CN114989250A - 一种来源于海水珍珠的血管紧张素转化酶抑制多肽及其应用 - Google Patents
一种来源于海水珍珠的血管紧张素转化酶抑制多肽及其应用 Download PDFInfo
- Publication number
- CN114989250A CN114989250A CN202210638560.4A CN202210638560A CN114989250A CN 114989250 A CN114989250 A CN 114989250A CN 202210638560 A CN202210638560 A CN 202210638560A CN 114989250 A CN114989250 A CN 114989250A
- Authority
- CN
- China
- Prior art keywords
- pearl
- converting enzyme
- angiotensin
- matrix protein
- enzyme inhibitory
- 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
- 108090000765 processed proteins & peptides Proteins 0.000 title claims abstract description 41
- 108090000882 Peptidyl-Dipeptidase A Proteins 0.000 title claims abstract description 39
- 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 title claims abstract description 37
- 230000002401 inhibitory effect Effects 0.000 title claims abstract description 32
- 102000004196 processed proteins & peptides Human genes 0.000 title claims abstract description 28
- 229920001184 polypeptide Polymers 0.000 title claims abstract description 27
- 239000013535 sea water Substances 0.000 title claims abstract description 7
- 102000004270 Peptidyl-Dipeptidase A Human genes 0.000 title claims description 38
- 239000011049 pearl Substances 0.000 claims abstract description 45
- 239000003814 drug Substances 0.000 claims abstract description 9
- 230000000694 effects Effects 0.000 claims abstract description 9
- 238000002360 preparation method Methods 0.000 claims description 5
- 230000036772 blood pressure Effects 0.000 claims description 4
- 239000005541 ACE inhibitor Substances 0.000 claims description 3
- 229940044094 angiotensin-converting-enzyme inhibitor Drugs 0.000 claims description 3
- 101710129690 Angiotensin-converting enzyme inhibitor Proteins 0.000 claims description 2
- 101710086378 Bradykinin-potentiating and C-type natriuretic peptides Proteins 0.000 claims description 2
- 230000001105 regulatory effect Effects 0.000 claims description 2
- 125000003275 alpha amino acid group Chemical group 0.000 claims 1
- 108060003393 Granulin Proteins 0.000 abstract description 24
- 241000490568 Pinctada fucata Species 0.000 abstract description 7
- 238000004128 high performance liquid chromatography Methods 0.000 abstract description 5
- 150000003384 small molecules Chemical class 0.000 abstract description 4
- 206010020772 Hypertension Diseases 0.000 abstract description 3
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 abstract description 2
- 102100030988 Angiotensin-converting enzyme Human genes 0.000 abstract 2
- 238000012795 verification Methods 0.000 abstract 1
- DTQVDTLACAAQTR-UHFFFAOYSA-N Trifluoroacetic acid Chemical compound OC(=O)C(F)(F)F DTQVDTLACAAQTR-UHFFFAOYSA-N 0.000 description 10
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical group CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 9
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 9
- 239000000843 powder Substances 0.000 description 8
- 239000000203 mixture Substances 0.000 description 7
- 239000000243 solution Substances 0.000 description 7
- QIAFMBKCNZACKA-UHFFFAOYSA-N N-benzoylglycine Chemical compound OC(=O)CNC(=O)C1=CC=CC=C1 QIAFMBKCNZACKA-UHFFFAOYSA-N 0.000 description 6
- 102000004142 Trypsin Human genes 0.000 description 6
- 108090000631 Trypsin Proteins 0.000 description 6
- 238000004108 freeze drying Methods 0.000 description 6
- 239000012071 phase Substances 0.000 description 6
- 239000000758 substrate Substances 0.000 description 6
- 239000012588 trypsin Substances 0.000 description 6
- 239000011159 matrix material Substances 0.000 description 5
- 238000010298 pulverizing process Methods 0.000 description 5
- 108010064733 Angiotensins Proteins 0.000 description 4
- 102000015427 Angiotensins Human genes 0.000 description 4
- BTBUEUYNUDRHOZ-UHFFFAOYSA-N Borate Chemical compound [O-]B([O-])[O-] BTBUEUYNUDRHOZ-UHFFFAOYSA-N 0.000 description 4
- 102000004190 Enzymes Human genes 0.000 description 4
- 108090000790 Enzymes Proteins 0.000 description 4
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 4
- 239000007853 buffer solution Substances 0.000 description 4
- 229940079593 drug Drugs 0.000 description 4
- 239000000706 filtrate Substances 0.000 description 4
- 239000000523 sample Substances 0.000 description 4
- 239000006228 supernatant Substances 0.000 description 4
- AAXWBCKQYLBQKY-IRXDYDNUSA-N (2s)-2-[[(2s)-2-[(2-benzamidoacetyl)amino]-3-(1h-imidazol-5-yl)propanoyl]amino]-4-methylpentanoic acid Chemical compound C([C@@H](C(=O)N[C@@H](CC(C)C)C(O)=O)NC(=O)CNC(=O)C=1C=CC=CC=1)C1=CN=CN1 AAXWBCKQYLBQKY-IRXDYDNUSA-N 0.000 description 3
- 235000005956 Cosmos caudatus Nutrition 0.000 description 3
- 244000293323 Cosmos caudatus Species 0.000 description 3
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 3
- ROHFNLRQFUQHCH-YFKPBYRVSA-N L-leucine Chemical compound CC(C)C[C@H](N)C(O)=O ROHFNLRQFUQHCH-YFKPBYRVSA-N 0.000 description 3
- 230000009286 beneficial effect Effects 0.000 description 3
- 239000008367 deionised water Substances 0.000 description 3
- 229910021641 deionized water Inorganic materials 0.000 description 3
- 229960001484 edetic acid Drugs 0.000 description 3
- ADFCQWZHKCXPAJ-GFCCVEGCSA-N equol Chemical compound C1=CC(O)=CC=C1[C@@H]1CC2=CC=C(O)C=C2OC1 ADFCQWZHKCXPAJ-GFCCVEGCSA-N 0.000 description 3
- 235000019126 equol Nutrition 0.000 description 3
- 108010016268 hippuryl-histidyl-leucine Proteins 0.000 description 3
- ADFCQWZHKCXPAJ-UHFFFAOYSA-N indofine Natural products C1=CC(O)=CC=C1C1CC2=CC=C(O)C=C2OC1 ADFCQWZHKCXPAJ-UHFFFAOYSA-N 0.000 description 3
- 230000005764 inhibitory process Effects 0.000 description 3
- 238000001840 matrix-assisted laser desorption--ionisation time-of-flight mass spectrometry Methods 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- 102000004169 proteins and genes Human genes 0.000 description 3
- 108090000623 proteins and genes Proteins 0.000 description 3
- 238000000746 purification Methods 0.000 description 3
- 239000012488 sample solution Substances 0.000 description 3
- 238000000926 separation method Methods 0.000 description 3
- 238000000108 ultra-filtration Methods 0.000 description 3
- CUKWUWBLQQDQAC-VEQWQPCFSA-N (3s)-3-amino-4-[[(2s)-1-[[(2s)-1-[[(2s)-1-[[(2s,3s)-1-[[(2s)-1-[(2s)-2-[[(1s)-1-carboxyethyl]carbamoyl]pyrrolidin-1-yl]-3-(1h-imidazol-5-yl)-1-oxopropan-2-yl]amino]-3-methyl-1-oxopentan-2-yl]amino]-3-(4-hydroxyphenyl)-1-oxopropan-2-yl]amino]-3-methyl-1-ox Chemical compound C([C@@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC=1NC=NC=1)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](C)C(O)=O)NC(=O)[C@@H](NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@@H](N)CC(O)=O)C(C)C)C1=CC=C(O)C=C1 CUKWUWBLQQDQAC-VEQWQPCFSA-N 0.000 description 2
- 102400000345 Angiotensin-2 Human genes 0.000 description 2
- 101800000733 Angiotensin-2 Proteins 0.000 description 2
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 description 2
- 101000984728 Chiropsoides quadrigatus Angiotensin-converting enzyme inhibitory peptide Proteins 0.000 description 2
- NTYJJOPFIAHURM-UHFFFAOYSA-N Histamine Chemical compound NCCC1=CN=CN1 NTYJJOPFIAHURM-UHFFFAOYSA-N 0.000 description 2
- ROHFNLRQFUQHCH-UHFFFAOYSA-N Leucine Natural products CC(C)CC(N)C(O)=O ROHFNLRQFUQHCH-UHFFFAOYSA-N 0.000 description 2
- 241001212699 Pinctada martensii Species 0.000 description 2
- 150000001413 amino acids Chemical group 0.000 description 2
- 229950006323 angiotensin ii Drugs 0.000 description 2
- 238000009835 boiling Methods 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 238000000605 extraction Methods 0.000 description 2
- 235000013305 food Nutrition 0.000 description 2
- 230000002779 inactivation Effects 0.000 description 2
- 230000014759 maintenance of location Effects 0.000 description 2
- 239000012982 microporous membrane Substances 0.000 description 2
- 231100000252 nontoxic Toxicity 0.000 description 2
- 230000003000 nontoxic effect Effects 0.000 description 2
- 239000011780 sodium chloride Substances 0.000 description 2
- 238000000967 suction filtration Methods 0.000 description 2
- 231100000331 toxic Toxicity 0.000 description 2
- 230000002588 toxic effect Effects 0.000 description 2
- 208000002874 Acne Vulgaris Diseases 0.000 description 1
- 102400000344 Angiotensin-1 Human genes 0.000 description 1
- 101800000734 Angiotensin-1 Proteins 0.000 description 1
- 102000005367 Carboxypeptidases Human genes 0.000 description 1
- 108010006303 Carboxypeptidases Proteins 0.000 description 1
- 229920002101 Chitin Polymers 0.000 description 1
- 206010010904 Convulsion Diseases 0.000 description 1
- 206010012735 Diarrhoea Diseases 0.000 description 1
- 108010016626 Dipeptides Proteins 0.000 description 1
- ZUELLZFHJUPFEC-PMVMPFDFSA-N His-Phe-Trp Chemical compound C([C@H](N)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(O)=O)C1=CN=CN1 ZUELLZFHJUPFEC-PMVMPFDFSA-N 0.000 description 1
- 206010062717 Increased upper airway secretion Diseases 0.000 description 1
- 108010007859 Lisinopril Proteins 0.000 description 1
- AHFOKDZWPPGJAZ-SRVKXCTJSA-N Lys-Lys-Cys Chemical compound C(CCN)C[C@@H](C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CS)C(=O)O)N AHFOKDZWPPGJAZ-SRVKXCTJSA-N 0.000 description 1
- 150000007945 N-acyl ureas Chemical class 0.000 description 1
- 206010059013 Nocturnal emission Diseases 0.000 description 1
- 206010033557 Palpitations Diseases 0.000 description 1
- ZVEQWRWMRFIVSD-HRCADAONSA-N Pro-Phe-Pro Chemical compound C1C[C@H](NC1)C(=O)N[C@@H](CC2=CC=CC=C2)C(=O)N3CCC[C@@H]3C(=O)O ZVEQWRWMRFIVSD-HRCADAONSA-N 0.000 description 1
- 238000012300 Sequence Analysis Methods 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 206010000496 acne Diseases 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 238000007605 air drying Methods 0.000 description 1
- ORWYRWWVDCYOMK-HBZPZAIKSA-N angiotensin I Chemical compound C([C@@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC=1NC=NC=1)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H](CC=1NC=NC=1)C(=O)N[C@@H](CC(C)C)C(O)=O)NC(=O)[C@@H](NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@@H](N)CC(O)=O)C(C)C)C1=CC=C(O)C=C1 ORWYRWWVDCYOMK-HBZPZAIKSA-N 0.000 description 1
- 239000002220 antihypertensive agent Substances 0.000 description 1
- 229940127088 antihypertensive drug Drugs 0.000 description 1
- 229910000019 calcium carbonate Inorganic materials 0.000 description 1
- 230000001914 calming effect Effects 0.000 description 1
- 229960000830 captopril Drugs 0.000 description 1
- FAKRSMQSSFJEIM-RQJHMYQMSA-N captopril Chemical compound SC[C@@H](C)C(=O)N1CCC[C@H]1C(O)=O FAKRSMQSSFJEIM-RQJHMYQMSA-N 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 238000004140 cleaning Methods 0.000 description 1
- 230000036461 convulsion Effects 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 230000001186 cumulative effect Effects 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 230000001815 facial effect Effects 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- 229960001340 histamine Drugs 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
- 238000000338 in vitro Methods 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 239000007791 liquid phase Substances 0.000 description 1
- 229960002394 lisinopril Drugs 0.000 description 1
- RLAWWYSOJDYHDC-BZSNNMDCSA-N lisinopril Chemical compound C([C@H](N[C@@H](CCCCN)C(=O)N1[C@@H](CCC1)C(O)=O)C(O)=O)CC1=CC=CC=C1 RLAWWYSOJDYHDC-BZSNNMDCSA-N 0.000 description 1
- 239000002398 materia medica Substances 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 210000002464 muscle smooth vascular Anatomy 0.000 description 1
- 238000013021 overheating Methods 0.000 description 1
- 208000026435 phlegm Diseases 0.000 description 1
- 230000001766 physiological effect Effects 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 230000029865 regulation of blood pressure Effects 0.000 description 1
- 238000001179 sorption measurement Methods 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 230000001502 supplementing effect Effects 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 108010084932 tryptophyl-proline Proteins 0.000 description 1
- 238000000825 ultraviolet detection Methods 0.000 description 1
- 230000002087 whitening effect Effects 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
- C07K7/04—Linear peptides containing only normal peptide links
- C07K7/06—Linear peptides containing only normal peptide links having 5 to 11 amino acids
-
- 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
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/14—Hydrolases (3)
- C12N9/48—Hydrolases (3) acting on peptide bonds (3.4)
- C12N9/485—Exopeptidases (3.4.11-3.4.19)
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P21/00—Preparation of peptides or proteins
- C12P21/06—Preparation of peptides or proteins produced by the hydrolysis of a peptide bond, e.g. hydrolysate products
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y304/00—Hydrolases acting on peptide bonds, i.e. peptidases (3.4)
- C12Y304/15—Peptidyl-dipeptidases (3.4.15)
- C12Y304/15001—Peptidyl-dipeptidase A (3.4.15.1)
-
- 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
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/80—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in fisheries management
- Y02A40/81—Aquaculture, e.g. of fish
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Biochemistry (AREA)
- Molecular Biology (AREA)
- Medicinal Chemistry (AREA)
- General Engineering & Computer Science (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Microbiology (AREA)
- Biotechnology (AREA)
- Biomedical Technology (AREA)
- Biophysics (AREA)
- Veterinary Medicine (AREA)
- Animal Behavior & Ethology (AREA)
- Heart & Thoracic Surgery (AREA)
- Pharmacology & Pharmacy (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Public Health (AREA)
- Cardiology (AREA)
- Mycology (AREA)
- Nutrition Science (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Peptides Or Proteins (AREA)
Abstract
本发明属于海洋生物小分子活性肽领域，具体涉及一种来源于海水珍珠的血管紧张素转化酶抑制多肽及其应用，其氨基酸序列为Lys‑Lys‑Cys‑His‑Phe‑Trp‑Pro‑Phe‑Pro‑Trp。本发明通过将合浦珠母贝所产无核珍珠粉碎，脱钙制备珍珠基质蛋白，酶解珍珠基质蛋白，将酶解物通过高效液相色谱分离纯化得到该小分子活性肽并进行活性验证。结果表明本发明的活性肽具有很好的ACE抑制活性，IC50值为4.17μM，可用于高血压疾病的保健和治疗，可用于保健品及生物医药制品的开发，具有广阔的应用前景。
Description
技术领域
本发明涉及一种从合浦珠母贝珍珠基质蛋白中分离纯化得到的血管紧张素转化酶抑制多肽，属于生物技术领域。
背景技术
血管紧张素转化酶(Angiotensin Converting Enzyme,ACE)是一种广泛存在于哺乳动物组织中的二肽羧基肽酶，在人体内参与血压的调节。ACE可以水解血管紧张素Ⅰ将其末端的组氨酸亮氨酸去除，转化为血管紧张素Ⅱ。血管紧张素Ⅱ作用于动脉血管平滑肌，使其收缩引起血压迅速升高。因此血管紧张素转化酶抑制剂成为临床上重要的降血压药物，如开托普利、赖诺普利等。但此类化学合成降压药物往往要长期服用，毒副作用的累积效应不容忽视。食物来源的蛋白制备的ACE抑制肽具有生理活性高和毒副作用小的优点成为研究的热点。
珍珠，由95％的碳酸钙和5％的有机基质组成，其中有机基质包括基质蛋白、几丁质等多糖和极少量的脂类，基质蛋白是有机基质的主要成分。在中国古代，珍珠不仅用作装饰品或首饰，还是一味珍贵的中药材，《本草纲目》记载：珍珠无毒，镇心点目珍珠涂面，令人润泽好颜色，可坠痰，除面斑，止泻除小儿惊热，安魂魄止遗精白浊，解痘疗毒，令光泽洁白。并且可以将珍珠磨成珍珠粉直接内服，如中国古代武则天、慈禧、和珅具有内服珍珠粉并对健康有益的历史记载，在古埃及也有埃及艳后将珍珠溶解服用的故事，可见珍珠粉可以看作一种特殊的保健食品，是安全无毒有益的，只是在古代比较奢侈昂贵而只有少数人能享用。目前珍珠基质蛋白ACE抑制肽未见报道。
发明内容
本发明针对上述问题、现状和发展前景，提供一种来源于海水珍珠的血管紧张素酶抑制活性肽及应用，该活性肽具有ACE抑制活性，分子量小，活性高，易于分离纯化，可用于高血压相关疾病的保健及治疗，在生物医药领域有广阔的应用前景。
本发明为实现上述目的提供的技术方案为：
一种从海水珍珠中分离纯化的新型血管紧张素酶抑制活性肽，其氨基酸序列为Lys-Lys-Cys-His-Phe-Trp-Pro-Phe-Pro-Trp。
以合浦珠母贝珍珠为例，一种从合浦珠母贝珍珠基质蛋白中分离纯化的新型血管紧张素酶抑制活性肽，其制备方法包括以下步骤：
(1)珍珠基质蛋白的提取：将合浦珠母贝无核珍珠粉碎，收集珍珠粉，置于烧杯中，加入五倍体积的1.5M乙二胺四乙酸(EDTA)脱钙，离心收集上清液透析，然后冷冻干燥获得珍珠基质蛋白。
(2)珍珠基质蛋白的酶解：取珍珠基质蛋白冻干粉加入胰蛋白酶(trypsin)进行酶解，胰蛋白酶的加入量为珍珠基质蛋白重量的5％，酶解体系的pH为8.0，反应温度为50℃，酶解时间为2小时，沸水浴灭活后离心收集上清液，抽滤后使用蛋白浓缩离心管进行离心超滤，截留分子量的规格为3KD，收集分子量小于3KD的滤过液，浓缩后冷冻干燥获得酶解好的珍珠基质蛋白多肽混合物。
(3)珍珠基质蛋白ACE抑制多肽的分离纯化：使用高效液相色谱对酶解好的珍珠基质蛋白多肽混合物进行纯化，色谱柱为C18，流动相A为含有0.1％三氟乙酸的去离子水，流动相B为含有0.1％三氟乙酸的乙腈，流速为1ml/min，紫外检测波长为280nm，收集保留时间为14min的样品，浓缩后冷冻干燥即得到序列为Lys-Lys-Cys-His-Phe-Trp-Pro-Phe-Pro-Trp的血管紧张素转化酶抑制多肽。
本发明提出的活性肽可以用于制备血管紧张素转化酶抑制剂、制备具有调节血压作用的药物或者制备保健品。
本发明的有益效果：
(1)本发明获得了一种从合浦珠母贝珍珠基质蛋白中分离纯化的新型血管紧张素酶抑制活性肽，该活性肽具有很好的ACE抑制活性，ACE抑制IC50值为4.17μM，可用于开发治疗高血压的相关保健品或者药物，在保健品及医药领域有广阔的应用前景。
(2)本发明为珍珠的进一步精深技工和医药开发提供了可靠依据。
附图说明
图1为本发明小分子活性肽的ACE抑制HPLC图，其中：A，对照；B，多肽终浓度0.025mg/ml(17.63μM)；C，多肽终浓度0.05mg/ml(35.26μM)；D，多肽终浓度0.1mg/ml(70.52μM)；E，多肽终浓度0.15mg/ml(105.78μM)；F，多肽终浓度0.2mg/ml(141.04μM)；G，多肽终浓度0.25mg/ml(176.3μM)。
图2为本发明小分子活性肽的分子式；
图3为本发明小分子活性肽的ACE抑制活性图。
具体实施方式
下面结合说明书附图和具体实施例进一步阐明本发明。
实施例1珍珠基质血管紧张素转化酶抑制多肽的制备
(1)珍珠基质蛋白的提取
将产自广东湛江的合浦珠母贝无核珍珠洗净、晾干，使用中药粉碎机充分粉碎，在粉碎过程中及时停顿，给粉碎容器降温，防止粉碎容器过热，破坏珍珠活性成分。收集珍珠粉，置于烧杯中，加入五倍体积的1.5M EDTA，置于4℃层析冷柜中搅拌脱钙，24小时后装入50ml高速离心管离心(10000rpm)。收集上清液，用0.2μm微孔滤膜抽滤，滤过液置于4℃层析冷柜中低温透析48h，期间多次换水，然后冷冻干燥获得珍珠基质蛋白。
(2)珍珠基质蛋白的酶解
取珍珠基质蛋白冻干粉，加入trypsin进行酶解。胰蛋白酶的活性为10000U/g，胰蛋白酶的加入量为珍珠基质蛋白重量的5％，酶解体系的pH调整为8.0，在50℃恒温水浴锅中酶解2小时，然后放入沸水浴中灭活，装入50ml高速离心管离心(10000rpm)。收集上清液，用0.2μm微孔滤膜抽滤，滤过液使用Millipore蛋白浓缩离心管进行离心超滤(4000rpm)，内置低吸附的Ultracel-PL超滤膜截留规格为3KD，收集分子量小于3KD的滤过液，浓缩后冷冻干燥获得酶解好的珍珠基质蛋白多肽混合物。
(3)珍珠基质蛋白ACE抑制多肽的分离纯化
使用高效液相色谱对酶解好的珍珠基质蛋白多肽混合物进行纯化，采用C18色谱柱，流动相A为含有0.1％三氟乙酸的去离子水，流动相B为含有0.1％三氟乙酸的乙腈。紫外检测波长为280nm，流速为1ml/min，收集保留时间为14min的样品，浓缩后冷冻干燥即得到序列为Lys-Lys-Cys-His-Phe-Trp-Pro-Phe-Pro-Trp的血管紧张素转化酶抑制多肽。
实施例2：珍珠基质血管紧张素转化酶抑制多肽的ACE抑制活性测定
使用高效液相色谱法测定ACE抑制多肽对ACE的抑制活性。
将得到的ACE抑制多肽溶于0.1M硼酸盐缓冲溶液(含0.3M NaCl，pH8.3)中制成1mg/ml的样品溶液，将血管紧张素转化酶及其底物马尿酰组胺酰亮氨酸HHL(Hip-His-Leu)溶于0.1M硼酸盐缓冲溶液(含0.3M NaCl，pH8.3)中，配成浓度200U/L的ACE酶液和浓度5mM的HHL底物溶液。
实验组共六组，取HHL底物溶液80μl，分别加入5μl、10μl、20μl、30μl、40μl和50μl样品溶液，用0.1M硼酸盐缓冲液补足至180μl，对照组用0.1M硼酸盐缓冲液代替样品液，混匀后放入37℃恒温水浴中保温5min，再加入20μlACE酶液启动反应，以120r/min频率37℃振荡水浴保育30min。加入0.2ml的1M HCl终止反应。将溶液置于高效液相色谱仪中测定其中马尿酸的含量。
液相条件如下：流速为每分钟0.5ml，流动相A为纯乙腈，流动相B为含0.5％三氟乙酸的去离子水，二者比例为1:3，检测波长为228nm。由于ACE可以稳定降解马尿酰组胺酰尿氨酸并生成马尿酸，一旦ACE的活性受到抑制，样品中马尿酸的生成量也随之减少。底物的ACE抑制活性可按以下公式计算：
ACE抑制率＝(As-Ad)/Ad
上式中，As为样品组中马尿酸检测峰面积；Ad为空白组中马尿酸检测峰面积，并计算IC50值。
实验结果见图1和图3，序列为Lys-Lys-Cys-His-Phe-Trp-Pro-Phe-Pro-Trp的多肽体外血管紧张素转化酶抑制IC50值为4.17μM，该多肽具有很好抑制血管紧张素转化酶的活性。
实施例3：珍珠基质血管紧张素转化酶抑制多肽的序列分析
取实施例1中所制备得到的珍珠基质血管紧张素转化酶抑制多肽，经基质辅助激光解吸电离飞行时间质谱(MALDI-TOF/TOF-MS)测定，其分子量为1417.5Da，其氨基酸序列为：Lys-Lys-Cys-His-Phe-Trp-Pro-Phe-Pro-Trp(如SEQ ID NO:1所示)。
以上所述，仅为本发明较佳的具体实施方式，但本发明的保护范围并不局限于此，任何熟悉本技术领域的技术人员在本发明披露的技术范围内，根据本发明的技术方案及其发明构思加以同等替换或改变，都应涵盖在本发明的保护范围内。
序列表
<110> 浙江清荣生物科技发展有限公司
浙江清华长三角研究院
<120> 一种来源于海水珍珠的血管紧张素转化酶抑制多肽及其应用
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 10
<212> PRT
<213> 合浦珠母贝(Pinctada fucata)
<400> 1
Lys Lys Cys His Phe Trp Pro Phe Pro Trp
1 5 10
Claims (4)
1.一种来源于海水珍珠的血管紧张素转化酶抑制多肽，其特征在于，所述血管紧张素转化酶抑制多肽的氨基酸顺序为Lys-Lys-Cys-His-Phe-Trp-Pro-Phe-Pro-Trp。
2.根据权利要求1所述的血管紧张素转化酶抑制多肽在制备血管紧张素转化酶抑制剂中的应用。
3.根据权利要求1所述的血管紧张素转化酶抑制多肽在制备具有调节血压作用的药物中的应用。
4.根据权利要求1所述的血管紧张素转化酶抑制多肽在制备保健品中的应用。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210638560.4A CN114989250B (zh) | 2022-06-07 | 2022-06-07 | 一种来源于海水珍珠的血管紧张素转化酶抑制多肽及其应用 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210638560.4A CN114989250B (zh) | 2022-06-07 | 2022-06-07 | 一种来源于海水珍珠的血管紧张素转化酶抑制多肽及其应用 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN114989250A true CN114989250A (zh) | 2022-09-02 |
| CN114989250B CN114989250B (zh) | 2023-08-15 |
Family
ID=83032938
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202210638560.4A Active CN114989250B (zh) | 2022-06-07 | 2022-06-07 | 一种来源于海水珍珠的血管紧张素转化酶抑制多肽及其应用 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN114989250B (zh) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN117143190A (zh) * | 2023-08-31 | 2023-12-01 | 浙江清荣生物科技发展有限公司 | Kkch多肽在抗冠状病毒中的应用 |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2001106699A (ja) * | 1999-10-05 | 2001-04-17 | Suetsuna Yoko | 新規なヘクサペプチドおよびアンジオテンシン変換酵素阻害剤 |
| CN101756237A (zh) * | 2009-11-27 | 2010-06-30 | 广东海洋大学 | 马氏珍珠贝肉抗高血压功能营养液生产方法 |
| CN113024633A (zh) * | 2021-03-12 | 2021-06-25 | 中国科学院南海海洋研究所 | 一种珍珠贝肉来源ace活性抑制肽及其制备筛选方法和应用 |
-
2022
- 2022-06-07 CN CN202210638560.4A patent/CN114989250B/zh active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2001106699A (ja) * | 1999-10-05 | 2001-04-17 | Suetsuna Yoko | 新規なヘクサペプチドおよびアンジオテンシン変換酵素阻害剤 |
| CN101756237A (zh) * | 2009-11-27 | 2010-06-30 | 广东海洋大学 | 马氏珍珠贝肉抗高血压功能营养液生产方法 |
| CN113024633A (zh) * | 2021-03-12 | 2021-06-25 | 中国科学院南海海洋研究所 | 一种珍珠贝肉来源ace活性抑制肽及其制备筛选方法和应用 |
Non-Patent Citations (2)
| Title |
|---|
| CHANG-BUM AHN ET AL.: "Angiotensin I converting enzyme (ACE) inhibitory peptides from salmon byproduct protein hydrolysate by Alcalase hydrolysis", 《PROCESS BIOCHEMISTRY》, pages 2240 - 2245 * |
| PENGRU LIU ET AL.: "Purification, Characterization and Evaluation of Inhibitory Mechanism of ACE Inhibitory Peptides from Pearl Oyster (Pinctada fucata martensii) Meat Protein Hydrolysate", 《MAR. DRUGS 》, pages 463 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN117143190A (zh) * | 2023-08-31 | 2023-12-01 | 浙江清荣生物科技发展有限公司 | Kkch多肽在抗冠状病毒中的应用 |
| CN117143190B (zh) * | 2023-08-31 | 2024-03-26 | 浙江清荣生物科技发展有限公司 | Kkch多肽在抗冠状病毒中的应用 |
Also Published As
| Publication number | Publication date |
|---|---|
| CN114989250B (zh) | 2023-08-15 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| JP5763203B2 (ja) | トウモロコシ降圧活性ペプチドを調製するための工業的方法 | |
| CN109400678A (zh) | 一种刺参来源的抗氧化和dpp-iv抑制活性肽 | |
| CN101037468A (zh) | 一种牡蛎活性肽的制备方法 | |
| WO2021082294A1 (zh) | 一种免疫调节肽的制备及方法 | |
| CN114989250A (zh) | 一种来源于海水珍珠的血管紧张素转化酶抑制多肽及其应用 | |
| CN112457387A (zh) | 一种厚壳贻贝寡肽及其用途 | |
| JP2009029772A (ja) | 酵素処理ローヤルゼリー、並びにそれを含有する抗酸化剤、保湿剤、皮膚繊維芽細胞の増殖促進剤、血圧降下剤、疲労回復剤、及びカルシウム吸収促進剤 | |
| CN108840909B (zh) | 一种紫菜降压肽和紫菜降压肽提取物及应用 | |
| CN105200108A (zh) | 一种海马血管紧张素转化酶抑制肽的制备方法 | |
| CN111995659B (zh) | 一种来源于牡丹籽粕的ace抑制肽 | |
| JP2009035525A (ja) | メラニン生成阻害剤 | |
| CN114032267B (zh) | 一种活性肽的制备方法和应用 | |
| CN113087773B (zh) | 一种具有降血糖和抗氧化功能的牦牛骨肽及其制备方法 | |
| CN112028970B (zh) | 一种牡丹籽粕ace抑制肽及其制备方法与应用 | |
| CN114989248A (zh) | 一种具有抗炎抗氧化活性的忧遁草多肽及其制备方法和应用 | |
| CN103805663A (zh) | 一种从海洋产品中分离纯化提取活性肽的方法 | |
| CN102787154A (zh) | 一种乌鸡低聚肽的制备及其活性肽段分离、鉴定方法 | |
| CN108101960B (zh) | 一种具有ace抑制活性和抗肿瘤的多肽分子及其制备方法 | |
| KR20120049047A (ko) | 굴 가수분해물을 유효성분으로 함유하는 항염증 조성물 | |
| CN113735957B (zh) | 一种新型促睡眠乳源活性肽ccl-4s及其制备方法和应用 | |
| CN113730549B (zh) | 乳源活性肽ccl-1s在制备促睡眠产品中的应用 | |
| CN113444145B (zh) | 一种聚球藻血管紧张素转化酶抑制肽及其制备方法与应用 | |
| CN112961212B (zh) | 一类ace抑制肽及其制备方法和应用 | |
| CN116789747A (zh) | 一种源于紫菜蛋白的降血脂寡肽及其用途 | |
| KR20120049046A (ko) | 바지락 가수분해물을 유효성분으로 함유하는 항염증 조성물 |
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 |