CN109400674A - Antihypertensive tripeptide derived from euphausia superba and application thereof - Google Patents
Antihypertensive tripeptide derived from euphausia superba and application thereof Download PDFInfo
- Publication number
- CN109400674A CN109400674A CN201811397409.6A CN201811397409A CN109400674A CN 109400674 A CN109400674 A CN 109400674A CN 201811397409 A CN201811397409 A CN 201811397409A CN 109400674 A CN109400674 A CN 109400674A
- Authority
- CN
- China
- Prior art keywords
- decompression
- krill
- tripeptides
- derived
- chromatography
- 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
- 230000003276 anti-hypertensive effect Effects 0.000 title abstract 4
- 241000239370 Euphausia superba Species 0.000 title abstract 3
- 241000239366 Euphausiacea Species 0.000 claims abstract description 52
- 230000006837 decompression Effects 0.000 claims abstract description 40
- 230000036772 blood pressure Effects 0.000 claims abstract description 36
- UHRNIXJAGGLKHP-DLOVCJGASA-N Phe-Ala-Ser Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](C)C(=O)N[C@@H](CO)C(O)=O UHRNIXJAGGLKHP-DLOVCJGASA-N 0.000 claims abstract description 23
- 235000013305 food Nutrition 0.000 claims abstract description 18
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 16
- 108090000765 processed proteins & peptides Proteins 0.000 claims abstract description 14
- 239000004365 Protease Substances 0.000 claims abstract description 13
- 238000004007 reversed phase HPLC Methods 0.000 claims abstract description 13
- 108091005804 Peptidases Proteins 0.000 claims abstract description 12
- 239000000047 product Substances 0.000 claims abstract description 11
- 238000000108 ultra-filtration Methods 0.000 claims abstract description 11
- 239000012153 distilled water Substances 0.000 claims abstract description 10
- 238000002360 preparation method Methods 0.000 claims abstract description 9
- 238000005571 anion exchange chromatography Methods 0.000 claims abstract description 8
- 238000002330 electrospray ionisation mass spectrometry Methods 0.000 claims abstract description 5
- 238000000746 purification Methods 0.000 claims abstract description 3
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 claims abstract 3
- 125000003275 alpha amino acid group Chemical group 0.000 claims abstract 2
- 230000000694 effects Effects 0.000 claims description 23
- 239000000243 solution Substances 0.000 claims description 22
- 239000007788 liquid Substances 0.000 claims description 17
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 16
- 239000003814 drug Substances 0.000 claims description 14
- 229940079593 drug Drugs 0.000 claims description 10
- 238000000034 method Methods 0.000 claims description 10
- 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 claims description 9
- 150000001413 amino acids Chemical class 0.000 claims description 9
- 238000010828 elution Methods 0.000 claims description 9
- 230000036541 health Effects 0.000 claims description 8
- 239000011780 sodium chloride Substances 0.000 claims description 8
- UUUHXMGGBIUAPW-UHFFFAOYSA-N 1-[1-[2-[[5-amino-2-[[1-[5-(diaminomethylideneamino)-2-[[1-[3-(1h-indol-3-yl)-2-[(5-oxopyrrolidine-2-carbonyl)amino]propanoyl]pyrrolidine-2-carbonyl]amino]pentanoyl]pyrrolidine-2-carbonyl]amino]-5-oxopentanoyl]amino]-3-methylpentanoyl]pyrrolidine-2-carbon Chemical compound C1CCC(C(=O)N2C(CCC2)C(O)=O)N1C(=O)C(C(C)CC)NC(=O)C(CCC(N)=O)NC(=O)C1CCCN1C(=O)C(CCCN=C(N)N)NC(=O)C1CCCN1C(=O)C(CC=1C2=CC=CC=C2NC=1)NC(=O)C1CCC(=O)N1 UUUHXMGGBIUAPW-UHFFFAOYSA-N 0.000 claims description 7
- 102000004190 Enzymes Human genes 0.000 claims description 7
- 108090000790 Enzymes Proteins 0.000 claims description 7
- 108090000882 Peptidyl-Dipeptidase A Proteins 0.000 claims description 7
- 229920005654 Sephadex Polymers 0.000 claims description 7
- 239000012507 Sephadex™ Substances 0.000 claims description 7
- 201000010099 disease Diseases 0.000 claims description 7
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims description 7
- 239000006228 supernatant Substances 0.000 claims description 7
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims description 6
- 239000000872 buffer Substances 0.000 claims description 6
- 238000005227 gel permeation chromatography Methods 0.000 claims description 6
- 230000001077 hypotensive effect Effects 0.000 claims description 6
- 206010020772 Hypertension Diseases 0.000 claims description 5
- 238000002835 absorbance Methods 0.000 claims description 5
- 238000001641 gel filtration chromatography Methods 0.000 claims description 5
- 238000004255 ion exchange chromatography Methods 0.000 claims description 5
- 238000005259 measurement Methods 0.000 claims description 5
- 238000005119 centrifugation Methods 0.000 claims description 4
- 239000003480 eluent Substances 0.000 claims description 4
- 230000004048 modification Effects 0.000 claims description 4
- 238000012986 modification Methods 0.000 claims description 4
- 239000007853 buffer solution Substances 0.000 claims description 3
- 238000004519 manufacturing process Methods 0.000 claims description 3
- 230000026731 phosphorylation Effects 0.000 claims description 3
- 238000006366 phosphorylation reaction Methods 0.000 claims description 3
- 229910021642 ultra pure water Inorganic materials 0.000 claims description 3
- 239000012498 ultrapure water Substances 0.000 claims description 3
- 150000002500 ions Chemical class 0.000 claims description 2
- 238000000825 ultraviolet detection Methods 0.000 claims description 2
- 102000004270 Peptidyl-Dipeptidase A Human genes 0.000 claims 1
- 230000008033 biological extinction Effects 0.000 claims 1
- 238000002086 displacement chromatography Methods 0.000 claims 1
- 108050009340 Endothelin Proteins 0.000 abstract description 11
- 102000002045 Endothelin Human genes 0.000 abstract description 11
- 210000004027 cell Anatomy 0.000 abstract description 11
- ZUBDGKVDJUIMQQ-UBFCDGJISA-N endothelin-1 Chemical compound C([C@@H](C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(O)=O)NC(=O)[C@H]1NC(=O)[C@H](CC=2C=CC=CC=2)NC(=O)[C@@H](CC=2C=CC(O)=CC=2)NC(=O)[C@H](C(C)C)NC(=O)[C@H]2CSSC[C@@H](C(N[C@H](CO)C(=O)N[C@@H](CO)C(=O)N[C@H](CC(C)C)C(=O)N[C@@H](CCSC)C(=O)N[C@H](CC(O)=O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCC(O)=O)C(=O)N2)=O)NC(=O)[C@@H](CO)NC(=O)[C@H](N)CSSC1)C1=CNC=N1 ZUBDGKVDJUIMQQ-UBFCDGJISA-N 0.000 abstract description 11
- 210000002889 endothelial cell Anatomy 0.000 abstract description 9
- 230000002401 inhibitory effect Effects 0.000 abstract description 9
- 210000003606 umbilical vein Anatomy 0.000 abstract description 9
- 238000011161 development Methods 0.000 abstract description 6
- 230000002040 relaxant effect Effects 0.000 abstract description 6
- 108010038807 Oligopeptides Proteins 0.000 abstract description 5
- 102000015636 Oligopeptides Human genes 0.000 abstract description 5
- 231100000331 toxic Toxicity 0.000 abstract description 5
- 230000002588 toxic effect Effects 0.000 abstract description 3
- 239000000413 hydrolysate Substances 0.000 abstract description 2
- 239000002220 antihypertensive agent Substances 0.000 abstract 1
- 229940127088 antihypertensive drug Drugs 0.000 abstract 1
- 238000004440 column chromatography Methods 0.000 abstract 1
- 230000002255 enzymatic effect Effects 0.000 abstract 1
- 102100030988 Angiotensin-converting enzyme Human genes 0.000 description 28
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 24
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 description 12
- 235000018102 proteins Nutrition 0.000 description 12
- 102000004169 proteins and genes Human genes 0.000 description 12
- 108090000623 proteins and genes Proteins 0.000 description 12
- 102000035195 Peptidases Human genes 0.000 description 9
- 235000019419 proteases Nutrition 0.000 description 9
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 6
- 229940088598 enzyme Drugs 0.000 description 6
- 230000007062 hydrolysis Effects 0.000 description 6
- 238000006460 hydrolysis reaction Methods 0.000 description 6
- 239000012086 standard solution Substances 0.000 description 6
- 239000008280 blood Substances 0.000 description 5
- 210000004369 blood Anatomy 0.000 description 5
- 238000005265 energy consumption Methods 0.000 description 5
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 4
- 102000004407 Lactalbumin Human genes 0.000 description 4
- 108090000942 Lactalbumin Proteins 0.000 description 4
- 102000007079 Peptide Fragments Human genes 0.000 description 4
- 108010033276 Peptide Fragments Proteins 0.000 description 4
- 229920002472 Starch Polymers 0.000 description 4
- 239000005862 Whey Substances 0.000 description 4
- 102000007544 Whey Proteins Human genes 0.000 description 4
- 108010046377 Whey Proteins Proteins 0.000 description 4
- 230000000052 comparative effect Effects 0.000 description 4
- 238000011156 evaluation Methods 0.000 description 4
- 230000006870 function Effects 0.000 description 4
- 229910052500 inorganic mineral Inorganic materials 0.000 description 4
- 150000002632 lipids Chemical class 0.000 description 4
- 239000011707 mineral Substances 0.000 description 4
- 239000003921 oil Substances 0.000 description 4
- 235000019198 oils Nutrition 0.000 description 4
- 239000012071 phase Substances 0.000 description 4
- 235000019698 starch Nutrition 0.000 description 4
- 239000008107 starch Substances 0.000 description 4
- 102000011632 Caseins Human genes 0.000 description 3
- 108010076119 Caseins Proteins 0.000 description 3
- 241001465754 Metazoa Species 0.000 description 3
- PVNIIMVLHYAWGP-UHFFFAOYSA-N Niacin Chemical compound OC(=O)C1=CC=CN=C1 PVNIIMVLHYAWGP-UHFFFAOYSA-N 0.000 description 3
- 239000003513 alkali Substances 0.000 description 3
- 230000008901 benefit Effects 0.000 description 3
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 description 3
- 239000006071 cream Substances 0.000 description 3
- 229910052757 nitrogen Inorganic materials 0.000 description 3
- 239000000843 powder Substances 0.000 description 3
- 230000008569 process Effects 0.000 description 3
- 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 2
- GHOKWGTUZJEAQD-ZETCQYMHSA-N (D)-(+)-Pantothenic acid Chemical compound OCC(C)(C)[C@@H](O)C(=O)NCCC(O)=O GHOKWGTUZJEAQD-ZETCQYMHSA-N 0.000 description 2
- GVJHHUAWPYXKBD-UHFFFAOYSA-N (±)-α-Tocopherol Chemical class OC1=C(C)C(C)=C2OC(CCCC(C)CCCC(C)CCCC(C)C)(C)CCC2=C1C GVJHHUAWPYXKBD-UHFFFAOYSA-N 0.000 description 2
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical class OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 description 2
- 101800004538 Bradykinin Proteins 0.000 description 2
- 102400000967 Bradykinin Human genes 0.000 description 2
- 241000238557 Decapoda Species 0.000 description 2
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 2
- QXZGBUJJYSLZLT-UHFFFAOYSA-N H-Arg-Pro-Pro-Gly-Phe-Ser-Pro-Phe-Arg-OH Natural products NC(N)=NCCCC(N)C(=O)N1CCCC1C(=O)N1C(C(=O)NCC(=O)NC(CC=2C=CC=CC=2)C(=O)NC(CO)C(=O)N2C(CCC2)C(=O)NC(CC=2C=CC=CC=2)C(=O)NC(CCCN=C(N)N)C(O)=O)CCC1 QXZGBUJJYSLZLT-UHFFFAOYSA-N 0.000 description 2
- OAKJQQAXSVQMHS-UHFFFAOYSA-N Hydrazine Chemical compound NN OAKJQQAXSVQMHS-UHFFFAOYSA-N 0.000 description 2
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 description 2
- 238000013019 agitation Methods 0.000 description 2
- 210000004712 air sac Anatomy 0.000 description 2
- 125000003277 amino group Chemical group 0.000 description 2
- 230000004872 arterial blood pressure Effects 0.000 description 2
- QXZGBUJJYSLZLT-FDISYFBBSA-N bradykinin Chemical compound NC(=N)NCCC[C@H](N)C(=O)N1CCC[C@H]1C(=O)N1[C@H](C(=O)NCC(=O)N[C@@H](CC=2C=CC=CC=2)C(=O)N[C@@H](CO)C(=O)N2[C@@H](CCC2)C(=O)N[C@@H](CC=2C=CC=CC=2)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O)CCC1 QXZGBUJJYSLZLT-FDISYFBBSA-N 0.000 description 2
- 150000001720 carbohydrates Chemical class 0.000 description 2
- 235000014633 carbohydrates Nutrition 0.000 description 2
- 230000035487 diastolic blood pressure Effects 0.000 description 2
- 230000007071 enzymatic hydrolysis Effects 0.000 description 2
- 238000006047 enzymatic hydrolysis reaction Methods 0.000 description 2
- 235000019197 fats Nutrition 0.000 description 2
- OVBPIULPVIDEAO-LBPRGKRZSA-N folic acid Chemical compound C=1N=C2NC(N)=NC(=O)C2=NC=1CNC1=CC=C(C(=O)N[C@@H](CCC(O)=O)C(O)=O)C=C1 OVBPIULPVIDEAO-LBPRGKRZSA-N 0.000 description 2
- 238000004108 freeze drying Methods 0.000 description 2
- 235000013402 health food Nutrition 0.000 description 2
- 235000001497 healthy food Nutrition 0.000 description 2
- 210000002216 heart Anatomy 0.000 description 2
- 229910052739 hydrogen Inorganic materials 0.000 description 2
- 239000001257 hydrogen Substances 0.000 description 2
- 230000002209 hydrophobic effect Effects 0.000 description 2
- 230000002779 inactivation Effects 0.000 description 2
- 229910052742 iron Inorganic materials 0.000 description 2
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 description 2
- 230000003902 lesion Effects 0.000 description 2
- 239000012528 membrane Substances 0.000 description 2
- QSHDDOUJBYECFT-UHFFFAOYSA-N mercury Chemical compound [Hg] QSHDDOUJBYECFT-UHFFFAOYSA-N 0.000 description 2
- 229910052753 mercury Inorganic materials 0.000 description 2
- 230000007935 neutral effect Effects 0.000 description 2
- 229960003512 nicotinic acid Drugs 0.000 description 2
- 235000001968 nicotinic acid Nutrition 0.000 description 2
- 239000011664 nicotinic acid Substances 0.000 description 2
- 150000007524 organic acids Chemical class 0.000 description 2
- 239000011574 phosphorus Substances 0.000 description 2
- 229910052698 phosphorus Inorganic materials 0.000 description 2
- 229920001184 polypeptide Polymers 0.000 description 2
- 102000004196 processed proteins & peptides Human genes 0.000 description 2
- 235000020183 skimmed milk Nutrition 0.000 description 2
- 241000894007 species Species 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 230000035488 systolic blood pressure Effects 0.000 description 2
- 239000011782 vitamin Substances 0.000 description 2
- 239000011701 zinc Substances 0.000 description 2
- BJEPYKJPYRNKOW-REOHCLBHSA-N (S)-malic acid Chemical compound OC(=O)[C@@H](O)CC(O)=O BJEPYKJPYRNKOW-REOHCLBHSA-N 0.000 description 1
- ODIGIKRIUKFKHP-UHFFFAOYSA-N (n-propan-2-yloxycarbonylanilino) acetate Chemical compound CC(C)OC(=O)N(OC(C)=O)C1=CC=CC=C1 ODIGIKRIUKFKHP-UHFFFAOYSA-N 0.000 description 1
- OWEGMIWEEQEYGQ-UHFFFAOYSA-N 100676-05-9 Natural products OC1C(O)C(O)C(CO)OC1OCC1C(O)C(O)C(O)C(OC2C(OC(O)C(O)C2O)CO)O1 OWEGMIWEEQEYGQ-UHFFFAOYSA-N 0.000 description 1
- FPIPGXGPPPQFEQ-UHFFFAOYSA-N 13-cis retinol Natural products OCC=C(C)C=CC=C(C)C=CC1=C(C)CCCC1(C)C FPIPGXGPPPQFEQ-UHFFFAOYSA-N 0.000 description 1
- WIGIZIANZCJQQY-UHFFFAOYSA-N 4-ethyl-3-methyl-N-[2-[4-[[[(4-methylcyclohexyl)amino]-oxomethyl]sulfamoyl]phenyl]ethyl]-5-oxo-2H-pyrrole-1-carboxamide Chemical compound O=C1C(CC)=C(C)CN1C(=O)NCCC1=CC=C(S(=O)(=O)NC(=O)NC2CCC(C)CC2)C=C1 WIGIZIANZCJQQY-UHFFFAOYSA-N 0.000 description 1
- ZCYVEMRRCGMTRW-UHFFFAOYSA-N 7553-56-2 Chemical compound [I] ZCYVEMRRCGMTRW-UHFFFAOYSA-N 0.000 description 1
- 108010064733 Angiotensins Proteins 0.000 description 1
- 102000015427 Angiotensins Human genes 0.000 description 1
- 239000002028 Biomass Substances 0.000 description 1
- ZUBDGKVDJUIMQQ-VVTNISDDSA-N CC[C@H](C)[C@H](NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](Cc1c[nH]cn1)NC(=O)[C@@H]1CSSC[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@H]2CSSC[C@H](NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CCCCN)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CCSC)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CO)NC(=O)[C@H](CO)NC2=O)C(=O)N[C@@H](C(C)C)C(=O)N[C@H](Cc2ccc(O)cc2)C(=O)N[C@H](Cc2ccccc2)C(=O)N1)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](Cc1c[nH]c2ccccc12)C(O)=O Chemical compound CC[C@H](C)[C@H](NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](Cc1c[nH]cn1)NC(=O)[C@@H]1CSSC[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@H]2CSSC[C@H](NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CCCCN)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CCSC)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CO)NC(=O)[C@H](CO)NC2=O)C(=O)N[C@@H](C(C)C)C(=O)N[C@H](Cc2ccc(O)cc2)C(=O)N[C@H](Cc2ccccc2)C(=O)N1)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](Cc1c[nH]c2ccccc12)C(O)=O ZUBDGKVDJUIMQQ-VVTNISDDSA-N 0.000 description 1
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 1
- 102100034279 Calcium-binding mitochondrial carrier protein Aralar2 Human genes 0.000 description 1
- GHOKWGTUZJEAQD-UHFFFAOYSA-N Chick antidermatitis factor Natural products OCC(C)(C)C(O)C(=O)NCCC(O)=O GHOKWGTUZJEAQD-UHFFFAOYSA-N 0.000 description 1
- ZAMOUSCENKQFHK-UHFFFAOYSA-N Chlorine atom Chemical compound [Cl] ZAMOUSCENKQFHK-UHFFFAOYSA-N 0.000 description 1
- RYGMFSIKBFXOCR-UHFFFAOYSA-N Copper Chemical compound [Cu] RYGMFSIKBFXOCR-UHFFFAOYSA-N 0.000 description 1
- ZZZCUOFIHGPKAK-UHFFFAOYSA-N D-erythro-ascorbic acid Natural products OCC1OC(=O)C(O)=C1O ZZZCUOFIHGPKAK-UHFFFAOYSA-N 0.000 description 1
- 239000004375 Dextrin Substances 0.000 description 1
- 229920001353 Dextrin Polymers 0.000 description 1
- 102000002322 Egg Proteins Human genes 0.000 description 1
- 108010000912 Egg Proteins Proteins 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- PXGOKWXKJXAPGV-UHFFFAOYSA-N Fluorine Chemical compound FF PXGOKWXKJXAPGV-UHFFFAOYSA-N 0.000 description 1
- 229930091371 Fructose Natural products 0.000 description 1
- 239000005715 Fructose Substances 0.000 description 1
- RFSUNEUAIZKAJO-ARQDHWQXSA-N Fructose Chemical compound OC[C@H]1O[C@](O)(CO)[C@@H](O)[C@@H]1O RFSUNEUAIZKAJO-ARQDHWQXSA-N 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- 206010019233 Headaches Diseases 0.000 description 1
- SQUHHTBVTRBESD-UHFFFAOYSA-N Hexa-Ac-myo-Inositol Natural products CC(=O)OC1C(OC(C)=O)C(OC(C)=O)C(OC(C)=O)C(OC(C)=O)C1OC(C)=O SQUHHTBVTRBESD-UHFFFAOYSA-N 0.000 description 1
- 101000925493 Homo sapiens Endothelin-1 Proteins 0.000 description 1
- 206010010164 Hypertension complications Diseases 0.000 description 1
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 1
- 102000008192 Lactoglobulins Human genes 0.000 description 1
- 108010060630 Lactoglobulins Proteins 0.000 description 1
- GUBGYTABKSRVRQ-QKKXKWKRSA-N Lactose Natural products OC[C@H]1O[C@@H](O[C@H]2[C@H](O)[C@@H](O)C(O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@H]1O GUBGYTABKSRVRQ-QKKXKWKRSA-N 0.000 description 1
- FYYHWMGAXLPEAU-UHFFFAOYSA-N Magnesium Chemical compound [Mg] FYYHWMGAXLPEAU-UHFFFAOYSA-N 0.000 description 1
- GUBGYTABKSRVRQ-PICCSMPSSA-N Maltose Natural products O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@@H](CO)OC(O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-PICCSMPSSA-N 0.000 description 1
- 102000005741 Metalloproteases Human genes 0.000 description 1
- 108010006035 Metalloproteases Proteins 0.000 description 1
- 239000004368 Modified starch Substances 0.000 description 1
- 229920000881 Modified starch Polymers 0.000 description 1
- OVBPIULPVIDEAO-UHFFFAOYSA-N N-Pteroyl-L-glutaminsaeure Natural products C=1N=C2NC(N)=NC(=O)C2=NC=1CNC1=CC=C(C(=O)NC(CCC(O)=O)C(O)=O)C=C1 OVBPIULPVIDEAO-UHFFFAOYSA-N 0.000 description 1
- 206010029164 Nephrotic syndrome Diseases 0.000 description 1
- 235000019482 Palm oil Nutrition 0.000 description 1
- 108090000526 Papain Proteins 0.000 description 1
- 102000057297 Pepsin A Human genes 0.000 description 1
- 108090000284 Pepsin A Proteins 0.000 description 1
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 1
- 108010009736 Protein Hydrolysates Proteins 0.000 description 1
- 235000019484 Rapeseed oil Nutrition 0.000 description 1
- 235000019774 Rice Bran oil Nutrition 0.000 description 1
- 235000019485 Safflower oil Nutrition 0.000 description 1
- BUGBHKTXTAQXES-UHFFFAOYSA-N Selenium Chemical compound [Se] BUGBHKTXTAQXES-UHFFFAOYSA-N 0.000 description 1
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 1
- XUIMIQQOPSSXEZ-UHFFFAOYSA-N Silicon Chemical compound [Si] XUIMIQQOPSSXEZ-UHFFFAOYSA-N 0.000 description 1
- 108010073771 Soybean Proteins Proteins 0.000 description 1
- 101000693530 Staphylococcus aureus Staphylokinase Proteins 0.000 description 1
- 229930006000 Sucrose Natural products 0.000 description 1
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 1
- 208000009205 Tinnitus Diseases 0.000 description 1
- 102000004142 Trypsin Human genes 0.000 description 1
- 108090000631 Trypsin Proteins 0.000 description 1
- FPIPGXGPPPQFEQ-BOOMUCAASA-N Vitamin A Natural products OC/C=C(/C)\C=C\C=C(\C)/C=C/C1=C(C)CCCC1(C)C FPIPGXGPPPQFEQ-BOOMUCAASA-N 0.000 description 1
- 229930003268 Vitamin C Natural products 0.000 description 1
- 229930003316 Vitamin D Chemical class 0.000 description 1
- QYSXJUFSXHHAJI-XFEUOLMDSA-N Vitamin D3 Chemical class C1(/[C@@H]2CC[C@@H]([C@]2(CCC1)C)[C@H](C)CCCC(C)C)=C/C=C1\C[C@@H](O)CCC1=C QYSXJUFSXHHAJI-XFEUOLMDSA-N 0.000 description 1
- 229930003427 Vitamin E Natural products 0.000 description 1
- 229930003448 Vitamin K Chemical class 0.000 description 1
- HCHKCACWOHOZIP-UHFFFAOYSA-N Zinc Chemical compound [Zn] HCHKCACWOHOZIP-UHFFFAOYSA-N 0.000 description 1
- 230000002159 abnormal effect Effects 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 235000020167 acidified milk Nutrition 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- FPIPGXGPPPQFEQ-OVSJKPMPSA-N all-trans-retinol Chemical compound OC\C=C(/C)\C=C\C=C(/C)\C=C\C1=C(C)CCCC1(C)C FPIPGXGPPPQFEQ-OVSJKPMPSA-N 0.000 description 1
- BJEPYKJPYRNKOW-UHFFFAOYSA-N alpha-hydroxysuccinic acid Natural products OC(=O)C(O)CC(O)=O BJEPYKJPYRNKOW-UHFFFAOYSA-N 0.000 description 1
- 238000005349 anion exchange Methods 0.000 description 1
- 150000001450 anions Chemical class 0.000 description 1
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 1
- 108010051210 beta-Fructofuranosidase Proteins 0.000 description 1
- SQVRNKJHWKZAKO-UHFFFAOYSA-N beta-N-Acetyl-D-neuraminic acid Natural products CC(=O)NC1C(O)CC(O)(C(O)=O)OC1C(O)C(O)CO SQVRNKJHWKZAKO-UHFFFAOYSA-N 0.000 description 1
- GUBGYTABKSRVRQ-QUYVBRFLSA-N beta-maltose Chemical compound OC[C@H]1O[C@H](O[C@H]2[C@H](O)[C@@H](O)[C@H](O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@@H]1O GUBGYTABKSRVRQ-QUYVBRFLSA-N 0.000 description 1
- 229960002685 biotin Drugs 0.000 description 1
- 235000020958 biotin Nutrition 0.000 description 1
- 239000011616 biotin Substances 0.000 description 1
- 235000015895 biscuits Nutrition 0.000 description 1
- 210000004204 blood vessel Anatomy 0.000 description 1
- 210000004556 brain Anatomy 0.000 description 1
- 235000008429 bread Nutrition 0.000 description 1
- 235000014121 butter Nutrition 0.000 description 1
- 239000011575 calcium Substances 0.000 description 1
- 229910052791 calcium Inorganic materials 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 230000003197 catalytic effect Effects 0.000 description 1
- 238000006555 catalytic reaction Methods 0.000 description 1
- 235000013351 cheese Nutrition 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 239000000460 chlorine Substances 0.000 description 1
- 229910052801 chlorine Inorganic materials 0.000 description 1
- OEYIOHPDSNJKLS-UHFFFAOYSA-N choline Chemical compound C[N+](C)(C)CCO OEYIOHPDSNJKLS-UHFFFAOYSA-N 0.000 description 1
- 229960001231 choline Drugs 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 235000015165 citric acid Nutrition 0.000 description 1
- 108010084210 citrin Proteins 0.000 description 1
- 238000004140 cleaning Methods 0.000 description 1
- 239000003240 coconut oil Substances 0.000 description 1
- 235000019864 coconut oil Nutrition 0.000 description 1
- ACTIUHUUMQJHFO-UPTCCGCDSA-N coenzyme Q10 Chemical compound COC1=C(OC)C(=O)C(C\C=C(/C)CC\C=C(/C)CC\C=C(/C)CC\C=C(/C)CC\C=C(/C)CC\C=C(/C)CC\C=C(/C)CC\C=C(/C)CC\C=C(/C)CCC=C(C)C)=C(C)C1=O ACTIUHUUMQJHFO-UPTCCGCDSA-N 0.000 description 1
- 239000012141 concentrate Substances 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 229910052802 copper Inorganic materials 0.000 description 1
- 239000010949 copper Substances 0.000 description 1
- 230000009849 deactivation Effects 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 238000007872 degassing Methods 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 235000019425 dextrin Nutrition 0.000 description 1
- 235000013325 dietary fiber Nutrition 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- 208000002173 dizziness Diseases 0.000 description 1
- 235000020776 essential amino acid Nutrition 0.000 description 1
- 239000003797 essential amino acid Substances 0.000 description 1
- 150000002148 esters Chemical class 0.000 description 1
- 235000021323 fish oil Nutrition 0.000 description 1
- 239000011737 fluorine Substances 0.000 description 1
- 229910052731 fluorine Inorganic materials 0.000 description 1
- 229960000304 folic acid Drugs 0.000 description 1
- 235000019152 folic acid Nutrition 0.000 description 1
- 239000011724 folic acid Substances 0.000 description 1
- 239000003205 fragrance Substances 0.000 description 1
- 235000015203 fruit juice Nutrition 0.000 description 1
- 235000013376 functional food Nutrition 0.000 description 1
- WIGCFUFOHFEKBI-UHFFFAOYSA-N gamma-tocopherol Natural products CC(C)CCCC(C)CCCC(C)CCCC1CCC2C(C)C(O)C(C)C(C)C2O1 WIGCFUFOHFEKBI-UHFFFAOYSA-N 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 239000004519 grease Substances 0.000 description 1
- 231100000869 headache Toxicity 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 239000003112 inhibitor Substances 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- CDAISMWEOUEBRE-GPIVLXJGSA-N inositol Chemical compound O[C@H]1[C@H](O)[C@@H](O)[C@H](O)[C@H](O)[C@@H]1O CDAISMWEOUEBRE-GPIVLXJGSA-N 0.000 description 1
- 229960000367 inositol Drugs 0.000 description 1
- 230000003834 intracellular effect Effects 0.000 description 1
- 239000001573 invertase Substances 0.000 description 1
- 235000011073 invertase Nutrition 0.000 description 1
- 239000011630 iodine Substances 0.000 description 1
- 229910052740 iodine Inorganic materials 0.000 description 1
- 239000010977 jade Substances 0.000 description 1
- 210000003734 kidney Anatomy 0.000 description 1
- 230000003907 kidney function Effects 0.000 description 1
- 239000004310 lactic acid Substances 0.000 description 1
- 235000014655 lactic acid Nutrition 0.000 description 1
- 239000008101 lactose Substances 0.000 description 1
- 239000007791 liquid phase Substances 0.000 description 1
- 235000004213 low-fat Nutrition 0.000 description 1
- 239000011777 magnesium Substances 0.000 description 1
- 229910052749 magnesium Inorganic materials 0.000 description 1
- 239000001630 malic acid Substances 0.000 description 1
- 235000011090 malic acid Nutrition 0.000 description 1
- WPBNNNQJVZRUHP-UHFFFAOYSA-L manganese(2+);methyl n-[[2-(methoxycarbonylcarbamothioylamino)phenyl]carbamothioyl]carbamate;n-[2-(sulfidocarbothioylamino)ethyl]carbamodithioate Chemical compound [Mn+2].[S-]C(=S)NCCNC([S-])=S.COC(=O)NC(=S)NC1=CC=CC=C1NC(=S)NC(=O)OC WPBNNNQJVZRUHP-UHFFFAOYSA-L 0.000 description 1
- 238000004949 mass spectrometry Methods 0.000 description 1
- 235000013372 meat Nutrition 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 235000019426 modified starch Nutrition 0.000 description 1
- 208000010125 myocardial infarction Diseases 0.000 description 1
- 208000009928 nephrosis Diseases 0.000 description 1
- 231100001027 nephrosis Toxicity 0.000 description 1
- 229920001542 oligosaccharide Polymers 0.000 description 1
- 150000002482 oligosaccharides Chemical class 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 239000001254 oxidized starch Substances 0.000 description 1
- 235000013808 oxidized starch Nutrition 0.000 description 1
- 239000002540 palm oil Substances 0.000 description 1
- 229940055726 pantothenic acid Drugs 0.000 description 1
- 235000019161 pantothenic acid Nutrition 0.000 description 1
- 239000011713 pantothenic acid Substances 0.000 description 1
- 235000019834 papain Nutrition 0.000 description 1
- 229940055729 papain Drugs 0.000 description 1
- 229940111202 pepsin Drugs 0.000 description 1
- 239000008194 pharmaceutical composition Substances 0.000 description 1
- SHUZOJHMOBOZST-UHFFFAOYSA-N phylloquinone Chemical class CC(C)CCCCC(C)CCC(C)CCCC(=CCC1=C(C)C(=O)c2ccccc2C1=O)C SHUZOJHMOBOZST-UHFFFAOYSA-N 0.000 description 1
- 235000013550 pizza Nutrition 0.000 description 1
- 239000011591 potassium Substances 0.000 description 1
- 229910052700 potassium Inorganic materials 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
- 239000003531 protein hydrolysate Substances 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 239000008165 rice bran oil Substances 0.000 description 1
- 235000005713 safflower oil Nutrition 0.000 description 1
- 239000003813 safflower oil Substances 0.000 description 1
- CDAISMWEOUEBRE-UHFFFAOYSA-N scyllo-inosotol Natural products OC1C(O)C(O)C(O)C(O)C1O CDAISMWEOUEBRE-UHFFFAOYSA-N 0.000 description 1
- 229910052711 selenium Inorganic materials 0.000 description 1
- 239000011669 selenium Substances 0.000 description 1
- SQVRNKJHWKZAKO-OQPLDHBCSA-N sialic acid Chemical compound CC(=O)N[C@@H]1[C@@H](O)C[C@@](O)(C(O)=O)OC1[C@H](O)[C@H](O)CO SQVRNKJHWKZAKO-OQPLDHBCSA-N 0.000 description 1
- 229910052710 silicon Inorganic materials 0.000 description 1
- 239000010703 silicon Substances 0.000 description 1
- 229910052708 sodium Inorganic materials 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 235000014214 soft drink Nutrition 0.000 description 1
- 235000019710 soybean protein Nutrition 0.000 description 1
- 239000004575 stone Substances 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- 150000005846 sugar alcohols Chemical class 0.000 description 1
- 230000001629 suppression Effects 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 208000011580 syndromic disease Diseases 0.000 description 1
- 230000009885 systemic effect Effects 0.000 description 1
- 238000010257 thawing Methods 0.000 description 1
- 231100000886 tinnitus Toxicity 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 230000001228 trophic effect Effects 0.000 description 1
- 239000012588 trypsin Substances 0.000 description 1
- 229940088594 vitamin Drugs 0.000 description 1
- 229930003231 vitamin Natural products 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
- 235000019155 vitamin A Nutrition 0.000 description 1
- 239000011719 vitamin A Substances 0.000 description 1
- 235000019154 vitamin C Nutrition 0.000 description 1
- 239000011718 vitamin C Substances 0.000 description 1
- 235000019166 vitamin D Nutrition 0.000 description 1
- 239000011710 vitamin D Chemical class 0.000 description 1
- 150000003710 vitamin D derivatives Chemical class 0.000 description 1
- 235000019165 vitamin E Nutrition 0.000 description 1
- 229940046009 vitamin E Drugs 0.000 description 1
- 239000011709 vitamin E Substances 0.000 description 1
- 235000019168 vitamin K Nutrition 0.000 description 1
- 239000011712 vitamin K Chemical class 0.000 description 1
- 150000003721 vitamin K derivatives Chemical class 0.000 description 1
- 229940045997 vitamin a Drugs 0.000 description 1
- 229940046008 vitamin d Drugs 0.000 description 1
- 229940046010 vitamin k Drugs 0.000 description 1
- 235000008939 whole milk Nutrition 0.000 description 1
- 235000014101 wine Nutrition 0.000 description 1
- 235000013618 yogurt Nutrition 0.000 description 1
- 229910052725 zinc Inorganic materials 0.000 description 1
- 235000021249 α-casein Nutrition 0.000 description 1
- 235000021241 α-lactalbumin Nutrition 0.000 description 1
- 235000021247 β-casein Nutrition 0.000 description 1
- 235000021246 κ-casein Nutrition 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
- C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
- C07K5/08—Tripeptides
- C07K5/0802—Tripeptides with the first amino acid being neutral
- C07K5/0812—Tripeptides with the first amino acid being neutral and aromatic or cycloaliphatic
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/17—Amino acids, peptides or proteins
- A23L33/18—Peptides; Protein hydrolysates
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P9/00—Drugs for disorders of the cardiovascular system
- A61P9/12—Antihypertensives
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P21/00—Preparation of peptides or proteins
- C12P21/06—Preparation of peptides or proteins produced by the hydrolysis of a peptide bond, e.g. hydrolysate products
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2200/00—Function of food ingredients
- A23V2200/30—Foods, ingredients or supplements having a functional effect on health
- A23V2200/326—Foods, ingredients or supplements having a functional effect on health having effect on cardiovascular health
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2250/00—Food ingredients
- A23V2250/54—Proteins
- A23V2250/55—Peptide, protein hydrolysate
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Molecular Biology (AREA)
- Wood Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Zoology (AREA)
- Genetics & Genomics (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biochemistry (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- Biophysics (AREA)
- Animal Behavior & Ethology (AREA)
- Pharmacology & Pharmacy (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Mycology (AREA)
- Nutrition Science (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Heart & Thoracic Surgery (AREA)
- Cardiology (AREA)
- General Engineering & Computer Science (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
The invention provides a blood pressure reducing tripeptide derived from euphausia superba and application thereof, belongs to the technical field of blood pressure reducing oligopeptides, and the blood pressure reducing tripeptide contains Phe-Ala-Ser as an amino acid sequence, and has the molecular weight of 323.35Da when measured by ESI-MS. The preparation method of the antihypertensive tripeptide comprises the following steps: homogenizing Antarctic krill, adding distilled water and protease, and performing water enzymolysis for 5.5-6.5 h in an environment at 45-55 ℃ to prepare an Antarctic krill enzymolysis solution; and (3) carrying out ultrafiltration, anion exchange chromatography, gel column chromatography and reversed-phase high performance liquid chromatography purification on the euphausia superba enzymatic hydrolysate to prepare the decompression tripeptide. The antihypertensive tripeptide has obvious inhibiting effect on ACE, has no obvious toxic effect on Human Umbilical Vein Endothelial Cells (HUVEC), can promote the release of endogenous relaxing factors in the HUVEC cells, inhibits endothelin, and can be used as an antihypertensive peptide segment with better development prospect to be applied to antihypertensive drugs, health-care products and special medical food.
Description
Technical field
The invention belongs to blood pressure lowering oligopeptides technical fields, and in particular to a kind of decompression tripeptides derived from krill and its
Using.
Background technique
Hypertension refer to systemic arterial blood pressure (systolic pressure and/or diastolic pressure) increase for main feature (systolic pressure >=
140 millimetress of mercury, diastolic pressure >=90 millimetress of mercury), it can be with the function of the organs such as the heart, brain, kidney or the clinic of organic lesion
Syndrome.Hypertension is a kind of common disease and frequently-occurring disease.The general onset of this disease is slow, and patient's early stage is often asymptomatic, or only head
The symptoms such as dizzy, headache, palpitaition, tinnitus, seemingly a kind of independent disease, actually causes the heart, the cerebrovascular and nephrosis
Become an important risk factor, if malpractice will lesion become more serious headstroke, myocardial infarction and kidney function
These common hypertension complications such as energy failure.Angiotensin converting enzyme (ACE) is the effect of this kind of inhibitor of blood pressure lowering peptide
Target spot, it is by inhibiting the catalysis reaction of angiotensin converting enzyme to have the function that blood pressure lowering.ACE is containing there are two Zn2+It is living
Property binding site, belongs to metallopeptidase, with Zn2+Binding site is the catalytic center that " must binding site " be ACE.By with
Zn in the activated centre ACE2+Have an effect power, its inhibiting effect occurs for the various Vel-Tyr-Pro-Trp-Thr-Gln-Arg-Phes such as blood pressure lowering peptide, ACE activity because
This inactivation.For inhibition level, blood pressure lowering peptide combines closely with the active region of ACE, generates than stronger competitiveness
Inhibit.Their tightness degrees in conjunction with ACE are firmer than angiotensin converting enzyme I or bradykinin, tightly fix
In the combined area ACE, such ACE is just difficult to play its activity, it is not easy to hydrolyze angiotensin converting enzyme I degradation and generate blood vessel
Angiotensin Converting Enzyme II, at the same can't catalyzing hydrolysis bradykinin, make its inactivation, to play the role of blood pressure lowering.From food
The blood pressure lowering peptide of product is the general name of a kind of micromolecule polypeptide that can reduce human blood-pressure.General active blood pressure lowering peptide
Molecular weight in 1000Da hereinafter, be usually by protease in comparatively gentle situation aminosal obtain.Due to its drop
Blood pressure effect it is obvious and on normal arterial pressure without influencing, being without side-effects the advantages that, it has also become the hot spot studied at present.
Krill is the key species of Antarctic ecosystems, is most successful animal on the earth for biomass energy
Species (about share 500,000,000 tons).Krill has the characteristics that typical high protein, low fat, and content is respectively 16.31%
With 1.3%.Rich amino acids in its protein hydrolysate contain 18 kinds of amino acid, wherein including 8 kinds of needed by human body
Essential amino acid.So krill resource is called the protein resource warehouse of human future.It is found by the applicant that with South Pole phosphorus
Shrimp is raw material, is in blank stage using the technical study that zymolysis technique prepares krill blood pressure lowering peptide.
Summary of the invention
Have the purpose of the present invention is to provide a kind of couple of ACE and significantly inhibit effect, to Human umbilical vein endothelial cells
(HUVEC) without overt toxicity effect, and the release of endogenous relaxing factor (NO) in HUVEC cell can be promoted, inhibits Endothelin
(ET-1) the decompression tripeptides derived from krill of generation, the decompression tripeptides can be used as the preferable blood pressure lowering peptide of development prospect
Section is applied to blood pressure lowering drug, health product and special medical food.
The technical solution that the present invention is taken to achieve the above object are as follows:
The present invention provides a kind of decompression tripeptides derived from krill, the drop for being Phe-Ala-Ser containing amino acid sequence
Press tripeptides.Krill blood pressure lowering tripeptides Phe-Ala-Ser (FAS) provided by the present invention, which has its ACE, significantly inhibits work
With half-inhibitory concentration (IC50) it is 0.152mg/mL, Phe-Ala-Ser (FAS) makees Human umbilical vein endothelial cells (HUVEC)
Evaluation of Functional the result shows that: under 100-500 μM of concentration, Phe-Ala-Ser (FAS) is for HUVEC cell without obvious
Toxic effect, and the release of endogenous relaxing factor (NO) in HUVEC cell can be promoted, inhibit the generation of Endothelin (ET-1).
Therefore, Phe-Ala-Ser (FAS) illustrates that it, with good activity, can be used as to the result of ACE and HUVEC cytosis
The preferable blood pressure lowering peptide fragment of development prospect is applied to blood pressure lowering drug, health product and special medical food.
Preferably, the ESI-MS measurement molecular weight of peptide is 323.35Da.
Preferably, decompression tripeptides one or more modifications chosen from the followings:
(I) N- end cap；
(II) C- end cap；
(III) phosphorylation；
(IV) it glycosylates；With
(V) amino acid of one or more non-genomic codings.
Preferably, decompression tripeptides is prepared by following phases,
(a) stage that krill is homogenized；
(b) after distilled water and protease being added in the homogenate, water digests 5.5~6.5h in 45~55 DEG C of environment
Prepare the stage of krill enzymolysis liquid；
(c) by the krill enzymolysis liquid through ultrafiltration, anion-exchange chromatography, gel filtration chromatography and reversed phase high performance liquid
Phase chromatogram purification, the stage of preparation decompression tripeptides；
Wherein, homogenate takes intermittent super-pressure to be homogenized, method particularly includes: under conditions of pressure is 165-175MPa
Homogeneous 20-40s, then pressure maintaining 60-80s, repeats 1-4 identical HIGH PRESSURE TREATMENT after pressure release；
Decompression tripeptides accounts for 20~70wt% of whole krill enzymolysis liquid.When ultra high pressure, homogenizing time, pressure maintaining
Between between have an obvious reciprocation, and with homogenate rate, be averagely homogenized between energy consumption and homogenate that there are quadratic nonlinearities
Relationship, the intermittent super-pressure homogenate of the present embodiment can seriously destroy the integrality of air bladder, then make during pressure release
It obtains the quick releases such as containing substance such as protein into the cell to come out, then continue under conditions of continuing boosting and super-pressure
Three strands of sufficiently stable superhelixes of the property of protein are acted on, hydrogen bond are destroyed, so that the specific space of protein
Conformation is changed to unordered loose stretch-like structure, is largely exposed to molecule in intramolecular hydrophobic grouping originally
The touch opportunity of subsequent enzymolysis process neutral and alkali protease and protein is improved on surface, repeatedly, can maximumlly mention
The yield of high reducing blood lipid hexapeptide, and low energy consumption.
For optimisation technique scheme, the measure taken further include:
Protease in step (b) is trypsase, enzyme activity >=2.0 × 105U/g。
The specific steps of anion-exchange chromatography in step (c) are as follows: match ultrafiltration enzymolysis liquid Tris-Hcl buffer solution
The solution for being 55~65mg/mL at concentration, centrifugation, supernatant are added to QFF anion according to volume ratio 1mL:20~25mL and hand over
It changes in chromatographic column, it is molten with Tris-HCl buffer, 0.1mol/L NaCl solution, 0.5mol/L NaCl solution, 1mol/L NaCl
Liquid is eluted respectively, flow velocity 3mL/min, collects elution fraction according to the absorbance curve under 280nm, wherein blood pressure lowering
Most strong group of activity is divided into ion-exchange chromatography zymolyte.
The specific steps of gel filtration chromatography in step (c) are as follows: above-mentioned ion-exchange chromatography zymolyte is dissolved in distilled water and is matched
The solution for being 45~55mg/mL at concentration, centrifugation, supernatant are added to sephadex according to volume ratio 1mL:15~20mL
It in Sephadex G-15 chromatographic column, is eluted with ultrapure water, flow velocity is 0.5~0.8mL/min, and eluent is examined in 280nm
It surveys, elution fraction is collected according to the absorbance curve under 280nm, wherein most strong group of hypotensive activity is divided into gel chromatography enzymatic hydrolysis
Object.
The specific steps that reversed-phase high performance liquid chromatography purifies in step (c) are as follows: steam above-mentioned gel chromatography zymolyte with double
Water is made into the solution of 45~55 μ g/mL, is purified using RP-HPLC, is screened according to each component hypotensive activity and obtains drop blood
It presses tripeptides Phe-Ala-Ser (FAS)；RP-HPLC condition are as follows: 15~20 μ L of sample volume；Chromatographic column Kromasil C18(250mm
× 4.6mm, 5 μm)；Mobile phase: 30% acetonitrile；0.6~1.0mL/min of elution speed；Ultraviolet detection wavelength 280nm.
The present invention also provides a kind of decompression tripeptides derived from krill in preparation for treating prehypertensive or height
The application of the blood-pressure drug and/or health product and/or special medical food of blood pressure.
The present invention also provides a kind of decompression tripeptides derived from krill in production for preventing and/or treating by blood
Application in the medicament of the extremely caused disease of angiotensin invertase.
Compared with prior art, the invention has the benefit that
Krill blood pressure lowering tripeptides Phe-Ala-Ser (FAS) provided by the present invention, which has its ACE, to be significantly inhibited
Effect, half-inhibitory concentration (IC50) it is 0.152mg/mL.Phe-Ala-Ser (FAS) is to Human umbilical vein endothelial cells (HUVEC)
The Evaluation of Functional of effect the result shows that: under 100-500 μM of concentration, Phe-Ala-Ser (FAS) is for HUVEC cell without bright
Aobvious toxic effect, and the release of endogenous relaxing factor (NO) in HUVEC cell can be promoted, inhibit the life of Endothelin (ET-1)
At.Therefore, Phe-Ala-Ser (FAS) illustrates that it, with good activity, can make to the result of ACE and HUVEC cytosis
It is applied to blood pressure lowering drug, health product and special medical food for the preferable blood pressure lowering peptide fragment of development prospect, furthermore this hair
The preparation method of the krill decompression tripeptides of bright offer can maximumlly improve the yield of reducing blood lipid hexapeptide, and low energy consumption.
It provides a kind of derived from the decompression tripeptides of krill and its application present invention employs above-mentioned technical proposal, makes up
The deficiencies in the prior art, reasonable design, easy operation.
Detailed description of the invention
Fig. 1 is the ACE inhibiting rate for the krill enzymolysis product that in the embodiment of the present invention 1 prepared by five kinds of protease；
Fig. 2 is the ACE inhibiting rate of krill zymolyte difference ultrafiltration component in the embodiment of the present invention 1；
Fig. 3 is the QFF anion exchange stepwise elution curve of ultrafiltration component in the embodiment of the present invention 1；
Fig. 4 is the sephadex G-15 chromatogram of 1 intermediate ion of embodiment of the present invention exchange zymolyte；
Fig. 5 is the spectrogram of the Zorbax C18 RP-HPLC of C3 in the embodiment of the present invention 1；
Fig. 6 is the mass spectrogram that tripeptides is depressured in the embodiment of the present invention 1；
Fig. 7 is to be depressured tripeptides in the embodiment of the present invention 1 to influence the proliferation activity of Human umbilical vein endothelial cells (HUVEC)；
Fig. 8 is that influence of the tripeptides to Human umbilical vein endothelial cells (HUVEC) NO content is depressured in the embodiment of the present invention 1；
Fig. 9 is that tripeptides is depressured in the embodiment of the present invention 1 to Human umbilical vein endothelial cells (HUVEC) human endothelin -1 (ET-
1) influence of content.
Specific embodiment
In the following, in conjunction with specific embodiments to a kind of decompression tripeptides derived from krill of an embodiment of the present invention and
Its application is described further.
Embodiment 1:
A kind of decompression tripeptides derived from krill, the decompression tripeptides for being Phe-Ala-Ser containing amino acid sequence.This
Krill blood pressure lowering tripeptides Phe-Ala-Ser (FAS) provided by inventing has the effect that significantly inhibits, half suppression to its ACE
Concentration (IC processed50) it is 0.152mg/mL, the function that Phe-Ala-Ser (FAS) acts on Human umbilical vein endothelial cells (HUVEC)
Property evaluation result show: under 100-500 μM of concentration, Phe-Ala-Ser (FAS) for HUVEC cell without overt toxicity make
With, and the release of endogenous relaxing factor (NO) in HUVEC cell can be promoted, inhibit the generation of Endothelin (ET-1).Therefore,
Phe-Ala-Ser (FAS) illustrates it with good activity, before can be used as development to the result of ACE and HUVEC cytosis
The preferable blood pressure lowering peptide fragment of scape is applied to blood pressure lowering drug, health product and special medical food.
The ESI-MS measurement molecular weight of peptide is 323.35Da.
It is depressured tripeptides one or more modifications chosen from the followings:
(I) N- end cap；
(II) C- end cap；
(III) phosphorylation；
(IV) it glycosylates；With
(V) amino acid of one or more non-genomic codings.
Above-mentioned decompression tripeptides is prepared by following phases,
(a) it is homogenized: taking frost krill, thaw, clean, homogeneous 20- under conditions of pressure is 165-175MPa
40s, then pressure maintaining 60-80s, repeat 1-4 identical HIGH PRESSURE TREATMENT to get homogenate after pressure release, when ultra high pressure, homogeneous
Between, have obvious reciprocation between the dwell time, and with homogenate rate, be averagely homogenized between energy consumption and homogenate and exist
Nonlinear relationship, the intermittent super-pressure homogenate of the present embodiment can seriously destroy the integrality of air bladder, then in pressure release
During the quick releases such as intracellular containing substance such as protein are come out, then continuing boosting and super-pressure
Under the conditions of continue to act on the sufficiently stable three strands of superhelixes of property of protein, hydrogen bond is destroyed, so that protein
Specific space conformation is changed to unordered loose stretch-like structure, originally a large amount of in intramolecular hydrophobic grouping
It is exposed to molecular surface, the touch opportunity for improving subsequent enzymolysis process neutral and alkali protease and protein repeatedly can
The maximized yield for improving reducing blood lipid hexapeptide, and low energy consumption；
(b) enzymatic hydrolysis of krill: taking frost krill, after thawing, cleaning homogenate, by solid-liquid ratio 1g:8~12mL
Distilled water is added, is separately added into different protease (such as table 1) according to the 2.0% of krill weight, is adjusted separately according to table 1
Hydrolysis temperature and pH, in 95~100 DEG C, 10~15min enzyme deactivation, are cooled to room temperature, 8,000~12,000r/ after digesting 6h
Min is centrifuged 15~25min, collects supernatant, measures ACE inhibiting rate of the enzymolysis liquid under 1mg/mL concentration, as a result (such as Fig. 1)
Show that the ACE inhibitory activity of trypsin digestion object is most strong.Select trypsase for hydrolysis enzyme, in pH 8.0, enzyme concentration
2.1%, solid-liquid ratio 1:10.1 and temperature 50 C hydrolyze krill 5.95h, obtain zymolyte, and decompression tripeptides accounts for whole South Pole phosphorus
20~70wt% of shrimp enzymolysis liquid；
The enzyme activity and optimum temperature and pH of 1 five kinds of protease of table
| Project | Optimum temperature/DEG C | Optimal pH | Enzyme activity/(u/g) |
| Trypsase | 50 | 8.0 | 200000 |
| Alkali protease | 50 | 8.0 | 160000 |
| Papain | 55 | 7.0 | 500000 |
| Neutral proteinase | 40 | 7.0 | 50000 |
| Pepsin | 37 | 2.1 | 300000 |
(c) preparation of krill blood pressure lowering oligopeptides: it is 5kDa that krill enzymolysis liquid, which is selected molecular cut off,
The ultrafiltration membrane of 3.5kDa and 1kDa carries out ultrafiltration classification to krill albumen, obtains four components: ES-I (< 1kDa), ES-
II (1-3.5kDa), ES-III (3.5-5kDa) and ES-IV (> 5kDa) ultrafiltration membrane is classified.ACE inhibitory activity (such as Fig. 2)
Show that ES-I has strongest inhibitory activity, freeze-drying.ES-I sample is made into certain density solution and uses gel column respectively after freeze-drying
Chromatography and reversed-phase high performance liquid chromatography (RP-HPLC) purifying, obtain the decompression tripeptides of krill.Utilize amino acid sequence point
Analyzer and mass spectroscopy its structure, detailed process are as follows:
1. anion-exchange chromatography: above-mentioned ultrafiltration enzymolysis liquid is dissolved in Tris-Hcl buffer solution (0.05mol/L, pH
8.4) it, is made into the solution that concentration is 60mg/mL, is centrifuged 15min in 4 DEG C, 12,000r/min, supernatant is according to volume ratio 1mL:
20~25mL is added in QFF anion exchange chromatography, uses Tris-HCl buffer, the 0.1mol/L of 150mL respectively
NaCl solution (being dissolved in Tris-HCl buffer), 0.5mol/L NaCl solution (being dissolved in Tris-HCl buffer), 1mol/
LNaCl solution (is dissolved in Tris-HCl buffer) solution and is eluted respectively, flow velocity 3mL/min, and the every 3min of eluent is received
One pipe of collection, collects elution fraction (Fig. 3) according to the absorbance curve under 280nm, wherein component C has strongest decompression energy
Power, as ion-exchange chromatography zymolyte；
2. gel filtration chromatography: it is 50mg/ that above-mentioned ion-exchange chromatography zymolyte component C, which is dissolved in distilled water to be made into concentration,
The solution of mL is centrifuged 20min in 4 DEG C, 10,000r/min, and supernatant is added to sephadex according to volume ratio 1mL:20mL
It in Sephadex G-15 chromatographic column, being eluted with ultrapure water, flow velocity 0.6mL/min, the every 3min of eluent collects a pipe,
And detected in 280nm, elution fraction (such as Fig. 4) is collected according to the absorbance curve under 280nm, wherein C3 component blood pressure lowering is living
Property is most strong, as gel chromatography zymolyte；
2. RP-HPLC is purified: above-mentioned gel chromatography zymolyte C3 being made into the solution of 40~50 μ g/mL with distilled water, benefit
With RP-HPLC (20 μ L of sample volume；Chromatographic column Kromasil C18(250mm × 4.6mm, 5 μm) is purified, according to blood pressure lowering
Activity prepares hypotensive activity oligopeptides (such as Fig. 5), is detected as simple spike through RP-HPLC, utilizes protein/polypeptide sequenator
Measurement amino acid sequence is Phe-Ala-Ser (FAS), and it is 323.35Da that ESI-MS, which measures molecular weight,.
Krill blood pressure lowering oligopeptides Phe-Ala-Ser (FAS) obtained is subjected to activity rating, as a result (such as Fig. 5-9)
Show: Phe-Ala-Ser (FAS), which has ACE, significantly inhibits effect, half-inhibitory concentration (IC50) it is 0.152mg/mL.
Evaluation of Functional that Phe-Ala-Ser (FAS) acts on Human umbilical vein endothelial cells (HUVEC) the result shows that: in 100-500 μ
Under M concentration, Phe-Ala-Ser (FAS) without overt toxicity effect (such as Fig. 6), and can promote HUVEC cell for HUVEC cell
The release of middle endogenous relaxing factor (NO) inhibits the generation of Endothelin (ET-1).Therefore, Phe-Ala-Ser (FAS) is right
The result of ACE and HUVEC cytosis illustrates that it, with good activity, can be used as the preferable blood pressure lowering peptide fragment of development prospect
Applied to blood pressure lowering drug, health product and special medical food.
A kind of decompression tripeptides derived from krill is in the hypotensor prepared for treating prehypertensive or hypertension
The application of object and/or health product and/or special medical food.
The decompression tripeptides can be used in different forms, such as food and drink, specific health food, trophic function
Property food, particular utility food, healthy food, quasi- drug (quasi-drug), Pharmaceutical composition etc..For example, it can be used as medicine
With composition or quasi- drug directly to or as the particular utility food or battalion of such as healthy food and specific health food
Feeding functional food directly absorbs；Or it is ingested and being added it in various food in advance, various food such as ox
Cream, soft drink, acidified milk, yogurt, cheese, bread, biscuit, cracknel and pizza slices.In order to produce above-mentioned food, can incite somebody to action
Water, protein, carbohydrate, lipid, vitamins, minerals class, organic acid, fruit juice, fragrance etc. merge as main component.Example
Attached bag includes of animal or plant nature albumen such as whole milk powder, skimmed milk power, partially skimmed milk powder, whey powder, lactalbumin, lactalbumin
Concentrate, lactalbumin isolate, alpha-casein, beta-casein, κ-casein, beta lactoglobulin, alpha lactalbumin, newborn iron
Albumen, soybean protein, egg protein, meat albumen and its hydrolysate；A variety of cream origin ingredients, as butter, whey mineral, cream,
Whey, whey mineral, nonprotein nitrogen, sialic acid, phosphatide and lactose；Carbohydrate, such as sucrose, glucose, fructose, sugar
Alcohols, maltose, oligosaccharides, modified starch (dextrin and soluble starch, British starch, oxidized starch, starch
Ester, starch ether etc.) and dietary fibre；Animal raw fat, such as lard and fish oil；Vegetative grease, such as palm oil, safflower oil, jade
Rice bran oil, rapeseed oil, coconut oil and its distillate oil (fractional oil) include hydrogenated oil and fat, ester-exchanged oil etc.；A variety of dimension lifes
Element, as vitamin A, vitamine B group, vitamin C, vitamin D class, vitamin E, vitamin K class, citrin, CoenzymeQ10,
Niacin (niacin), niacin (nicotinic acid), pantothenic acid, biotin, inositol, choline, folic acid；Minerals such as calcium,
Potassium, magnesium, sodium, chlorine, copper, iron, manganese, zinc, selenium, fluorine, silicon, iodine；Organic acid and acylate, such as malic acid, citric acid, lactic acid, wine
Stone acid etc.；And the one or two or more for optionally selecting and being added thereto.If it is desired, in addition to sintetics it
Outside, it is further preferably added using them as containing they a large amount of food.
A kind of decompression tripeptides derived from krill is in production for preventing and/or treating by angiotensin converting enzyme
Application in the medicament of abnormal caused disease.
Comparative example 1:
The preparation method of the decompression tripeptides of krill:
Wherein homogenate uses conventional method, remaining and embodiment 1 are identical.
Measure the amino nitrogen content that dissociates in enzymolysis liquid prepared by embodiment 1 and comparative example 1:
The measurement bibliography method of free amine group are as follows: take formaldehyde 200ml, use 0.01mol/l's during electromagnetic agitation
NaOH standard solution adjusting pH value is spare to 8.2.
Solution 5ml to be measured is taken, 60ml water degasification is added, adjust pH to 8.2 with the NaOH standard solution of 0.01mol/l and is protected
1min is held, the formaldehyde 20ml of configuration is slowly added to, after magnetic agitation 3min, is titrated to the NaOH standard solution of 0.01mol/l
PH is 9.2, and record NaOH consumes volume, is denoted as V1；Using distilled water as blank, according to above-mentioned steps, record consumed NaOH's
Volume is denoted as V2。
Free amine group nitrogen content (%) calculation formula:
In formula, the concentration (mol/l) of c:NaOH standard solution；
V1: hydrolyzate is titrated to the volume (ml) of the consumed NaOH standard solution of pH 9.2 after formaldehyde is added；
V2: the volume (ml) of the consumed NaOH standard solution of pH 9.2 is titrated to after blank addition formaldehyde；
M: for the volume (ml) of taken degree of hydrolysis liquid；
0.014 is mM quality (g/mmol) of nitrogen；
The amino nitrogen content that dissociates in the enzymolysis liquid that embodiment 1 and comparative example 1 obtain is respectively as follows: 78.1% and 69.8%, this
Illustrate that the hydrolysis result of embodiment 1 is better than comparative example 1, homogenate takes intermittent super-pressure homogenate to can be improved hydrolysis result, most
The yield of the decompression tripeptides of krill is improved eventually.
The prior art of routine techniques dawn known to those skilled in the art in above-described embodiment, therefore herein no longer in detail
Carefully repeat.
The above embodiments are only used to illustrate the present invention, and not limitation of the present invention, the ordinary skill people of this field
Member can also make a variety of changes and modification without departing from the spirit and scope of the present invention.Therefore, all equivalent
Technical solution also belong to scope of the invention, scope of patent protection of the invention should be defined by the claims.
Claims (10)
1. a kind of decompression tripeptides derived from krill, it is characterised in that: the decompression for being Phe-Ala-Ser containing amino acid sequence
Tripeptides.
2. a kind of decompression tripeptides derived from krill according to claim 1, it is characterised in that: the ESI-MS of the peptide
Measurement molecular weight is 323.35Da.
3. a kind of decompression tripeptides derived from krill according to claim 1, it is characterised in that: the decompression tripeptides choosing
From following one or more modifications:
(I) N- end cap；
(II) C- end cap；
(III) phosphorylation；
(IV) it glycosylates；With
(V) amino acid of one or more non-genomic codings.
4. a kind of decompression tripeptides derived from krill according to claim 1 or 2, it is characterised in that: the decompression three
Peptide is prepared by following phases,
(a) stage that krill is homogenized；
(b) after distilled water and protease being added in the homogenate, water digests 5.5~6.5h preparation south in 45~55 DEG C of environment
The stage of pole krill enzymolysis liquid；
(c) by the krill enzymolysis liquid through ultrafiltration, anion-exchange chromatography, gel filtration chromatography and reversed-phase high performance liquid chromatography
Purifying, the stage of preparation decompression tripeptides；
Wherein, homogenate takes intermittent super-pressure to be homogenized, method particularly includes: homogeneous under conditions of pressure is 165-175MPa
20-40s, then pressure maintaining 60-80s, repeats 1-4 identical HIGH PRESSURE TREATMENT after pressure release.
5. a kind of decompression tripeptides derived from krill according to claim 4, it is characterised in that: in the step (b)
Protease be trypsase, enzyme activity >=2.0 × 105U/g。
6. a kind of decompression tripeptides derived from krill according to claim 4, it is characterised in that: in the step (c)
The specific steps of anion-exchange chromatography are as follows: it is 55~65mg/ that ultrafiltration enzymolysis liquid, which is made into concentration with Tris-Hcl buffer solution,
The solution of mL, centrifugation, supernatant are added in QFF anion exchange chromatography according to volume ratio 1mL:20~25mL, use Tris-
HCl buffer, 0.1mol/L NaCl solution, 0.5mol/L NaCl solution, 1mol/L NaCl solution are eluted respectively, stream
Speed is 3mL/min, collects elution fraction according to the absorbance curve under 280nm, wherein most strong group of hypotensive activity is divided into ion
Displacement chromatography zymolyte.
7. a kind of decompression tripeptides derived from krill according to claim 4, it is characterised in that: in the step (c)
The specific steps of gel filtration chromatography are as follows: being dissolved in distilled water to be made into concentration by the ion-exchange chromatography zymolyte is 45~55mg/
The solution of mL, centrifugation, supernatant are added to sephadex Sephadex G-15 chromatographic column according to volume ratio 1mL:15~20mL
In, it is eluted with ultrapure water, flow velocity is 0.5~0.8mL/min, and eluent is detected in 280nm, according to the extinction under 280nm
Line of writing music collects elution fraction, wherein most strong group of hypotensive activity is divided into gel chromatography zymolyte.
8. a kind of decompression tripeptides derived from krill according to claim 4, it is characterised in that: in the step (c)
The specific steps of reversed-phase high performance liquid chromatography purifying are as follows: the gel chromatography zymolyte is made into 45~55 μ g/mL with distilled water
Solution, purified using reversed-phase high performance liquid chromatography, according to each component hypotensive activity screen obtain blood pressure lowering tripeptides；
The purification condition are as follows: sample volume is 15~20 μ L；Mobile phase is 30% acetonitrile；Elution speed is 0.6~1.0mL/
min；Ultraviolet detection wavelength 280nm.
9. a kind of described in any item decompression tripeptides derived from krill of claim 1-8 are before preparation is for treating hypertension
The application of the blood-pressure drug and/or health product and/or special medical food of phase or hypertension.
10. a kind of described in any item decompression tripeptides derived from krill of claim 1-8 are in production for preventing and/or controlling
It treats by the application in the medicament of the extremely caused disease of angiotensin converting enzyme.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811397409.6A CN109400674B (en) | 2018-11-22 | 2018-11-22 | Antihypertensive tripeptide derived from euphausia superba and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811397409.6A CN109400674B (en) | 2018-11-22 | 2018-11-22 | Antihypertensive tripeptide derived from euphausia superba and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN109400674A true CN109400674A (en) | 2019-03-01 |
| CN109400674B CN109400674B (en) | 2021-03-16 |
Family
ID=65474655
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201811397409.6A Active CN109400674B (en) | 2018-11-22 | 2018-11-22 | Antihypertensive tripeptide derived from euphausia superba and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN109400674B (en) |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110760554A (en) * | 2019-10-14 | 2020-02-07 | 浙江海洋大学 | Method for preparing ACE inhibitory peptide by enzymolysis of euphausia superba protein |
| CN110563808B (en) * | 2019-09-26 | 2021-04-20 | 浙江海洋大学 | Euphausia superba antioxidant oligopeptide and preparation method thereof |
| CN113698453A (en) * | 2021-08-31 | 2021-11-26 | 浙江海洋大学 | Euphausia superba blood lipid reducing peptide and application thereof in treating hyperlipidemia |
| CN115368440A (en) * | 2022-08-18 | 2022-11-22 | 山东鲁华海洋生物科技有限公司 | Euphausia superba oligomeric composite peptide |
| CN117567562A (en) * | 2023-11-24 | 2024-02-20 | 中国水产科学研究院黄海水产研究所 | Antarctic krill ACE (angiotensin converting enzyme) inhibitory peptide as well as preparation method and application thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1769072A1 (en) * | 2004-07-22 | 2007-04-04 | University of Chile | Protein and nucleic acid sequence encoding a krill-derived cold adapted trypsin-like activity enzyme |
| CN102559825A (en) * | 2012-01-18 | 2012-07-11 | 辽宁省大连海洋渔业集团公司 | Method for preparing antarctic krill low-fluorine hydrolysis polypeptide |
| CN107586319A (en) * | 2017-10-26 | 2018-01-16 | 浙江海洋大学 | A kind of brown croaker air bladder anti-oxidation peptide and its application |
-
2018
- 2018-11-22 CN CN201811397409.6A patent/CN109400674B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1769072A1 (en) * | 2004-07-22 | 2007-04-04 | University of Chile | Protein and nucleic acid sequence encoding a krill-derived cold adapted trypsin-like activity enzyme |
| CN102559825A (en) * | 2012-01-18 | 2012-07-11 | 辽宁省大连海洋渔业集团公司 | Method for preparing antarctic krill low-fluorine hydrolysis polypeptide |
| CN107586319A (en) * | 2017-10-26 | 2018-01-16 | 浙江海洋大学 | A kind of brown croaker air bladder anti-oxidation peptide and its application |
Non-Patent Citations (3)
| Title |
|---|
| JING-YA LI等: "Mutations at the S1 Sites of Methionine Aminopeptidases from Escherichia coli and Homo sapiens Reveal the Residues Critical for Substrate Specificity", 《THE JOURNAL OF BIOLOGICAL CHEMISTRY》 * |
| NAKANO, D等: "ntihypertensive Effect of Angiotensin I-Converting Enzyme Inhibitory Peptides from a Sesame Protein Hydrolysate in Spontaneously Hypertensive Rats", 《BIOSCIENCE, BIOTECHNOLOGY, AND BIOCHEMISTRY》 * |
| 李明杰: "南极大磷虾多肽制备工艺优化、脱氟及其体外活性的研究", 《中国优秀硕士学位论文全文数据库 工程科技I辑》 * |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110563808B (en) * | 2019-09-26 | 2021-04-20 | 浙江海洋大学 | Euphausia superba antioxidant oligopeptide and preparation method thereof |
| CN110760554A (en) * | 2019-10-14 | 2020-02-07 | 浙江海洋大学 | Method for preparing ACE inhibitory peptide by enzymolysis of euphausia superba protein |
| CN113698453A (en) * | 2021-08-31 | 2021-11-26 | 浙江海洋大学 | Euphausia superba blood lipid reducing peptide and application thereof in treating hyperlipidemia |
| CN113698453B (en) * | 2021-08-31 | 2023-04-25 | 浙江海洋大学 | Antarctic krill hypolipidemic peptide and application thereof in treating hyperlipidemia |
| CN115368440A (en) * | 2022-08-18 | 2022-11-22 | 山东鲁华海洋生物科技有限公司 | Euphausia superba oligomeric composite peptide |
| CN117567562A (en) * | 2023-11-24 | 2024-02-20 | 中国水产科学研究院黄海水产研究所 | Antarctic krill ACE (angiotensin converting enzyme) inhibitory peptide as well as preparation method and application thereof |
| CN117567562B (en) * | 2023-11-24 | 2024-06-04 | 中国水产科学研究院黄海水产研究所 | Antarctic krill ACE (angiotensin converting enzyme) inhibitory peptide as well as preparation method and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN109400674B (en) | 2021-03-16 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN109400674A (en) | Antihypertensive tripeptide derived from euphausia superba and application thereof | |
| Kaur et al. | Recently isolated food-derived antihypertensive hydrolysates and peptides: A review | |
| Liu et al. | Purification and identification of an ACE inhibitory peptide from walnut protein | |
| US10414795B2 (en) | Techniques of preparing collagen active peptides | |
| Aluko | Antihypertensive peptides from food proteins | |
| JP5048487B2 (en) | Antihypertensive functional food | |
| Saleh et al. | Recent research in antihypertensive activity of food protein-derived hydrolyzates and peptides | |
| CN101305017B (en) | Bioactive peptides identified in enzymatic hydrolyzates of milk caseins and method of obtaining same | |
| EP2253324A1 (en) | Use of a casein-derived peptide and compositions thereof as antihypertensive | |
| de Oliveira et al. | Anti-hypertensive peptides derived from caseins: mechanism of physiological action, production bioprocesses, and challenges for food applications | |
| KR101449763B1 (en) | Phaseolus vulgaris extracts, their use, and formulations containing them | |
| JP2008056645A (en) | Anti-oxidant peptide obtained by reaction of protein in enzymatically treated royal jelly with polypeptide and method for producing the same | |
| CN101022737B (en) | Anti-hypertensive functional food products | |
| CN1377590A (en) | Method for producing soybean protein hydrolyate and use | |
| CN104480176A (en) | Silver carp DPP-IV inhibitory polypeptide and applications thereof | |
| CN102030817B (en) | Peptide for inhibiting angiontensin converting enzyme | |
| CA2810641A1 (en) | Skin collagen production-promoting agent | |
| TWI590766B (en) | Thrombotic disease prevention preparation | |
| CN104877007A (en) | Yak milk lactalbumin-source ACE inhibitory peptides and preparation method thereof | |
| CN110499351A (en) | A kind of soybean peptide and the preparation method and application thereof with high ACE inhibitory activity | |
| CA2195053C (en) | Agents for inhibiting accumulation of visceral fat | |
| CN115124591A (en) | Spirulina platensis phycocyanin angiotensin converting enzyme inhibitory peptide and preparation method and application thereof | |
| JP2011036241A (en) | Method for preparation of angiotensin-converting enzyme inhibitor peptide | |
| KR20100096442A (en) | Methods for fermentative production of fibrinolytic enzyme from auricularia auricula-judae and uses thereof | |
| KR20120049043A (en) | Antiinflammatory composition comprising enzymatic hydrolysates of mytilus coruscus |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| TR01 | Transfer of patent right | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20220523 Address after: 056899 plant 4, hi tech Incubation Park, Weixian Economic Development Zone, Handan City, Hebei Province Patentee after: Zhongkelijun Co.,Ltd. Address before: 710026 No. B49, Xinda Zhongchuang space, 26th Street, block C, No. 2 Trading Plaza, South China City, international port district, Xi'an, Shaanxi Province Patentee before: Xi'an Huaqi Zhongxin Technology Development Co.,Ltd. Effective date of registration: 20220523 Address after: 710026 No. B49, Xinda Zhongchuang space, 26th Street, block C, No. 2 Trading Plaza, South China City, international port district, Xi'an, Shaanxi Province Patentee after: Xi'an Huaqi Zhongxin Technology Development Co.,Ltd. Address before: 316000 plot C2-10, Xiaohui Industrial Zone, Putuo Zhanmao, Putuo marine science and Technology Industrial Park, Zhoushan City, Zhejiang Province Patentee before: Zhejiang Ocean University |