JP5931738B2 - 新規のペプチジルアルファ−ヒドロキシグリシンアルファ−アミド化リアーゼ - Google Patents
新規のペプチジルアルファ−ヒドロキシグリシンアルファ−アミド化リアーゼ Download PDFInfo
- Publication number
- JP5931738B2 JP5931738B2 JP2012541487A JP2012541487A JP5931738B2 JP 5931738 B2 JP5931738 B2 JP 5931738B2 JP 2012541487 A JP2012541487 A JP 2012541487A JP 2012541487 A JP2012541487 A JP 2012541487A JP 5931738 B2 JP5931738 B2 JP 5931738B2
- Authority
- JP
- Japan
- Prior art keywords
- enzyme
- xaa
- polypeptide
- peptide
- seq
- 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.)
- Expired - Fee Related
Links
- 108090000856 Lyases Proteins 0.000 title claims description 39
- 102000004317 Lyases Human genes 0.000 title claims description 39
- -1 peptidyl alpha-hydroxyglycine Chemical compound 0.000 title claims description 10
- 108090000765 processed proteins & peptides Proteins 0.000 claims description 384
- 102000004190 Enzymes Human genes 0.000 claims description 344
- 108090000790 Enzymes Proteins 0.000 claims description 344
- 102000004196 processed proteins & peptides Human genes 0.000 claims description 257
- 229920001184 polypeptide Polymers 0.000 claims description 222
- 238000006243 chemical reaction Methods 0.000 claims description 82
- 238000000034 method Methods 0.000 claims description 82
- 230000000694 effects Effects 0.000 claims description 74
- 230000004048 modification Effects 0.000 claims description 66
- 238000012986 modification Methods 0.000 claims description 66
- 235000001014 amino acid Nutrition 0.000 claims description 63
- 150000001413 amino acids Chemical class 0.000 claims description 61
- 210000004899 c-terminal region Anatomy 0.000 claims description 61
- 241000588724 Escherichia coli Species 0.000 claims description 41
- 241000894006 Bacteria Species 0.000 claims description 31
- 238000004519 manufacturing process Methods 0.000 claims description 28
- 125000003630 glycyl group Chemical group [H]N([H])C([H])([H])C(*)=O 0.000 claims description 27
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 claims description 20
- 102100027467 Pro-opiomelanocortin Human genes 0.000 claims description 17
- 108010041872 Islet Amyloid Polypeptide Proteins 0.000 claims description 15
- 102000036770 Islet Amyloid Polypeptide Human genes 0.000 claims description 15
- 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 13
- 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 claims description 13
- 108010088847 Peptide YY Proteins 0.000 claims description 13
- 102100029909 Peptide YY Human genes 0.000 claims description 13
- 125000000151 cysteine group Chemical group N[C@@H](CS)C(=O)* 0.000 claims description 13
- 102000055006 Calcitonin Human genes 0.000 claims description 12
- 108060001064 Calcitonin Proteins 0.000 claims description 12
- 101800001751 Melanocyte-stimulating hormone alpha Proteins 0.000 claims description 12
- 108010074633 Mixed Function Oxygenases Proteins 0.000 claims description 12
- 102000008109 Mixed Function Oxygenases Human genes 0.000 claims description 12
- 108010069820 Pro-Opiomelanocortin Proteins 0.000 claims description 12
- 239000000683 Pro-Opiomelanocortin Substances 0.000 claims description 12
- 229960004015 calcitonin Drugs 0.000 claims description 12
- BBBFJLBPOGFECG-VJVYQDLKSA-N calcitonin Chemical compound N([C@H](C(=O)N[C@@H](CC(C)C)C(=O)NCC(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC=1NC=NC=1)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)NCC(=O)N[C@@H](CO)C(=O)NCC(=O)N[C@@H]([C@@H](C)O)C(=O)N1[C@@H](CCC1)C(N)=O)C(C)C)C(=O)[C@@H]1CSSC[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)O)C(=O)N1 BBBFJLBPOGFECG-VJVYQDLKSA-N 0.000 claims description 12
- WHNFPRLDDSXQCL-UAZQEYIDSA-N α-msh Chemical compound C([C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC=1NC=NC=1)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)NCC(=O)N[C@@H](CCCCN)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](C(C)C)C(N)=O)NC(=O)[C@H](CO)NC(C)=O)C1=CC=C(O)C=C1 WHNFPRLDDSXQCL-UAZQEYIDSA-N 0.000 claims description 12
- 108010088406 Glucagon-Like Peptides Proteins 0.000 claims description 11
- 229920001192 peptidylglycine Polymers 0.000 claims description 11
- 235000010323 ascorbic acid Nutrition 0.000 claims description 10
- 229960005070 ascorbic acid Drugs 0.000 claims description 10
- 239000011668 ascorbic acid Substances 0.000 claims description 10
- 230000002255 enzymatic effect Effects 0.000 claims description 10
- 102400000921 Gastrin Human genes 0.000 claims description 7
- 108090001053 Gastrin releasing peptide Proteins 0.000 claims description 7
- 102000004862 Gastrin releasing peptide Human genes 0.000 claims description 7
- 108010052343 Gastrins Proteins 0.000 claims description 7
- HEAUFJZALFKPBA-YRVBCFNBSA-N Neurokinin A Chemical compound C([C@@H](C(=O)N[C@H](C(=O)NCC(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCSC)C(N)=O)C(C)C)NC(=O)[C@H](CO)NC(=O)[C@H](CC(O)=O)NC(=O)[C@@H](NC(=O)[C@H](CCCCN)NC(=O)[C@@H](N)CC=1NC=NC=1)C(C)O)C1=CC=CC=C1 HEAUFJZALFKPBA-YRVBCFNBSA-N 0.000 claims description 7
- 101800000399 Neurokinin A Proteins 0.000 claims description 7
- 102400000097 Neurokinin A Human genes 0.000 claims description 7
- 101800000989 Oxytocin Proteins 0.000 claims description 7
- 102400000050 Oxytocin Human genes 0.000 claims description 7
- XNOPRXBHLZRZKH-UHFFFAOYSA-N Oxytocin Natural products N1C(=O)C(N)CSSCC(C(=O)N2C(CCC2)C(=O)NC(CC(C)C)C(=O)NCC(N)=O)NC(=O)C(CC(N)=O)NC(=O)C(CCC(N)=O)NC(=O)C(C(C)CC)NC(=O)C1CC1=CC=C(O)C=C1 XNOPRXBHLZRZKH-UHFFFAOYSA-N 0.000 claims description 7
- 108010086019 Secretin Proteins 0.000 claims description 7
- 102100037505 Secretin Human genes 0.000 claims description 7
- GXBMIBRIOWHPDT-UHFFFAOYSA-N Vasopressin Natural products N1C(=O)C(CC=2C=C(O)C=CC=2)NC(=O)C(N)CSSCC(C(=O)N2C(CCC2)C(=O)NC(CCCN=C(N)N)C(=O)NCC(N)=O)NC(=O)C(CC(N)=O)NC(=O)C(CCC(N)=O)NC(=O)C1CC1=CC=CC=C1 GXBMIBRIOWHPDT-UHFFFAOYSA-N 0.000 claims description 7
- 108010004977 Vasopressins Proteins 0.000 claims description 7
- 102000002852 Vasopressins Human genes 0.000 claims description 7
- KBZOIRJILGZLEJ-LGYYRGKSSA-N argipressin Chemical compound C([C@H]1C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CSSC[C@@H](C(N[C@@H](CC=2C=CC(O)=CC=2)C(=O)N1)=O)N)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CCCN=C(N)N)C(=O)NCC(N)=O)C1=CC=CC=C1 KBZOIRJILGZLEJ-LGYYRGKSSA-N 0.000 claims description 7
- AOXOCDRNSPFDPE-UKEONUMOSA-N chembl413654 Chemical compound C([C@H](C(=O)NCC(=O)N[C@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@H](CCSC)C(=O)N[C@H](CC(O)=O)C(=O)N[C@H](CC=1C=CC=CC=1)C(N)=O)NC(=O)[C@@H](C)NC(=O)[C@@H](CCC(O)=O)NC(=O)[C@@H](CCC(O)=O)NC(=O)[C@@H](CCC(O)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CC=1C2=CC=CC=C2NC=1)NC(=O)[C@H]1N(CCC1)C(=O)CNC(=O)[C@@H](N)CCC(O)=O)C1=CC=C(O)C=C1 AOXOCDRNSPFDPE-UKEONUMOSA-N 0.000 claims description 7
- PUBCCFNQJQKCNC-XKNFJVFFSA-N gastrin-releasingpeptide Chemical compound C([C@@H](C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCSC)C(N)=O)NC(=O)CNC(=O)[C@@H](NC(=O)[C@H](C)NC(=O)[C@H](CC=1C2=CC=CC=C2NC=1)NC(=O)[C@H](CC=1N=CNC=1)NC(=O)[C@H](CC(N)=O)NC(=O)CNC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H]1N(CCC1)C(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CCSC)NC(=O)[C@H](CCCCN)NC(=O)[C@@H](NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](NC(=O)[C@@H](NC(=O)CNC(=O)CNC(=O)CNC(=O)[C@H](C)NC(=O)[C@H]1N(CCC1)C(=O)[C@H](CC(C)C)NC(=O)[C@H]1N(CCC1)C(=O)[C@@H](N)C(C)C)[C@@H](C)O)C(C)C)[C@@H](C)O)C(C)C)C1=CNC=N1 PUBCCFNQJQKCNC-XKNFJVFFSA-N 0.000 claims description 7
- XNOPRXBHLZRZKH-DSZYJQQASA-N oxytocin Chemical compound C([C@H]1C(=O)N[C@H](C(N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CSSC[C@H](N)C(=O)N1)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CC(C)C)C(=O)NCC(N)=O)=O)[C@@H](C)CC)C1=CC=C(O)C=C1 XNOPRXBHLZRZKH-DSZYJQQASA-N 0.000 claims description 7
- 229960001723 oxytocin Drugs 0.000 claims description 7
- 230000008569 process Effects 0.000 claims description 7
- 229960002101 secretin Drugs 0.000 claims description 7
- OWMZNFCDEHGFEP-NFBCVYDUSA-N secretin human Chemical compound C([C@@H](C(=O)N[C@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(O)=O)C(=O)NCC(=O)N[C@@H](C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(N)=O)C(=O)NCC(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C(C)C)C(N)=O)[C@@H](C)O)NC(=O)[C@@H](NC(=O)CNC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](N)CC=1NC=NC=1)[C@@H](C)O)C1=CC=CC=C1 OWMZNFCDEHGFEP-NFBCVYDUSA-N 0.000 claims description 7
- 229960003726 vasopressin Drugs 0.000 claims description 7
- 230000003197 catalytic effect Effects 0.000 claims description 6
- 102100038518 Calcitonin Human genes 0.000 claims description 5
- 108090000932 Calcitonin Gene-Related Peptide Proteins 0.000 claims description 5
- 102000018886 Pancreatic Polypeptide Human genes 0.000 claims description 5
- 108700020479 Pancreatic hormone Proteins 0.000 claims description 5
- 150000001408 amides Chemical class 0.000 claims description 5
- BPGXUIVWLQTVLZ-OFGSCBOVSA-N neuropeptide y(npy) Chemical compound C([C@@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(O)=O)NC(=O)[C@H](CC=1N=CNC=1)NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](C)NC(=O)[C@H](CO)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](C)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](C)NC(=O)[C@H]1N(CCC1)C(=O)[C@H](C)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)CNC(=O)[C@H]1N(CCC1)C(=O)[C@H](CC(N)=O)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H]1N(CCC1)C(=O)[C@H](CCCCN)NC(=O)[C@H](CO)NC(=O)[C@H]1N(CCC1)C(=O)[C@@H](N)CC=1C=CC(O)=CC=1)C1=CC=C(O)C=C1 BPGXUIVWLQTVLZ-OFGSCBOVSA-N 0.000 claims description 5
- 101800000520 Melanotropin gamma Proteins 0.000 claims description 4
- 150000002500 ions Chemical class 0.000 claims description 4
- 229940088598 enzyme Drugs 0.000 description 342
- 108090000623 proteins and genes Proteins 0.000 description 168
- 102000004169 proteins and genes Human genes 0.000 description 147
- 235000018102 proteins Nutrition 0.000 description 142
- 210000004027 cell Anatomy 0.000 description 121
- ZHWLPDIRXJCEJY-UHFFFAOYSA-N alpha-hydroxyglycine Chemical compound NC(O)C(O)=O ZHWLPDIRXJCEJY-UHFFFAOYSA-N 0.000 description 97
- 238000000746 purification Methods 0.000 description 77
- 125000003275 alpha amino acid group Chemical group 0.000 description 71
- 230000014509 gene expression Effects 0.000 description 68
- 102000039446 nucleic acids Human genes 0.000 description 60
- 108020004707 nucleic acids Proteins 0.000 description 60
- 150000007523 nucleic acids Chemical class 0.000 description 60
- 229940024606 amino acid Drugs 0.000 description 57
- KXDAEFPNCMNJSK-UHFFFAOYSA-N Benzamide Chemical compound NC(=O)C1=CC=CC=C1 KXDAEFPNCMNJSK-UHFFFAOYSA-N 0.000 description 38
- 241000190844 Erythrobacter Species 0.000 description 37
- 239000013612 plasmid Substances 0.000 description 36
- 108010076504 Protein Sorting Signals Proteins 0.000 description 35
- 239000013598 vector Substances 0.000 description 33
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 32
- 238000003776 cleavage reaction Methods 0.000 description 32
- 230000007017 scission Effects 0.000 description 32
- 239000000872 buffer Substances 0.000 description 31
- 108020001507 fusion proteins Proteins 0.000 description 31
- 102000037865 fusion proteins Human genes 0.000 description 31
- 238000004458 analytical method Methods 0.000 description 27
- 230000004927 fusion Effects 0.000 description 24
- 108010029667 pramlintide Proteins 0.000 description 22
- 239000002243 precursor Substances 0.000 description 21
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 20
- GCWCVCCEIQXUQU-UHFFFAOYSA-N alpha-hydroxyhippuric acid Chemical compound OC(=O)C(O)NC(=O)C1=CC=CC=C1 GCWCVCCEIQXUQU-UHFFFAOYSA-N 0.000 description 20
- ONJSZLXSECQROL-UHFFFAOYSA-N alpha-hydroxyhippuric acid Natural products OC(=O)CNC(=O)C1=CC=CC=C1O ONJSZLXSECQROL-UHFFFAOYSA-N 0.000 description 20
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 20
- 239000000758 substrate Substances 0.000 description 20
- 108091005804 Peptidases Proteins 0.000 description 18
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 18
- 235000014680 Saccharomyces cerevisiae Nutrition 0.000 description 18
- 239000012634 fragment Substances 0.000 description 18
- 239000002773 nucleotide Substances 0.000 description 18
- 125000003729 nucleotide group Chemical group 0.000 description 18
- NRKVKVQDUCJPIZ-MKAGXXMWSA-N pramlintide acetate Chemical compound C([C@@H](C(=O)NCC(=O)N1CCC[C@H]1C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC(C)C)C(=O)N1[C@@H](CCC1)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](C(C)C)C(=O)NCC(=O)N[C@@H](CO)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(N)=O)NC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CO)NC(=O)[C@H](CO)NC(=O)[C@H](CC=1NC=NC=1)NC(=O)[C@@H](NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CC=1C=CC=CC=1)NC(=O)[C@H](CC(N)=O)NC(=O)[C@H](C)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@@H](NC(=O)[C@H](C)NC(=O)[C@H](CS)NC(=O)[C@@H](NC(=O)[C@H](C)NC(=O)[C@@H](NC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CS)NC(=O)[C@@H](N)CCCCN)[C@@H](C)O)[C@@H](C)O)[C@@H](C)O)C(C)C)C1=CC=CC=C1 NRKVKVQDUCJPIZ-MKAGXXMWSA-N 0.000 description 18
- 239000004365 Protease Substances 0.000 description 17
- 101000959116 Rattus norvegicus Peptidyl-alpha-hydroxyglycine alpha-amidating lyase Proteins 0.000 description 17
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 description 17
- 235000019419 proteases Nutrition 0.000 description 17
- 108091028043 Nucleic acid sequence Proteins 0.000 description 16
- 229960003611 pramlintide Drugs 0.000 description 16
- 241000894007 species Species 0.000 description 16
- 238000011282 treatment Methods 0.000 description 16
- 238000007112 amidation reaction Methods 0.000 description 15
- 230000001580 bacterial effect Effects 0.000 description 15
- 108020004414 DNA Proteins 0.000 description 14
- 229920002684 Sepharose Polymers 0.000 description 14
- 241000700159 Rattus Species 0.000 description 13
- 239000007983 Tris buffer Substances 0.000 description 13
- 230000001747 exhibiting effect Effects 0.000 description 13
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 description 13
- ROHFNLRQFUQHCH-YFKPBYRVSA-N L-leucine Chemical compound CC(C)C[C@H](N)C(O)=O ROHFNLRQFUQHCH-YFKPBYRVSA-N 0.000 description 12
- 239000013604 expression vector Substances 0.000 description 12
- 239000002609 medium Substances 0.000 description 12
- 108091033319 polynucleotide Proteins 0.000 description 12
- 102000040430 polynucleotide Human genes 0.000 description 12
- 239000002157 polynucleotide Substances 0.000 description 12
- 238000004895 liquid chromatography mass spectrometry Methods 0.000 description 11
- 238000001228 spectrum Methods 0.000 description 11
- 241000233866 Fungi Species 0.000 description 10
- 201000001421 hyperglycemia Diseases 0.000 description 10
- 239000011780 sodium chloride Substances 0.000 description 10
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 9
- 241000269370 Xenopus <genus> Species 0.000 description 9
- 238000005341 cation exchange Methods 0.000 description 9
- 238000010367 cloning Methods 0.000 description 9
- 239000002299 complementary DNA Substances 0.000 description 9
- 230000006870 function Effects 0.000 description 9
- 238000004128 high performance liquid chromatography Methods 0.000 description 9
- 238000002360 preparation method Methods 0.000 description 9
- 238000001195 ultra high performance liquid chromatography Methods 0.000 description 9
- 230000002862 amidating effect Effects 0.000 description 8
- 238000005119 centrifugation Methods 0.000 description 8
- 239000000523 sample Substances 0.000 description 8
- 238000000926 separation method Methods 0.000 description 8
- 239000006228 supernatant Substances 0.000 description 8
- 238000012360 testing method Methods 0.000 description 8
- 230000035897 transcription Effects 0.000 description 8
- 238000013518 transcription Methods 0.000 description 8
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 7
- 208000008589 Obesity Diseases 0.000 description 7
- 102100039087 Peptidyl-alpha-hydroxyglycine alpha-amidating lyase Human genes 0.000 description 7
- 239000000356 contaminant Substances 0.000 description 7
- 238000012258 culturing Methods 0.000 description 7
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 7
- 239000003814 drug Substances 0.000 description 7
- 239000000463 material Substances 0.000 description 7
- 235000020824 obesity Nutrition 0.000 description 7
- 230000003248 secreting effect Effects 0.000 description 7
- 238000006467 substitution reaction Methods 0.000 description 7
- YBJHBAHKTGYVGT-ZKWXMUAHSA-N (+)-Biotin Chemical compound N1C(=O)N[C@@H]2[C@H](CCCCC(=O)O)SC[C@@H]21 YBJHBAHKTGYVGT-ZKWXMUAHSA-N 0.000 description 6
- 239000004382 Amylase Substances 0.000 description 6
- 108010065511 Amylases Proteins 0.000 description 6
- 241000228212 Aspergillus Species 0.000 description 6
- 108091026890 Coding region Proteins 0.000 description 6
- YWAQATDNEKZFFK-BYPYZUCNSA-N Gly-Gly-Ser Chemical compound NCC(=O)NCC(=O)N[C@@H](CO)C(O)=O YWAQATDNEKZFFK-BYPYZUCNSA-N 0.000 description 6
- CKLJMWTZIZZHCS-REOHCLBHSA-N L-aspartic acid Chemical compound OC(=O)[C@@H](N)CC(O)=O CKLJMWTZIZZHCS-REOHCLBHSA-N 0.000 description 6
- 230000009435 amidation Effects 0.000 description 6
- 125000000539 amino acid group Chemical group 0.000 description 6
- 230000006378 damage Effects 0.000 description 6
- 206010012601 diabetes mellitus Diseases 0.000 description 6
- 201000010099 disease Diseases 0.000 description 6
- 239000000499 gel Substances 0.000 description 6
- 238000011534 incubation Methods 0.000 description 6
- 238000003780 insertion Methods 0.000 description 6
- 230000037431 insertion Effects 0.000 description 6
- 210000004962 mammalian cell Anatomy 0.000 description 6
- 239000000203 mixture Substances 0.000 description 6
- 239000000047 product Substances 0.000 description 6
- 150000003839 salts Chemical class 0.000 description 6
- 208000001072 type 2 diabetes mellitus Diseases 0.000 description 6
- 102000013142 Amylases Human genes 0.000 description 5
- 208000024172 Cardiovascular disease Diseases 0.000 description 5
- 208000002705 Glucose Intolerance Diseases 0.000 description 5
- XQJCEKXQUJQNNK-ZLUOBGJFSA-N Ser-Ser-Ser Chemical compound OC[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@@H](CO)C(O)=O XQJCEKXQUJQNNK-ZLUOBGJFSA-N 0.000 description 5
- 208000002847 Surgical Wound Diseases 0.000 description 5
- 206010067584 Type 1 diabetes mellitus Diseases 0.000 description 5
- 208000027418 Wounds and injury Diseases 0.000 description 5
- 229960000723 ampicillin Drugs 0.000 description 5
- 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 5
- 235000019418 amylase Nutrition 0.000 description 5
- 238000003556 assay Methods 0.000 description 5
- 238000005277 cation exchange chromatography Methods 0.000 description 5
- 208000029078 coronary artery disease Diseases 0.000 description 5
- 208000014674 injury Diseases 0.000 description 5
- BPHPUYQFMNQIOC-NXRLNHOXSA-N isopropyl beta-D-thiogalactopyranoside Chemical compound CC(C)S[C@@H]1O[C@H](CO)[C@H](O)[C@H](O)[C@H]1O BPHPUYQFMNQIOC-NXRLNHOXSA-N 0.000 description 5
- 108020004999 messenger RNA Proteins 0.000 description 5
- 208000010125 myocardial infarction Diseases 0.000 description 5
- 201000009104 prediabetes syndrome Diseases 0.000 description 5
- 230000002265 prevention Effects 0.000 description 5
- 230000006798 recombination Effects 0.000 description 5
- 238000005215 recombination Methods 0.000 description 5
- 230000005030 transcription termination Effects 0.000 description 5
- 210000005253 yeast cell Anatomy 0.000 description 5
- PGUYEUCYVNZGGV-QWRGUYRKSA-N Asp-Gly-Tyr Chemical compound OC(=O)C[C@H](N)C(=O)NCC(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 PGUYEUCYVNZGGV-QWRGUYRKSA-N 0.000 description 4
- 241000228245 Aspergillus niger Species 0.000 description 4
- 241000193830 Bacillus <bacterium> Species 0.000 description 4
- 241000328900 Candidatus Solibacter Species 0.000 description 4
- 108020004705 Codon Proteins 0.000 description 4
- 241000190842 Erythrobacter sp. Species 0.000 description 4
- 101001081479 Homo sapiens Islet amyloid polypeptide Proteins 0.000 description 4
- KZNQNBZMBZJQJO-UHFFFAOYSA-N N-glycyl-L-proline Natural products NCC(=O)N1CCCC1C(O)=O KZNQNBZMBZJQJO-UHFFFAOYSA-N 0.000 description 4
- 101710189920 Peptidyl-alpha-hydroxyglycine alpha-amidating lyase Proteins 0.000 description 4
- 208000006011 Stroke Diseases 0.000 description 4
- 108020005038 Terminator Codon Proteins 0.000 description 4
- 230000001195 anabolic effect Effects 0.000 description 4
- 230000004071 biological effect Effects 0.000 description 4
- 230000015572 biosynthetic process Effects 0.000 description 4
- 238000012217 deletion Methods 0.000 description 4
- 230000037430 deletion Effects 0.000 description 4
- 238000006911 enzymatic reaction Methods 0.000 description 4
- KZNQNBZMBZJQJO-YFKPBYRVSA-N glyclproline Chemical compound NCC(=O)N1CCC[C@H]1C(O)=O KZNQNBZMBZJQJO-YFKPBYRVSA-N 0.000 description 4
- 108010077515 glycylproline Proteins 0.000 description 4
- 239000001963 growth medium Substances 0.000 description 4
- 238000000338 in vitro Methods 0.000 description 4
- 239000011159 matrix material Substances 0.000 description 4
- 230000007935 neutral effect Effects 0.000 description 4
- 239000000813 peptide hormone Substances 0.000 description 4
- 108010007262 peptidylglycine monooxygenase Proteins 0.000 description 4
- 230000008488 polyadenylation Effects 0.000 description 4
- 238000012545 processing Methods 0.000 description 4
- 238000003259 recombinant expression Methods 0.000 description 4
- 108090000907 ribosomal protein L9 Proteins 0.000 description 4
- 102000004346 ribosomal protein L9 Human genes 0.000 description 4
- 239000001488 sodium phosphate Substances 0.000 description 4
- 229910000162 sodium phosphate Inorganic materials 0.000 description 4
- 238000000527 sonication Methods 0.000 description 4
- RYFMWSXOAZQYPI-UHFFFAOYSA-K trisodium phosphate Chemical compound [Na+].[Na+].[Na+].[O-]P([O-])([O-])=O RYFMWSXOAZQYPI-UHFFFAOYSA-K 0.000 description 4
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 3
- 240000006439 Aspergillus oryzae Species 0.000 description 3
- 241000193744 Bacillus amyloliquefaciens Species 0.000 description 3
- 244000063299 Bacillus subtilis Species 0.000 description 3
- 235000014469 Bacillus subtilis Nutrition 0.000 description 3
- 241001453380 Burkholderia Species 0.000 description 3
- 208000028399 Critical Illness Diseases 0.000 description 3
- 102000053602 DNA Human genes 0.000 description 3
- 241000206602 Eukaryota Species 0.000 description 3
- 108010073178 Glucan 1,4-alpha-Glucosidase Proteins 0.000 description 3
- 102100022624 Glucoamylase Human genes 0.000 description 3
- 108010070675 Glutathione transferase Proteins 0.000 description 3
- 102000005720 Glutathione transferase Human genes 0.000 description 3
- 239000004471 Glycine Substances 0.000 description 3
- 241001465754 Metazoa Species 0.000 description 3
- 241000235648 Pichia Species 0.000 description 3
- 206010036105 Polyneuropathy Diseases 0.000 description 3
- 108020005067 RNA Splice Sites Proteins 0.000 description 3
- 108020004511 Recombinant DNA Proteins 0.000 description 3
- 241001532577 Sorangium Species 0.000 description 3
- 108010090804 Streptavidin Proteins 0.000 description 3
- 239000002253 acid Substances 0.000 description 3
- 125000003295 alanine group Chemical group N[C@@H](C)C(=O)* 0.000 description 3
- 108090000637 alpha-Amylases Proteins 0.000 description 3
- 102000004139 alpha-Amylases Human genes 0.000 description 3
- 229940024171 alpha-amylase Drugs 0.000 description 3
- 238000005349 anion exchange Methods 0.000 description 3
- 230000008901 benefit Effects 0.000 description 3
- 229960002685 biotin Drugs 0.000 description 3
- 235000020958 biotin Nutrition 0.000 description 3
- 239000011616 biotin Substances 0.000 description 3
- 238000004113 cell culture Methods 0.000 description 3
- 210000000349 chromosome Anatomy 0.000 description 3
- 150000001875 compounds Chemical class 0.000 description 3
- 238000012790 confirmation Methods 0.000 description 3
- 229940079593 drug Drugs 0.000 description 3
- 230000006698 induction Effects 0.000 description 3
- 238000004811 liquid chromatography Methods 0.000 description 3
- 239000006166 lysate Substances 0.000 description 3
- 239000003550 marker Substances 0.000 description 3
- 229910052751 metal Inorganic materials 0.000 description 3
- 239000002184 metal Substances 0.000 description 3
- 230000035772 mutation Effects 0.000 description 3
- 125000001151 peptidyl group Chemical group 0.000 description 3
- 239000012071 phase Substances 0.000 description 3
- 230000007824 polyneuropathy Effects 0.000 description 3
- 239000011541 reaction mixture Substances 0.000 description 3
- 230000002441 reversible effect Effects 0.000 description 3
- 230000028327 secretion Effects 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 230000009466 transformation Effects 0.000 description 3
- 238000011144 upstream manufacturing Methods 0.000 description 3
- DGVVWUTYPXICAM-UHFFFAOYSA-N β‐Mercaptoethanol Chemical compound OCCS DGVVWUTYPXICAM-UHFFFAOYSA-N 0.000 description 3
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 2
- 102100034042 Alcohol dehydrogenase 1C Human genes 0.000 description 2
- 241001135756 Alphaproteobacteria Species 0.000 description 2
- 235000002247 Aspergillus oryzae Nutrition 0.000 description 2
- 241000193422 Bacillus lentus Species 0.000 description 2
- 241000194103 Bacillus pumilus Species 0.000 description 2
- 241000589173 Bradyrhizobium Species 0.000 description 2
- 241000193764 Brevibacillus brevis Species 0.000 description 2
- 101000796894 Coturnix japonica Alcohol dehydrogenase 1 Proteins 0.000 description 2
- 108010013369 Enteropeptidase Proteins 0.000 description 2
- 102100029727 Enteropeptidase Human genes 0.000 description 2
- 241000193385 Geobacillus stearothermophilus Species 0.000 description 2
- FGPLUIQCSKGLTI-WDSKDSINSA-N Gly-Ser-Glu Chemical compound NCC(=O)N[C@@H](CO)C(=O)N[C@H](C(O)=O)CCC(O)=O FGPLUIQCSKGLTI-WDSKDSINSA-N 0.000 description 2
- SOEGEPHNZOISMT-BYPYZUCNSA-N Gly-Ser-Gly Chemical compound NCC(=O)N[C@@H](CO)C(=O)NCC(O)=O SOEGEPHNZOISMT-BYPYZUCNSA-N 0.000 description 2
- 241000238631 Hexapoda Species 0.000 description 2
- 101000780463 Homo sapiens Alcohol dehydrogenase 1C Proteins 0.000 description 2
- 101000801742 Homo sapiens Triosephosphate isomerase Proteins 0.000 description 2
- 108010000521 Human Growth Hormone Proteins 0.000 description 2
- 241000701109 Human adenovirus 2 Species 0.000 description 2
- 206010020772 Hypertension Diseases 0.000 description 2
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 description 2
- 241000235058 Komagataella pastoris Species 0.000 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 2
- KQFZKDITNUEVFJ-JYJNAYRXSA-N Leu-Phe-Gln Chemical compound NC(=O)CC[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)[C@@H](N)CC(C)C)CC1=CC=CC=C1 KQFZKDITNUEVFJ-JYJNAYRXSA-N 0.000 description 2
- 102000004882 Lipase Human genes 0.000 description 2
- 108090001060 Lipase Proteins 0.000 description 2
- 239000004367 Lipase Substances 0.000 description 2
- 241000124008 Mammalia Species 0.000 description 2
- 101710151321 Melanostatin Proteins 0.000 description 2
- 102400000064 Neuropeptide Y Human genes 0.000 description 2
- 108010019160 Pancreatin Proteins 0.000 description 2
- 241000589952 Planctomyces Species 0.000 description 2
- 241000700157 Rattus norvegicus Species 0.000 description 2
- 102000002278 Ribosomal Proteins Human genes 0.000 description 2
- 108010000605 Ribosomal Proteins Proteins 0.000 description 2
- 241000235070 Saccharomyces Species 0.000 description 2
- 241000426682 Salinispora Species 0.000 description 2
- 108091081024 Start codon Proteins 0.000 description 2
- 108010022394 Threonine synthase Proteins 0.000 description 2
- 241000723792 Tobacco etch virus Species 0.000 description 2
- 241000223259 Trichoderma Species 0.000 description 2
- 241000012469 Trimerotropis maritima Species 0.000 description 2
- 102100033598 Triosephosphate isomerase Human genes 0.000 description 2
- IXKSXJFAGXLQOQ-XISFHERQSA-N WHWLQLKPGQPMY Chemical group C([C@@H](C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(C)C)C(=O)N1CCC[C@H]1C(=O)NCC(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(O)=O)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(O)=O)NC(=O)[C@@H](N)CC=1C2=CC=CC=C2NC=1)C1=CNC=N1 IXKSXJFAGXLQOQ-XISFHERQSA-N 0.000 description 2
- 238000002835 absorbance Methods 0.000 description 2
- 238000010521 absorption reaction Methods 0.000 description 2
- 230000002378 acidificating effect Effects 0.000 description 2
- 150000007513 acids Chemical class 0.000 description 2
- 235000004279 alanine Nutrition 0.000 description 2
- 238000005571 anion exchange chromatography Methods 0.000 description 2
- 230000001588 bifunctional effect Effects 0.000 description 2
- 230000000975 bioactive effect Effects 0.000 description 2
- 108020001778 catalytic domains Proteins 0.000 description 2
- 238000006555 catalytic reaction Methods 0.000 description 2
- 239000013592 cell lysate Substances 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 238000012512 characterization method Methods 0.000 description 2
- 239000005547 deoxyribonucleotide Substances 0.000 description 2
- 125000002637 deoxyribonucleotide group Chemical group 0.000 description 2
- 238000011033 desalting Methods 0.000 description 2
- 102000004419 dihydrofolate reductase Human genes 0.000 description 2
- 238000011143 downstream manufacturing Methods 0.000 description 2
- 238000010828 elution Methods 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 230000009144 enzymatic modification Effects 0.000 description 2
- 210000003527 eukaryotic cell Anatomy 0.000 description 2
- 238000011156 evaluation Methods 0.000 description 2
- 238000000855 fermentation Methods 0.000 description 2
- 230000004151 fermentation Effects 0.000 description 2
- 230000002538 fungal effect Effects 0.000 description 2
- 238000001641 gel filtration chromatography Methods 0.000 description 2
- 238000010353 genetic engineering Methods 0.000 description 2
- 101150021650 gluA gene Proteins 0.000 description 2
- 238000004191 hydrophobic interaction chromatography Methods 0.000 description 2
- 230000010354 integration Effects 0.000 description 2
- 235000019421 lipase Nutrition 0.000 description 2
- 238000011068 loading method Methods 0.000 description 2
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 description 2
- 244000005700 microbiome Species 0.000 description 2
- URPYMXQQVHTUDU-OFGSCBOVSA-N nucleopeptide y Chemical compound C([C@@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)CC)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)NC(=O)[C@H](CC=1NC=NC=1)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](C)NC(=O)[C@H](CO)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H](C)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](C)NC(=O)[C@H]1N(CCC1)C(=O)[C@H](C)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)CNC(=O)[C@H]1N(CCC1)C(=O)[C@H](CC(N)=O)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H]1N(CCC1)C(=O)[C@H](CCCCN)NC(=O)[C@H](CO)NC(=O)[C@H]1N(CCC1)C(=O)[C@@H](N)CC=1C=CC(O)=CC=1)C1=CC=C(O)C=C1 URPYMXQQVHTUDU-OFGSCBOVSA-N 0.000 description 2
- 235000015097 nutrients Nutrition 0.000 description 2
- 229940055695 pancreatin Drugs 0.000 description 2
- 230000037361 pathway Effects 0.000 description 2
- 230000006228 peptide amidation Effects 0.000 description 2
- 238000010635 peptide amidation reaction Methods 0.000 description 2
- 108010040793 peptidylamidoglycolate lyase Proteins 0.000 description 2
- 150000008300 phosphoramidites Chemical class 0.000 description 2
- 230000004481 post-translational protein modification Effects 0.000 description 2
- 238000001742 protein purification Methods 0.000 description 2
- 238000010188 recombinant method Methods 0.000 description 2
- 230000010076 replication Effects 0.000 description 2
- 102200067010 rs587621539 Human genes 0.000 description 2
- 102220045199 rs587781908 Human genes 0.000 description 2
- 102220058908 rs751646468 Human genes 0.000 description 2
- 210000004739 secretory vesicle Anatomy 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- 239000002904 solvent Substances 0.000 description 2
- 238000010561 standard procedure Methods 0.000 description 2
- 238000013519 translation Methods 0.000 description 2
- 230000014616 translation Effects 0.000 description 2
- OUYCCCASQSFEME-UHFFFAOYSA-N tyrosine Natural products OC(=O)C(N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-UHFFFAOYSA-N 0.000 description 2
- 241000701161 unidentified adenovirus Species 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- 239000003643 water by type Substances 0.000 description 2
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-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
- UHPMCKVQTMMPCG-UHFFFAOYSA-N 5,8-dihydroxy-2-methoxy-6-methyl-7-(2-oxopropyl)naphthalene-1,4-dione Chemical compound CC1=C(CC(C)=O)C(O)=C2C(=O)C(OC)=CC(=O)C2=C1O UHPMCKVQTMMPCG-UHFFFAOYSA-N 0.000 description 1
- 102100025601 60S ribosomal protein L27 Human genes 0.000 description 1
- 229920001817 Agar Polymers 0.000 description 1
- 108010021809 Alcohol dehydrogenase Proteins 0.000 description 1
- 241000024188 Andala Species 0.000 description 1
- 241000203069 Archaea Species 0.000 description 1
- 101710082738 Aspartic protease 3 Proteins 0.000 description 1
- 241001513093 Aspergillus awamori Species 0.000 description 1
- 241000351920 Aspergillus nidulans Species 0.000 description 1
- 101150071434 BAR1 gene Proteins 0.000 description 1
- 241000193752 Bacillus circulans Species 0.000 description 1
- 241000193749 Bacillus coagulans Species 0.000 description 1
- 241000194108 Bacillus licheniformis Species 0.000 description 1
- 108010029675 Bacillus licheniformis alpha-amylase Proteins 0.000 description 1
- 241000194107 Bacillus megaterium Species 0.000 description 1
- 101100007857 Bacillus subtilis (strain 168) cspB gene Proteins 0.000 description 1
- 241000193388 Bacillus thuringiensis Species 0.000 description 1
- 108091005658 Basic proteases Proteins 0.000 description 1
- UHOVQNZJYSORNB-UHFFFAOYSA-N Benzene Chemical class C1=CC=CC=C1 UHOVQNZJYSORNB-UHFFFAOYSA-N 0.000 description 1
- 101000959112 Bos taurus Peptidyl-alpha-hydroxyglycine alpha-amidating lyase Proteins 0.000 description 1
- 101100280051 Brucella abortus biovar 1 (strain 9-941) eryH gene Proteins 0.000 description 1
- 125000001433 C-terminal amino-acid group Chemical group 0.000 description 1
- 102000005367 Carboxypeptidases Human genes 0.000 description 1
- 108010006303 Carboxypeptidases Proteins 0.000 description 1
- 102100035882 Catalase Human genes 0.000 description 1
- 108010053835 Catalase Proteins 0.000 description 1
- 241001176476 Chthoniobacter Species 0.000 description 1
- 241000627991 Chthoniobacter flavus Ellin428 Species 0.000 description 1
- 108020004638 Circular DNA Proteins 0.000 description 1
- 238000001712 DNA sequencing Methods 0.000 description 1
- BWGNESOTFCXPMA-UHFFFAOYSA-N Dihydrogen disulfide Chemical compound SS BWGNESOTFCXPMA-UHFFFAOYSA-N 0.000 description 1
- 206010059866 Drug resistance Diseases 0.000 description 1
- 241000701832 Enterobacteria phage T3 Species 0.000 description 1
- YQYJSBFKSSDGFO-UHFFFAOYSA-N Epihygromycin Natural products OC1C(O)C(C(=O)C)OC1OC(C(=C1)O)=CC=C1C=C(C)C(=O)NC1C(O)C(O)C2OCOC2C1O YQYJSBFKSSDGFO-UHFFFAOYSA-N 0.000 description 1
- 241000588722 Escherichia Species 0.000 description 1
- 241001198387 Escherichia coli BL21(DE3) Species 0.000 description 1
- 241000701959 Escherichia virus Lambda Species 0.000 description 1
- 241001468125 Exiguobacterium Species 0.000 description 1
- 108010074860 Factor Xa Proteins 0.000 description 1
- 241000192125 Firmicutes Species 0.000 description 1
- 241000223218 Fusarium Species 0.000 description 1
- 241000223221 Fusarium oxysporum Species 0.000 description 1
- 108700028146 Genetic Enhancer Elements Proteins 0.000 description 1
- 108700039691 Genetic Promoter Regions Proteins 0.000 description 1
- 108700007698 Genetic Terminator Regions Proteins 0.000 description 1
- VIPDPMHGICREIS-GVXVVHGQSA-N Glu-Val-Leu Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(C)C)C(O)=O VIPDPMHGICREIS-GVXVVHGQSA-N 0.000 description 1
- AEMRFAOFKBGASW-UHFFFAOYSA-M Glycolate Chemical compound OCC([O-])=O AEMRFAOFKBGASW-UHFFFAOYSA-M 0.000 description 1
- HVLSXIKZNLPZJJ-TXZCQADKSA-N HA peptide Chemical compound C([C@@H](C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](C(C)C)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)N[C@@H](C)C(O)=O)NC(=O)[C@H]1N(CCC1)C(=O)[C@@H](N)CC=1C=CC(O)=CC=1)C1=CC=C(O)C=C1 HVLSXIKZNLPZJJ-TXZCQADKSA-N 0.000 description 1
- 241000125500 Hedypnois rhagadioloides Species 0.000 description 1
- 101000741885 Homo sapiens Protection of telomeres protein 1 Proteins 0.000 description 1
- 102000002265 Human Growth Hormone Human genes 0.000 description 1
- 239000000854 Human Growth Hormone Substances 0.000 description 1
- 241000223198 Humicola Species 0.000 description 1
- 108700005091 Immunoglobulin Genes Proteins 0.000 description 1
- 102000004195 Isomerases Human genes 0.000 description 1
- 108090000769 Isomerases Proteins 0.000 description 1
- 241001138401 Kluyveromyces lactis Species 0.000 description 1
- 150000008575 L-amino acids Chemical class 0.000 description 1
- FFEARJCKVFRZRR-BYPYZUCNSA-N L-methionine Chemical compound CSCC[C@H](N)C(O)=O FFEARJCKVFRZRR-BYPYZUCNSA-N 0.000 description 1
- FBOZXECLQNJBKD-ZDUSSCGKSA-N L-methotrexate Chemical compound C=1N=C2N=C(N)N=C(N)C2=NC=1CN(C)C1=CC=C(C(=O)N[C@@H](CCC(O)=O)C(O)=O)C=C1 FBOZXECLQNJBKD-ZDUSSCGKSA-N 0.000 description 1
- 125000000174 L-prolyl group Chemical group [H]N1C([H])([H])C([H])([H])C([H])([H])[C@@]1([H])C(*)=O 0.000 description 1
- 241000235087 Lachancea kluyveri Species 0.000 description 1
- ROHFNLRQFUQHCH-UHFFFAOYSA-N Leucine Natural products CC(C)CC(N)C(O)=O ROHFNLRQFUQHCH-UHFFFAOYSA-N 0.000 description 1
- 108091027974 Mature messenger RNA Proteins 0.000 description 1
- 208000009018 Medullary thyroid cancer Diseases 0.000 description 1
- 241000970829 Mesorhizobium Species 0.000 description 1
- 108090000157 Metallothionein Proteins 0.000 description 1
- 241000205276 Methanosarcina Species 0.000 description 1
- 101710181812 Methionine aminopeptidase Proteins 0.000 description 1
- 101100235161 Mycolicibacterium smegmatis (strain ATCC 700084 / mc(2)155) lerI gene Proteins 0.000 description 1
- SITLTJHOQZFJGG-UHFFFAOYSA-N N-L-alpha-glutamyl-L-valine Natural products CC(C)C(C(O)=O)NC(=O)C(N)CCC(O)=O SITLTJHOQZFJGG-UHFFFAOYSA-N 0.000 description 1
- 108010002311 N-glycylglutamic acid Proteins 0.000 description 1
- 229930193140 Neomycin Natural products 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 108090000189 Neuropeptides Proteins 0.000 description 1
- 108091034117 Oligonucleotide Proteins 0.000 description 1
- 108700026244 Open Reading Frames Proteins 0.000 description 1
- 101100378536 Ovis aries ADRB1 gene Proteins 0.000 description 1
- 241000194109 Paenibacillus lautus Species 0.000 description 1
- 102000035195 Peptidases Human genes 0.000 description 1
- 101800001442 Peptide pr Proteins 0.000 description 1
- 102100038745 Protection of telomeres protein 1 Human genes 0.000 description 1
- 108010029485 Protein Isoforms Proteins 0.000 description 1
- 102000001708 Protein Isoforms Human genes 0.000 description 1
- 241000589517 Pseudomonas aeruginosa Species 0.000 description 1
- 101100084022 Pseudomonas aeruginosa (strain ATCC 15692 / DSM 22644 / CIP 104116 / JCM 14847 / LMG 12228 / 1C / PRS 101 / PAO1) lapA gene Proteins 0.000 description 1
- 239000012614 Q-Sepharose Substances 0.000 description 1
- 102000007056 Recombinant Fusion Proteins Human genes 0.000 description 1
- 108010008281 Recombinant Fusion Proteins Proteins 0.000 description 1
- 108020005091 Replication Origin Proteins 0.000 description 1
- 241000235402 Rhizomucor Species 0.000 description 1
- 241000235403 Rhizomucor miehei Species 0.000 description 1
- 101000968489 Rhizomucor miehei Lipase Proteins 0.000 description 1
- 241000235343 Saccharomycetales Species 0.000 description 1
- 241000235346 Schizosaccharomyces Species 0.000 description 1
- 241000235347 Schizosaccharomyces pombe Species 0.000 description 1
- OHKFXGKHSJKKAL-NRPADANISA-N Ser-Glu-Val Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C(C)C)C(O)=O OHKFXGKHSJKKAL-NRPADANISA-N 0.000 description 1
- SRSPTFBENMJHMR-WHFBIAKZSA-N Ser-Ser-Gly Chemical compound OC[C@H](N)C(=O)N[C@@H](CO)C(=O)NCC(O)=O SRSPTFBENMJHMR-WHFBIAKZSA-N 0.000 description 1
- 241000187747 Streptomyces Species 0.000 description 1
- 241000187398 Streptomyces lividans Species 0.000 description 1
- 241001468239 Streptomyces murinus Species 0.000 description 1
- PZBFGYYEXUXCOF-UHFFFAOYSA-N TCEP Chemical compound OC(=O)CCP(CCC(O)=O)CCC(O)=O PZBFGYYEXUXCOF-UHFFFAOYSA-N 0.000 description 1
- 101150033985 TPI gene Proteins 0.000 description 1
- 239000004098 Tetracycline Substances 0.000 description 1
- PRNGXSILMXSWQQ-OEAJRASXSA-N Thr-Leu-Phe Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O PRNGXSILMXSWQQ-OEAJRASXSA-N 0.000 description 1
- 108090000190 Thrombin Proteins 0.000 description 1
- 108020005202 Viral DNA Proteins 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- 241000269368 Xenopus laevis Species 0.000 description 1
- 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 1
- 108010048241 acetamidase Proteins 0.000 description 1
- 230000003044 adaptive effect Effects 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- 108091005598 amidated proteins Proteins 0.000 description 1
- 125000003368 amide group Chemical group 0.000 description 1
- VZTDIZULWFCMLS-UHFFFAOYSA-N ammonium formate Chemical compound [NH4+].[O-]C=O VZTDIZULWFCMLS-UHFFFAOYSA-N 0.000 description 1
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 description 1
- 229910052921 ammonium sulfate Inorganic materials 0.000 description 1
- 235000011130 ammonium sulphate Nutrition 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 229940009098 aspartate Drugs 0.000 description 1
- 229930194563 aspergillus acid Natural products 0.000 description 1
- 229940054340 bacillus coagulans Drugs 0.000 description 1
- 230000010310 bacterial transformation Effects 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 1
- 239000003054 catalyst Substances 0.000 description 1
- 210000000170 cell membrane Anatomy 0.000 description 1
- 210000002421 cell wall Anatomy 0.000 description 1
- 210000003850 cellular structure Anatomy 0.000 description 1
- 239000003638 chemical reducing agent Substances 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 210000004978 chinese hamster ovary cell Anatomy 0.000 description 1
- 229960005091 chloramphenicol Drugs 0.000 description 1
- WIIZWVCIJKGZOK-RKDXNWHRSA-N chloramphenicol Chemical compound ClC(Cl)C(=O)N[C@H](CO)[C@H](O)C1=CC=C([N+]([O-])=O)C=C1 WIIZWVCIJKGZOK-RKDXNWHRSA-N 0.000 description 1
- 238000011210 chromatographic step Methods 0.000 description 1
- 230000002759 chromosomal effect Effects 0.000 description 1
- 238000010835 comparative analysis Methods 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- NKLPQNGYXWVELD-UHFFFAOYSA-M coomassie brilliant blue Chemical compound [Na+].C1=CC(OCC)=CC=C1NC1=CC=C(C(=C2C=CC(C=C2)=[N+](CC)CC=2C=C(C=CC=2)S([O-])(=O)=O)C=2C=CC(=CC=2)N(CC)CC=2C=C(C=CC=2)S([O-])(=O)=O)C=C1 NKLPQNGYXWVELD-UHFFFAOYSA-M 0.000 description 1
- 101150110403 cspA gene Proteins 0.000 description 1
- 101150068339 cspLA gene Proteins 0.000 description 1
- 230000001086 cytosolic effect Effects 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 239000007857 degradation product Substances 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- VHJLVAABSRFDPM-QWWZWVQMSA-N dithiothreitol Chemical compound SC[C@@H](O)[C@H](O)CS VHJLVAABSRFDPM-QWWZWVQMSA-N 0.000 description 1
- 230000009977 dual effect Effects 0.000 description 1
- 210000002472 endoplasmic reticulum Anatomy 0.000 description 1
- 239000003623 enhancer Substances 0.000 description 1
- 238000007824 enzymatic assay Methods 0.000 description 1
- 239000000706 filtrate Substances 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 235000019253 formic acid Nutrition 0.000 description 1
- 238000001502 gel electrophoresis Methods 0.000 description 1
- 108010061330 glucan 1,4-alpha-maltohydrolase Proteins 0.000 description 1
- 230000002414 glycolytic effect Effects 0.000 description 1
- 210000002288 golgi apparatus Anatomy 0.000 description 1
- 230000012010 growth Effects 0.000 description 1
- HNDVDQJCIGZPNO-UHFFFAOYSA-N histidine Natural products OC(=O)C(N)CC1=CN=CN1 HNDVDQJCIGZPNO-UHFFFAOYSA-N 0.000 description 1
- 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 1
- 238000002744 homologous recombination Methods 0.000 description 1
- 230000006801 homologous recombination Effects 0.000 description 1
- 229940088597 hormone Drugs 0.000 description 1
- 239000005556 hormone Substances 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- 210000003000 inclusion body Anatomy 0.000 description 1
- 238000010348 incorporation Methods 0.000 description 1
- 238000009776 industrial production Methods 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 229960000318 kanamycin Drugs 0.000 description 1
- 229930027917 kanamycin Natural products 0.000 description 1
- SBUJHOSQTJFQJX-NOAMYHISSA-N kanamycin Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CN)O[C@@H]1O[C@H]1[C@H](O)[C@@H](O[C@@H]2[C@@H]([C@@H](N)[C@H](O)[C@@H](CO)O2)O)[C@H](N)C[C@@H]1N SBUJHOSQTJFQJX-NOAMYHISSA-N 0.000 description 1
- 229930182823 kanamycin A Natural products 0.000 description 1
- 101150109249 lacI gene Proteins 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 230000002934 lysing effect Effects 0.000 description 1
- 238000004949 mass spectrometry Methods 0.000 description 1
- 208000023356 medullary thyroid gland carcinoma Diseases 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 229930182817 methionine Natural products 0.000 description 1
- 229960000485 methotrexate Drugs 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 238000002703 mutagenesis Methods 0.000 description 1
- 231100000350 mutagenesis Toxicity 0.000 description 1
- 239000006225 natural substrate Substances 0.000 description 1
- 239000013642 negative control Substances 0.000 description 1
- 229960004927 neomycin Drugs 0.000 description 1
- 238000007857 nested PCR Methods 0.000 description 1
- 230000000955 neuroendocrine Effects 0.000 description 1
- 239000000712 neurohormone Substances 0.000 description 1
- 239000002581 neurotoxin Substances 0.000 description 1
- 231100000618 neurotoxin Toxicity 0.000 description 1
- 101150095344 niaD gene Proteins 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 108090000021 oryzin Proteins 0.000 description 1
- 239000008188 pellet Substances 0.000 description 1
- 238000010647 peptide synthesis reaction Methods 0.000 description 1
- 229920002616 peptidyl amide Polymers 0.000 description 1
- 210000001322 periplasm Anatomy 0.000 description 1
- 239000008194 pharmaceutical composition Substances 0.000 description 1
- 101150009573 phoA gene Proteins 0.000 description 1
- 238000002264 polyacrylamide gel electrophoresis Methods 0.000 description 1
- 238000003752 polymerase chain reaction Methods 0.000 description 1
- 230000001124 posttranscriptional effect Effects 0.000 description 1
- 229960004457 pramlintide acetate Drugs 0.000 description 1
- 230000001376 precipitating effect Effects 0.000 description 1
- 238000002953 preparative HPLC Methods 0.000 description 1
- 235000019833 protease Nutrition 0.000 description 1
- 108010042388 protease C Proteins 0.000 description 1
- 210000001938 protoplast Anatomy 0.000 description 1
- 101150054232 pyrG gene Proteins 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 230000003362 replicative effect Effects 0.000 description 1
- 108091008146 restriction endonucleases Proteins 0.000 description 1
- 230000000717 retained effect Effects 0.000 description 1
- 238000010839 reverse transcription Methods 0.000 description 1
- 108010067528 ribosomal proteins L27 Proteins 0.000 description 1
- 239000012723 sample buffer Substances 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 239000007790 solid phase Substances 0.000 description 1
- 230000002269 spontaneous effect Effects 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 230000000638 stimulation Effects 0.000 description 1
- 239000013589 supplement Substances 0.000 description 1
- 229940099093 symlin Drugs 0.000 description 1
- 230000008685 targeting Effects 0.000 description 1
- 229960002180 tetracycline Drugs 0.000 description 1
- 229930101283 tetracycline Natural products 0.000 description 1
- 235000019364 tetracycline Nutrition 0.000 description 1
- 150000003522 tetracyclines Chemical class 0.000 description 1
- 238000002560 therapeutic procedure Methods 0.000 description 1
- 229960004072 thrombin Drugs 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 101150080369 tpiA gene Proteins 0.000 description 1
- 230000002103 transcriptional effect Effects 0.000 description 1
- 230000001131 transforming effect Effects 0.000 description 1
- 230000001052 transient effect Effects 0.000 description 1
- 230000009452 underexpressoin Effects 0.000 description 1
- 241000701447 unidentified baculovirus Species 0.000 description 1
- 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 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
- C12N9/88—Lyases (4.)
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P3/00—Drugs for disorders of the metabolism
- A61P3/04—Anorexiants; Antiobesity agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P3/00—Drugs for disorders of the metabolism
- A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
- A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/575—Hormones
-
- 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
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y403/00—Carbon-nitrogen lyases (4.3)
- C12Y403/02—Amidine-lyases (4.3.2)
- C12Y403/02005—Peptidylamidoglycolate lyase (4.3.2.5)
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Wood Science & Technology (AREA)
- Genetics & Genomics (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Biochemistry (AREA)
- Molecular Biology (AREA)
- Diabetes (AREA)
- Microbiology (AREA)
- Biotechnology (AREA)
- General Engineering & Computer Science (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Biomedical Technology (AREA)
- Endocrinology (AREA)
- Public Health (AREA)
- Obesity (AREA)
- Hematology (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Veterinary Medicine (AREA)
- Animal Behavior & Ethology (AREA)
- Pharmacology & Pharmacy (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Biophysics (AREA)
- Gastroenterology & Hepatology (AREA)
- Toxicology (AREA)
- Emergency Medicine (AREA)
- Child & Adolescent Psychology (AREA)
- Enzymes And Modification Thereof (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Applications Claiming Priority (5)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| EP09177593.2 | 2009-12-01 | ||
| EP09177593 | 2009-12-01 | ||
| US26671109P | 2009-12-04 | 2009-12-04 | |
| US61/266,711 | 2009-12-04 | ||
| PCT/EP2010/068630 WO2011067283A1 (en) | 2009-12-01 | 2010-12-01 | Novel peptidyl alpha-hydroxyglycine alpha-amidating lyases |
Publications (3)
| Publication Number | Publication Date |
|---|---|
| JP2013511997A JP2013511997A (ja) | 2013-04-11 |
| JP2013511997A5 JP2013511997A5 (cg-RX-API-DMAC7.html) | 2014-01-16 |
| JP5931738B2 true JP5931738B2 (ja) | 2016-06-08 |
Family
ID=44114627
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP2012541487A Expired - Fee Related JP5931738B2 (ja) | 2009-12-01 | 2010-12-01 | 新規のペプチジルアルファ−ヒドロキシグリシンアルファ−アミド化リアーゼ |
Country Status (6)
| Country | Link |
|---|---|
| US (2) | US9096843B2 (cg-RX-API-DMAC7.html) |
| EP (1) | EP2507258B1 (cg-RX-API-DMAC7.html) |
| JP (1) | JP5931738B2 (cg-RX-API-DMAC7.html) |
| CN (1) | CN102666570B (cg-RX-API-DMAC7.html) |
| ES (1) | ES2583259T3 (cg-RX-API-DMAC7.html) |
| WO (1) | WO2011067283A1 (cg-RX-API-DMAC7.html) |
Families Citing this family (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20160098278A (ko) * | 2013-12-20 | 2016-08-18 | 에프. 호프만-라 로슈 아게 | Ｃ-말단 아미드화를 위한 펩티딜글라이신 알파-아미드화 모노옥시게나제 (ｐａｍ) 의 용도 |
| CN105985995A (zh) * | 2015-01-29 | 2016-10-05 | 暨南大学 | 一种利用家蚕生物反应器生产普兰林肽(pa)的方法 |
| CN113302310B (zh) * | 2018-12-18 | 2024-05-03 | 帕西昂奥地利有限两合公司 | 单步生物催化酰胺化 |
| CN110034855B (zh) * | 2019-04-10 | 2021-12-14 | 国网辽宁省电力有限公司 | 一种信息传输校验方法及系统 |
| MX2022010556A (es) * | 2020-02-26 | 2022-11-30 | Pam Theragnostics Gmbh | Uso de peptidilglicina alfa-amidante monooxigenasa (pam) para proposito terapeutico. |
| EP4111201A1 (en) * | 2020-02-26 | 2023-01-04 | PAM Theragnostics GmbH | Methods for determining peptidylglycine alpha-amidating monooxygenase (pam) and its use for diagnostic purpose |
| EP4142792A4 (en) | 2020-05-01 | 2024-11-27 | Kashiv Biosciences, LLC | An improved process of purification of protein |
| WO2023180977A1 (en) * | 2022-03-23 | 2023-09-28 | Kashiv Biosciences, Llc | An improved process for the separation and analysis of pancreatic enzymes |
| WO2025133225A1 (en) * | 2023-12-22 | 2025-06-26 | Pam Theragnostics Gmbh | Pharmaceutical combination of peptide-gly and peptidylglycine alpha-amidating monooxygenase (pam) |
Family Cites Families (37)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4203570A (en) | 1978-08-23 | 1980-05-20 | The Western States Machine Company | Power-operated loading gate for centrifugal machines incorporating an auxiliary drive device |
| US4546082A (en) | 1982-06-17 | 1985-10-08 | Regents Of The Univ. Of California | E. coli/Saccharomyces cerevisiae plasmid cloning vector containing the alpha-factor gene for secretion and processing of hybrid proteins |
| US4599311A (en) | 1982-08-13 | 1986-07-08 | Kawasaki Glenn H | Glycolytic promotersfor regulated protein expression: protease inhibitor |
| JPS60501140A (ja) | 1983-04-22 | 1985-07-25 | アムジエン | 酵母による外因性ポリペプチドの分泌 |
| NZ207926A (en) | 1983-04-25 | 1988-04-29 | Genentech Inc | Use of yeast #a#-factor to assist in expression of proteins heterologus to yeast |
| US4870008A (en) | 1983-08-12 | 1989-09-26 | Chiron Corporation | Secretory expression in eukaryotes |
| US4931373A (en) | 1984-05-25 | 1990-06-05 | Zymogenetics, Inc. | Stable DNA constructs for expression of α-1 antitrypsin |
| DK58285D0 (da) | 1984-05-30 | 1985-02-08 | Novo Industri As | Peptider samt fremstilling og anvendelse deraf |
| US4708934A (en) | 1984-09-27 | 1987-11-24 | Unigene Laboratories, Inc. | α-amidation enzyme |
| US4766073A (en) | 1985-02-25 | 1988-08-23 | Zymogenetics Inc. | Expression of biologically active PDGF analogs in eucaryotic cells |
| US4885249A (en) | 1984-12-05 | 1989-12-05 | Allelix, Inc. | Aspergillus niger transformation system |
| US4683202A (en) | 1985-03-28 | 1987-07-28 | Cetus Corporation | Process for amplifying nucleic acid sequences |
| DE3650202T3 (de) | 1985-08-29 | 2004-06-03 | Genencor International, Inc., South San Francisco | Expression von heterologen Polypeptiden in filamentösen Pilzen, Verfahren zu deren Herstellung und Vektoren zu deren Herstellung. |
| US4882279A (en) | 1985-10-25 | 1989-11-21 | Phillips Petroleum Company | Site selective genomic modification of yeast of the genus pichia |
| WO1987002670A1 (en) | 1985-10-25 | 1987-05-07 | Mackay Vivian L | Method of using bar1 for secreting foreign proteins |
| US4935349A (en) | 1986-01-17 | 1990-06-19 | Zymogenetics, Inc. | Expression of higher eucaryotic genes in aspergillus |
| DK122686D0 (da) | 1986-03-17 | 1986-03-17 | Novo Industri As | Fremstilling af proteiner |
| GB8610600D0 (en) | 1986-04-30 | 1986-06-04 | Novo Industri As | Transformation of trichoderma |
| GB8720115D0 (en) * | 1987-08-26 | 1987-09-30 | Cooper G J S | Treatment of diabetes mellitus |
| US5789234A (en) | 1987-08-14 | 1998-08-04 | Unigene Laboratories, Inc. | Expression systems for amidating enzyme |
| DE3886221T3 (de) | 1987-09-04 | 2000-12-21 | Novo Nordisk A/S, Bagsvaerd | VERFAHREN ZUR HERSTELLUNG VON PROTEINPRODUKTEN IN -i(ASPERGILLUS) UND PROMOTOREN ZUR VERWENDUNG IN -i(ASPERGILLUS). |
| DK463887D0 (da) | 1987-09-07 | 1987-09-07 | Novo Industri As | Gaerleader |
| US6255067B1 (en) | 1987-09-15 | 2001-07-03 | The Johns Hopkins University | cDNA encoding peptidyl-glycine alpha-amidating monooxygenase (PAM) |
| US5037743A (en) | 1988-08-05 | 1991-08-06 | Zymogenetics, Inc. | BAR1 secretion signal |
| JP2535398B2 (ja) | 1989-01-17 | 1996-09-18 | サントリー株式会社 | アミド化ペプチドの製造方法 |
| JP2845468B2 (ja) * | 1989-01-17 | 1999-01-13 | サントリー株式会社 | ヒト甲状腺由来Ｃ―末端α―アミド化酵素 |
| EP0448513A3 (en) | 1990-03-21 | 1991-12-27 | Japat Ltd | Process for production of peptidylglycine alpha-hydroxylating monooxygenase and use thereof |
| ES2079054T3 (es) * | 1990-06-01 | 1996-01-01 | Ciba Geigy Ag | Peptidilhidroxiglicina n-c-liasa y adn codificante de la misma. |
| DK300090D0 (da) | 1990-12-19 | 1990-12-19 | Novo Nordisk As | Fremgangsmaade til fremstilling af leadersekvenser |
| US6495514B1 (en) | 1998-01-21 | 2002-12-17 | Mercer University | Method for reducing inflammation and inducing an analgesic effect and compounds thereof |
| AU3274101A (en) * | 2000-01-06 | 2001-07-16 | Genoptera, Llc | Nucleic acids and polypeptides of invertebrate enzymes and methods of use |
| AU2002228608A1 (en) | 2000-12-13 | 2002-06-24 | Eli Lilly And Company | Amidated glucagon-like peptide-1 |
| PL1651249T3 (pl) * | 2003-07-23 | 2013-04-30 | Novartis Ag | Zastosowanie kalcytoniny w zapaleniu kości i stawów |
| US8137929B2 (en) | 2005-04-15 | 2012-03-20 | Novo Nordisk Health Care Ag | Basic protein purification tags from thermophilic bacteria |
| EP1907561B1 (en) | 2005-06-24 | 2013-08-28 | Unigene Laboratories, Inc. | Cell lines for expressing enzyme useful in the preparation of amidated products |
| ES2531372T3 (es) | 2006-10-13 | 2015-03-13 | Novo Nordisk Healthcare Ag | Enzimas de procesamiento fusionadas a marcadores de proteínas básicas |
| US8222016B2 (en) | 2007-06-29 | 2012-07-17 | Daiichi Sankyo Company, Limited | Recombinant C-terminal α-amidating enzyme derivative |
-
2010
- 2010-12-01 EP EP10788057.7A patent/EP2507258B1/en not_active Not-in-force
- 2010-12-01 WO PCT/EP2010/068630 patent/WO2011067283A1/en not_active Ceased
- 2010-12-01 ES ES10788057.7T patent/ES2583259T3/es active Active
- 2010-12-01 US US13/513,049 patent/US9096843B2/en not_active Expired - Fee Related
- 2010-12-01 CN CN201080054717.4A patent/CN102666570B/zh not_active Expired - Fee Related
- 2010-12-01 JP JP2012541487A patent/JP5931738B2/ja not_active Expired - Fee Related
-
2015
- 2015-06-30 US US14/755,183 patent/US20160083713A1/en not_active Abandoned
Also Published As
| Publication number | Publication date |
|---|---|
| US20160083713A1 (en) | 2016-03-24 |
| US20120322733A1 (en) | 2012-12-20 |
| CN102666570A (zh) | 2012-09-12 |
| US9096843B2 (en) | 2015-08-04 |
| EP2507258B1 (en) | 2016-04-20 |
| WO2011067283A1 (en) | 2011-06-09 |
| CN102666570B (zh) | 2015-12-02 |
| JP2013511997A (ja) | 2013-04-11 |
| ES2583259T3 (es) | 2016-09-20 |
| EP2507258A1 (en) | 2012-10-10 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| JP5931738B2 (ja) | 新規のペプチジルアルファ−ヒドロキシグリシンアルファ−アミド化リアーゼ | |
| JP4857279B2 (ja) | カルボキシ末端をアミド化したペプチドの製造方法 | |
| JP6714902B2 (ja) | ペプチドおよびタンパク質の発現のための方法 | |
| Liew et al. | Preparation of recombinant thioredoxin fused N-terminal proCNP: analysis of enterokinase cleavage products reveals new enterokinase cleavage sites | |
| IL190229A (en) | Method for adding amide to their C-tip polypeptides with basic amino acid using specific endoproteases | |
| US8426566B2 (en) | Basic protein purification tags from thermophilic bacteria | |
| EP2794875B1 (en) | Modified enterokinase light chain | |
| US9670257B2 (en) | Methods for producing peptides using engineered inteins | |
| CN103998606B (zh) | 修饰的肠激酶轻链 | |
| KR102643064B1 (ko) | 발현 수준 및 가용성이 증대된 인간 엔테로키나제 융합 단백질 및 그 제조방법 | |
| Nataraj et al. | Truncated Thioredoxin Peptides Serves as an Efficient Fusion Tag for Production of Proinsulin | |
| WO2017174760A1 (en) | Novel multiply backbone n-methyl transferases and uses thereof | |
| WO2022263559A1 (en) | Production of cross-reactive material 197 fusion proteins | |
| Zhu et al. | Annex 1: Published version of the paper Escherichia coli ‘TatExpress’ strains export over 5g/L human growth hormone to the periplasm by the Tat pathway | |
| Jin-Hua et al. | Refolding of the fusion protein of recombinant enterokinase light chain rEKL | |
| CN117795062A (zh) | 作为支架蛋白的过氧化物还原酶家族的多聚体蛋白 | |
| WO2004113382A1 (en) | Recombinant igf expression systems | |
| HK1111426B (en) | Method for producing carboxy-terminal amidified peptides | |
| Guerrero‐Montero et al. | F., Peswani, Amber R., Labrit, Mickael, Allen, Matthew, Aubry, Cedric, Davé, Emma, Humphreys, David P. and others (2019) Escherichia coli ‘TatExpress’ strains export several g/L human growth hormone to the periplasm by the Tat pathway. Biotechnology and Bioengineering, 116 (12). pp. 3282-3291. ISSN | |
| WO2016034534A1 (en) | Lysine rich basic pre-sequences |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20131120 |
|
| A621 | Written request for application examination |
Free format text: JAPANESE INTERMEDIATE CODE: A621 Effective date: 20131120 |
|
| A977 | Report on retrieval |
Free format text: JAPANESE INTERMEDIATE CODE: A971007 Effective date: 20141126 |
|
| A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20150105 |
|
| A601 | Written request for extension of time |
Free format text: JAPANESE INTERMEDIATE CODE: A601 Effective date: 20150406 |
|
| A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20150430 |
|
| A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20150914 |
|
| A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20151204 |
|
| 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: 20160404 |
|
| A61 | First payment of annual fees (during grant procedure) |
Free format text: JAPANESE INTERMEDIATE CODE: A61 Effective date: 20160427 |
|
| R150 | Certificate of patent or registration of utility model |
Ref document number: 5931738 Country of ref document: JP Free format text: JAPANESE INTERMEDIATE CODE: R150 |
|
| LAPS | Cancellation because of no payment of annual fees |