0.000 description 2
- 241000187398 Streptomyces lividans Species 0.000 description 2
- 239000007983 Tris buffer Substances 0.000 description 2
- 230000007815 allergy Effects 0.000 description 2
- 238000009635 antibiotic susceptibility testing Methods 0.000 description 2
- 239000000872 buffer Substances 0.000 description 2
- YZBQHRLRFGPBSL-RXMQYKEDSA-N carbapenem Chemical compound C1C=CN2C(=O)C[C@H]21 YZBQHRLRFGPBSL-RXMQYKEDSA-N 0.000 description 2
- HVFLCNVBZFFHBT-ZKDACBOMSA-N cefepime Chemical compound S([C@@H]1[C@@H](C(N1C=1C([O-])=O)=O)NC(=O)\C(=N/OC)C=2N=C(N)SC=2)CC=1C[N+]1(C)CCCC1 HVFLCNVBZFFHBT-ZKDACBOMSA-N 0.000 description 2
- 229960002100 cefepime Drugs 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 238000010367 cloning Methods 0.000 description 2
- 239000002299 complementary DNA Substances 0.000 description 2
- 150000001875 compounds Chemical class 0.000 description 2
- 239000000287 crude extract Substances 0.000 description 2
- 239000012228 culture supernatant Substances 0.000 description 2
- 238000012258 culturing Methods 0.000 description 2
- 238000012217 deletion Methods 0.000 description 2
- 230000037430 deletion Effects 0.000 description 2
- 230000001419 dependent effect Effects 0.000 description 2
- 108091007735 digestive proteases Proteins 0.000 description 2
- 238000010790 dilution Methods 0.000 description 2
- 239000012895 dilution Substances 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 239000013613 expression plasmid Substances 0.000 description 2
- 238000000605 extraction Methods 0.000 description 2
- 230000002779 inactivation Effects 0.000 description 2
- 230000002401 inhibitory effect Effects 0.000 description 2
- 244000000074 intestinal pathogen Species 0.000 description 2
- 238000002955 isolation Methods 0.000 description 2
- 230000000670 limiting effect Effects 0.000 description 2
- 208000030159 metabolic disease Diseases 0.000 description 2
- 238000002703 mutagenesis Methods 0.000 description 2
- 231100000350 mutagenesis Toxicity 0.000 description 2
- 229910052757 nitrogen Inorganic materials 0.000 description 2
- 230000007030 peptide scission Effects 0.000 description 2
- 230000004481 post-translational protein modification Effects 0.000 description 2
- 230000001124 posttranscriptional effect Effects 0.000 description 2
- 230000001376 precipitating effect Effects 0.000 description 2
- 238000001556 precipitation Methods 0.000 description 2
- 238000001742 protein purification Methods 0.000 description 2
- 230000002829 reductive effect Effects 0.000 description 2
- 238000005067 remediation Methods 0.000 description 2
- 230000028327 secretion Effects 0.000 description 2
- 230000036962 time dependent Effects 0.000 description 2
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 description 2
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 1
- 241000193744 Bacillus amyloliquefaciens Species 0.000 description 1
- 241000193752 Bacillus circulans Species 0.000 description 1
- 241001328122 Bacillus clausii Species 0.000 description 1
- 241000193749 Bacillus coagulans Species 0.000 description 1
- 241000194108 Bacillus licheniformis Species 0.000 description 1
- 244000063299 Bacillus subtilis Species 0.000 description 1
- 235000014469 Bacillus subtilis Nutrition 0.000 description 1
- 241000193388 Bacillus thuringiensis Species 0.000 description 1
- 125000001433 C-terminal amino-acid group Chemical group 0.000 description 1
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- GNWUOVJNSFPWDD-XMZRARIVSA-M Cefoxitin sodium Chemical compound [Na+].N([C@]1(OC)C(N2C(=C(COC(N)=O)CS[C@@H]21)C([O-])=O)=O)C(=O)CC1=CC=CS1 GNWUOVJNSFPWDD-XMZRARIVSA-M 0.000 description 1
- 102000005575 Cellulases Human genes 0.000 description 1
- 108010084185 Cellulases Proteins 0.000 description 1
- 229930186147 Cephalosporin Natural products 0.000 description 1
- 108090000317 Chymotrypsin Proteins 0.000 description 1
- 102000016928 DNA-directed DNA polymerase Human genes 0.000 description 1
- 108010014303 DNA-directed DNA polymerase Proteins 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 241001198387 Escherichia coli BL21(DE3) Species 0.000 description 1
- 108091092566 Extrachromosomal DNA Proteins 0.000 description 1
- 241000192125 Firmicutes Species 0.000 description 1
- 241000193385 Geobacillus stearothermophilus Species 0.000 description 1
- 102000005720 Glutathione transferase Human genes 0.000 description 1
- 108010070675 Glutathione transferase Proteins 0.000 description 1
- 241000238631 Hexapoda Species 0.000 description 1
- 102220474121 Inorganic pyrophosphatase 2, mitochondrial_I19T_mutation Human genes 0.000 description 1
- 241000235058 Komagataella pastoris Species 0.000 description 1
- 108010029541 Laccase Proteins 0.000 description 1
- 102000003960 Ligases Human genes 0.000 description 1
- 108090000364 Ligases Proteins 0.000 description 1
- 108090001060 Lipase Proteins 0.000 description 1
- 102000004882 Lipase Human genes 0.000 description 1
- 239000004367 Lipase Substances 0.000 description 1
- 101710175625 Maltose/maltodextrin-binding periplasmic protein Proteins 0.000 description 1
- 102000016943 Muramidase Human genes 0.000 description 1
- 108010014251 Muramidase Proteins 0.000 description 1
- 108010062010 N-Acetylmuramoyl-L-alanine Amidase Proteins 0.000 description 1
- 108700026244 Open Reading Frames Proteins 0.000 description 1
- 206010073261 Ovarian theca cell tumour Diseases 0.000 description 1
- 241000194109 Paenibacillus lautus Species 0.000 description 1
- 229930182555 Penicillin Natural products 0.000 description 1
- JGSARLDLIJGVTE-MBNYWOFBSA-N Penicillin G Chemical compound N([C@H]1[C@H]2SC([C@@H](N2C1=O)C(O)=O)(C)C)C(=O)CC1=CC=CC=C1 JGSARLDLIJGVTE-MBNYWOFBSA-N 0.000 description 1
- 108020004511 Recombinant DNA Proteins 0.000 description 1
- 102000007056 Recombinant Fusion Proteins Human genes 0.000 description 1
- 108010008281 Recombinant Fusion Proteins Proteins 0.000 description 1
- 241000187747 Streptomyces Species 0.000 description 1
- 241001468239 Streptomyces murinus Species 0.000 description 1
- 241000282887 Suidae Species 0.000 description 1
- 101710183280 Topoisomerase Proteins 0.000 description 1
- GSEJCLTVZPLZKY-UHFFFAOYSA-N Triethanolamine Chemical compound OCCN(CCO)CCO GSEJCLTVZPLZKY-UHFFFAOYSA-N 0.000 description 1
- 108090000631 Trypsin Proteins 0.000 description 1
- 102000004142 Trypsin Human genes 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 239000012190 activator Substances 0.000 description 1
- 238000000246 agarose gel electrophoresis Methods 0.000 description 1
- 238000012870 ammonium sulfate precipitation Methods 0.000 description 1
- AVKUERGKIZMTKX-NJBDSQKTSA-N ampicillin Chemical compound C1([C@@H](N)C(=O)N[C@H]2[C@H]3SC([C@@H](N3C2=O)C(O)=O)(C)C)=CC=CC=C1 AVKUERGKIZMTKX-NJBDSQKTSA-N 0.000 description 1
- 229960000723 ampicillin Drugs 0.000 description 1
- 238000005349 anion exchange Methods 0.000 description 1
- 238000005571 anion exchange chromatography Methods 0.000 description 1
- 230000000844 anti-bacterial effect Effects 0.000 description 1
- 229940054340 bacillus coagulans Drugs 0.000 description 1
- 229940097012 bacillus thuringiensis Drugs 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- XIURVHNZVLADCM-IUODEOHRSA-N cefalotin Chemical compound N([C@H]1[C@@H]2N(C1=O)C(=C(CS2)COC(=O)C)C(O)=O)C(=O)CC1=CC=CS1 XIURVHNZVLADCM-IUODEOHRSA-N 0.000 description 1
- 229960000603 cefalotin Drugs 0.000 description 1
- 229960004261 cefotaxime Drugs 0.000 description 1
- AZZMGZXNTDTSME-JUZDKLSSSA-M cefotaxime sodium Chemical compound [Na+].N([C@@H]1C(N2C(=C(COC(C)=O)CS[C@@H]21)C([O-])=O)=O)C(=O)\C(=N/OC)C1=CSC(N)=N1 AZZMGZXNTDTSME-JUZDKLSSSA-M 0.000 description 1
- SRZNHPXWXCNNDU-RHBCBLIFSA-N cefotetan Chemical compound N([C@]1(OC)C(N2C(=C(CSC=3N(N=NN=3)C)CS[C@@H]21)C(O)=O)=O)C(=O)C1SC(=C(C(N)=O)C(O)=O)S1 SRZNHPXWXCNNDU-RHBCBLIFSA-N 0.000 description 1
- 229960005495 cefotetan Drugs 0.000 description 1
- 229960002682 cefoxitin Drugs 0.000 description 1
- VAAUVRVFOQPIGI-SPQHTLEESA-N ceftriaxone Chemical compound S([C@@H]1[C@@H](C(N1C=1C(O)=O)=O)NC(=O)\C(=N/OC)C=2N=C(N)SC=2)CC=1CSC1=NC(=O)C(=O)NN1C VAAUVRVFOQPIGI-SPQHTLEESA-N 0.000 description 1
- 229960004755 ceftriaxone Drugs 0.000 description 1
- 239000006143 cell culture medium Substances 0.000 description 1
- 239000013592 cell lysate Substances 0.000 description 1
- 230000006037 cell lysis Effects 0.000 description 1
- 230000004663 cell proliferation Effects 0.000 description 1
- 210000002421 cell wall Anatomy 0.000 description 1
- 229940124587 cephalosporin Drugs 0.000 description 1
- 150000001780 cephalosporins Chemical class 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- 239000002738 chelating agent Substances 0.000 description 1
- 210000004978 chinese hamster ovary cell Anatomy 0.000 description 1
- 230000002759 chromosomal effect Effects 0.000 description 1
- 210000000349 chromosome Anatomy 0.000 description 1
- 229960002376 chymotrypsin Drugs 0.000 description 1
- 238000002983 circular dichroism Methods 0.000 description 1
- 239000012459 cleaning agent Substances 0.000 description 1
- 230000005757 colony formation Effects 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 125000004122 cyclic group Chemical group 0.000 description 1
- 230000009089 cytolysis Effects 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 230000000593 degrading effect Effects 0.000 description 1
- 238000004925 denaturation Methods 0.000 description 1
- 230000036425 denaturation Effects 0.000 description 1
- 238000009795 derivation Methods 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- 230000001079 digestive effect Effects 0.000 description 1
- 210000003717 douglas' pouch Anatomy 0.000 description 1
- 239000003937 drug carrier Substances 0.000 description 1
- 238000012377 drug delivery Methods 0.000 description 1
- 210000001198 duodenum Anatomy 0.000 description 1
- 238000001962 electrophoresis Methods 0.000 description 1
- 238000004520 electroporation Methods 0.000 description 1
- 238000007824 enzymatic assay Methods 0.000 description 1
- 210000003527 eukaryotic cell Anatomy 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 238000001704 evaporation Methods 0.000 description 1
- 230000008020 evaporation Effects 0.000 description 1
- 230000007717 exclusion Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 230000004927 fusion Effects 0.000 description 1
- 230000002496 gastric effect Effects 0.000 description 1
- 210000004051 gastric juice Anatomy 0.000 description 1
- 210000001035 gastrointestinal tract Anatomy 0.000 description 1
- 238000001502 gel electrophoresis Methods 0.000 description 1
- 238000002523 gelfiltration Methods 0.000 description 1
- 230000002068 genetic effect Effects 0.000 description 1
- 238000010353 genetic engineering Methods 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- HNDVDQJCIGZPNO-UHFFFAOYSA-N histidine Natural products OC(=O)C(N)CC1=CN=CN1 HNDVDQJCIGZPNO-UHFFFAOYSA-N 0.000 description 1
- 230000003301 hydrolyzing effect Effects 0.000 description 1
- 238000010348 incorporation Methods 0.000 description 1
- 230000006698 induction Effects 0.000 description 1
- 238000001361 intraarterial administration Methods 0.000 description 1
- 230000003834 intracellular effect Effects 0.000 description 1
- 238000007918 intramuscular administration Methods 0.000 description 1
- 238000007912 intraperitoneal administration Methods 0.000 description 1
- 238000001990 intravenous administration Methods 0.000 description 1
- 238000005342 ion exchange Methods 0.000 description 1
- 230000002427 irreversible effect Effects 0.000 description 1
- 238000001155 isoelectric focusing Methods 0.000 description 1
- 235000019421 lipase Nutrition 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 230000004807 localization Effects 0.000 description 1
- 229960000274 lysozyme Drugs 0.000 description 1
- 239000004325 lysozyme Substances 0.000 description 1
- 235000010335 lysozyme Nutrition 0.000 description 1
- 239000011159 matrix material Substances 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 238000002844 melting Methods 0.000 description 1
- 230000008018 melting Effects 0.000 description 1
- 239000002184 metal Substances 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 230000036961 partial effect Effects 0.000 description 1
- 244000052769 pathogen Species 0.000 description 1
- 229940049954 penicillin Drugs 0.000 description 1
- 239000008194 pharmaceutical composition Substances 0.000 description 1
- 238000000053 physical method Methods 0.000 description 1
- 229960002292 piperacillin Drugs 0.000 description 1
- WCMIIGXFCMNQDS-IDYPWDAWSA-M piperacillin sodium Chemical compound [Na+].O=C1C(=O)N(CC)CCN1C(=O)N[C@H](C=1C=CC=CC=1)C(=O)N[C@@H]1C(=O)N2[C@@H](C([O-])=O)C(C)(C)S[C@@H]21 WCMIIGXFCMNQDS-IDYPWDAWSA-M 0.000 description 1
- 239000013600 plasmid vector Substances 0.000 description 1
- 230000008488 polyadenylation Effects 0.000 description 1
- 229920000642 polymer Polymers 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 210000001236 prokaryotic cell Anatomy 0.000 description 1
- 238000003259 recombinant expression Methods 0.000 description 1
- 238000010188 recombinant method Methods 0.000 description 1
- 230000006798 recombination Effects 0.000 description 1
- 238000005215 recombination Methods 0.000 description 1
- 230000008929 regeneration Effects 0.000 description 1
- 238000011069 regeneration method Methods 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 230000010076 replication Effects 0.000 description 1
- 102220325989 rs1555376587 Human genes 0.000 description 1
- 102220008853 rs193922308 Human genes 0.000 description 1
- 102200142069 rs2577154 Human genes 0.000 description 1
- 102200114514 rs535274413 Human genes 0.000 description 1
- 102200132327 rs769653717 Human genes 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 230000003248 secreting effect Effects 0.000 description 1
- 230000011664 signaling Effects 0.000 description 1
- 238000002741 site-directed mutagenesis Methods 0.000 description 1
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 1
- 238000010563 solid-state fermentation Methods 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 238000000527 sonication Methods 0.000 description 1
- 238000001179 sorption measurement Methods 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 238000002798 spectrophotometry method Methods 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 239000007921 spray Substances 0.000 description 1
- 238000001694 spray drying Methods 0.000 description 1
- 210000002784 stomach Anatomy 0.000 description 1
- 238000007920 subcutaneous administration Methods 0.000 description 1
- 150000003457 sulfones Chemical class 0.000 description 1
- 230000002194 synthesizing effect Effects 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 208000001644 thecoma Diseases 0.000 description 1
- OHKOGUYZJXTSFX-KZFFXBSXSA-N ticarcillin Chemical compound C=1([C@@H](C(O)=O)C(=O)N[C@H]2[C@H]3SC([C@@H](N3C2=O)C(O)=O)(C)C)C=CSC=1 OHKOGUYZJXTSFX-KZFFXBSXSA-N 0.000 description 1
- 229960004659 ticarcillin Drugs 0.000 description 1
- 239000003053 toxin Substances 0.000 description 1
- 231100000765 toxin Toxicity 0.000 description 1
- 108700012359 toxins Proteins 0.000 description 1
- 230000005026 transcription initiation Effects 0.000 description 1
- 230000005030 transcription termination Effects 0.000 description 1
- 238000010361 transduction Methods 0.000 description 1
- 230000026683 transduction Effects 0.000 description 1
- 238000001890 transfection Methods 0.000 description 1
- 238000000844 transformation Methods 0.000 description 1
- 239000012588 trypsin Substances 0.000 description 1
- 238000000108 ultra-filtration Methods 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
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/14—Hydrolases (3)
- C12N9/78—Hydrolases (3) acting on carbon to nitrogen bonds other than peptide bonds (3.5)
- C12N9/86—Hydrolases (3) acting on carbon to nitrogen bonds other than peptide bonds (3.5) acting on amide bonds in cyclic amides, e.g. penicillinase (3.5.2)
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/40—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil
- A61K31/407—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil condensed with other heterocyclic ring systems, e.g. ketorolac, physostigmine
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
- A61K31/496—Non-condensed piperazines containing further heterocyclic rings, e.g. rifampin, thiothixene or sparfloxacin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/54—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one sulfur as the ring hetero atoms, e.g. sulthiame
- A61K31/542—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one sulfur as the ring hetero atoms, e.g. sulthiame ortho- or peri-condensed with heterocyclic ring systems
- A61K31/545—Compounds containing 5-thia-1-azabicyclo [4.2.0] octane ring systems, i.e. compounds containing a ring system of the formula:, e.g. cephalosporins, cefaclor, or cephalexine
- A61K31/546—Compounds containing 5-thia-1-azabicyclo [4.2.0] octane ring systems, i.e. compounds containing a ring system of the formula:, e.g. cephalosporins, cefaclor, or cephalexine containing further heterocyclic rings, e.g. cephalothin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/43—Enzymes; Proenzymes; Derivatives thereof
- A61K38/46—Hydrolases (3)
- A61K38/50—Hydrolases (3) acting on carbon-nitrogen bonds, other than peptide bonds (3.5), e.g. asparaginase
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y305/00—Hydrolases acting on carbon-nitrogen bonds, other than peptide bonds (3.5)
- C12Y305/02—Hydrolases acting on carbon-nitrogen bonds, other than peptide bonds (3.5) in cyclic amides (3.5.2)
- C12Y305/02006—Beta-lactamase (3.5.2.6)
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Medicinal Chemistry (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Genetics & Genomics (AREA)
- Veterinary Medicine (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Pharmacology & Pharmacy (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- Epidemiology (AREA)
- Microbiology (AREA)
- Biotechnology (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- Immunology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Gastroenterology & Hepatology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Oncology (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Communicable Diseases (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Enzymes And Modification Thereof (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Peptides Or Proteins (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
Description
V10A、及び
Q22H,N、及び
Q34R、及び
E130D
を含む。
(i)配列番号1の残基236〜240のC末端トランケーション、及び
(ii)位置10及び22の各々における置換、
位置10及び34の各々における置換、
位置10及び130の各々における置換、
位置10、22、及び34の各々における置換、
位置10、22、及び130の各々における置換、
位置10、34、及び130の各々における置換、又は
位置10、22、34、及び130の各々における置換
を含む。
VIM-2変異体の生産及び精製
VIM-2変異体は、T7プロモーターベースの発現系(pET-9発現プラスミドを使用)のいずれかを使用して、大腸菌において生産された。簡潔に言えば、NdeI及びBamHI制限部位を用いて突然変異体blaVIM-2遺伝子をプラスミドベクターpET-9にクローニングし、得られたプラスミドを大腸菌BL21(DE3)細胞に導入した。得られた宿主細胞を、ZnSO4を添加した富栄養自動誘導細胞培養培地ZYP-5052(Studier, F. W.、2005、Protein production by auto-induction in high density shaking cultures.、Protein Expr. Purif.、41:207〜234頁)中で24時間増殖させた。細菌細胞と培養上清とを遠心分離により分離した。次いで、清澄化した培養上清を物理的方法(例えば限外濾過)又は化学的方法(沈殿)を用いて濃縮した。或いは、細胞溶解を誘導するために物理的(超音波処理、フレンチプレス)又は化学的(リゾチーム、洗浄剤)薬剤で処理する前に、100〜200mlの20mMトリス(pH8.0)中に再懸濁させた細菌細胞からタンパク質を抽出できる。遠心分離により細胞抽出物を清澄化した。得られた清澄化試料を陰イオン交換クロマトグラフィーカラムに装填した。タンパク質を、20mMトリス緩衝液(pH8.0)中のNaClの直線勾配を用いて溶出し、活性画分をプールし、濃縮した。次いで、タンパク質試料を第2の陰イオン交換カラムに装填し、20mMトリエタノールアミン(pH7.2)中のNaClの直線勾配を用いて溶出した。得られた試料をゲル濾過カラムに装填し、50μΜ ZnSO4及び150mM NaClを添加した50mM HEPES(pH7.5)でタンパク質を溶出した。次いで、精製されたタンパク質を濃縮して保存した。
VIM-2変異体の腸管培地中での安定性の向上の決定
VIM-2変異体の安定性を測定するために、以下の手法を適用した:精製されたタンパク質試料のイミペネム加水分解活性を、ヒト回腸抽出物中でインキュベーション後に決定した。変異体に対する比活性(Sp.Act.)を、インキュベーション中の異なる時点(0、60、120及び240分)で測定した。腸管抽出物中の変異体の活性低下を評価するために、時刻tにおける比活性を、時刻t=0における比活性と比較した。経時変化は、開始時の活性(t=0の時)と後に測定された活性との比で表された((Sp.Act.変異体)t/(Sp.Act.変異体)t=0。1未満の値は、腸管抽出物中でインキュベートした場合の活性低下を示す。異なる時点での残存活性を比較し、野生型酵素と比較したいくつかの変異体の腸管抽出物でのより大きな安定性を評価する。
さまざまなβ-ラクタムに対する所与のVIM-2変異体の特異的な酵素活性及び触媒活性の決定
実施例1で述べたように生産した変異体酵素については、β-ラクタム系抗生物質のような特定のβ-ラクタム化合物に対する比活性や触媒特性を測定することが可能である。
腸管培地で4時間のインキュベーション後のさまざまなβ-ラクタムに対する所与のVIM-2変異体の酵素活性の決定
酵素活性の評価は、以下に記載するように行った。ヒト回腸抽出物中でVIM-2変異体を4時間のインキュベーション後、β-ラクタムの加水分解を適当な波長で分光光度法でモニターした。酵素活性は、加水分解されたβ-ラクタム基質のnmolを分単位で、試料中の総タンパク質1mgあたりで表してもよい。
Claims (16)
- β-ラクタマーゼ活性を有するポリペプチドであって、前記ポリペプチドは、配列番号1に対して少なくとも70%の配列同一性を有するアミノ酸配列を含み、前記ポリペプチドは、
- 位置10及び22の各々における置換、
- 位置10及び34の各々における置換、
- 位置10及び130の各々における置換、
- 位置10、22、及び34の各々における置換、
- 位置10、22、及び130の各々における置換、
- 位置10、34、及び130の各々における置換、又は
- 位置10、22、34、及び130の各々における置換
を含み、
前記位置は、配列番号1に示される配列中の位置に対応し、前記ポリペプチドは、配列番号1の野生型VIM-2と比較して、安定性に関して向上した特性を有する、ポリペプチド。 - 以下:
V10A、
Q22H又はQ22N、
Q34R、及び
E130D
から選択される少なくとも1つの改変を含む、請求項1に記載のポリペプチド。 - 配列番号1の第1の残基に対応する残基が、メチオニン残基に置き換えられる、請求項1又は2に記載のポリペプチド。
- そのN末端にシグナルペプチドを更に含む、請求項1から3のいずれか一項に記載のポリペプチド。
- 配列番号1に示される配列と比較して、そのN末端又はC末端にトランケーションを含む、請求項1から4のいずれか一項に記載のポリペプチド。
- - 配列番号1の残基237〜240のC末端トランケーション
- 配列番号1の残基236〜240のC末端トランケーション、又は
- 配列番号1の残基235〜240のC末端トランケーション
を含む、請求項5に記載のポリペプチド。 - 配列番号4〜23のいずれか1つのアミノ酸配列、又はβ-ラクタマーゼ活性を有するそのフラグメントを含む又はこれからなる、請求項1から5のいずれか一項に記載のポリペプチド。
- 請求項1から7のいずれか一項に記載のポリペプチドをコードする核酸配列を含む、単離核酸配列。
- 適切な発現宿主における前記ポリペプチドの発現を指示する1つ又は複数の制御配列に作動可能に連結されている、請求項8に記載の核酸配列を含む、核酸コンストラクト。
- 請求項9に記載の核酸コンストラクトを含む、組換え宿主細胞。
- 請求項1から7のいずれか一項に記載のポリペプチドを含む、組成物。
- 経口投与可能であり、腸の所望の部分において、特に空腸、回腸、盲腸、又は結腸において前記ポリペプチドを放出することができる、請求項11に記載の組成物。
- 個別投与、連続投与、又は同時投与のための、
(a)請求項1から7のいずれか一項に記載のポリペプチド、又は請求項11若しくは12に記載の組成物、及び
(b)(a)の又は(a)の組成物に含有される前記ポリペプチドに感受性を有するβ-ラクタム系抗生物質
を含む、キットオブパーツ。 - 医薬として使用するための、請求項1から7のいずれか一項に記載のポリペプチド。
- それを必要とする対象においてβ-ラクタム系抗生物質を不活性化するための方法において使用するための、請求項1から7のいずれか一項に記載のポリペプチド、請求項11若しくは12に記載の組成物、請求項13に記載のキットオブパーツ、又は請求項10に記載の組換え宿主細胞。
- 前記ポリペプチド又は前記組成物又は前記宿主細胞が、前記ポリペプチドに感受性を有するβ-ラクタム系抗生物質と組み合わせて使用するためのものである、細菌感染を治療するための方法において使用するための、請求項1から7のいずれか一項に記載のポリペプチド、請求項11若しくは12に記載の組成物、請求項13に記載のキットオブパーツ、又は請求項10に記載の宿主細胞。
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
EP17198414 | 2017-10-25 | ||
EP17198414.9 | 2017-10-25 | ||
PCT/EP2018/079223 WO2019081614A1 (en) | 2017-10-25 | 2018-10-24 | VARIANTS OF BETA-LACTAMASE |
Publications (2)
Publication Number | Publication Date |
---|---|
JP2021501603A true JP2021501603A (ja) | 2021-01-21 |
JP7223015B2 JP7223015B2 (ja) | 2023-02-15 |
Family
ID=60331400
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP2020543711A Active JP7223015B2 (ja) | 2017-10-25 | 2018-10-24 | β-ラクタマーゼ変異体 |
Country Status (10)
Country | Link |
---|---|
US (2) | US11365403B2 (ja) |
EP (1) | EP3701022B1 (ja) |
JP (1) | JP7223015B2 (ja) |
KR (1) | KR20200074982A (ja) |
CN (1) | CN111247246A (ja) |
BR (1) | BR112020007974A2 (ja) |
CA (1) | CA3077603A1 (ja) |
IL (1) | IL273728A (ja) |
MX (1) | MX2020004319A (ja) |
WO (1) | WO2019081614A1 (ja) |
Families Citing this family (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP7223015B2 (ja) * | 2017-10-25 | 2023-02-15 | ダ・ヴォルテッラ | β-ラクタマーゼ変異体 |
Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2016057744A1 (en) * | 2014-10-08 | 2016-04-14 | Synthetic Biologics, Inc. | Beta-lactamase formulations and uses thereof |
WO2017144495A1 (en) * | 2016-02-23 | 2017-08-31 | Da Volterra | Beta-lactamase variants |
WO2017144496A1 (en) * | 2016-02-23 | 2017-08-31 | Da Volterra | Beta-lactamase variants |
Family Cites Families (8)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
FI920206A0 (fi) | 1992-01-17 | 1992-01-17 | Pekka Untamo Heino | Medicinsk anvaendning, medicinskt foerfarande och preparat. |
US5605793A (en) | 1994-02-17 | 1997-02-25 | Affymax Technologies N.V. | Methods for in vitro recombination |
DE4422198C2 (de) | 1994-06-24 | 1997-08-28 | Audi Ag | Verfahren zum Steuern der elektrischen Beheizung eines Katalysators |
NZ330940A (en) | 1997-07-24 | 2000-02-28 | F | Production of consensus phytases from fungal origin using computer programmes |
FR2843302B1 (fr) | 2002-08-09 | 2004-10-22 | Centre Nat Rech Scient | Forme galenique pour la delivrance colique de principes actifs |
EP2407433A1 (en) | 2010-07-15 | 2012-01-18 | Da Volterra | Methods for the inactivation of antibiotics |
CN102757950A (zh) * | 2011-04-25 | 2012-10-31 | 天津市国际生物医药联合研究院 | 超级细菌新德里金属依赖型β-内酰胺酶NDM-1的表达纯化及β-内酰胺酶NDM-1的晶体结构 |
JP7223015B2 (ja) * | 2017-10-25 | 2023-02-15 | ダ・ヴォルテッラ | β-ラクタマーゼ変異体 |
-
2018
- 2018-10-24 JP JP2020543711A patent/JP7223015B2/ja active Active
- 2018-10-24 US US16/757,968 patent/US11365403B2/en active Active
- 2018-10-24 CA CA3077603A patent/CA3077603A1/en active Pending
- 2018-10-24 CN CN201880068725.0A patent/CN111247246A/zh active Pending
- 2018-10-24 BR BR112020007974-0A patent/BR112020007974A2/pt unknown
- 2018-10-24 KR KR1020207014812A patent/KR20200074982A/ko not_active Application Discontinuation
- 2018-10-24 MX MX2020004319A patent/MX2020004319A/es unknown
- 2018-10-24 WO PCT/EP2018/079223 patent/WO2019081614A1/en active Application Filing
- 2018-10-24 EP EP18789169.2A patent/EP3701022B1/en active Active
-
2020
- 2020-03-31 IL IL273728A patent/IL273728A/en unknown
-
2022
- 2022-05-16 US US17/745,205 patent/US20220356459A1/en not_active Abandoned
Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2016057744A1 (en) * | 2014-10-08 | 2016-04-14 | Synthetic Biologics, Inc. | Beta-lactamase formulations and uses thereof |
WO2017144495A1 (en) * | 2016-02-23 | 2017-08-31 | Da Volterra | Beta-lactamase variants |
WO2017144496A1 (en) * | 2016-02-23 | 2017-08-31 | Da Volterra | Beta-lactamase variants |
Also Published As
Publication number | Publication date |
---|---|
BR112020007974A2 (pt) | 2020-10-27 |
KR20200074982A (ko) | 2020-06-25 |
EP3701022A1 (en) | 2020-09-02 |
EP3701022B1 (en) | 2023-03-01 |
US20200318094A1 (en) | 2020-10-08 |
MX2020004319A (es) | 2020-10-12 |
CA3077603A1 (en) | 2019-05-02 |
US11365403B2 (en) | 2022-06-21 |
CN111247246A (zh) | 2020-06-05 |
WO2019081614A1 (en) | 2019-05-02 |
US20220356459A1 (en) | 2022-11-10 |
IL273728A (en) | 2020-05-31 |
JP7223015B2 (ja) | 2023-02-15 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
EP3210997B1 (en) | Agents and methods for the expression and secretion of peptides and proteins | |
CN111172129B (zh) | 一种提高热稳定、扩增均一性和扩增效率的Phi29 DNA聚合酶突变体及其应用 | |
KR20110106286A (ko) | 친화성 태그가 결합된 융합 콜라게나제 및 그의 제조방법 | |
JP6845258B2 (ja) | ベータ−ラクタマーゼ変異体 | |
US20220356459A1 (en) | Beta-lactamase variants | |
KR20140092835A (ko) | 히단토이나제의 돌연변이체 | |
US9732331B2 (en) | Codon modified amylase from Bacillus akibai | |
US10988749B2 (en) | Beta-lactamase variants | |
US20230183712A1 (en) | Methods for engineering amino acid ammonia lyase enzymes and enzymes thereby obtained | |
US20100137563A1 (en) | Cysteine Protease Autoprocessing of Fusion Proteins | |
KR20220154221A (ko) | 용해성 재조합 단백질의 생산 | |
JP4714848B2 (ja) | Dnaポリメラーゼ変異体 | |
CN114480345B (zh) | MazF突变体、重组载体、重组工程菌及其应用 | |
US20090259035A1 (en) | Method for producing recombinant RNase A | |
JP6218512B2 (ja) | アユ冷水病菌由来毒素 | |
Novikov et al. | The highly efficient expression of the aspartase gene (L-aspartate ammonia-lyase) in Escherichia coli cells | |
Grishin et al. | Preparation and Characterization of a New Mutant Homolog of the Chemotaxis Protein CheY from the Anaerobic Hyperthermophilic Microorganism Thermotoga Naphthophila | |
KR100755727B1 (ko) | β-갈락토시다제 유래의 융합 단편 펩타이드 및 이를 융합파트너로 이용하는 재조합 단백질의 제조방법 | |
CN117187202A (zh) | 高稳定性超氧化物歧化酶突变体及其应用 | |
Feng | Metal ion-mediated inactivation and small molecule inhibition of the metalloprotease in botulinum neurotoxin A | |
JP2008220196A (ja) | タンパク質の欠失変異体の作製方法 | |
JP2003219892A (ja) | 耐熱性システイン合成酵素 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
A621 | Written request for application examination |
Free format text: JAPANESE INTERMEDIATE CODE: A621 Effective date: 20210816 |
|
A977 | Report on retrieval |
Free format text: JAPANESE INTERMEDIATE CODE: A971007 Effective date: 20220622 |
|
A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20220627 |
|
A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20220927 |
|
TRDD | Decision of grant or rejection written | ||
A01 | Written decision to grant a patent or to grant a registration (utility model) |
Free format text: JAPANESE INTERMEDIATE CODE: A01 Effective date: 20221205 |
|
A601 | Written request for extension of time |
Free format text: JAPANESE INTERMEDIATE CODE: A601 Effective date: 20230104 |
|
A61 | First payment of annual fees (during grant procedure) |
Free format text: JAPANESE INTERMEDIATE CODE: A61 Effective date: 20230203 |
|
R150 | Certificate of patent or registration of utility model |
Ref document number: 7223015 Country of ref document: JP Free format text: JAPANESE INTERMEDIATE CODE: R150 |