CN117143191A - Corn active peptide and glycopeptide with ADH activating activity and antioxidant activity, and preparation methods and applications thereof - Google Patents
Corn active peptide and glycopeptide with ADH activating activity and antioxidant activity, and preparation methods and applications thereof Download PDFInfo
- Publication number
- CN117143191A CN117143191A CN202311162420.5A CN202311162420A CN117143191A CN 117143191 A CN117143191 A CN 117143191A CN 202311162420 A CN202311162420 A CN 202311162420A CN 117143191 A CN117143191 A CN 117143191A
- Authority
- CN
- China
- Prior art keywords
- active peptide
- corn
- glycopeptide
- adh
- activity
- 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.)
- Pending
Links
- 240000008042 Zea mays Species 0.000 title claims abstract description 106
- 235000002017 Zea mays subsp mays Nutrition 0.000 title claims abstract description 106
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 title claims abstract description 103
- 235000005822 corn Nutrition 0.000 title claims abstract description 103
- 108090000765 processed proteins & peptides Proteins 0.000 title claims abstract description 97
- 108010015899 Glycopeptides Proteins 0.000 title claims abstract description 57
- 102000002068 Glycopeptides Human genes 0.000 title claims abstract description 57
- DQJCDTNMLBYVAY-ZXXIYAEKSA-N (2S,5R,10R,13R)-16-{[(2R,3S,4R,5R)-3-{[(2S,3R,4R,5S,6R)-3-acetamido-4,5-dihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy}-5-(ethylamino)-6-hydroxy-2-(hydroxymethyl)oxan-4-yl]oxy}-5-(4-aminobutyl)-10-carbamoyl-2,13-dimethyl-4,7,12,15-tetraoxo-3,6,11,14-tetraazaheptadecan-1-oic acid Chemical compound NCCCC[C@H](C(=O)N[C@@H](C)C(O)=O)NC(=O)CC[C@H](C(N)=O)NC(=O)[C@@H](C)NC(=O)C(C)O[C@@H]1[C@@H](NCC)C(O)O[C@H](CO)[C@H]1O[C@H]1[C@H](NC(C)=O)[C@@H](O)[C@H](O)[C@@H](CO)O1 DQJCDTNMLBYVAY-ZXXIYAEKSA-N 0.000 title claims abstract description 44
- 230000003213 activating effect Effects 0.000 title claims abstract description 34
- 230000003078 antioxidant effect Effects 0.000 title claims abstract description 31
- 238000002360 preparation method Methods 0.000 title claims abstract description 10
- 102000004169 proteins and genes Human genes 0.000 claims abstract description 20
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 20
- 102000004196 processed proteins & peptides Human genes 0.000 claims abstract description 16
- 239000000843 powder Substances 0.000 claims abstract description 8
- 206010067125 Liver injury Diseases 0.000 claims abstract description 5
- 230000001476 alcoholic effect Effects 0.000 claims abstract description 5
- 231100000753 hepatic injury Toxicity 0.000 claims abstract description 5
- 239000000047 product Substances 0.000 claims description 18
- 238000000034 method Methods 0.000 claims description 12
- 238000006243 chemical reaction Methods 0.000 claims description 11
- 239000007788 liquid Substances 0.000 claims description 10
- MSWZFWKMSRAUBD-IVMDWMLBSA-N 2-amino-2-deoxy-D-glucopyranose Chemical compound N[C@H]1C(O)O[C@H](CO)[C@@H](O)[C@@H]1O MSWZFWKMSRAUBD-IVMDWMLBSA-N 0.000 claims description 9
- 108060008539 Transglutaminase Proteins 0.000 claims description 9
- MSWZFWKMSRAUBD-UHFFFAOYSA-N beta-D-galactosamine Natural products NC1C(O)OC(CO)C(O)C1O MSWZFWKMSRAUBD-UHFFFAOYSA-N 0.000 claims description 9
- 230000029087 digestion Effects 0.000 claims description 9
- 229960002442 glucosamine Drugs 0.000 claims description 9
- 102000003601 transglutaminase Human genes 0.000 claims description 9
- 108091005658 Basic proteases Proteins 0.000 claims description 8
- 238000001819 mass spectrum Methods 0.000 claims description 8
- 239000013553 cell monolayer Substances 0.000 claims description 7
- 239000006228 supernatant Substances 0.000 claims description 7
- 238000006555 catalytic reaction Methods 0.000 claims description 6
- 238000000338 in vitro Methods 0.000 claims description 5
- 238000010521 absorption reaction Methods 0.000 claims description 4
- 210000001035 gastrointestinal tract Anatomy 0.000 claims description 4
- 125000003275 alpha amino acid group Chemical group 0.000 claims description 3
- 239000012528 membrane Substances 0.000 claims description 3
- 230000007071 enzymatic hydrolysis Effects 0.000 claims 3
- 238000006047 enzymatic hydrolysis reaction Methods 0.000 claims 3
- 230000002265 prevention Effects 0.000 claims 1
- 206010061218 Inflammation Diseases 0.000 abstract description 2
- 230000004054 inflammatory process Effects 0.000 abstract description 2
- 206010036067 polydipsia Diseases 0.000 abstract description 2
- 230000011506 response to oxidative stress Effects 0.000 abstract description 2
- 102000007698 Alcohol dehydrogenase Human genes 0.000 description 38
- 108010021809 Alcohol dehydrogenase Proteins 0.000 description 38
- 235000018102 proteins Nutrition 0.000 description 16
- 238000000926 separation method Methods 0.000 description 15
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 13
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical group CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 12
- 102000004190 Enzymes Human genes 0.000 description 12
- 108090000790 Enzymes Proteins 0.000 description 12
- 229940088598 enzyme Drugs 0.000 description 12
- 239000000243 solution Substances 0.000 description 12
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 12
- 229920001184 polypeptide Polymers 0.000 description 10
- -1 DPPH free radical Chemical class 0.000 description 9
- 239000003480 eluent Substances 0.000 description 9
- 239000000523 sample Substances 0.000 description 9
- 239000000872 buffer Substances 0.000 description 8
- 230000000694 effects Effects 0.000 description 8
- 238000002835 absorbance Methods 0.000 description 7
- 230000004913 activation Effects 0.000 description 7
- OHDRQQURAXLVGJ-HLVWOLMTSA-N azane;(2e)-3-ethyl-2-[(e)-(3-ethyl-6-sulfo-1,3-benzothiazol-2-ylidene)hydrazinylidene]-1,3-benzothiazole-6-sulfonic acid Chemical compound [NH4+].[NH4+].S/1C2=CC(S([O-])(=O)=O)=CC=C2N(CC)C\1=N/N=C1/SC2=CC(S([O-])(=O)=O)=CC=C2N1CC OHDRQQURAXLVGJ-HLVWOLMTSA-N 0.000 description 7
- 238000010828 elution Methods 0.000 description 7
- 238000001514 detection method Methods 0.000 description 6
- 238000004255 ion exchange chromatography Methods 0.000 description 6
- 238000011068 loading method Methods 0.000 description 6
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 description 5
- 208000022309 Alcoholic Liver disease Diseases 0.000 description 5
- 239000007853 buffer solution Substances 0.000 description 5
- 238000004949 mass spectrometry Methods 0.000 description 5
- 239000000203 mixture Substances 0.000 description 5
- 150000003254 radicals Chemical class 0.000 description 5
- 230000002000 scavenging effect Effects 0.000 description 5
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 4
- 108010068370 Glutens Proteins 0.000 description 4
- 230000002292 Radical scavenging effect Effects 0.000 description 4
- 125000002252 acyl group Chemical group 0.000 description 4
- 230000015572 biosynthetic process Effects 0.000 description 4
- 230000009849 deactivation Effects 0.000 description 4
- 238000011033 desalting Methods 0.000 description 4
- HHEAADYXPMHMCT-UHFFFAOYSA-N dpph Chemical compound [O-][N+](=O)C1=CC([N+](=O)[O-])=CC([N+]([O-])=O)=C1[N]N(C=1C=CC=CC=1)C1=CC=CC=C1 HHEAADYXPMHMCT-UHFFFAOYSA-N 0.000 description 4
- 238000005227 gel permeation chromatography Methods 0.000 description 4
- 235000021312 gluten Nutrition 0.000 description 4
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 description 4
- 239000010931 gold Substances 0.000 description 4
- 229910052737 gold Inorganic materials 0.000 description 4
- 238000004811 liquid chromatography Methods 0.000 description 4
- 239000000463 material Substances 0.000 description 4
- 235000012054 meals Nutrition 0.000 description 4
- 238000000746 purification Methods 0.000 description 4
- QEVHRUUCFGRFIF-MDEJGZGSSA-N reserpine Chemical compound O([C@H]1[C@@H]([C@H]([C@H]2C[C@@H]3C4=C(C5=CC=C(OC)C=C5N4)CCN3C[C@H]2C1)C(=O)OC)OC)C(=O)C1=CC(OC)=C(OC)C(OC)=C1 QEVHRUUCFGRFIF-MDEJGZGSSA-N 0.000 description 4
- 238000003786 synthesis reaction Methods 0.000 description 4
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 3
- 239000012614 Q-Sepharose Substances 0.000 description 3
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 235000016383 Zea mays subsp huehuetenangensis Nutrition 0.000 description 3
- 239000004480 active ingredient Substances 0.000 description 3
- 238000003556 assay Methods 0.000 description 3
- 210000000170 cell membrane Anatomy 0.000 description 3
- 238000013375 chromatographic separation Methods 0.000 description 3
- MGJZITXUQXWAKY-UHFFFAOYSA-N diphenyl-(2,4,6-trinitrophenyl)iminoazanium Chemical compound [O-][N+](=O)C1=CC([N+](=O)[O-])=CC([N+]([O-])=O)=C1N=[N+](C=1C=CC=CC=1)C1=CC=CC=C1 MGJZITXUQXWAKY-UHFFFAOYSA-N 0.000 description 3
- 230000002496 gastric effect Effects 0.000 description 3
- 239000000413 hydrolysate Substances 0.000 description 3
- 235000009973 maize Nutrition 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 239000000758 substrate Substances 0.000 description 3
- 238000009777 vacuum freeze-drying Methods 0.000 description 3
- KPYXMALABCDPGN-HYOZMBHHSA-N (4s)-5-[[(2s)-6-amino-1-[[(2s,3s)-1-[[(2s)-1-[[(2s)-1-[[(2s)-1-[[(2s)-1-[[(2r)-1-[[2-[[2-[[(1s)-3-amino-1-carboxy-3-oxopropyl]amino]-2-oxoethyl]amino]-2-oxoethyl]amino]-1-oxo-3-sulfanylpropan-2-yl]amino]-4-methyl-1-oxopentan-2-yl]amino]-1-oxopropan-2-yl]a Chemical compound NC(=O)C[C@@H](C(O)=O)NC(=O)CNC(=O)CNC(=O)[C@H](CS)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](C)NC(=O)[C@H](C(C)C)NC(=O)[C@H](CC(C)C)NC(=O)[C@H]([C@@H](C)CC)NC(=O)[C@H](CCCCN)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@@H](NC(=O)[C@H](CC(O)=O)NC(=O)CNC(=O)[C@@H](N)CCCCN)CC1=CC=C(O)C=C1 KPYXMALABCDPGN-HYOZMBHHSA-N 0.000 description 2
- IKHGUXGNUITLKF-UHFFFAOYSA-N Acetaldehyde Chemical compound CC=O IKHGUXGNUITLKF-UHFFFAOYSA-N 0.000 description 2
- BAWFJGJZGIEFAR-NNYOXOHSSA-N NAD zwitterion Chemical compound NC(=O)C1=CC=C[N+]([C@H]2[C@@H]([C@H](O)[C@@H](COP([O-])(=O)OP(O)(=O)OC[C@@H]3[C@H]([C@@H](O)[C@@H](O3)N3C4=NC=NC(N)=C4N=C3)O)O2)O)=C1 BAWFJGJZGIEFAR-NNYOXOHSSA-N 0.000 description 2
- 108010033276 Peptide Fragments Proteins 0.000 description 2
- 102000007079 Peptide Fragments Human genes 0.000 description 2
- 101800001386 Peptide II Proteins 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 229920002472 Starch Polymers 0.000 description 2
- 229920002494 Zein Polymers 0.000 description 2
- 108010081577 aldehyde dehydrogenase (NAD(P)+) Proteins 0.000 description 2
- 239000007864 aqueous solution Substances 0.000 description 2
- 238000009835 boiling Methods 0.000 description 2
- 229910052799 carbon Inorganic materials 0.000 description 2
- 238000005119 centrifugation Methods 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 239000008367 deionised water Substances 0.000 description 2
- 229910021641 deionized water Inorganic materials 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 239000012153 distilled water Substances 0.000 description 2
- 230000002255 enzymatic effect Effects 0.000 description 2
- 235000013305 food Nutrition 0.000 description 2
- 230000007760 free radical scavenging Effects 0.000 description 2
- 238000004108 freeze drying Methods 0.000 description 2
- ZDXPYRJPNDTMRX-UHFFFAOYSA-N glutamine Natural products OC(=O)C(N)CCC(N)=O ZDXPYRJPNDTMRX-UHFFFAOYSA-N 0.000 description 2
- 229910002804 graphite Inorganic materials 0.000 description 2
- 239000010439 graphite Substances 0.000 description 2
- 230000007062 hydrolysis Effects 0.000 description 2
- 238000006460 hydrolysis reaction Methods 0.000 description 2
- 210000004185 liver Anatomy 0.000 description 2
- 230000004060 metabolic process Effects 0.000 description 2
- 238000002156 mixing Methods 0.000 description 2
- 239000013642 negative control Substances 0.000 description 2
- 230000037361 pathway Effects 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 230000035484 reaction time Effects 0.000 description 2
- 235000019698 starch Nutrition 0.000 description 2
- 239000008107 starch Substances 0.000 description 2
- 239000000725 suspension Substances 0.000 description 2
- 239000005019 zein Substances 0.000 description 2
- 229940093612 zein Drugs 0.000 description 2
- 102000057297 Pepsin A Human genes 0.000 description 1
- 108090000284 Pepsin A Proteins 0.000 description 1
- 102000004142 Trypsin Human genes 0.000 description 1
- 108090000631 Trypsin Proteins 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 230000006978 adaptation Effects 0.000 description 1
- 208000029650 alcohol withdrawal Diseases 0.000 description 1
- 125000000539 amino acid group Chemical group 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 150000001450 anions Chemical class 0.000 description 1
- 230000003064 anti-oxidating effect Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000000975 bioactive effect Effects 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000006227 byproduct Substances 0.000 description 1
- 238000004364 calculation method Methods 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 238000011067 equilibration Methods 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 238000007710 freezing Methods 0.000 description 1
- 230000008014 freezing Effects 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 230000000968 intestinal effect Effects 0.000 description 1
- 238000001294 liquid chromatography-tandem mass spectrometry Methods 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 238000000691 measurement method Methods 0.000 description 1
- 230000001404 mediated effect Effects 0.000 description 1
- 230000037353 metabolic pathway Effects 0.000 description 1
- 239000012982 microporous membrane Substances 0.000 description 1
- 239000011259 mixed solution Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 239000012452 mother liquor Substances 0.000 description 1
- 239000010413 mother solution Substances 0.000 description 1
- 229950006238 nadide Drugs 0.000 description 1
- 229940101270 nicotinamide adenine dinucleotide (nad) Drugs 0.000 description 1
- 229930027945 nicotinamide-adenine dinucleotide Natural products 0.000 description 1
- 229940111202 pepsin Drugs 0.000 description 1
- USHAGKDGDHPEEY-UHFFFAOYSA-L potassium persulfate Chemical compound [K+].[K+].[O-]S(=O)(=O)OOS([O-])(=O)=O USHAGKDGDHPEEY-UHFFFAOYSA-L 0.000 description 1
- 239000002244 precipitate Substances 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 230000000750 progressive effect Effects 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 239000012488 sample solution Substances 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 238000012163 sequencing technique Methods 0.000 description 1
- 239000002356 single layer Substances 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 239000003270 steroid hormone Substances 0.000 description 1
- 210000002784 stomach Anatomy 0.000 description 1
- 238000002305 strong-anion-exchange chromatography Methods 0.000 description 1
- 229940126585 therapeutic drug Drugs 0.000 description 1
- 239000012588 trypsin Substances 0.000 description 1
- 238000000108 ultra-filtration Methods 0.000 description 1
- 238000012795 verification Methods 0.000 description 1
- 239000002699 waste material Substances 0.000 description 1
- 239000012224 working solution Substances 0.000 description 1
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
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/16—Drugs for disorders of the alimentary tract or the digestive system for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
-
- 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
-
- 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
- C12P21/005—Glycopeptides, glycoproteins
-
- 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
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
Abstract
The invention provides corn active peptide and glycopeptide with ADH activating activity and antioxidant activity, and a preparation method and application thereof, and belongs to the technical field of active peptides. The corn active peptide SSNCQPF, TGCPVLQ and QPQQPW and the glycopeptides thereof with double functions of ADH activating activity and antioxidant activity are separated from corn protein powder for the first time. The corn active peptide and the glycopeptide can be used for preparing a product with ADH activating and/or antioxidant effects, and also can be used for preparing a product for preventing alcoholic liver injury, and relieving oxidative stress and inflammatory reaction caused by excessive drinking of an organism.
Description
Technical Field
The invention belongs to the technical field of active peptides, and particularly relates to a corn active peptide and glycopeptide with ADH activating activity and antioxidant activity, and a preparation method and application thereof.
Background
There are a number of alcohol metabolic pathways in the human liver, of which catalysis by Alcohol Dehydrogenase (ADH) is one of the most critical pathways. When ADH is activated and reacts with Nicotinamide Adenine Dinucleotide (NAD) + ) In response, ADH and acetaldehyde dehydrogenase (ALDH) metabolize about 90% of the ethanol in this pathway to acetaldehyde and acetic acid, most of which is transferred to blood and extrahepatic tissue. Short term overdose reduces the activity of ADH in the liver, resulting in excessive alcohol accumulation and Alcoholic Liver Disease (ALD). In addition to alcohol withdrawal and sebaceous steroid hormone as the primary intervention, there is currently no effective ALD therapeutic drug. Therefore, the search for safe and effective natural ingredients and the development of substances capable of promoting alcohol metabolism and activating ADH activity are significant in preventing alcoholic liver diseases.
Corn gluten meal is a wet processed starch byproduct of corn with high yield and high protein content. However, the corn gluten meal is poor in solubility and strong in hydrophobicity, is not easy to be effectively utilized in the food industry, so that the corn gluten meal is mostly used as feed, and resource waste is caused. Therefore, if the zein can be modified, the development of substances with ADH activating activity to promote alcohol metabolism so as to prevent alcoholic liver diseases, improve the added value of the substances and have important significance for the deep processing of the zein.
Disclosure of Invention
Accordingly, the present invention is directed to a corn active peptide and glycopeptide having ADH activating activity and antioxidant activity.
In order to achieve the above object, the present invention provides the following technical solutions:
the invention provides a corn active peptide with ADH activating activity and antioxidant activity, which comprises at least one corn active peptide with an amino acid sequence shown as SEQ ID NO. 1-3.
The invention also provides a glycopeptide with ADH activating activity and antioxidant activity, which is obtained by the catalytic reaction of the corn active peptide and glucosamine through transglutaminase.
Preferably, the glycopeptide comprises at least one of TGCPVLgQ, SSNCgQPF, gQPQQPW, QPgQQPW, QPQgQPW, gQPgQQPW, gQPQgQPW, QPgQgQPW and gQPgQgQPW.
Preferably, the time of the catalytic reaction is 90min, and the pH value of the reaction is 7.
Preferably, the amount of the transglutaminase is 0.6U/. Mu.mol of the corn active peptide based on the amount of the corn active peptide.
The invention also provides an application of the corn active peptide or the glycopeptide in preparing a functional product with ADH activating activity and/or antioxidant activity.
The invention also provides application of the corn active peptide or the glycopeptide in preparing a product for preventing and treating alcoholic liver injury.
The invention also provides a preparation method of the corn active peptide, which comprises the following steps: adopting alkaline protease to carry out enzymolysis on corn protein powder, and taking supernatant as corn protein enzymolysis liquid; separating and purifying the corn protein enzymolysis liquid to obtain corn active peptide smaller than 1 kDa; then simulating in vitro gastrointestinal tract digestion and Caco-2 cell monolayer membrane absorption, and obtaining the corn active peptide with ADH activating activity and antioxidant activity through mass spectrum identification.
Preferably, the pH value of the enzymolysis is 8.0, the temperature of the enzymolysis is 55 ℃, and the time of the enzymolysis is 180min.
The invention has the beneficial effects that:
the corn active peptide and the glycopeptide thereof with double functions of ADH activating activity and antioxidant activity are separated from corn protein powder. The corn active peptide and the glycopeptide thereof can be used for preparing products with ADH activating and/or antioxidant effects, and also can be used for preparing products for preventing alcoholic liver injury, and relieving oxidative stress and inflammatory reaction caused by excessive drinking of organisms.
Drawings
FIG. 1 is a technical scheme for preparing corn active peptide and glycopeptide of the present invention;
FIG. 2 is a mass spectrum of corn active peptide I (SSNCQPF) of the present invention;
FIG. 3 is a mass spectrum of corn active peptide II (TGCPVLQ) of the present invention;
FIG. 4 is a mass spectrum of corn active peptide III (QPQQPW) of the present invention;
FIG. 5 is a mass spectrum of the glycopeptide SSNCgQPF of corn active peptide I of the present invention;
FIG. 6 is a TGCPVLgQ mass spectrum analysis of glycopeptides of corn active peptide II of the present invention;
FIGS. 7-13 are mass spectrometric detection patterns of glycopeptides gQPQQPW, QPgQQPW, QPQgQPW, gQPgQQPW, gQPQgQPW, QPgQgQPW and gQPgQgQPW, respectively, of corn active peptide III of the present invention.
Detailed Description
The invention provides a corn active peptide with ADH activating activity and antioxidant activity, which comprises at least one of SSNCQPF (SEQ ID NO. 1), TGCPVLQ (SEQ ID NO. 2) and QPQQPW (SEQ ID NO. 3).
The invention also provides a preparation method of the corn active peptide, which comprises the following steps: adopting alkaline protease to carry out enzymolysis on corn protein powder, and taking supernatant as corn protein enzymolysis liquid; separating and purifying the corn protein enzymolysis liquid to obtain corn active peptide smaller than 1 kDa; then simulating in vitro gastrointestinal tract digestion and Caco-2 cell monolayer membrane absorption, and obtaining the corn active peptide with ADH activating activity and antioxidant activity through mass spectrum identification.
The specific sources of the alkaline protease and the corn gluten meal are not particularly limited, and the conventional commercial products in the field can be adopted. In the invention, before enzymolysis, the corn protein powder is preferably prepared into a suspension with the mass concentration of 5% (w/v), and then is mixed with alkaline protease for enzymolysis, wherein the usage amount of the alkaline protease is preferably 400U/g protein based on the weight of protein in the corn protein powder, the pH of the enzymolysis is preferably 8.0, the temperature of the enzymolysis is preferably 55 ℃, and the time of the enzymolysis is preferably 180min. After the enzymolysis is finished, the method preferably further comprises the step of carrying out enzyme deactivation treatment on the enzymolysis product to obtain the enzyme-deactivated enzymolysis product. The temperature of the enzyme deactivation treatment is preferably 100 ℃; the time for the enzyme deactivation treatment is preferably 10 minutes. After the enzyme deactivation treatment is completed, the invention preferably further comprises centrifuging the enzyme-deactivated enzymolysis product, and the obtained supernatant is the enzymolysis liquid comprising the corn active peptide. The rotational speed of the centrifugation is preferably 4000r/min and the time is preferably 10min in the present invention.
In the invention, after the enzymatic hydrolysate containing the corn active peptide is obtained, the enzymatic hydrolysate is preferably separated and purified, and the separation and purification method preferably comprises gel chromatography separation, nano ultrafiltration concentration centrifuge tube progressive separation and ion exchange chromatography separation. In the present invention, the gel chromatography pre-packed column for gel chromatography separation is preferably Superdexeptide 10/300GL, and the conditions for gel chromatography separation preferably comprise: the loading concentration is 50mg/L, and the loading amount is 1mL; the eluent was 20mM PBS buffer containing 0.15mol/LNaCl at pH 7.0; the flow rate of the eluent is 0.25mL/min; the detection wavelength is 214nm, and the active peptide components with the molecular weight of less than 3kDa are collected. Ultrafiltering the active peptide component with molecular weight cut off of 1kDa in a Nanosep ultrafiltering concentration centrifuge tube to obtain corn active peptide with molecular weight less than 1kDa, and freeze drying. After obtaining the fraction of corn active peptide of less than 1kDa, it is preferably subjected to ion exchange chromatography. In the present invention, the ion exchange chromatography separation chromatography is preferably Q-Sepharose High Performance ion exchange chromatography, and the conditions for separation using the Q-Sepharose High Performance ion exchange chromatography preferably include: the loading amount is 50mL; eluent A is preferably Tris-HCl buffer, the concentration of the Tris-HCl buffer is preferably 20mM, and the pH value is preferably 7.5; eluent B was 1mol/LNaCl in 20mM Tris-HCl buffer pH 7.5; the flow rate of the eluent was 2mL/min, the detection wavelength was 214nm, the ladder-wash volume was 60mL, and the peak fraction collection volume was 6 mL/tube. The invention preferably further comprises measuring the ADH activating activity and the antioxidant activity of each component obtained by collection to obtain a component with relatively stronger ADH activating activity and antioxidant activity, namely a component I. After the first component is obtained, the invention preferably simulates in-vitro gastrointestinal digestion of the first component to obtain the peptide for resisting gastrointestinal digestion, namely the second component. After the second component is obtained, the second component is absorbed by the Caco-2 cell monolayer film, so that the corn peptide which is completely absorbed by the Caco-2 cell monolayer film can be obtained, namely the third component. After the third component is obtained, the corn peptide which is completely absorbed by the Caco-2 cell monolayer film can be obtained by comparing the first component with the second component through the identification of a liquid chromatograph/mass spectrometer. The process and steps of mass spectrometry are not particularly limited in the present invention, and conventional mass spectrometry sequencing steps in the art may be employed. Prior to mass spectrometry, the sample is preferably subjected to zip-tip desalting and vacuum centrifugation for concentration. The steps of desalting and centrifugal concentration are not particularly limited, and conventional desalting and centrifugal concentration processes and steps in the field are adopted.
The invention also provides a glycopeptide with ADH activating activity and antioxidant activity, which is obtained by the catalytic reaction of the corn active peptide and glucosamine through transglutaminase.
In the invention, the corn active peptide is used as an acyl donor, and the glucosamine is used as an acyl acceptor for reaction. The concentration of the corn active peptide is preferably 10 mu mol/mL, the concentration of the glucosamine is controlled according to the number (n) of glutamine in the corn active peptide, preferably 15n mu mol/mL, the enzyme adding amount of the transglutaminase is preferably 0.6U/mu mol of the corn active peptide based on the mass of the corn active peptide, the reaction time is preferably 90min, and the pH of the reaction is preferably 7, so that the glycopeptide is obtained. After obtaining the glycopeptides prepared as described above, the following separation and purification steps are preferably further performed: through liquid chromatography multi-stage separation, the model of the chromatographic column is Thermo Scientific TM Hypersil GOLD chromatographic column (1.9 μm. Times.2.1 mm. Times.100 mm), mobile phase preferably comprises mobile phase A and mobile phase B, mobile phase A is water, mobile phase B is acetonitrile, preferably at a flow rate of 0.3mL/min, and column temperature preferably at 30deg.C. Using Thermo Scientific TM The elution procedure for a Hypersil GOLD column (1.9 μm x 2.1mm x 100 mm) is preferably a gradient elution, the gradient elution conditions are preferably 30min, and the mobile phase conditions for the gradient elution are preferably 0-2 min:2% of B, 2-20 min:2% -20% of B, 20-25 min:20% B,25-30min:20% -2% of the solution returns to the initial equilibrium condition, and the substituted glycopeptides at different positions can not be separated and also need to pass through Thermo Scientific TM The Hypercarb porous graphite carbon chromatographic column (3 mu m multiplied by 2.1mm multiplied by 150 mm) is separated, the mobile phase preferably comprises a mobile phase A and a mobile phase B, the mobile phase A is water, the mobile phase B is acetonitrile, the preferred flow rate is 0.3mL/min, the column temperature is preferably 30 ℃, the preferred condition of the mobile phase is 45% B, and finally the glycopeptide pure product is obtained through liquid chromatography separation.
In the present invention, the glycopeptide preferably comprises at least one of TGCPVLgQ, SSNCgQPF, gQPQQPW, QPgQQPW, QPQgQPW, gQPgQQPW, gQPQgQPW, QPgQgQPW and gQPgQgQPW, wherein "g" in the glycopeptide represents glucosamine (GlcN) and the compound has epsilon-amino group, and the "g" can be positioned and connected to the amino acid residue Q of the corn active peptide under the action of transglutaminase.
The invention also provides an application of the corn active peptide or the glycopeptide in preparing a functional product with ADH activating activity and/or antioxidant activity. The invention also provides application of the corn active peptide or the glycopeptide in preparing a product for preventing and treating alcoholic liver injury.
In the present invention, the product preferably includes a pharmaceutical and/or food product. In the invention, the product preferably further comprises auxiliary materials and/or other active ingredients, the types of the auxiliary materials and/or other active ingredients are not particularly limited, and the conventional auxiliary materials or other active ingredients of the product to be prepared can be adopted.
The technical solutions provided by the present invention are described in detail below with reference to examples, but they should not be construed as limiting the scope of the present invention.
In the following examples, conventional methods are used unless otherwise specified.
Materials, reagents and the like used in the examples described below are commercially available unless otherwise specified.
Example 1
The preparation of the corn active peptide and the glycopeptide thereof comprises the following steps (technical scheme is shown in figure 1):
1. preparation of corn protein enzymolysis liquid
Taking a certain amount of corn protein powder (purchased from zizihatherum vernix biological technology Co., ltd.) which is extruded, puffed and removed with starch, adding water to prepare a suspension with a substrate concentration of 5% (w/v), and then carrying out enzymolysis by using alkaline protease under the following enzymolysis conditions: the pH is 8.0, the temperature is 55 ℃, the dosage of alkaline protease is 400U/g protein, and the hydrolysis time is 180min. Heating boiling water to deactivate enzyme for 10min after enzymolysis is finished, centrifuging the enzymolysis product at 4000r/min for 10min, discarding the precipitate, obtaining supernatant which is corn protein enzymolysis liquid, and determining ADH activating activity and antioxidant activity to confirm that the supernatant is a polypeptide mixture with high ADH activating activity and antioxidant activity. The method for measuring ADH activating activity and antioxidant activity comprises the following steps:
ADH activation Activity assay: mu.L of the sample solution was mixed with 150. Mu.L of a detection reagent (containing buffer, NAD + And ethanol) and after equilibration at 37℃for 5min, 50. Mu.LADH (0.2U/mL) was added to initiate the reaction. Absorbance values were measured using a microplate reader at 340nm and recorded every 10 seconds for 10 minutes. Distilled water was used as a negative control instead of the sample. Fitting a reaction dynamics curve, and obtaining a first derivative of the curve at 0min, namely the initial reaction rate. The initial reaction rate of the sample was recorded as V s While the record of negative control is V 0 . The ADH activation rate of the sample can be calculated according to the following equation:
measurement of antioxidant Activity: (1) determination of the ability to scavenge DPPH free radicals: 100. Mu.L of polypeptide aqueous solution with different concentrations is sucked into a 96-well plate, 100. Mu.L of 0.1mmol/LDPPH absolute ethanol solution is added, the mixture is uniformly mixed, the reaction is carried out for 30min in the absence of light, and the absorbance at 517nm (A i ) The method comprises the steps of carrying out a first treatment on the surface of the 100. Mu.L of the polypeptide solution was added to a 96-well plate, 100. Mu.L of absolute ethanol was added thereto, and the absorbance at 517nm was determined to be (A j ) The method comprises the steps of carrying out a first treatment on the surface of the 100. Mu.L of a 0.1mmol/LDPPH absolute ethanol solution was reacted with 100. Mu.L of water as a reference to determine the absorbance at 517nm (A 0 ). The calculation formula of the DPPH free radical clearance K of the sample is as follows:
(2) Determination of ABTS radical scavenging ability: accurately preparing 7.0 mmol/LABSS solution and 2.45mmol/L potassium persulfate aqueous solution, uniformly mixing, and standing for 12 hours at room temperature in dark condition to obtain the ABTS free radical mother solution. Diluting ABTS free radical mother liquor with deionized water to make its absorbance at 734nm be 0.70+ -0.02, and balancing at 30deg.C for 30min to obtain ABTS +· And (5) working fluid. Respectively taking 100 mu L of polypeptide solution with different concentrations and 100 mu L of ABTS ﹢· Adding the working solution into 96-well plate, mixing, standing at room temperature in dark place for 20min, and measuring absorbance at 734nm to obtain absorbance A Sample of . The deionized water is used for replacing the reaction mixed solution with the same volume of the polypeptide solution as a comparison, 3 repeats are set, and the average value is A Blank space . The clearance of ABTS free radicals by the polypeptide solution was calculated according to the following formula.
2. Stepwise separation of maize peptides of different molecular weights
And (2) performing gel chromatographic separation on the polypeptide mixture with high ADH activating activity and antioxidant activity in the step (1), wherein a gel chromatographic pre-packed column for the gel chromatographic separation is Superdexeptide 10/300GL. The conditions for performing the gel chromatographic separation are: the loading concentration is 50mg/L, and the loading amount is 1mL; the eluent is 20mM PBS buffer solution containing 0.15mol/L NaCl with pH 7.0; the flow rate of the eluent is 0.25mL/min; the detection wavelength is 214nm, and the active peptide components with the molecular weight of less than 3kDa are collected; ultrafiltering the active peptide component with molecular weight cut off of 1kDa in a Nanosep ultrafiltering concentration centrifuge tube to obtain corn active peptide with molecular weight less than 1kDa, and freeze drying.
3. Ion exchange chromatography separation
The lyophilized fraction of step 2, i.e., the fraction having a molecular weight of less than 1kDa and relatively high ADH activating and antioxidant activities, was formulated into a solution having a protein concentration of 5mg/mL and passed through a microporous membrane of 0.22 μm, and separated using a strong anion exchanger of Q-Sepharose High Performance. Eluent a of the strong anion exchange chromatography: 20mM Tris-HCl buffer, pH 7.5, eluent B: pH 7.5 of 20mM Tris-HCl buffer solution containing 1mol/LNaCl, loading amount of 50mL, flow rate of 2mL/min, detection wavelength of 214nm, ladder-wash volume of 60mL, and peak component collection of 6mL per tube; the ADH activating activity and the antioxidant activity of each tube collection were measured (the specific measurement method is the same as step 1), and the components having relatively high ADH activating activity and antioxidant activity were collected.
4. Simulating in vitro gastrointestinal digestion
The corn active peptide in step 3 was dissolved in distilled water (3%, w/v), adjusted to pH 2.0 with 1mol/L HCl, pepsin (enzyme activity 3000 units/mg) was added, and the enzyme: the substrate ratio was 1:50 (w/w), and the digestion in the stomach was simulated by hydrolysis for 90min in a thermostated shaker at 37 ℃. Then, pH 7.0 was adjusted with 1mol/L NaOH, trypsin (enzyme activity 300 units/mg) was added, and the enzyme: the substrate ratio is 1:50 (w/w), and the mixture is subjected to enzymolysis for 4 hours at 37 ℃ again to simulate intestinal digestion. After digestion, the sample is placed in a boiling water bath for 10min to inactivate enzyme, and cooled to room temperature. Centrifuging the hydrolysate at 4000r/min for 10min, collecting supernatant, vacuum freeze drying, and storing at-20deg.C. The samples were reconstituted to 1mg/mL and assayed for ADH activating activity (specific assay procedure is as in step 1).
5. Absorption through Caco-2 cell membranes
The corn peptide digested by the simulated gastrointestinal tract in the step 4 is absorbed by a Caco-2 cell monolayer film. The specific operation is as follows: first, caco-2 cell membranes were washed 2 times with Hank's buffer, and 0.5mL and 1.5mL of fresh Hank's buffer were added to the AP side and BL side, respectively, at 37℃with 5% CO 2 Culturing for 30min under the condition. Then, the Hank's buffer solution on the AP side and BL side was removed, 0.5mL of a corn peptide solution (5 mg/mL concentration dissolved in Hank's buffer solution) was added to the AP side, 1.5mL of fresh Hank's buffer solution was added to the BL side, and the mixture was heated to 37℃and 5% CO 2 After 2h incubation, AP-and BL-side samples were collected, and stored at-20℃after vacuum freeze-drying. The BL-side sample is corn active peptide that can be absorbed by Caco-2 monolayer film, i.e., in the BL-side sample.
6. LC-MS/MS mass spectrometry
And (3) desalting the components obtained in the steps (3), (4) and (5) by zip-tip, freezing and spin drying, dissolving by using an initial mobile phase, performing De Novo mass spectrometry, identifying the three components to obtain a common peptide fragment which is the corn active peptide capable of being absorbed by a Caco-2 cell membrane, performing computer-aided analysis and screening, taking the amino acid sequences of the common peptide fragment as SSNCQPF (polypeptide I, SEQ ID NO. 1), TGCPVLQ (polypeptide II, SEQ ID NO. 2) and QPQQPW (polypeptide III, SEQ ID NO. 3) as potential bioactive peptides, and performing subsequent synthesis verification activity, wherein the structure identification result is shown in the figures (2-4).
7. Synthesis of corn peptide and glycopeptide
After the corn active peptide (SEQ ID NO. 1-3) identified in the step 6 is chemically synthesized (entrusted to Shanghai blaze Biotechnology Co., ltd.), the subsequent synthesis, separation and purification of the corn glycopeptide are carried out.
7.1 transglutaminase (TGase) mediated Synthesis of maize glycopeptides
The chemically synthesized corn active peptide (SEQ ID NO. 1-3) is used as an acyl donor, the glucosamine is used as an acyl acceptor for reaction, wherein the concentration of the corn active peptide is 10 mu mol/mL, the concentration of the glucosamine is 15n mu mol/mL according to the number (n) of the glutamine in the active peptide, the reaction time is 90min, the pH is 7, the enzyme adding amount of transglutaminase is 0.6U/mu mol of the corn active peptide, and the corn active peptide is stored at-20 ℃ after vacuum freeze drying.
7.2 separation and purification of pure corn glycopeptide
The glycopeptide prepared in step 7.1 is separated by liquid chromatography, and the chromatographic column model is Thermo Scientific TM Hypersil GOLD chromatographic column (1.9 μm×2.1mm×100 mm), mobile phase comprising mobile phase A and mobile phase B, mobile phase A being water, mobile phase B being acetonitrile, flow rate being 0.3mL/min, column temperature being 30deg.C. Using Thermo Scientific TM The elution program is gradient elution when the Hypersil GOLD chromatographic column (1.9 μm multiplied by 2.1mm multiplied by 100 mm), the gradient elution condition is 30min, and the mobile phase condition of gradient elution is 0-2 min:2% of B, 2-20 min:2% -20% of B, 20-25 min:20% B,25-30min:20% -2% returns to the initial flatThe constant conditions, for the different positions of the substituted glycopeptides which can not be separated by the chromatographic column, also need to pass through Thermo Scientific TM The Hypercarb porous graphite carbon chromatographic column (3 μm. Times.2.1 mm. Times.150 mm) was separated, and the mobile phase included mobile phase A and mobile phase B, mobile phase A was water, mobile phase B was acetonitrile, the flow rate was 0.3mL/min, the column temperature was 30 ℃, and the mobile phase condition was 45% B. The glycopeptides prepared by the method are separated by liquid chromatography to obtain glycopeptides pure products, and the results are shown in figures 5-13.
Example 2
The corn active peptides SSNCQPF, TGCPVLQ and QPQQPW identified in step 6 and the purified corn glycopeptides TGCPVLgQ, SSNCgQPF, gQPQQPW, QPgQQPW, QPQgQPW, gQPgQQPW, gQPQgQPW, QPgQgQPW and gQPgQgQPW in step 7 were assayed for ADH activating activity and scavenging ability to DPPH and ABTS free radicals, respectively, and the specific assay was the same as that of step 1 of example 1. The results were as follows:
(1) When the corn active peptide SSNCQPF and the glycopeptide SSNCgQPF thereof are 1 mu mol/mL, the ADH activation rates are 25.68% and 34.39%, respectively.
When the concentration of the corn active peptide TGCPVLQ and the glycopeptide TGCPVLgQ are 1 mu mol/mL, the ADH activation rate is 21.32% and 27.96% respectively.
The activation rate of QPQQPW was 18.57% for the corn active peptide QPQQPW and the corresponding glycopeptide at 1. Mu. Mol/mL, and the ADH activation rate for the corresponding glycopeptide is shown in Table 1.
TABLE 1 ADH Activity of maize active peptide QPQQPW corresponding to glycopeptides at different sites
(2) IC of corn active peptide SSNCQPF on DPPH free radical scavenging ability 50 IC with a value of 0.265. Mu. Mol/mL, its glycopeptide (SSNCgQPF) to DPPH radical scavenging rate 50 The value was 0.191. Mu. Mol/mL.
Corn active peptideTGCPVLQ and IC of its glycopeptide (TGCPVLgQ) for scavenging DPPH free radical 50 The values were 0.336. Mu. Mol/mL and 0.272. Mu. Mol/mL, respectively.
IC of corn active peptide QPQQPW on DPPH free radical scavenging ability 50 IC with a value of 0.853. Mu. Mol/mL and its corresponding glycopeptide having DPPH radical scavenging ability 50 The values are shown in Table 2.
TABLE 2 IC of the DPPH radical scavenging ability of the various glycopeptides at the various sites of the active peptide QPQQPW 50 Value of
(3) IC of corn active peptide SSNCQPF on scavenging ability of ABTS free radical 50 IC with a value of 0.113. Mu. Mol/mL, its glycopeptide (SSNCgQPF) ability to scavenge ABTS free radicals 50 The value was 0.087. Mu. Mol/mL.
IC of corn active peptide TGCPVLQ and its glycopeptide (TGCPVLgQ) for scavenging ABTS free radical 50 The values were 0.188. Mu. Mol/mL and 0.127. Mu. Mol/mL, respectively.
IC of the ability of corn active peptide QPQQPW to scavenge ABTS free radical 50 IC with value of 0.262 mu mol/mL and corresponding glycopeptide with ability of scavenging ABTS free radical 50 The values are shown in Table 3.
TABLE 3 IC of the ability of the glycopeptides at different sites of the active peptide QPQQPW to scavenge ABTS free radicals 50 Value of
The results show that the corn active peptide and the glycopeptide thereof provided by the invention have double activities of ADH activation and antioxidation.
The foregoing is merely a preferred embodiment of the present invention and it should be noted that modifications and adaptations to those skilled in the art may be made without departing from the principles of the present invention, which are intended to be comprehended within the scope of the present invention.
Claims (9)
1. The corn active peptide with ADH activating activity and antioxidant activity is characterized by comprising at least one of corn active peptides with amino acid sequences shown in SEQ ID NO. 1-3.
2. A glycopeptide having ADH activating activity and antioxidant activity, which is obtained by a transglutaminase-catalyzed reaction of the corn active peptide of claim 1 with glucosamine.
3. The glycopeptide of claim 2, wherein the glycopeptide comprises at least one of TGCPVLgQ, SSNCgQPF, gQPQQPW, QPgQQPW, QPQgQPW, gQPgQQPW, gQPQgQPW, QPgQgQPW and gqpgqgpw.
4. The glycopeptide of claim 2, wherein the catalytic reaction takes 90 minutes and the pH of the reaction is 7.
5. The glycopeptide of claim 2, wherein the amount of transglutaminase is 0.6U/. Mu.mol of corn active peptide based on the amount of corn active peptide.
6. Use of a corn active peptide according to claim 1 or a glycopeptide according to any one of claims 2 to 5 for the preparation of a functional product having ADH activating activity and/or antioxidant activity.
7. Use of the corn active peptide of claim 1 or the glycopeptide of any one of claims 2 to 5 for the preparation of a product for the prevention and treatment of alcoholic liver injury.
8. The method for preparing corn active peptide according to claim 1, comprising the steps of: adopting alkaline protease to carry out enzymolysis on corn protein powder, and taking supernatant as corn protein enzymolysis liquid; separating and purifying the corn protein enzymolysis liquid to obtain corn active peptide smaller than 1 kDa; then simulating in vitro gastrointestinal tract digestion and Caco-2 cell monolayer membrane absorption, and obtaining the corn active peptide with ADH activating activity and antioxidant activity through mass spectrum identification.
9. The method according to claim 8, wherein the pH of the enzymatic hydrolysis is 8.0, the temperature of the enzymatic hydrolysis is 55 ℃, and the time of the enzymatic hydrolysis is 180 minutes.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202311162420.5A CN117143191A (en) | 2023-09-11 | 2023-09-11 | Corn active peptide and glycopeptide with ADH activating activity and antioxidant activity, and preparation methods and applications thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202311162420.5A CN117143191A (en) | 2023-09-11 | 2023-09-11 | Corn active peptide and glycopeptide with ADH activating activity and antioxidant activity, and preparation methods and applications thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN117143191A true CN117143191A (en) | 2023-12-01 |
Family
ID=88907810
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202311162420.5A Pending CN117143191A (en) | 2023-09-11 | 2023-09-11 | Corn active peptide and glycopeptide with ADH activating activity and antioxidant activity, and preparation methods and applications thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN117143191A (en) |
-
2023
- 2023-09-11 CN CN202311162420.5A patent/CN117143191A/en active Pending
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Matheson | The determinatio of tryptophan in purified proteins and in feeding-stuffs | |
| CN104664039B (en) | A kind of saury Mei Lade peptides and its preparation method and application with anti-trioxypurine activity | |
| JP3195937B2 (en) | Method for obtaining amylase inhibitor | |
| Fontaine | Amino acid analysis of feeds. | |
| JP2020531627A (en) | Chinese giant salamander salamander cartilage preparation | |
| CN115960167A (en) | Corn anti-adhesion peptide and preparation method and application thereof | |
| CN116589533A (en) | Euphausia superba small molecule active peptide, and preparation method and application thereof | |
| US20120208214A1 (en) | Rapid saccharide biomarker assay | |
| CN117143191A (en) | Corn active peptide and glycopeptide with ADH activating activity and antioxidant activity, and preparation methods and applications thereof | |
| CA3071190C (en) | Spent turmeric - methods and compositions thereof | |
| CN109486886B (en) | Corn oligopeptide selenium and preparation method and application thereof | |
| JP6190999B2 (en) | Method for producing oligopeptide effective as alcohol metabolism promoter | |
| CN114891065B (en) | Blood sugar-reducing sea cucumber peptide with alpha-amylase inhibitory activity and preparation method and application thereof | |
| Yoshioka et al. | Bifidus factors in carrot. II. The structure of the factor in fraction IV | |
| Kølvraa et al. | D-glyceric acidemia: Biochemical studies of a new syndrome | |
| JPS61171431A (en) | Amylase-inhibiting substance and preparation thereof | |
| JP2006254876A (en) | Method for producing l-tryptophan | |
| CN112741199B (en) | Oyster peptide with sexual function improving effect and preparation method thereof | |
| CN117402215B (en) | Corn peptide for inhibiting alpha-glucosidase activity and preparation method and application thereof | |
| Masamune et al. | Chemical Nature of Toxohormone (Nakahara) Third Report KIK-inactive Toxohormones from Urines, Cancerous and Normal | |
| JP6308510B2 (en) | Alcohol metabolism promoter | |
| CN101852788B (en) | Method for quick derivatization of leucine in blood sample | |
| CN117186182A (en) | Pentapeptide with alcohol dehydrogenase activating capability, preparation method and application thereof | |
| AU638267B2 (en) | Dithiochrome, an insulin-binding molecule with glucose metabolism-related pharmaceutical utility | |
| Ortega et al. | Development and Validation of a Method for Hydrolysis and Analysis of Amino Acids in Ruminant Feeds, Tissue, and Milk Using Isotope Dilution Z-HILIC Coupled with Electrospray Ionization Triple Quadrupole LC-MS/MS |
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 |