CN108794604A - A kind of biologically active polypeptide SVAPAAAGIN and its preparation method and application - Google Patents
A kind of biologically active polypeptide SVAPAAAGIN and its preparation method and application Download PDFInfo
- Publication number
- CN108794604A CN108794604A CN201810717782.9A CN201810717782A CN108794604A CN 108794604 A CN108794604 A CN 108794604A CN 201810717782 A CN201810717782 A CN 201810717782A CN 108794604 A CN108794604 A CN 108794604A
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- CN
- China
- Prior art keywords
- svapaaagin
- biologically active
- active polypeptide
- ala
- polypeptide
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- QNTBGBCOEYNAPV-CWRNSKLLSA-N Trp-Asn-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CC(=O)N)NC(=O)[C@H](CC2=CNC3=CC=CC=C32)N)C(=O)O QNTBGBCOEYNAPV-CWRNSKLLSA-N 0.000 description 1
- GLNADSQYFUSGOU-GPTZEZBUSA-J Trypan blue Chemical compound [Na+].[Na+].[Na+].[Na+].C1=C(S([O-])(=O)=O)C=C2C=C(S([O-])(=O)=O)C(/N=N/C3=CC=C(C=C3C)C=3C=C(C(=CC=3)\N=N\C=3C(=CC4=CC(=CC(N)=C4C=3O)S([O-])(=O)=O)S([O-])(=O)=O)C)=C(O)C2=C1N GLNADSQYFUSGOU-GPTZEZBUSA-J 0.000 description 1
- 102000004142 Trypsin Human genes 0.000 description 1
- 108090000631 Trypsin Proteins 0.000 description 1
- QPBJXNYYQTUTDD-KKUMJFAQSA-N Tyr-Met-Gln Chemical compound CSCC[C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)O)NC(=O)[C@H](CC1=CC=C(C=C1)O)N QPBJXNYYQTUTDD-KKUMJFAQSA-N 0.000 description 1
- VKYDVKAKGDNZED-STECZYCISA-N Tyr-Val-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)[C@H](C(C)C)NC(=O)[C@H](CC1=CC=C(C=C1)O)N VKYDVKAKGDNZED-STECZYCISA-N 0.000 description 1
- 210000001015 abdomen Anatomy 0.000 description 1
- 210000000683 abdominal cavity Anatomy 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 230000010933 acylation Effects 0.000 description 1
- 238000005917 acylation reaction Methods 0.000 description 1
- 239000000654 additive Substances 0.000 description 1
- 230000000996 additive effect Effects 0.000 description 1
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- 230000003064 anti-oxidating effect Effects 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- UHOVQNZJYSORNB-UHFFFAOYSA-N benzene Substances C1=CC=CC=C1 UHOVQNZJYSORNB-UHFFFAOYSA-N 0.000 description 1
- 230000000975 bioactive effect Effects 0.000 description 1
- 238000005842 biochemical reaction Methods 0.000 description 1
- 230000008033 biological extinction Effects 0.000 description 1
- 235000010290 biphenyl Nutrition 0.000 description 1
- 239000004305 biphenyl Substances 0.000 description 1
- 230000017531 blood circulation Effects 0.000 description 1
- 210000000481 breast Anatomy 0.000 description 1
- 238000009395 breeding Methods 0.000 description 1
- 230000001488 breeding effect Effects 0.000 description 1
- 238000004364 calculation method Methods 0.000 description 1
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 1
- 229940021722 caseins Drugs 0.000 description 1
- 239000006285 cell suspension Substances 0.000 description 1
- 230000003833 cell viability Effects 0.000 description 1
- 238000005352 clarification Methods 0.000 description 1
- 210000003022 colostrum Anatomy 0.000 description 1
- 235000021277 colostrum Nutrition 0.000 description 1
- 238000000205 computational method Methods 0.000 description 1
- 238000009833 condensation Methods 0.000 description 1
- 230000005494 condensation Effects 0.000 description 1
- 238000012790 confirmation Methods 0.000 description 1
- 235000013365 dairy product Nutrition 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- 238000004043 dyeing Methods 0.000 description 1
- 238000000855 fermentation Methods 0.000 description 1
- 230000004151 fermentation Effects 0.000 description 1
- 230000035558 fertility Effects 0.000 description 1
- 238000011010 flushing procedure Methods 0.000 description 1
- 210000001035 gastrointestinal tract Anatomy 0.000 description 1
- 108010027668 glycyl-alanyl-valine Proteins 0.000 description 1
- 108010050848 glycylleucine Proteins 0.000 description 1
- 108010081551 glycylphenylalanine Proteins 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 230000002209 hydrophobic effect Effects 0.000 description 1
- 229940088592 immunologic factor Drugs 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 229910052500 inorganic mineral Inorganic materials 0.000 description 1
- 210000002490 intestinal epithelial cell Anatomy 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- 125000001449 isopropyl group Chemical group [H]C([H])([H])C([H])(*)C([H])([H])[H] 0.000 description 1
- 238000009533 lab test Methods 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 239000007791 liquid phase Substances 0.000 description 1
- 230000014759 maintenance of location Effects 0.000 description 1
- 238000004949 mass spectrometry Methods 0.000 description 1
- 238000001819 mass spectrum Methods 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 230000001035 methylating effect Effects 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 239000011707 mineral Substances 0.000 description 1
- 238000010369 molecular cloning Methods 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 239000003399 opiate peptide Substances 0.000 description 1
- 229940126701 oral medication Drugs 0.000 description 1
- 230000017448 oviposition Effects 0.000 description 1
- 230000001590 oxidative effect Effects 0.000 description 1
- 230000000242 pagocytic effect Effects 0.000 description 1
- 229940055695 pancreatin Drugs 0.000 description 1
- 229940111202 pepsin Drugs 0.000 description 1
- 230000000737 periodic effect Effects 0.000 description 1
- 108010051242 phenylalanylserine Proteins 0.000 description 1
- ZUOUZKKEUPVFJK-UHFFFAOYSA-N phenylbenzene Natural products C1=CC=CC=C1C1=CC=CC=C1 ZUOUZKKEUPVFJK-UHFFFAOYSA-N 0.000 description 1
- 230000000770 proinflammatory effect Effects 0.000 description 1
- 230000002062 proliferating effect Effects 0.000 description 1
- 108010079317 prolyl-tyrosine Proteins 0.000 description 1
- 125000006239 protecting group Chemical group 0.000 description 1
- 150000003254 radicals Chemical class 0.000 description 1
- 239000011435 rock Substances 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 210000002966 serum Anatomy 0.000 description 1
- 229910000077 silane Inorganic materials 0.000 description 1
- 210000000952 spleen Anatomy 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 238000010998 test method Methods 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 231100000041 toxicology testing Toxicity 0.000 description 1
- 230000032258 transport Effects 0.000 description 1
- 150000003852 triazoles Chemical class 0.000 description 1
- 239000012588 trypsin Substances 0.000 description 1
- 230000007306 turnover Effects 0.000 description 1
- 238000000825 ultraviolet detection Methods 0.000 description 1
- 108010030651 valyl-glutamyl-prolyl-isoleucyl-prolyl-tyrosine Proteins 0.000 description 1
- 108010009962 valyltyrosine Proteins 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
- 210000004885 white matter Anatomy 0.000 description 1
- 235000013618 yogurt Nutrition 0.000 description 1
- 235000021247 β-casein Nutrition 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/195—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria
- C07K14/335—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria from Lactobacillus (G)
-
- 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
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/64—Proteins; Peptides; Derivatives or degradation products thereof
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P37/00—Drugs for immunological or allergic disorders
- A61P37/02—Immunomodulators
- A61P37/04—Immunostimulants
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P39/00—General protective or antinoxious agents
- A61P39/06—Free radical scavengers or antioxidants
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
- A61Q19/08—Anti-ageing preparations
-
- 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
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- Animal Behavior & Ethology (AREA)
- Organic Chemistry (AREA)
- Immunology (AREA)
- Medicinal Chemistry (AREA)
- Molecular Biology (AREA)
- Pharmacology & Pharmacy (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Biochemistry (AREA)
- Engineering & Computer Science (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Polymers & Plastics (AREA)
- Genetics & Genomics (AREA)
- Food Science & Technology (AREA)
- Mycology (AREA)
- Gastroenterology & Hepatology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biophysics (AREA)
- Nutrition Science (AREA)
- Birds (AREA)
- Epidemiology (AREA)
- Toxicology (AREA)
- Gerontology & Geriatric Medicine (AREA)
- Dermatology (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Peptides Or Proteins (AREA)
Abstract
The present invention relates to albumen fields, and in particular to its amino acid sequence of a kind of biologically active polypeptide SVAPAAAGIN and its preparation method and application, biologically active polypeptide SVAPAAAGIN is Ser-Val-Ala-Pro-Ala-Ala-Ala-Gly-Ile-Asn.By the experiment of ion vitro immunization function point analysis, internal Antisenility Experiment, demonstrating polypeptide SVAPAAAGIN has preferable immunoloregulation function and activity of fighting against senium, on the one hand, the biologically active polypeptide SVAPAAAGIN of the present invention can enhance the in-vitro multiplication ability of lymphocyte and macrophage, the ability that body resists extraneous pathogenic infection is improved, body incidence is reduced；On the other hand, the vigor of internal anti-peroxidation enzyme system can be improved, enhance the function that body resists external source sexual stimulus, to reduce organism aging process, aging and sick probability, there is immunoloregulation function, the food of anti-senescence function, health products and drug to have a very important significance exploitation.
Description
Technical field
The present invention relates to albumen field, more particularly, to a kind of biologically active polypeptide SVAPAAAGIN and preparation method thereof and
Using.
Background technology
During cow's milk is through lactobacillus-fermented, a part of protein in cow's milk is metabolized by lactic acid bacteria to be utilized, concurrently
A series of biochemical reactions have been given birth to, so that protein is become polypeptide or free amino acid, is digested or passes through
The absorption and transport of intestinal epithelial cell is directly entered the blood circulation of human body.There is also the eggs of some itself synthesis for lactic acid bacteria thalline
White matter polypeptide fragment is utilized for bacterial growth.In these polypeptides, some is referred to as " raw with special physiological function
Object active peptide ".
It is particularly important that safe biologically active peptide is found in natural food source.In recent years, it has been found that some foods
The polypeptides matter in object source has good bioactivity, such as corn small peptide, soybean peptide, cow's milk polypeptide.These polypeptides can
To be obtained by number of ways such as microbial fermentation, digestion enzymolysis, and biologically active polypeptide is by 2~20 mostly
Amino acid residue forms, and molecular weight is less than 6000Da, contains a certain amount of hydrophobic amino acid, aromatic amino acid.
Immune-active peptides are to obtain and prove that one kind biology of its physiological activity is living from breast for the first time after opioid peptides discovery
Property polypeptide.Jolles in 1981 et al. has found for the first time, using trypsin hydrolysis people lactoprotein, can obtain an amino acid sequence
It is classified as the hexapeptide of Val-Glu-Pro-Ile-Pro-Tyr, experiment in vitro proves that the peptide can enhance Turnover of Mouse Peritoneal Macrophages pair
The phagocytosis of sheep red blood cell (SRBC).Migliore-Samour et al. has found the hexapeptide Thr-Thr-Met-Pro- from casein
Leu-Trp can stimulate sheep erythrocyte to the phagocytosis of mouse peritoneal macrophages and enhancing for kerekou pneumonia primary
The resistance of bacterium.Newborn source immunomodulatory peptides (PGPIPN) the feeding rat of Li Su duckweeds et al. synthesis finds rat peritoneal macrophages
Phagocytosis and the relevant immunoloregulation function of red blood cell have significant enhancing.
Studies have shown that immune-active peptides can not only enhance immunity of organisms, the proliferation of body lymphocyte, enhancing are stimulated
The phagocytic function of macrophage promotes the release of cell factor, improves the ability that body resists extraneous pathogenic infection, reduces machine
Body incidence, and the immunological rejection of body will not be caused.
Aging is a natural phenomena, and process is often accompanied by the variation of antioxidant levels, organ-tissue, immune factor,
Complicated variation occurs for middle cell factor, such as the trend that proinflammatory cytokine IL-6, IL-4, TNF-α presentation increase, IL-6
It is all considered to play an important role in the generating process of geriatric disease with TNF-a.With science of heredity and molecular biology
Development, the research of biological decay mechanism achieve gratifying progress.Researcher by using some model organisms, as mouse,
The term single gene mutating experiment of drosophila and caenorhabditis elegan etc. finds that some genes can dramatically increase service life of these organisms and reach
As many as 6 times.
Anti-aging peptide as a kind of emerging antidotal agent, in terms of physiological function have amino acid cannot compare it is excellent
Gesture can generate promotion or inhibiting effect to the enzyme in organism, improve the absorption to minerals and other nutrients and profit
With removing interior free yl enhances the resistance to oxidation of body itself, to slow down aging.Therefore, the nutrition and health care of biologically active peptide
Effect has become the emphasis of domestic and foreign scholars' subject study.Qiu Juan et al. pass through experimental studies have found that, milk-derived bioactive micro peptide
Life span of drosophila melanogaster can effectively be extended, delay its aging, and also there is preferable antioxidation, thus it is speculated that may be wherein to be rich in coloured glaze
Base peptides.SOD vigor in serum, reduces its lipid in discovery bovine colostrum extract energy conspicuousness raising the elderly's body such as the brightness in week
Peroxide and enhancing body resistance to oxidation, have certain anti-senescence function.
Have much about the research of biologically active polypeptide at present, for example Chinese patent CN105254738A discloses one kind and comes
Derived from the milk-derived biologically active polypeptide DELQDKIH of beta-casein, Chinese patent CN105254739A discloses one kind and derives from
Milk-derived the biologically active polypeptide GTQYTD, Chinese patent CN105254740A of α s1- caseins disclose a kind of from α s2-
The milk-derived biologically active polypeptide NQFYQKF of casein.
Invention content
The purpose of the present invention is to provide a kind of biologically active polypeptide SVAPAAAGIN and its preparation method and application.
The purpose of the present invention can be achieved through the following technical solutions：
First aspect present invention provides a kind of biologically active polypeptide SVAPAAAGIN, amino acid sequence Ser-Val-
Ala-Pro-Ala-Ala-Ala-Gly-Ile-Asn, such as SEQ ID NO：Shown in 1.
Preferably, the biologically active polypeptide derives from Lactobacillus helveticus mycoprotein.It is derive specifically from LBH_1221 |
m.1131 LBH_1221|g.1131 ORF LBH_1221|g.1131 LBH_1221|m.1131 type:complete len:
119(+)LBH_1221:598-954 (+) albumen, and the amino acid residue of the 104th~113, albumen thus.LBH_1221|
m.1131 LBH_1221|g.1131 ORF LBH_1221|g.1131 LBH_1221|m.1131type:complete len:
119(+)LBH_1221:598-954 (+) protein amino acid sequence such as SEQ ID NO：Shown in 3.
LBH_1221|m.1131 LBH_1221|g.1131 ORF LBH_1221|g.1131 LBH_1221|
m.1131type:complete len:119(+)LBH_1221:The amino acid sequence and corresponding core of 598-954 (+) albumen
Nucleotide sequence is existing technology, encodes the life of the nucleotide fragments energy encoding mature of this 104th~113 amino acids residue of albumen
Object active peptides SVAPAAAGIN.
Preferably, the biologically active polypeptide has immunoloregulation function and anti-senescence function.
Second aspect of the present invention provides the nucleotide fragments for encoding the biologically active polypeptide SVAPAAAGIN, sequence
It is classified as：5 '-ctg ttg ctc ctg cag ctg ctg gca taa att-3 ', such as SEQ ID NO：Shown in 2.
Third aspect present invention provides the preparation method of the biologically active polypeptide SVAPAAAGIN, can pass through base
Because the method for engineering is artificial synthesized, can be directly obtained from Lactobacillus helveticus thalline by the method that clasmatosis isolates and purifies,
It can directly be prepared by chemical synthesis.
Fourth aspect present invention provides the biologically active polypeptide SVAPAAAGIN and is preparing with immunoloregulation function
Food, health products, the application in drug or cosmetics.
Fifth aspect present invention provides the biologically active polypeptide SVAPAAAGIN and is preparing with anti-senescence function
Application in food, health products or drug.
Sixth aspect present invention provides the biologically active polypeptide SVAPAAAGIN and is preparing while having immunological regulation
Application in the food of function and anti-senescence function, health products or drug.
Specifically, the biologically active polypeptide SVAPAAAGIN of the present invention, which can be used for preparing, reduces free radical to skin wound
The harmful drug of cosmetics, preparation with immunological regulation and/or anti-aging；And due to the biologically active polypeptide of the present invention
Product after SVAPAAAGIN is degraded by gastrointestinal tract still has bioactivity, therefore can be also used for preparing the foods such as Yoghourt
Product, the health products for adjusting immunity, and the oral drug being used to prepare with immunological regulation and/or anti-aging.
Seventh aspect present invention provides a kind of immunological regulation product, including the biologically active polypeptide SVAPAAAGIN
Or the derivative of the biologically active polypeptide SVAPAAAGIN；The immunological regulation product includes immunological regulation food, is immunized
Adjust health products, immunoregulation medicament or immunological regulation cosmetics；The derivative of the biologically active polypeptide SVAPAAAGIN is
Refer on the amino acid side groups of biologically active polypeptide SVAPAAAGIN, aminoterminal or c-terminus carry out hydroxylating, carboxylated,
Be carbonylated, methylate, acetylation, phosphorylation, the modifications such as esterification or glycosylation, obtained polypeptide derivative.
Eighth aspect present invention, provides a kind of anti-aging product, including the biologically active polypeptide SVAPAAAGIN or
The derivative of the biologically active polypeptide SVAPAAAGIN；The anti-aging product includes antisenility cistanche food, anti-ageing healthcare
Product or antiaging agent；The derivative of the biologically active polypeptide SVAPAAAGIN, refers in biologically active polypeptide
On the amino acid side groups of SVAPAAAGIN, aminoterminal or c-terminus progress hydroxylating, carboxylated, it is carbonylated, methylates, second
The modifications such as acylation, phosphorylation, esterification or glycosylation, obtained polypeptide derivative.
Ninth aspect present invention provides product that is a kind of while having immunoloregulation function and anti-senescence function, including
The derivative of the biologically active polypeptide SVAPAAAGIN or described biologically active polypeptides SVAPAAAGIN；With immunological regulation work(
Can include food, health products or drug with the product of anti-senescence function；The derivative of the biologically active polypeptide SVAPAAAGIN,
Refer on the amino acid side groups of biologically active polypeptide SVAPAAAGIN, aminoterminal or c-terminus progress hydroxylating, carboxyl
Change, be carbonylated, methylating, acetylation, phosphorylation, the modifications such as esterification or glycosylation, obtained polypeptide derivative.
Biologically active polypeptide SVAPAAAGIN's of the present invention has the beneficial effect that：The biologically active polypeptide of the present invention
SVAPAAAGIN has preferable adjusting immunity of organisms activity and activity of fighting against senium；On the one hand, bioactivity of the invention is more
Peptide SVAPAAAGIN can enhance the in-vitro multiplication ability of lymphocyte and macrophage, improve body and resist extraneous cause of disease body-sensing
The ability of dye reduces body incidence；On the other hand, the vigor of internal anti-peroxidation enzyme system can be improved, enhancing body is resisted
The function of external source sexual stimulus with immunoloregulation function and resists exploitation to reduce organism aging process, aging and sick probability
The dairy products and health products of aging function have a very important significance.
Description of the drawings
Fig. 1：Mass chromatography extraction figure (m/z=870.4579)；
Fig. 2：The second order ms figure for the segment that mass-to-charge ratio is 870.4579；
Fig. 3：Polypeptide az, by crack conditions that mass-to-charge ratio is 870.4579；
Fig. 4：Influence situations of the biologically active polypeptide SVAPAAAGIN to female Drosophila survival rate；
Fig. 5：Hydrogen peroxide (H2O2) acute experiment.
Specific implementation mode
Before further describing specific embodiments of the present invention, it should be appreciated that protection scope of the present invention is not limited to down
State specific specific embodiment；It is also understood that the term used in the embodiment of the present invention is specific specific in order to describe
Embodiment, the protection domain being not intended to be limiting of the invention.
When embodiment provides numberical range, it should be appreciated that except non-present invention is otherwise noted, two ends of each numberical range
Any one numerical value can be selected between point and two endpoints.Unless otherwise defined, in the present invention all technologies for using and
Scientific terminology is identical as the normally understood meaning of those skilled in the art of the present technique.Except used in embodiment specific method, equipment,
Outside material, the record according to those skilled in the art to the grasp of the prior art and the present invention can also use and this
Any method, equipment and the material of the similar or equivalent prior art of method, equipment described in inventive embodiments, material come real
The existing present invention.
Unless otherwise stated, disclosed in this invention experimental method, detection method, preparation method be all made of this technology neck
Molecular biology, biochemistry, chromatin Structure and the analysis of domain routine, analytical chemistry, cell culture, recombinant DNA technology and
The routine techniques of related field.These technologies existing perfect explanation in the prior art, for details, reference can be made to Sambrook etc.
MOLECULAR CLONING：A LABORATORY MANUAL, Second edition, Cold Spring Harbor
Laboratory Press, 1989and Third edition, 2001；Ausubel etc., CURRENT PROTOCOLS IN
MOLECULAR BIOLOGY, John Wiley&Sons, New York, 1987and periodic updates；the
Series METHODS IN ENZYMOLOGY, Academic Press, San Diego；Wolffe, CHROMATIN
STRUCTURE AND FUNCTION, Third edition, Academic Press, San Diego, 1998；METHODS IN
ENZYMOLOGY, Vol.304, Chromatin (P.M.Wassarman and A.P.Wolffe, eds.), Academic
Press, San Diego, 1999；With METHODS IN MOLECULAR BIOLOGY, Vol.119, Chromatin
Protocols (P.B.Becker, ed.) Humana Press, Totowa, 1999 etc..
The present invention is described in detail with specific embodiment below in conjunction with the accompanying drawings.
1 active peptide SVAPAAAGIN's of embodiment is artificial synthesized
One, the synthesis of biologically active peptide
1. RINK resin 3g (degree of substitution 0.3mmol/g) are weighed in the reactor of 150ml, with the dichloromethane of 50ml
(DCM) it impregnates.
After 2.2 hours, resin is washed with nitrogen-dimethylformamide (DMF) of 3 times of resin volumes, is then drained, so weight
It is four times multiple, it is for use after resin is drained.
3. a certain amount of 20% piperidines (piperidines/DMF=1 is added into reactor:4,v:V), it is placed on decolorization swinging table and shakes
20min is shaken, the Fmoc blocking groups on resin are sloughed with this.It is washed four times with the DMF of 3 times of resin volumes after having taken off protection,
Then it drains.
4. taking the detection of a small amount of resin ninhydrin (nine well ninhydrins) method (each two drop of inspection A, inspection B, 100 DEG C of reactions
1min), resin has color, illustrates to be deprotected successfully.
5. it weighs amino acid Ser in right amount and 1- hydroxyls-benzene a pair of horses going side by side triazole (HOBT) is in right amount in the centrifuge tube of 50ml, addition
The DMF of 20ml is dissolved, and the N of 3ml is then added, and N diisopropylcarbodiimide (DIC) oscillation shakes up 1min, waits for that solution is clear
It is added in reactor after clear, then reactor is placed in 30 DEG C of shaking table and is reacted.
After 6.2 hours, with a certain amount of acetic anhydride end socket (acetic anhydride:DIEA:DCM=1:1:2,v:v:V) half an hour, so
It is washed four times, is drained for use with the DMF of 3 times of resin volumes afterwards.
7. a certain amount of 20% piperidines (piperidines/DMF=1 is added into reactor:4, v:V), it is placed on decolorization swinging table and shakes
20min is shaken, the Fmoc blocking groups on resin are sloughed with this.It is washed four times with DMF after having taken off protection, is then drained.
8. taking the detection of a small amount of resin ninhydrin (nine well ninhydrins) method (each two drop of inspection A, inspection B, 100 DEG C of reactions
1min), resin has color, illustrates to be deprotected successfully.
9. weighing second amino acid next in right amount and HOBT being in right amount in the centrifuge tube of 50ml, the DMF generals of 25ml are added
It is dissolved, and the DIC oscillations that 2.5ml is then added shake up 1min, are added in reactor after solution clarification, then by reactor
It is placed in 30 DEG C of shaking table and reacts.
After 10.1 hours, a small amount of resin is taken to detect, with ninhydrin method detection (each two drop of inspection A, inspection B, 100 DEG C of reactions
1min), if resin is colourless, illustrate that the reaction was complete；If resin has color, illustrate that condensation is incomplete, the reaction was continued.
11. after complete reaction, washing resin four times with DMF, then drains, a certain amount of 20% is added into reactor
Piperidines (piperidines/DMF=1:4,v:V), it is placed on decolorization swinging table and rocks 20min, the Fmoc protecting groups on resin are sloughed with this
Group.It is washed four times with DMF after having taken off protection, then drains whether detection protection sloughs.
12. connecting amino acid Val, Ala, Pro, Ala, Ala, Ala, Gly, Ile and Asn successively according to step 9-11.
13. after connecting the last one amino acid, protection is sloughed, is washed four times with DMF, is then taken out resin with methanol
It is dry.Then with 95 cutting liquid (trifluoroacetic acids:1,2 dithioglycols:3, isopropyl base silane:Water=95:2:2:1, v:v:V) by polypeptide
(every gram of resin adds 10ml cutting liquids) is cut down from resin, and ice ether (cutting liquid is used in combination：Ether=1:9,v:V) centrifugation is heavy
Drop four times.
So far, artificial synthesized biologically active peptide SVAPAAAGIN.
Two, the confirmation of biologically active peptide
1) UPLC is analyzed
UPLC conditions are as follows：
Instrument：Waters ACQUITY UPLC ultra high efficiencies liquid phase-electron spray-level four bars-time of-flight mass spectrometer
Chromatographic column specification：BEH C18 chromatographic columns
Flow velocity：0.4mL/min
Temperature：50℃
Ultraviolet detection wavelength：210nm
Sample size：2μL
Gradient condition：A liquid：Water containing 0.1% formic acid (v/v), B liquid：Acetonitrile containing 0.1% formic acid (v/v)
2) mass spectral analysis
Mass Spectrometry Conditions are as follows：
Ionic means：ES+
Mass range (m/z)：100-1000
Capillary voltage (Capillary) (kV)：3.0
Sampling spiroid (V)：35.0
Ion source temperature (DEG C)：115
Remove solvent temperature (DEG C)：350
Go solvent stream (L/hr)：700.0
Collision energy (eV)：4.0
Sweep time (sec)：0.25
Interior sweep time (sec)：0.02
According to the above analysis method, using ultra high efficiency liquid phase-electron spray-level four bars-flight time mass spectrum, to bioactivity
Peptide SVAPAAAGIN carries out chromatography and mass spectral analysis, and mass chromatography extraction figure is as shown in Figure 1, extract the two level matter at this peak
Spectrogram and az, by crack conditions are as shown in Figures 2 and 3, and the polypeptide mass-to-charge ratio that can obtain this peak is 870.4579Da, and retention time is
48.3min。
3) result
From the figure 3, it may be seen that the case where being broken according to az, by, analyzes by Mascot softwares and calculates, obtain mass-to-charge ratio
The fragment sequence of 870.4579Da is Ser-Val-Ala-Pro-Ala-Ala-Ala-Gly-Ile-Asn (SVAPAAAGIN), note
For SEQ ID NO：1.The segment and LBH_1221 | m.1131 LBH_1221 | g.1131 ORF LBH_1221 | g.1131
LBH_1221|m.1131 type:complete len:119(+)LBH_1221:The residue that 598-954 (+) is the 104th~113
Sequence is corresponding, and sequence is shown in SEQ ID NO：3.
The adjusting immunity of organisms activity experiment of 2 biologically active peptide of embodiment
One, mtt assay measures the vitro lymphocyte proliferation capacity experimental of biologically active polypeptide SVAPAAAGIN
1. experiment material and instrument：
Reagent and material：Experimental animal balb/c mouse (male 6-8 week old, Shanghai Communications University's agricultural and biological institute
Animal experimental center)；The biologically active polypeptide SVAPAAAGIN that embodiment 1 obtains；Mouse lymphocyte extracting solution is (purchased from rope
Precious company)；RPMI1640 culture mediums (are purchased from GIBCO companies)；3- (4,5- dimethylthiazole -2) -2,5- diphenyltetrazolium bromide bromines
Salt (MTT is purchased from Amresco companies)；ConA (ConA is purchased from Sigma companies)；(BSA is purchased from bovine serum albumin(BSA)
Genebase companies)；Pepsin (is purchased from Sigma companies)；Pancreatin (Corolase PP are purchased from AB companies).
Instrument and equipment：LRH-250F biochemical cultivation cases, Shanghai perseverance Science and Technology Ltd.；GL-22M high speed freezing centrifuges,
Shanghai Lu Xiang instrument centrifuges Instrument Ltd.；150 CO2 incubators of Hera cell, Heraeus companies；Dragon
Wellscan MK3 microplate reader, Labsystems companies；ALPHA 1-2-LD vacuum freeze driers, Christ companies；Superelevation
Effect liquid phase chromatogram-quadrupole rod time of-flight mass spectrometer, waters companies.
2. experimental method：
Mouse spleen is taken under aseptic condition, mouse lymphocyte is extracted with lymphocyte extracting solution, carries out Yuan Dynasty's culture.With
Cell density is adjusted to 2.5 × 10 by complete RPMI1640 culture solutions6A/mL.It is sequentially added in 96 porocyte culture plates：
100 μ L mouse lymphocyte suspensions, 100 μ L RPMI1640 complete culture solutions, 20 μ L ConAs, 100 μ L samples.In addition,
Blank control group (PBS of pH7.2~7.4,3mol/L) and negative control group (500 μ g/mL BSA) are set, research shows that its is right
It is not influenced in vitro lymphocyte proliferation.Every group of 3 parallel laboratory test samples.In 5%CO2After 68h being cultivated in 37 DEG C of incubators,
20 μ L MTT are added under aseptic condition per hole, continues to cultivate 4h, carefully discards supernatant liquid, 100 μ L dimethyl sulfoxide (DMSO)s are added per hole,
37 DEG C of biochemical cultivation cases hatch 10min, shake up, light absorption value is measured at 570nm with microplate reader.
Vitro lymphocyte proliferation ability indicates that computational methods are as follows with stimulus index：
In formula：A1For blank control at the 570nm under light absorption value；A2For negative control group at the 570nm under extinction
Value, A3For experimental group at the 570nm under light absorption value.
3. experimental result and analysis：
Influences of the 1 biologically active polypeptide SVAPAAAGIN of table to vitro lymphocyte proliferation
| Experiment packet | Stimulus index SI |
| Negative control group | 1 |
| SVAPAAAGIN | 1.172±0.036* |
Note：* labelled notation is the significant difference (P < 0.05) compared with negative control.
Experimental result is shown in Table 1.As shown in Table 1, it is 100 μ g/mL's in the mass concentration of biologically active peptide SVAPAAAGIN
Under the conditions of, the stimulus index of biologically active peptide SVAPAAAGIN is more than BSA, illustrates that SVAPAAAGIN can stimulate body to a certain extent
The proliferation of outer mouse lymphocyte.And the stimulus index of SVAPAAAGIN reached 1.172 and negative control group have it is notable
Difference (P<0.05).Therefore, it can be assumed that active peptides SVAPAAAGIN, which has, remarkably promotes mouse lymphocyte proliferation
Ability can be used as a kind of health products or additive edible, can improve the immunity of animal and human body.
Two, mtt assay measures the macrophages in vitro proliferative capacity experiment of biologically active polypeptide SVAPAAAGIN
1) experiment reagent and instrument
Reagent：Experimental animal balb/c mouse (male 6-8 week old) Shanghai Communications University's agricultural and biological institute animal reality
Test center；The biologically active polypeptide SVAPAAAGIN that embodiment 1 obtains；3- (4,5- dimethylthiazole -2) -2,5- diphenyl four
Nitrogen azoles bromide (MTT) Amresco companies；LPS (lipopolysaccharides) Sigma companies；Bovine serum albumin(BSA) (Bovine Serum
Albumin, BSA) Genebase companies；Three lysates, the water containing 10%SDS, 5% isobutanol and 0.012mol/L HCl
Solution.
Instrument and equipment：LRH-250F biochemical cultivation cases Shanghai perseverance Science and Technology Ltd.；On GL-22M high speed freezing centrifuges
Hai Luxiang instrument centrifuges Instrument Ltd.；Hera cell 150 CO2Incubator Heraeus companies；Dragon Wellscan
MK3 microplate reader Labsystems companies.
2) test method：
Balb/c mouse peritoneals inject 2% (w/w) the sterilizing starch solutions of 2ml, and continuous injection three days, last time is injected
The neck that breaks after 24 hours is put to death.Skin of abdomen is peelled off, 4 DEG C of phosphate buffers (PBS) is drawn with syringe and rinses abdominal cavity repeatedly, from
After heart pipe collects flushing liquor, supernatant is abandoned in centrifugation (1000rpm, 4 DEG C) after ten minutes, (is contained with 4 DEG C of RPMI1640 complete culture solutions
It 10%FBS) washes twice, cell viability examination is done in the dyeing of 0.2% trypan blue solution, confirms collected vibrant macrophage
Account for 95% or more.After cell counting board reading, adjustment cell concentration to suitable concentration.
It will blow and beat to the cell suspension to suspend completely and 96 porocyte culture plates, 37 DEG C, 5%CO be added with suitable volumes2
After being cultivated 4 hours under environment, liquid in hole is abandoned in suction, and cell culture plate well is carefully cleaned with 37 DEG C of RPMI1640 complete culture solutions
Bottom washes away not adherent cell and cell fragment, obtains adherent peritoneal macrophage after purification.0.2ml is added per hole
RPMI1640 complete mediums, experiment small peptide sample and LPS are added after being dissolved in culture medium in advance, start cell culture.
After obtaining adherent peritoneal macrophage after purification, experimental group adds per hole dissolved with biologically active polypeptide
200 holes μ l/ of RPMI1640 complete culture solutions (10%FBS) of SVAPAAAGIN (1mg/ml), continuously cultivate 48h；Negative control
Group adds 200 holes μ l/ of RPMI1640 complete culture solutions (10%FBS) dissolved with BSA (500 μ g/mL) per hole；Blank group is added
200 holes μ l/ of RPMI1640 complete culture solutions (10%FBS), continuously cultivate 48h.Also, experimental group, negative control group and blank
Group, which is divided into, does not set normal group and inflammation group；LPS to final concentration of 100ng/ml is added when culture is arrived for 24 hours for inflammation group；Normal group
It is not added with LPS；And normally 20 holes μ l/ 5%MTT are added in 44h for group and inflammation group；100 μ are added after reaching 48h in cell culture
Three lysates in the holes l/ after dissolving overnight, survey the absorbance value in each hole at wavelength 570nm to terminate culture with microplate reader
(OD570), the calculation formula of growth index (Growth Indices) is as follows：
Wherein, blank culture solution is the RPMI1640 complete culture solutions containing 10%FBS.
3) experimental result and analysis
The influence that 2 biologically active polypeptide SVAPAAAGIN of table is proliferated macrophages in vitro
| Experiment packet | Normal group GI | Inflammation group GI |
| Negative control group | 1 | 1 |
| SVAPAAAGIN(1mg/ml) | 1.0854±0.0612** | 1.1498±0.0264** |
Note：* it indicates compared with negative control, significant difference (P < 0.05)；* indicate compared with negative control group,
Significant difference (P < 0.01)
Experimental result is shown in Table 2, as shown in Table 2, under conditions of adding 1mg/ml biologically active polypeptide SVAPAAAGIN, just
Often the macrophage of group and inflammation group has proliferation.And compared with negative control group, significant difference (P < 0.01).
Illustrate that biologically active polypeptide SVAPAAAGIN has significant proliferation function to macrophages in vitro.
The activity of fighting against senium of 3 biologically active peptide of embodiment is tested
One, biologically active polypeptide SVAPAAAGIN improves the experiment of drosophila survival ability
1. experiment reagent and instrument：
Reagent：Oregon K wild type Drosophila melanogasters, agricultural college of Shanghai Communications University genetics experiments room；Agar powder, state
Chemical reagent Co., Ltd of medicine group；The biologically active polypeptide SVAPAAAGIN that embodiment 1 obtains.
Instrument and equipment：The ultra-clean water of CM-230 types mole, Shanghai Moller scientific instrument Co., Ltd；G136T type Zealway intelligence
Energy high-temperature sterilization pot, Xiamen Zhi Wei instruments Science and Technology Ltd.；BJ-CD SERIES bio-incubators, Shanghai is rich to prove to be true after interrogation industry public affairs
Department；GRX-9073 type hot air sterilizers, the permanent Science and Technology Ltd. in Shanghai one.
2. experimental method：
Using drosophila as experimental model：The drosophila adult newly to sprout wings in 8 hours is collected, divides male and female random transferring to respectively after anesthesia
In experimental group, every group of each gender 100, every group of setting 3 is parallel, and control group gives conventional corn powder culture medium, experimental group
SVAPAAAGIN biologically active peptides-corn culture medium respectively containing 0.05mg/ml, 0.5mg/ml, 1mg/ml.Every 2 days more
It changes that fresh culture is primary, observes and records the death toll of different sexes drosophila daily, until drosophila is all dead.It draws
Drosophila survival curve, and calculate the average life span of different sexes drosophila and maximum life span (takes 5 drosophilas of last death to carry out
Statistics).
3. experimental result and analysis：
This experiment is as follows to the result of study of the life span of drosophila melanogaster of feeding various concentration biologically active peptide：It can be sent out from Fig. 4
Existing, for blank control group Male Drosophila, the SVAPAAAGIN of a concentration of 0.05mg/ml of feeding is not significantly changed
The survival rate of Male Drosophila, and when peptide concentration reaches 0.5mg/ml and 1mg/ml, same time point, the survival rate of female Drosophila
It increases, but result difference unobvious.
Influence situations of the table 3-1 SVAPAAAGIN to the Male Drosophila service life
Note：* mark is compared with blank control group, significant difference (P<0.05)；Similarly hereinafter.
Influence situations of the table 3-2 SVAPAAAGIN to the female Drosophila service life
From table 3-1 it is found that relative to blank control group, low dose group Male Drosophila average life span does not have significant change,
But middle dose group and advanced amount group Male Drosophila average life span are improved, and respectively 16.51% and 11.77%, but only middle agent
Amount group produces significant difference (p<0.05), illustrate that the average life span conspicuousness of middle dose group Male Drosophila improves.In meanwhile
The half death time of dosage group and high dose group drosophila is improved, but does not have notable difference in terms of MaLS.By
Table 3-2 is it is found that female Drosophila low dose group, middle dose group and high dose group increase in terms of average life span, but not
Generate significant difference.But the MaLS of middle dose group and high dose group increases, and extends 5 respectively compared with blank control group
It and 4 days, and produce significant difference (P<0.05).
This is the experiment results show that biologically active polypeptide SVAPAAAGIN can improve the average longevity of drosophila under a certain concentration
Life and MaLS, but it is related with concentration and gender.This phenomenon related to tested material concentration, strain may be because
SVAPAAAGIN participates in the part biological metabolism of drosophila, or the antioxidant system by improving drosophila tissue extends to reach
The effect of life span of drosophila melanogaster.Since the metabolism of different lines drosophila can have any different, to cause the difference of result.And the difference of gender
It is different, it may be possible to since female Drosophila is inherently with certain conservative and to the resistance of external environment, so
SVAPAAAGIN, which extends the female Drosophila service life, to be not obvious.
Two, biologically active polypeptide SVAPAAAGIN improves the experiment of drosophila fertility
1. experiment reagent and instrument：
Reagent：Oregon K wild type Drosophila melanogasters, agricultural college of Shanghai Communications University genetics experiments room；Agar powder, state
Chemical reagent Co., Ltd of medicine group；The biologically active polypeptide SVAPAAAGIN that embodiment 1 obtains.
Instrument and equipment：The ultra-clean water of CM-230 types mole, Shanghai Moller scientific instrument Co., Ltd；G136T type Zealway intelligence
Energy high-temperature sterilization pot, Xiamen Zhi Wei instruments Science and Technology Ltd.；BJ-CD SERIES bio-incubators, Shanghai is rich to prove to be true after interrogation industry public affairs
Department；GRX-9073 type hot air sterilizers, the permanent Science and Technology Ltd. in Shanghai one.
2. experimental method：
The drosophila adult newly to sprout wings in 8 hours is collected, its male and female is separately fed, is separately added into culture medium a concentration of
The SVAPAAAGIN solution of 0mg/ml, 0.05mg/ml, 0.5mg/ml, 1mg/ml, continuous culture 12 days.It collects within 13rd day identical
The adult drosophila cultivated under concentration is simultaneously transferred in new Nostoc commune Vanch bottle, and each culture bottle ensures 1 female and 2 males
(every group 5 bottles) are given accurate 24 hours for every bottle and lay eggs.Parent drosophila is transferred to new Nostoc commune Vanch bottle after oviposition
In, old culture bottle continues breeding culture, counts progeny size, METHOD FOR CONTINUOUS DETERMINATION 7 days after larva sprouts wings, and be repeated 3 times.
3. experimental result and analysis：
4 reproductive capacity measurement result of table
From table 4, it can be seen that low concentration experimental group reproductive capacity does not generate conspicuousness variation, but middle dosage compared with the control group
Experimental group and the reproductive capacity of high dose experimental group drosophila are significantly increased (P compared with blank control group<0.05).Illustrate certain dense
The SVAPAAAGIN of degree can promote the reproductive capacity of drosophila.Originally the experimental results showed that, the extension of life span of drosophila melanogaster is SVAPAAAGIN
It is directly acting on as a result, rather than SVAPAAAGIN pass through reduce reproductive capacity caused by two level physiological effect.Also illustrate simultaneously
SVAPAAAGIN is safe to drosophila, without toxic hazard.
Three, biologically active polypeptide SVAPAAAGIN hydrogen peroxide Acute oxidative is tested
1. experiment reagent and instrument：
Reagent：Oregon K wild type Drosophila melanogasters, agricultural college of Shanghai Communications University genetics experiments room；Agar powder, state
Chemical reagent Co., Ltd of medicine group；Hydrogen peroxide, Shanghai Ling Feng chemical reagent Co., Ltd；The biology that embodiment 1 obtains is living
Property polypeptide SVAPAAAGIN.
Instrument and equipment：The ultra-clean water of CM-230 types mole, Shanghai Moller scientific instrument Co., Ltd；Mi Libo Millipore
MILLEX GP0.22 μm filter membranes, Millipore Corp. of the U.S.；GL-22M high speed freezing centrifuges, Shanghai Lu Xiang instrument centrifuge instruments
Co., Ltd.
2. experimental method：
The drosophila adult newly to sprout wings in 8 hours is collected, is divided after anesthesia in male and female random transferring to each experimental group, takes the service life real
The preferable peptide concentration culture medium of middle result is tested, blank control group is set and experimental group, control group give conventional corn powder culture medium.
Every group of male and female gender drosophila is 50, is cultivated three weeks drosophila.Then 5 males and 5 female Drosophilas is taken to be transferred to every time
In one new container, new container is included there are one papery disk, disk contain the sucrose solution of 300 μ L a concentration of 5% with
And a concentration of 30% hydrogen peroxide 1ml, blank group and experimental group are exposed to the toxicity peroxide that this hydrogen peroxide generates
In environment, 10 Duplicate Samples of every group of setting observe its oxidation resistance.Every 4 hour record drosophila The dead quantity and gender, until
Drosophila is all dead.
3. experimental result and analysis：
It is male in Each point in time after SVAPAAAGIN feedings from Fig. 5 (A) as can be seen that for Male Drosophila
The survival rate of property drosophila is above the drosophila without SVAPAAAGIN feedings, and the time-to-live increases compared with blank control group,
After illustrating feeding SVAPAAAGIN, Male Drosophila oxidation resistance increases.In Fig. 5 (B), the female of feeding SVAPAAAGIN
Drosophila, the apparent high and control group of survival rate in 15h in the hydrogen peroxide environment of high concentration, illustrates that this period, female Drosophila resisted
Oxidability increases.But later experiments group and control group survival curve essentially coincide, and illustrate that feeding SVAPAAAGIN's is female
The oxidation resistance of property drosophila gradually weakens, by not having difference with control group after a certain period of time.This experimental results showed that,
SVAPAAAGIN can improve the oxidation resistance of drosophila.According to H2O2Acute toxicity testing is as a result, can speculate SVAPAAAGIN
Drosophila may be improved to H by adjusting cat catalase activity2O2The resistivity of damage.
The above description of the embodiments is intended to facilitate ordinary skill in the art to understand and use the invention.
Person skilled in the art obviously easily can make various modifications to these embodiments, and described herein general
Principle is applied in other embodiment without having to go through creative labor.Therefore, the present invention is not limited to the above embodiments, ability
Field technique personnel announcement according to the present invention, improvement and modification made without departing from the scope of the present invention all should be the present invention's
Within protection domain.
Sequence table
<110>Zhejiang Hui Tai life and healths Science and Technology Ltd.；The Shanghai bio tech ltd Bo Hui
<120>A kind of biologically active polypeptide SVAPAAAGIN and its preparation method and application
<160> 3
<170> SIPOSequenceListing 1.0
<210> 1
<211> 10
<212> PRT
<213>Artificial sequence (Artificial Sequence)
<400> 1
Ser Val Ala Pro Ala Ala Ala Gly Ile Asn
1 5 10
<210> 2
<211> 30
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 2
ctgttgctcc tgcagctgct ggcataaatt 30
<210> 3
<211> 118
<212> PRT
<213>Artificial sequence (Artificial Sequence)
<400> 3
Met Arg Tyr Ile Gly Val Leu Ile Ser Gly Phe Met Ala Gly Leu Gly
1 5 10 15
Gly Ala Val Phe Ala Gln Ser Ile Ser Gly Asn Phe Ser Ile Ser Thr
20 25 30
Ile Val Gly Gln Gly Phe Ile Ala Leu Ala Ala Val Ile Phe Gly Lys
35 40 45
Trp Asn Pro Ile Gly Ala Met Leu Ser Ser Leu Phe Phe Gly Phe Ala
50 55 60
Gln Ser Leu Ser Ile Ile Gly Asn Gln Leu Pro Gly Phe Glu Lys Ile
65 70 75 80
Pro Thr Val Tyr Met Gln Ile Thr Pro Tyr Val Ile Thr Ile Val Val
85 90 95
Leu Val Ile Phe Leu Gly Lys Ser Val Ala Pro Ala Ala Ala Gly Ile
100 105 110
Asn Tyr Ile Lys Ser Lys
115
Claims (10)
1. a kind of biologically active polypeptide SVAPAAAGIN, which is characterized in that its amino acid sequence is Ser-Val-Ala-Pro-
Ala-Ala-Ala-Gly-Ile-Asn。
2. a kind of biologically active polypeptide SVAPAAAGIN according to claim 1, which is characterized in that the bioactivity is more
Peptide derives from Lactobacillus helveticus mycoprotein.
3. encoding the nucleotide fragments of biologically active polypeptide SVAPAAAGIN described in claim 1, which is characterized in that the nucleosides
The sequence of acid fragment such as SEQ ID NO：Shown in 2.
4. the preparation method of biologically active polypeptide SVAPAAAGIN as described in claim 1, which is characterized in that pass through genetic engineering
Method it is artificial synthesized or Lactobacillus helveticus thalline is directly obtained by the method that clasmatosis isolates and purifies, or directly pass through
It is prepared by chemical synthesis.
5. the application of biologically active polypeptide SVAPAAAGIN as described in claim 1, which is characterized in that the biologically active polypeptide
Applications of the SVAPAAAGIN in preparing the food with immunoloregulation function, health products, drug or cosmetics.
6. the application of biologically active polypeptide SVAPAAAGIN as described in claim 1, which is characterized in that the biologically active polypeptide
Applications of the SVAPAAAGIN in preparing the food with anti-senescence function, health products or drug.
7. the application of biologically active polypeptide SVAPAAAGIN as described in claim 1, which is characterized in that the biologically active polypeptide
Applications of the SVAPAAAGIN in preparing the food with immunoloregulation function and anti-senescence function, health products or drug.
8. a kind of immunological regulation product, which is characterized in that including biologically active polypeptide SVAPAAAGIN as described in claim 1 or
The derivative of the biologically active polypeptide SVAPAAAGIN；The immunological regulation product includes immunological regulation food, immune tune
Save health products, immunoregulation medicament or immunological regulation cosmetics；The derivative of the biologically active polypeptide SVAPAAAGIN refers to
On the amino acid side groups of biologically active polypeptide SVAPAAAGIN, aminoterminal or c-terminus progress hydroxylating, carboxylated, carbonyl
Base, methylate, acetylation, phosphorylation, esterification or glycosylation modified, obtained polypeptide derivative.
9. a kind of anti-aging product, which is characterized in that including biologically active polypeptide SVAPAAAGIN as described in claim 1 or institute
State the derivative of biologically active polypeptide SVAPAAAGIN；The anti-aging product includes antisenility cistanche food, antisenescence health product
Or antiaging agent；The derivative of the biologically active polypeptide SVAPAAAGIN refers in biologically active polypeptide SVAPAAAGIN
Amino acid side groups on, aminoterminal or c-terminus carry out hydroxylating, carboxylated, be carbonylated, methylate, acetylation, phosphoric acid
Change, esterification or glycosylation modified, obtained polypeptide derivative.
10. a kind of product with immunoloregulation function and anti-senescence function, which is characterized in that including as described in claim 1
The derivative of biologically active polypeptide SVAPAAAGIN or described biologically active polypeptides SVAPAAAGIN；With immunoloregulation function and
The product of anti-senescence function includes food, health products or drug；The derivative of the biologically active polypeptide SVAPAAAGIN refers to
On the amino acid side groups of biologically active polypeptide SVAPAAAGIN, aminoterminal or c-terminus progress hydroxylating, carboxylated, carbonyl
Base, methylate, acetylation, phosphorylation, esterification or glycosylation modified, obtained polypeptide derivative.
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| CN112679598A (en) * | 2021-01-19 | 2021-04-20 | 浙江辉肽生命健康科技有限公司 | Bioactive peptide SGVSLAALKKALAAAGYDVEK, and preparation method and application thereof |
| CN116444611A (en) * | 2022-11-30 | 2023-07-18 | 内蒙古伊利实业集团股份有限公司 | Milk active peptide TDPLFKG, preparation method and application thereof |
| CN116444611B (en) * | 2022-11-30 | 2024-06-04 | 内蒙古伊利实业集团股份有限公司 | Milk active peptide TDPLFKG and preparation method and application thereof |
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