CN109680030A - A kind of inoxidizability coconut polypeptide and the preparation method and application thereof - Google Patents
A kind of inoxidizability coconut polypeptide and the preparation method and application thereof Download PDFInfo
- Publication number
- CN109680030A CN109680030A CN201910080379.4A CN201910080379A CN109680030A CN 109680030 A CN109680030 A CN 109680030A CN 201910080379 A CN201910080379 A CN 201910080379A CN 109680030 A CN109680030 A CN 109680030A
- Authority
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- China
- Prior art keywords
- coconut
- polypeptide
- inoxidizability
- dregs
- rice
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- 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.)
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- 244000060011 Cocos nucifera Species 0.000 title claims abstract description 178
- 235000013162 Cocos nucifera Nutrition 0.000 title claims abstract description 178
- 108090000765 processed proteins & peptides Proteins 0.000 title claims abstract description 110
- 102000004196 processed proteins & peptides Human genes 0.000 title claims abstract description 109
- 229920001184 polypeptide Polymers 0.000 title claims abstract description 107
- 238000002360 preparation method Methods 0.000 title claims abstract description 41
- 241000209094 Oryza Species 0.000 claims abstract description 93
- 235000007164 Oryza sativa Nutrition 0.000 claims abstract description 93
- 235000009566 rice Nutrition 0.000 claims abstract description 93
- 239000004365 Protease Substances 0.000 claims abstract description 84
- 108091005804 Peptidases Proteins 0.000 claims abstract description 44
- 102000004190 Enzymes Human genes 0.000 claims abstract description 38
- 108090000790 Enzymes Proteins 0.000 claims abstract description 38
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 claims abstract description 28
- 230000003078 antioxidant effect Effects 0.000 claims abstract description 27
- 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 claims abstract description 26
- 230000002000 scavenging effect Effects 0.000 claims abstract description 22
- 230000000694 effects Effects 0.000 claims abstract description 19
- 150000001875 compounds Chemical class 0.000 claims abstract description 17
- 235000013305 food Nutrition 0.000 claims abstract description 11
- 239000003963 antioxidant agent Substances 0.000 claims abstract description 10
- 239000002537 cosmetic Substances 0.000 claims abstract description 10
- -1 hydroxyl radical free radical Chemical class 0.000 claims abstract description 7
- 238000012545 processing Methods 0.000 claims abstract description 5
- 239000003513 alkali Substances 0.000 claims description 48
- 229940088598 enzyme Drugs 0.000 claims description 37
- 239000000843 powder Substances 0.000 claims description 37
- 239000007788 liquid Substances 0.000 claims description 36
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 33
- 239000012153 distilled water Substances 0.000 claims description 28
- 230000009849 deactivation Effects 0.000 claims description 25
- 239000006228 supernatant Substances 0.000 claims description 25
- 229940055729 papain Drugs 0.000 claims description 24
- 235000019419 proteases Nutrition 0.000 claims description 24
- 238000000108 ultra-filtration Methods 0.000 claims description 24
- 239000012528 membrane Substances 0.000 claims description 22
- 102000035195 Peptidases Human genes 0.000 claims description 16
- 230000007935 neutral effect Effects 0.000 claims description 16
- 235000019833 protease Nutrition 0.000 claims description 14
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 9
- 238000000926 separation method Methods 0.000 claims description 7
- 108090000526 Papain Proteins 0.000 claims description 6
- 238000005238 degreasing Methods 0.000 claims description 6
- 108010004032 Bromelains Proteins 0.000 claims description 5
- 101000693530 Staphylococcus aureus Staphylokinase Proteins 0.000 claims description 5
- 238000005119 centrifugation Methods 0.000 claims description 5
- QCVGEOXPDFCNHA-UHFFFAOYSA-N 5,5-dimethyl-2,4-dioxo-1,3-oxazolidine-3-carboxamide Chemical compound CC1(C)OC(=O)N(C(N)=O)C1=O QCVGEOXPDFCNHA-UHFFFAOYSA-N 0.000 claims description 4
- 102000002322 Egg Proteins Human genes 0.000 claims description 4
- 108010000912 Egg Proteins Proteins 0.000 claims description 4
- 235000014103 egg white Nutrition 0.000 claims description 4
- 210000000969 egg white Anatomy 0.000 claims description 4
- 235000019441 ethanol Nutrition 0.000 claims description 4
- 235000019834 papain Nutrition 0.000 claims description 4
- 235000019835 bromelain Nutrition 0.000 claims description 3
- 238000006243 chemical reaction Methods 0.000 claims description 3
- 230000001376 precipitating effect Effects 0.000 claims description 3
- 238000010992 reflux Methods 0.000 claims description 3
- 238000001976 enzyme digestion Methods 0.000 claims description 2
- 239000000706 filtrate Substances 0.000 claims description 2
- 238000002156 mixing Methods 0.000 claims description 2
- 101710093543 Probable non-specific lipid-transfer protein Proteins 0.000 claims 1
- 238000000034 method Methods 0.000 abstract description 9
- 235000006708 antioxidants Nutrition 0.000 abstract description 8
- 230000007071 enzymatic hydrolysis Effects 0.000 abstract description 4
- 238000006047 enzymatic hydrolysis reaction Methods 0.000 abstract description 4
- 239000002994 raw material Substances 0.000 abstract description 3
- 230000001360 synchronised effect Effects 0.000 abstract description 3
- 238000011156 evaluation Methods 0.000 abstract description 2
- 239000013538 functional additive Substances 0.000 abstract description 2
- 230000007062 hydrolysis Effects 0.000 abstract description 2
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- 238000010998 test method Methods 0.000 description 34
- 238000012360 testing method Methods 0.000 description 21
- 230000007760 free radical scavenging Effects 0.000 description 20
- 238000004108 freeze drying Methods 0.000 description 19
- TUJKJAMUKRIRHC-UHFFFAOYSA-N hydroxyl Chemical compound [OH] TUJKJAMUKRIRHC-UHFFFAOYSA-N 0.000 description 19
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 description 6
- 238000002835 absorbance Methods 0.000 description 6
- 230000003064 anti-oxidating effect Effects 0.000 description 6
- 102000004169 proteins and genes Human genes 0.000 description 6
- 108090000623 proteins and genes Proteins 0.000 description 6
- 238000005259 measurement Methods 0.000 description 4
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 3
- 230000000052 comparative effect Effects 0.000 description 3
- 150000003254 radicals Chemical class 0.000 description 3
- 241000196324 Embryophyta Species 0.000 description 2
- 239000008280 blood Substances 0.000 description 2
- 210000004369 blood Anatomy 0.000 description 2
- HVYWMOMLDIMFJA-DPAQBDIFSA-N cholesterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 HVYWMOMLDIMFJA-DPAQBDIFSA-N 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
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- 238000002474 experimental method Methods 0.000 description 2
- 150000002632 lipids Chemical class 0.000 description 2
- 230000035790 physiological processes and functions Effects 0.000 description 2
- OCZVHBZNPVABKX-UHFFFAOYSA-N 1,1-diphenyl-2-(2,4,6-trinitrophenyl)hydrazine;ethanol Chemical compound CCO.[O-][N+](=O)C1=CC([N+](=O)[O-])=CC([N+]([O-])=O)=C1NN(C=1C=CC=CC=1)C1=CC=CC=C1 OCZVHBZNPVABKX-UHFFFAOYSA-N 0.000 description 1
- 244000099147 Ananas comosus Species 0.000 description 1
- 235000007119 Ananas comosus Nutrition 0.000 description 1
- 108010039627 Aprotinin Proteins 0.000 description 1
- 239000004475 Arginine Substances 0.000 description 1
- 244000189799 Asimina triloba Species 0.000 description 1
- 235000006264 Asimina triloba Nutrition 0.000 description 1
- 108010001478 Bacitracin Proteins 0.000 description 1
- NLZUEZXRPGMBCV-UHFFFAOYSA-N Butylhydroxytoluene Chemical compound CC1=CC(C(C)(C)C)=C(O)C(C(C)(C)C)=C1 NLZUEZXRPGMBCV-UHFFFAOYSA-N 0.000 description 1
- 235000009467 Carica papaya Nutrition 0.000 description 1
- 108010059892 Cellulase Proteins 0.000 description 1
- 241000195649 Chlorella <Chlorellales> Species 0.000 description 1
- GYCKQBWUSACYIF-UHFFFAOYSA-N Ethyl salicylate Chemical group CCOC(=O)C1=CC=CC=C1O GYCKQBWUSACYIF-UHFFFAOYSA-N 0.000 description 1
- 244000068988 Glycine max Species 0.000 description 1
- 235000010469 Glycine max Nutrition 0.000 description 1
- 241000237502 Ostreidae Species 0.000 description 1
- 244000170916 Paeonia officinalis Species 0.000 description 1
- 235000006484 Paeonia officinalis Nutrition 0.000 description 1
- 108010019160 Pancreatin Proteins 0.000 description 1
- 102100025292 Stress-induced-phosphoprotein 1 Human genes 0.000 description 1
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 description 1
- 230000002411 adverse Effects 0.000 description 1
- 230000032683 aging Effects 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 229960004405 aprotinin Drugs 0.000 description 1
- 229960003071 bacitracin Drugs 0.000 description 1
- 229930184125 bacitracin Natural products 0.000 description 1
- CLKOFPXJLQSYAH-ABRJDSQDSA-N bacitracin A Chemical compound C1SC([C@@H](N)[C@@H](C)CC)=N[C@@H]1C(=O)N[C@@H](CC(C)C)C(=O)N[C@H](CCC(O)=O)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H]1C(=O)N[C@H](CCCN)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@H](CC=2C=CC=CC=2)C(=O)N[C@@H](CC=2N=CNC=2)C(=O)N[C@H](CC(O)=O)C(=O)N[C@@H](CC(N)=O)C(=O)NCCCC1 CLKOFPXJLQSYAH-ABRJDSQDSA-N 0.000 description 1
- 239000002585 base Substances 0.000 description 1
- 235000010290 biphenyl Nutrition 0.000 description 1
- 239000004305 biphenyl Substances 0.000 description 1
- 235000010354 butylated hydroxytoluene Nutrition 0.000 description 1
- 239000006227 byproduct Substances 0.000 description 1
- 238000004364 calculation method Methods 0.000 description 1
- 238000011088 calibration curve Methods 0.000 description 1
- 239000004202 carbamide Substances 0.000 description 1
- 108010089934 carbohydrase Proteins 0.000 description 1
- YCIMNLLNPGFGHC-UHFFFAOYSA-N catechol Chemical compound OC1=CC=CC=C1O YCIMNLLNPGFGHC-UHFFFAOYSA-N 0.000 description 1
- 230000005779 cell damage Effects 0.000 description 1
- 229940106157 cellulase Drugs 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 235000012000 cholesterol Nutrition 0.000 description 1
- 235000020197 coconut milk Nutrition 0.000 description 1
- 235000009508 confectionery Nutrition 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
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- 230000035622 drinking Effects 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
- 235000013601 eggs Nutrition 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- 201000005577 familial hyperlipidemia Diseases 0.000 description 1
- 238000000855 fermentation Methods 0.000 description 1
- 230000004151 fermentation Effects 0.000 description 1
- 235000003891 ferrous sulphate Nutrition 0.000 description 1
- 239000011790 ferrous sulphate Substances 0.000 description 1
- 239000003337 fertilizer Substances 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 229960002449 glycine Drugs 0.000 description 1
- 239000004519 grease Substances 0.000 description 1
- 235000008216 herbs Nutrition 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- ZPNFWUPYTFPOJU-LPYSRVMUSA-N iniprol Chemical compound C([C@H]1C(=O)NCC(=O)NCC(=O)N[C@H]2CSSC[C@H]3C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@H](C(N[C@H](C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC=4C=CC(O)=CC=4)C(=O)N[C@@H](CC=4C=CC=CC=4)C(=O)N[C@@H](CC=4C=CC(O)=CC=4)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C)C(=O)NCC(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CSSC[C@H](NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](C)NC(=O)[C@H](CO)NC(=O)[C@H](CCCCN)NC(=O)[C@H](CC=4C=CC=CC=4)NC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H](CCCCN)NC(=O)[C@H](C)NC(=O)[C@H](CCCNC(N)=N)NC2=O)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CSSC[C@H](NC(=O)[C@H](CC=2C=CC=CC=2)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H]2N(CCC2)C(=O)[C@@H](N)CCCNC(N)=N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(=O)N2[C@@H](CCC2)C(=O)N2[C@@H](CCC2)C(=O)N[C@@H](CC=2C=CC(O)=CC=2)C(=O)N[C@@H]([C@@H](C)O)C(=O)NCC(=O)N2[C@@H](CCC2)C(=O)N3)C(=O)NCC(=O)NCC(=O)N[C@@H](C)C(O)=O)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@H](C(=O)N[C@@H](CC=2C=CC=CC=2)C(=O)N[C@H](C(=O)N1)C(C)C)[C@@H](C)O)[C@@H](C)CC)=O)[C@@H](C)CC)C1=CC=C(O)C=C1 ZPNFWUPYTFPOJU-LPYSRVMUSA-N 0.000 description 1
- 230000010354 integration Effects 0.000 description 1
- BAUYGSIQEAFULO-UHFFFAOYSA-L iron(2+) sulfate (anhydrous) Chemical compound [Fe+2].[O-]S([O-])(=O)=O BAUYGSIQEAFULO-UHFFFAOYSA-L 0.000 description 1
- 229910000359 iron(II) sulfate Inorganic materials 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 230000031700 light absorption Effects 0.000 description 1
- 230000003859 lipid peroxidation Effects 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 210000004072 lung Anatomy 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 231100000252 nontoxic Toxicity 0.000 description 1
- 230000003000 nontoxic effect Effects 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 230000000050 nutritive effect Effects 0.000 description 1
- 210000003463 organelle Anatomy 0.000 description 1
- 235000020636 oyster Nutrition 0.000 description 1
- 235000019629 palatability Nutrition 0.000 description 1
- 229940055695 pancreatin Drugs 0.000 description 1
- ZUOUZKKEUPVFJK-UHFFFAOYSA-N phenylbenzene Natural products C1=CC=CC=C1C1=CC=CC=C1 ZUOUZKKEUPVFJK-UHFFFAOYSA-N 0.000 description 1
- 235000015277 pork Nutrition 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 238000007348 radical reaction Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 210000000952 spleen Anatomy 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 125000000999 tert-butyl group Chemical group [H]C([H])([H])C(*)(C([H])([H])[H])C([H])([H])[H] 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 230000002110 toxicologic effect Effects 0.000 description 1
- 231100000027 toxicology Toxicity 0.000 description 1
- YNJBWRMUSHSURL-UHFFFAOYSA-N trichloroacetic acid Chemical compound OC(=O)C(Cl)(Cl)Cl YNJBWRMUSHSURL-UHFFFAOYSA-N 0.000 description 1
- 238000000870 ultraviolet spectroscopy Methods 0.000 description 1
- 235000013311 vegetables Nutrition 0.000 description 1
- 239000002699 waste material Substances 0.000 description 1
- 239000003643 water by type Substances 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P21/00—Preparation of peptides or proteins
- C12P21/06—Preparation of peptides or proteins produced by the hydrolysis of a peptide bond, e.g. hydrolysate products
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- 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
- A61K8/645—Proteins of vegetable origin; Derivatives or degradation products thereof
-
- 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
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/14—Extraction; Separation; Purification
- C07K1/34—Extraction; Separation; Purification by filtration, ultrafiltration or reverse osmosis
-
- 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
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- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Molecular Biology (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Animal Behavior & Ethology (AREA)
- Genetics & Genomics (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Biochemistry (AREA)
- Water Supply & Treatment (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Dermatology (AREA)
- Polymers & Plastics (AREA)
- Analytical Chemistry (AREA)
- Food Science & Technology (AREA)
- Biophysics (AREA)
- Nutrition Science (AREA)
- Medicinal Chemistry (AREA)
- Mycology (AREA)
- Biotechnology (AREA)
- Gerontology & Geriatric Medicine (AREA)
- General Chemical & Material Sciences (AREA)
- Microbiology (AREA)
- Epidemiology (AREA)
- Birds (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Medicines Containing Plant Substances (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
Abstract
The present invention discloses a kind of inoxidizability coconut polypeptide and the preparation method and application thereof, belongs to food and cosmetic technical field.The present invention is raw material using the coconut dregs of rice, obtain the optimised process that anti-oxidant coconut polypeptide is prepared by the synchronous enzymatic hydrolysis of compound protease, deep processing for the coconut dregs of rice provides theoretical foundation, and preparation process is simple, polypeptide yield is more single, and protease is higher, and all has stronger Scavenging activity to DPPH and hydroxyl radical free radical；The present invention carries out inoxidizability evaluation to resulting coconut polypeptide, and DPPH Scavenging activity is 57.3~84.5%, and hydroxyl radical free radical clearance rate is 53.5~74.2%.Complex enzyme hydrolysis technique through the invention, obtains the high income of coconut polypeptide, and molecular weight is low, inoxidizability effect is obvious, and it is safe and non-stimulating, it can be used as functional additive and be widely used in food and cosmetic field, the integrated application value for improving coconut resource is of great significance.
Description
Technical field
The invention belongs to food and cosmetic technical fields, and in particular to a kind of inoxidizability coconut polypeptide and its preparation side
Method and application.
Background technique
Free radical is the mesostate of various chemical reactions in human life activity, the chemical activity with height,
It will affect the vital movement of human body if it cannot maintain certain level, but free radical can excessively attack man-machine intracorporal life
Macromolecular and organelle lead to lipid peroxidation and the active decrease of antioxidant enzyme, to cause cellular damage, accelerate people
The aging course of body simultaneously induces various diseases.Therefore antioxidant has to human free radical and the prevention various diseases of the mankind is removed
Significance.Currently used in the market is mainly with chemically synthesized 2,6 di tert butyl 4 methyl phenol and tert-butyl pair
Benzenediol is the most extensive, but toxicological experiment shows that chemical synthesis antioxidant toxic side effect is larger, equal to human liver, spleen, lung etc.
It adversely affects.So the antioxidant of exploitation Nantural non-toxic has become the research hotspot of recent domestic scholar.At present
Passed through the technologies such as enzymatic hydrolysis, fermentation from a variety of food-borne albumen such as pork, egg, oyster, soybean, the peony seeds dregs of rice, chlorella
Obtain the polypeptide with antioxidant activity.Wherein enzymatic isolation method cost is lower, it is easier to the biologically active peptide for operating, while preparing
With good dissolubility, acidproof and heat-resistant stability.
Coconut is integration of drinking and medicinal herbs vegetables and fruits, is a kind of economic value and all very high plant of nutritive value, in addition to producing coconut
It is oily outer, it also can be processed into the Speciality Foods full of nutrition such as coconut milk, coconut candy, coconut cake, cocoa-nut cake.The coconut dregs of rice are that coconut mentions
By-product after taking grease, wherein protein content is up to 20%, and amino acid classes are complete in albumen, have certain reducing blood lipid,
It reduces cholesterol and inhibits the physiological functions such as hyperlipemia, be a kind of good protein resource.It is long but since palatability is poor
It is since phase that feed is taken as to use or discharge naturally more, cause the huge waste of protein resource.Compared with coconut protein, coconut palm
Sub- polypeptide is small due to molecular weight, it is easier to and be absorbed by the body utilization, and has the physiological functions such as inoxidizability and reducing blood lipid,
It has broad application prospects in the fields such as foods and cosmetics.Application No. is 201210041195.5 Chinese patents to disclose
It is a kind of to hydrolyze coconut protein in two steps using coloured glaze base protease and two kinds of protease of compound protease and prepare coconut polypeptide technique, Shen
Please number for 201710946924.4 a Chinese patent disclose it is a kind of using cellulase+carbohydrase+pancreatin and papain+
Compound fertilizer production secondary enzymolysis coconut protein prepares the technique of coconut polypeptide, but all synchronizes enzyme without reference to using complex enzyme
Solution prepares the technology of antioxidation active peptides, therefore studies a kind of preparation process of inoxidizability coconut polypeptide for improving coconut
The application value of the dregs of rice is of great significance.
Summary of the invention
In order to overcome the disadvantages and deficiencies of the prior art, the primary purpose of the present invention is that providing a kind of inoxidizability coconut
The preparation method of polypeptide.
Another object of the present invention is to provide the inoxidizability coconut polypeptides being prepared by above-mentioned preparation method.
A further object of the present invention is to provide the applications of above-mentioned inoxidizability coconut polypeptide.
The present invention is provided by the synchronous enzymatic hydrolysis of compound protease and a kind of prepares coconut dregs of rice antioxygen simple, that antioxidant activity is strong
Change active peptides, and antioxidant activity polypeptide can be applied to the development and exploitation of foods and cosmetics, realizes the coconut dregs of rice
The comprehensive utilization of resource improves its application value.
The purpose of the invention is achieved by the following technical solution:
A kind of preparation method of inoxidizability coconut polypeptide, includes the following steps:
(1) the coconut dregs of rice are preprocessed, obtain smart coconut dregs of rice powder；
(2) after mixing smart coconut dregs of rice powder according to mass ratio 1:15~25 with distilled water, the pH to 6~9 of mixed liquor is adjusted,
1~4h of enzyme digestion reaction is carried out under the conditions of 50~60 DEG C according to 2~3% addition compound proteases of smart coconut dregs of rice silty amount, is obtained
To enzymolysis liquid；
The compound protease includes in alkali protease, neutral proteinase, bromelain and papain
At least two；
(3) enzymolysis liquid that step (2) obtains is living through enzyme deactivation, is centrifugated, and supernatant uses molecule interception for 5000Da
Ultrafiltration membrane ultra-filtration and separation below, filtrate are freeze-dried to obtain inoxidizability coconut polypeptide.
Preferably, smart coconut dregs of rice powder described in step (1) is mixed with distilled water according to mass ratio 1:20；
Preferably, the dosage of compound protease described in step (1) is to be added again according to the 2% of smart coconut dregs of rice silty amount
Hop protein enzyme.
Preferably, the preparation method of smart coconut dregs of rice powder described in step (1), includes the following steps:
The coconut dregs of rice are added according to mass volume ratio 1:20 into ethyl alcohol, 130 DEG C of reflux 3h, separation is dried to obtain degreasing coconut palm
Sub- dregs of rice powder；Then degreasing coconut dregs of rice powder and distilled water is added by solid-liquid ratio 1:25, adjusts PH to 9, the lower 50 DEG C of processing of high-speed stirred
Then 2h, centrifuging and taking supernatant adjust PH to 3.5~3.6, precipitating are collected by centrifugation, is freeze-dried after being washed to neutrality, obtains smart coconut palm
Sub- dregs of rice powder.
Preferably, compound protease described in step (1) includes alkali protease-neutral proteinase, alkali protease-
Bromelain, neutral proteinase-papain, alkali protease-papain, neutrality -- protease-bromelain
Enzyme, papain-bromelain, alkali protease-papain-bromelain, alkali protease-pawpaw property egg
White enzyme-neutral proteinase.
It is furthermore preferred that compound protease described in step (1) includes alkali protease-neutral protein by quality ratio
Enzyme=1:0.5~2, alkali protease-bromelain=1:0.2~0.8, neutral proteinase-papain=1:0.25
~1, alkali protease-papain=1:0.25~1, neutral proteinase-bromelain=1:0.2~0.8, pawpaw egg
White enzyme-bromelain=1:0.4~1.6, alkali protease-papain-bromelain=1:0.5:0.2~0.8,
Alkali protease-papain-neutral proteinase=1:0.5:0.5~2.
It is further preferred that the compound protease includes alkali protease-neutral proteinase=1 by quality ratio:
1~2, alkali protease-bromelain=1:0.2~0.4, neutral proteinase-papain=1:0.25~1, alkalinity
Protease-papain=1:0.25~1, neutral proteinase-bromelain=1:0.4~0.8, papain-spinach
Trailing plants protease=1:0.8~1.6, alkali protease-papain-bromelain=1:0.5:0.8, alkali protease-
Papain-neutral proteinase=1:0.5:2.
Alkali protease enzyme activity >=the 200000U/g, neutral proteinase enzyme activity are >=200000U/g, pineapple
Proteinase activity power >=500000U/g, papain enzyme activity >=400000U/g.
Preferably, the condition living of enzyme deactivation described in step (3) is 5~10min of enzyme deactivation at 90~95 DEG C；More preferably 90
Enzyme deactivation 10min at DEG C.
Preferably, the condition of centrifugation described in step (3) is that 5~10min is centrifugated under 8000~10000rpm；More
10min is centrifugated under preferably 8000rpm.
Preferably, in step (3), use molecule interception for the ultrafiltration membrane ultra-filtration and separation of 4000Da, polypeptide yield is essence
The 9.6~25.8% of coconut dregs of rice silty amount, polypeptide molecular weight are 2547~3126Da.
It is furthermore preferred that polypeptide yield is the 17.3~25.8% of smart coconut dregs of rice silty amount, polypeptide molecular weight is 2547~
2879Da。
Preferably, the DPPH Scavenging activity of prepared coconut polypeptide is 57.3~84.5%, hydroxyl radical free radical clearance rate
It is 53.5~74.2%.
It is furthermore preferred that the DPPH Scavenging activity of prepared coconut polypeptide is 70.5~84.5%, hydroxyl radical free radical is removed
Rate is 60.4~74.2%.
A kind of inoxidizability coconut polypeptide, is prepared by above-mentioned preparation method.
Application of the inoxidizability coconut polypeptide as food or the antioxidant of cosmetics.
Further, the inoxidizability coconut polypeptide is preparing the application in food or cosmetics.
The present invention has the following advantages and effects with respect to the prior art:
(1) present invention is raw material using the coconut dregs of rice, obtains and prepares anti-oxidant coconut by the synchronous enzymatic hydrolysis of compound protease
The optimised process of polypeptide provides theoretical foundation for the deep processing of the coconut dregs of rice, and preparation process is simple, polypeptide yield (9.6~
25.8%) more single protease (5.3~8.7%) is higher, and all has stronger removing energy to DPPH and hydroxyl radical free radical
Power provides a kind of new raw material for the antioxidant of food and cosmetics, and safe and non-stimulating.
(2) solution that 2mg/mL is made to resulting coconut polypeptide in the present invention carries out inoxidizability evaluation, and DPPH removes energy
Power is 57.3~84.5%, and hydroxyl radical free radical clearance rate is 53.5~74.2%.Complex enzyme hydrolysis technique through the invention, obtains
The high income of coconut polypeptide, molecular weight is low, and inoxidizability effect is obvious, can be used as functional additive and is widely used in food
And cosmetic field, the integrated application value for improving coconut resource are of great significance.
Specific embodiment
Below with reference to embodiment, the present invention is described in further detail, and embodiments of the present invention are not limited thereto.
The preparation method of essence coconut dregs of rice powder, includes the following steps: used in embodiment
The 0g coconut dregs of rice are put into cable type extractor according, 130 DEG C of reflux 3h of 200mL ethyl alcohol are added, separation is dried to obtain degreasing coconut palm
Sub- dregs of rice powder.Degreasing coconut dregs of rice powder and distilled water is added by solid-liquid ratio 1:25, adjusts PH to 9, the lower 50 DEG C of processing 2h of high-speed stirred, from
The heart takes supernatant, then adjusts PH to 3.5~3.6, precipitating is collected by centrifugation, is freeze-dried after being washed to neutrality, obtains the smart coconut dregs of rice
Powder.
Embodiment 1
After 1g essence coconut dregs of rice powder is mixed with distilled water according to mass ratio 1:20, the PH to 9 of mixed liquor is adjusted, smart coconut palm is added
Alkali protease-neutral proteinase of sub- dregs of rice silty amount 2%, alkali protease-neutral proteinase ratio is 1:2, in temperature 55
4h is stirred to react at DEG C, enzymolysis liquid is centrifugated 10min at 8000rpm by enzyme deactivation 10min at 90 DEG C, takes supernatant to use super
Filter membrane (molecule interception is 4000Da) carries out separating-purifying, obtains antioxidant activity coconut polypeptide solution, obtains after freeze-drying
Inoxidizability coconut polypeptide.
Test: the measuring method of antioxidant activity:
(1) measurement of DPPH free radical scavenging ability
The solution for using distilled water to be configured to 2mg/mL inoxidizability coconut polypeptide obtained is measured as sample liquid.
The sample liquid of 2mL is added to 2mL 0.1mmol/LDPPH (1,1- diphenyl -2- trinitrophenyl-hydrazine) Fresh
In ethanol solution, after firmly shaking up, 30min is placed in the dark, measures suction at 517nm with ultraviolet-visible spectrophotometer
Luminosity uses the DPPH ethanol solution of not sample adding liquid as blank control, and clearance rate can be indicated with following formula: clearance rate
(%)=(A0-Ai)*100/A0；
In formula, AiFor the absorbance after placing 30min, A is added after sample liquid0For the absorbance of blank control.
(2) measurement of hydroxyl radical free radical Scavenging activity
The solution for using distilled water to be configured to 2mg/mL inoxidizability coconut polypeptide obtained is measured as sample liquid.
After ferrous sulfate 1mL, the 6mmol/L salicylic acid-ethyl alcohol 1mL for sequentially adding 6mmol/L in 10mL test tube, sample
Liquid 1mL is added in test tube, and 0.1% hydrogen peroxide 1mL is then added, and total volume 5mL supplies volume with distilled water.It is wherein right
Not sample adding liquid is looked after, hydrogen peroxide is not added in sample bottom tube, keeps the temperature 30min after shaking up in 37 DEG C of water-baths, surveys the absorbance at 510nm
Value.Clearance rate calculation formula are as follows:
Clearance rate (%)=[B0-(B-Bi)]*100/B0
In formula, B0The absorbance value of hydroxy radical system when for not sample adding liquid；B is the hydroxy radical system that sample liquid is added
Absorbance value；BiFor the absorbance value that hydrogen peroxide and sample adding liquid are not added in hydroxy radical reaction system.
(3) measurement of polypeptide yield
Appropriate antioxidant activity coconut polypeptide solution is taken, 10min is centrifuged with the speed of 5000r/min, supernatant is taken to be added
10% trichloroacetic acid of 5mL reacts 20min, then is centrifuged 10min with the speed of 5000r/min, then takes supernatant 2mL, is added double
Contracting urea reagent 6mL reacts 30min in 37 DEG C of water-baths, light absorption value is measured under 540nm wavelength, according to standard curve regression equation
Calculate the content of polypeptide.
(4) measurement of polypeptide molecular weight
Polypeptide powder resulting after freeze-drying is dissolved in water, is filtered with micropore filtering film, is surveyed using high performance liquid chromatography
Determine enzymolysis product molecular weight distribution.Experiment (works in 600 high performance liquid chromatograph of Waters with 2487 UV detector and M32
Stand) on carry out, operating condition is as follows: chromatographic column: TSKgel G2000SW × L (300 × 7.8mm)；Mobile phase: acetonitrile/water/
TFA=45/55/0.1 (V:V:V)；Detection wavelength is 220nm；30 DEG C of column temperature；Flow velocity 0.5mL/min；
Standard items used in molecular weight calibration curve: cromoci (MW 12500)；Aprotinin (MW 6500)；Bacitracin
(MW 1450)；Second glutamine-second glutamine-arginine (MW 451)；Second glutamine-second glutamine-aminoacetic acid (MW 189).
The present embodiment test result are as follows: 23.8%, the DPPH for digesting the polypeptide yield of preparation as smart coconut dregs of rice silty amount is clear
Removing solid capacity is 79.2%, and hydroxyl radical free radical Scavenging activity is 74.2%, molecular weight 2736Da.
Embodiment 2
After 1g essence coconut dregs of rice powder is mixed with distilled water according to mass ratio 1:20, the PH to 8 of mixed liquor is adjusted, smart coconut palm is added
Alkali protease-neutral proteinase of sub- dregs of rice silty amount 2%, alkali protease-neutral proteinase ratio is 1:1, in temperature 55
3h is stirred to react at DEG C, enzymolysis liquid is centrifugated 10min at 8000rpm by enzyme deactivation 10min at 90 DEG C, takes supernatant to use super
Filter membrane (molecule interception is 4000Da) carries out separating-purifying, obtains antioxidant activity coconut polypeptide solution, obtains after freeze-drying
Inoxidizability coconut polypeptide.
Test method of the test method with embodiment 1, the present embodiment test result are as follows: digest the polypeptide yield of preparation as essence
20.6%, the DPPH Scavenging activity of coconut dregs of rice silty amount is 72.1%, and hydroxyl radical free radical Scavenging activity is 69.4%, and molecular weight is
2865Da。
Embodiment 3
After the smart coconut dregs of rice powder of 1g is mixed with distilled water according to mass ratio 1:20, the PH to 9 of mixed liquor is adjusted, essence is added
Alkali protease-bromelain of coconut dregs of rice silty amount 2%, alkali protease-bromelain ratio is 1:0.4, in temperature
It is stirred to react 4h at 50 DEG C of degree, enzymolysis liquid is centrifugated 10min at 8000rpm, takes supernatant by enzyme deactivation 10min at 90 DEG C
Separating-purifying is carried out with ultrafiltration membrane (molecule interception is 4000Da), antioxidant activity coconut polypeptide solution is obtained, after freeze-drying
Obtain inoxidizability coconut polypeptide.
Test method of the test method with embodiment 1, the present embodiment test result are as follows: digest the polypeptide yield of preparation as essence
19.2%, the DPPH Scavenging activity of coconut dregs of rice silty amount is 75.2%, and hydroxyl radical free radical Scavenging activity is 65.3%, and molecular weight is
2742Da。
Embodiment 4
After the smart coconut dregs of rice powder of 1g is mixed with distilled water according to mass ratio 1:20, the PH to 7 of mixed liquor is adjusted, essence is added
Alkali protease-bromelain of coconut dregs of rice silty amount 2%, alkali protease-bromelain ratio is 1:0.2, in temperature
It is stirred to react 2h at 60 DEG C of degree, enzymolysis liquid is centrifugated 10min at 8000rpm, takes supernatant by enzyme deactivation 10min at 90 DEG C
Separating-purifying is carried out with ultrafiltration membrane (molecule interception is 4000Da), antioxidant activity coconut polypeptide solution is obtained, after freeze-drying
Obtain inoxidizability coconut polypeptide.
Test method of the test method with embodiment 1, the present embodiment test result are as follows: digest the polypeptide yield of preparation as essence
17.3%, the DPPH Scavenging activity of coconut dregs of rice silty amount is 70.5%, and hydroxyl radical free radical Scavenging activity is 60.4%, and molecular weight is
2879Da。
Embodiment 5
After the smart coconut dregs of rice powder of 1g is mixed with distilled water according to mass ratio 1:20, the PH to 6 of mixed liquor is adjusted, essence is added
Neutral proteinase-papain of coconut dregs of rice silty amount 2%, neutral proteinase-papain ratio is 1:1, in temperature
1h is stirred to react at 50 DEG C, enzymolysis liquid is centrifugated 10min at 8000rpm, supernatant is taken to use by enzyme deactivation 10min at 90 DEG C
Ultrafiltration membrane (molecule interception is 4000Da) carries out separating-purifying, obtains antioxidant activity coconut polypeptide solution, after freeze-drying
To inoxidizability coconut polypeptide.
Test method of the test method with embodiment 1, the present embodiment test result are as follows: digest the polypeptide yield of preparation as essence
10.8%, the DPPH Scavenging activity of coconut dregs of rice silty amount is 59.4%, and hydroxyl radical free radical Scavenging activity is 62.8%, and molecular weight is
3094Da。
Embodiment 6
After the smart coconut dregs of rice powder of 1g is mixed with distilled water according to mass ratio 1:20, the PH to 8 of mixed liquor is adjusted, essence is added
Neutral proteinase-papain of coconut dregs of rice silty amount 2%, neutral proteinase-papain ratio is 1:0.25, in temperature
It is stirred to react 4h at 55 DEG C of degree, enzymolysis liquid is centrifugated 10min at 8000rpm, takes supernatant by enzyme deactivation 10min at 90 DEG C
Separating-purifying is carried out with ultrafiltration membrane (molecule interception is 4000Da), antioxidant activity coconut polypeptide solution is obtained, after freeze-drying
Obtain inoxidizability coconut polypeptide.
Test method of the test method with embodiment 1, the present embodiment test result are as follows: digest the polypeptide yield of preparation as essence
16.5%, the DPPH Scavenging activity of coconut dregs of rice silty amount is 68.2%, and hydroxyl radical free radical Scavenging activity is 67.9%, and molecular weight is
2981Da。
Embodiment 7
After the smart coconut dregs of rice powder of 1g is mixed with distilled water according to mass ratio 1:20, the PH to 9 of mixed liquor is adjusted, essence is added
Alkali protease-papain of coconut dregs of rice silty amount 2%, alkali protease-papain ratio is 1:0.5, in temperature
It is stirred to react 4h at 55 DEG C of degree, enzymolysis liquid is centrifugated 10min at 8000rpm, takes supernatant by enzyme deactivation 10min at 90 DEG C
Separating-purifying is carried out with ultrafiltration membrane (molecule interception is 4000Da), antioxidant activity coconut polypeptide solution is obtained, after freeze-drying
Obtain inoxidizability coconut polypeptide.
Test method of the test method with embodiment 1, the present embodiment test result are as follows: digest the polypeptide yield of preparation as essence
25.8%, the DPPH Scavenging activity of coconut dregs of rice silty amount is 84.5%, and hydroxyl radical free radical Scavenging activity is 71.6%, and molecular weight is
2547Da。
Embodiment 8
After the smart coconut dregs of rice powder of 1g is mixed with distilled water according to mass ratio 1:20, the PH to 8 of mixed liquor is adjusted, essence is added
Alkali protease-papain of coconut dregs of rice silty amount 2%, alkali protease-papain ratio is 1:1, in temperature
3h is stirred to react at 60 DEG C, enzymolysis liquid is centrifugated 10min at 8000rpm, supernatant is taken to use by enzyme deactivation 10min at 90 DEG C
Ultrafiltration membrane (molecule interception is 4000Da) carries out separating-purifying, obtains antioxidant activity coconut polypeptide solution, after freeze-drying
To inoxidizability coconut polypeptide.
Test method of the test method with embodiment 1, the present embodiment test result are as follows: digest the polypeptide yield of preparation as essence
24.9%, the DPPH Scavenging activity of coconut dregs of rice silty amount is 82.7%, and hydroxyl radical free radical Scavenging activity is 69.8%, and molecular weight is
2683Da。
Embodiment 9
After the smart coconut dregs of rice powder of 1g is mixed with distilled water according to mass ratio 1:20, the PH to 9 of mixed liquor is adjusted, essence is added
Alkali protease-papain of coconut dregs of rice silty amount 2%, alkali protease-papain ratio is 1:0.25, in temperature
It is stirred to react 2h at 50 DEG C of degree, enzymolysis liquid is centrifugated 10min at 8000rpm, takes supernatant by enzyme deactivation 10min at 90 DEG C
Separating-purifying is carried out with ultrafiltration membrane (molecule interception is 4000Da), antioxidant activity coconut polypeptide solution is obtained, after freeze-drying
Obtain inoxidizability coconut polypeptide.
Test method of the test method with embodiment 1, the present embodiment test result are as follows: digest the polypeptide yield of preparation as essence
19.9%, the DPPH Scavenging activity of coconut dregs of rice silty amount is 74.9%, and hydroxyl radical free radical Scavenging activity is 62.7%, and molecular weight is
2806Da。
Embodiment 10
After the smart coconut dregs of rice powder of 1g is mixed with distilled water according to mass ratio 1:20, the PH to 6 of mixed liquor is adjusted, essence is added
Neutral proteinase-bromelain of coconut dregs of rice silty amount 2%, neutral proteinase-bromelain ratio is 1:0.8, in temperature
It is stirred to react 1h at 55 DEG C of degree, enzymolysis liquid is centrifugated 10min at 8000rpm, takes supernatant by enzyme deactivation 10min at 90 DEG C
Separating-purifying is carried out with ultrafiltration membrane (molecule interception is 4000Da), antioxidant activity coconut polypeptide solution is obtained, after freeze-drying
Obtain inoxidizability coconut polypeptide.
Test method of the test method with embodiment 1, the present embodiment test result are as follows: digest the polypeptide yield of preparation as essence
9.6%, the DPPH Scavenging activity of coconut dregs of rice silty amount is 57.3%, and hydroxyl radical free radical Scavenging activity is 53.5%, and molecular weight is
3126Da。
Embodiment 11
After the smart coconut dregs of rice powder of 1g is mixed with distilled water according to mass ratio 1:20, the PH to 6 of mixed liquor is adjusted, essence is added
Neutral proteinase-bromelain of coconut dregs of rice silty amount 2%, neutral proteinase-bromelain ratio is 1:0.4, in temperature
It is stirred to react 4h at 50 DEG C of degree, enzymolysis liquid is centrifugated 10min at 8000rpm, takes supernatant by enzyme deactivation 10min at 90 DEG C
Separating-purifying is carried out with ultrafiltration membrane (molecule interception is 4000Da), anti-oxidant Coconut activated polypeptide solution is obtained, after freeze-drying
Obtain inoxidizability coconut polypeptide.
Test method of the test method with embodiment 1, the present embodiment test result are as follows: digest the polypeptide yield of preparation as essence
12.1%, the DPPH Scavenging activity of coconut dregs of rice silty amount is 64.5%, and hydroxyl radical free radical Scavenging activity is 59.2%, and molecular weight is
3058Da。
Embodiment 12
After the smart coconut dregs of rice powder of 1g is mixed with distilled water according to mass ratio 1:20, the PH to 7 of mixed liquor is adjusted, essence is added
Papain-bromelain of coconut dregs of rice silty amount 2%, papain-bromelain ratio is 1:1.6, in temperature
It is stirred to react 2h at 55 DEG C of degree, enzymolysis liquid is centrifugated 10min at 8000rpm, takes supernatant by enzyme deactivation 10min at 90 DEG C
Separating-purifying is carried out with ultrafiltration membrane (molecule interception is 4000Da), antioxidant activity coconut polypeptide solution is obtained, after freeze-drying
Obtain inoxidizability coconut polypeptide.
Test method of the test method with embodiment 1, the present embodiment test result are as follows: digest the polypeptide yield of preparation as essence
14.7%, the DPPH Scavenging activity of coconut dregs of rice silty amount is 73.2%, and hydroxyl radical free radical Scavenging activity is 64.8%, and molecular weight is
2864Da。
Embodiment 13
After the smart coconut dregs of rice powder of 1g is mixed with distilled water according to mass ratio 1:20, the PH to 8 of mixed liquor is adjusted, essence is added
Papain-bromelain of coconut dregs of rice silty amount 2%, papain-bromelain ratio is 1:0.8, in temperature
It is stirred to react 3h at 50 DEG C of degree, enzymolysis liquid is centrifugated 10min at 8000rpm, takes supernatant by enzyme deactivation 10min at 90 DEG C
Separating-purifying is carried out with ultrafiltration membrane (molecule interception is 4000Da), antioxidant activity coconut polypeptide solution is obtained, after freeze-drying
Obtain inoxidizability coconut polypeptide.
Test method of the test method with embodiment 1, the present embodiment test result are as follows: digest the polypeptide yield of preparation as essence
13.6%, the DPPH Scavenging activity of coconut dregs of rice silty amount is 69.4%, and hydroxyl radical free radical Scavenging activity is 61.3%, and molecular weight is
2942Da。
Embodiment 14
After the smart coconut dregs of rice powder of 1g is mixed with distilled water according to mass ratio 1:20, the PH to 9 of mixed liquor is adjusted, essence is added
Alkali protease-papain-bromelain of coconut dregs of rice silty amount 2%, alkali protease-papain-pineapple
Protease ratio is 1:0.5:0.8,4h is stirred to react at 55 DEG C of temperature, enzyme deactivation 10min, enzymolysis liquid is existed at 90 DEG C
It is centrifugated 10min under 8000rpm, takes supernatant to carry out separating-purifying with ultrafiltration membrane (molecule interception is 4000Da), obtains anti-
Oxidation activity coconut polypeptide solution obtains inoxidizability coconut polypeptide after freeze-drying.
Test method of the test method with embodiment 1, the present embodiment test result are as follows: digest the polypeptide yield of preparation as essence
23.5%, the DPPH Scavenging activity of coconut dregs of rice silty amount is 82.6%, and hydroxyl radical free radical Scavenging activity is 64.3%, and molecular weight is
2673Da。
Embodiment 15
After the smart coconut dregs of rice powder of 1g is mixed with distilled water according to mass ratio 1:20, the PH to 9 of mixed liquor is adjusted, essence is added
The alkali protease of coconut dregs of rice silty amount 2%-pawpaw property protease-neutral proteinase, alkali protease-pawpaw property protease-
Neutral proteinase ratio is 1:0.5:2,4h is stirred to react under temperature 60 C, enzyme deactivation 10min, enzymolysis liquid is existed at 90 DEG C
It is centrifugated 10min under 8000rpm, takes supernatant to carry out separating-purifying with ultrafiltration membrane (molecule interception is 4000Da), obtains anti-
Oxidation activity coconut polypeptide solution obtains inoxidizability coconut polypeptide after freeze-drying.
Test method of the test method with embodiment 1, the present embodiment test result are as follows: digest the polypeptide yield of preparation as essence
22.6%, the DPPH Scavenging activity of coconut dregs of rice silty amount is 76.2%, and hydroxyl radical free radical Scavenging activity is 71.6%, and molecular weight is
2749Da。
Comparative example 1
After the smart coconut dregs of rice powder of 1g is mixed with distilled water according to mass ratio 1:20, the PH to 9 of mixed liquor is adjusted, essence is added
The alkali protease of coconut dregs of rice silty amount 2%, is stirred to react 1h, enzyme deactivation 10min, enzymolysis liquid is existed at 90 DEG C at 55 DEG C of temperature
It is centrifugated 10min under 8000rpm, takes supernatant to carry out separating-purifying with ultrafiltration membrane (molecule interception is 4000Da), obtains anti-
Oxidation activity coconut polypeptide solution obtains inoxidizability coconut polypeptide after freeze-drying.
Test method of the test method with embodiment 1, the present embodiment test result are as follows: digest the polypeptide yield of preparation as essence
5.3%, the DPPH Scavenging activity of coconut dregs of rice silty amount is 40.6%, and hydroxyl radical free radical Scavenging activity is 38.3%, and molecular weight is
3379Da。
Comparative example 2
After the smart coconut dregs of rice powder of 1g is mixed with distilled water according to mass ratio 1:20, the PH to 9 of mixed liquor is adjusted, essence is added
The alkali protease of coconut dregs of rice silty amount 2%, is stirred to react 2h, enzyme deactivation 10min, enzymolysis liquid is existed at 90 DEG C at 55 DEG C of temperature
It is centrifugated 10min under 8000rpm, takes supernatant to carry out separating-purifying with ultrafiltration membrane (molecule interception is 4000Da), obtains anti-
Oxidation activity coconut polypeptide solution obtains inoxidizability coconut polypeptide after freeze-drying.
Test method of the test method with embodiment 1, the present embodiment test result are as follows: digest the polypeptide yield of preparation as essence
8.1%, the DPPH Scavenging activity of coconut dregs of rice silty amount is 51.4%, and hydroxyl radical free radical Scavenging activity is 45.1%, and molecular weight is
3182Da。
Comparative example 3
After the smart coconut dregs of rice powder of 1g is mixed with distilled water according to mass ratio 1:20, the PH to 9 of mixed liquor is adjusted, essence is added
The alkali protease of coconut dregs of rice silty amount 2%, is stirred to react 4h, enzyme deactivation 10min, enzymolysis liquid is existed at 90 DEG C at 55 DEG C of temperature
It is centrifugated 10min under 8000rpm, takes supernatant to carry out separating-purifying with ultrafiltration membrane (molecule interception is 4000Da), obtains anti-
Oxidation activity coconut polypeptide solution obtains inoxidizability coconut polypeptide after freeze-drying.
Test method of the test method with embodiment 1, the present embodiment test result are as follows: digest the polypeptide yield of preparation as essence
8.7%, the DPPH Scavenging activity of coconut dregs of rice silty amount is 56.2%, and hydroxyl radical free radical Scavenging activity is 49.6%, and molecular weight is
3041Da。
The above embodiment is a preferred embodiment of the present invention, but embodiments of the present invention are not by above-described embodiment
Limitation, other any changes, modifications, substitutions, combinations, simplifications made without departing from the spirit and principles of the present invention,
It should be equivalent substitute mode, be included within the scope of the present invention.
Claims (10)
1. a kind of preparation method of inoxidizability coconut polypeptide, which comprises the steps of:
(1) the coconut dregs of rice are preprocessed, obtain smart coconut dregs of rice powder；
(2) after mixing smart coconut dregs of rice powder according to mass ratio 1:15~25 with distilled water, the pH to 6~9 of mixed liquor is adjusted, according to
2~3% addition compound proteases of smart coconut dregs of rice silty amount carry out 1~4h of enzyme digestion reaction under the conditions of 50~60 DEG C, obtain enzyme
Solve liquid；
The compound protease include in alkali protease, neutral proteinase, bromelain and papain at least
Two kinds；
(3) enzymolysis liquid that step (2) obtains is living through enzyme deactivation, is centrifugated, and supernatant uses molecule interception for 5000Da or less
Ultrafiltration membrane ultra-filtration and separation, filtrate is freeze-dried to obtain inoxidizability coconut polypeptide.
2. the preparation method of inoxidizability coconut polypeptide according to claim 1, it is characterised in that:
Smart coconut dregs of rice powder described in step (1) is mixed with distilled water according to mass ratio 1:20；
The dosage of compound protease described in step (1) is that compound protease is added according to the 2% of smart coconut dregs of rice silty amount.
3. the preparation method of inoxidizability coconut polypeptide according to claim 1 or 2, it is characterised in that:
Compound protease described in step (1) include alkali protease-neutral proteinase, alkali protease-bromelain,
Neutral proteinase-papain, alkali protease-papain, neutrality -- proteases Bromelain, Papain
Enzyme-bromelain, alkali protease-papain-bromelain, alkali protease-pawpaw property protease-neutrality egg
White enzyme.
4. the preparation method of inoxidizability coconut polypeptide according to claim 3, it is characterised in that:
Compound protease described in step (1) include by quality ratio, alkali protease-neutral proteinase=1:0.5~2,
Alkali protease-bromelain=1:0.2~0.8, neutral proteinase-papain=1:0.25~1, basic protein
Enzyme-papain=1:0.25~1, neutral proteinase-bromelain=1:0.2~0.8, papain-pineapple egg
White enzyme=1:0.4~1.6, alkali protease-papain-bromelain=1:0.5:0.2~0.8, alkali protease-
Papain-neutral proteinase=1:0.5:0.5~2.
5. the preparation method of inoxidizability coconut polypeptide according to claim 1 or 2, it is characterised in that:
The preparation method of smart coconut dregs of rice powder, includes the following steps: described in step (1)
The coconut dregs of rice are added according to mass volume ratio 1:20 into ethyl alcohol, 130 DEG C of reflux 3h, separation is dried to obtain the degreasing coconut dregs of rice
Powder；Then degreasing coconut dregs of rice powder and distilled water is added by solid-liquid ratio 1:25, adjusts PH to 9, the lower 50 DEG C of processing 2h of high-speed stirred, from
The heart takes supernatant, then adjusts PH to 3.5~3.6, precipitating is collected by centrifugation, is freeze-dried after being washed to neutrality, obtains the smart coconut dregs of rice
Powder.
6. the preparation method of inoxidizability coconut polypeptide according to claim 1 or 2, it is characterised in that:
Alkali protease enzyme activity >=the 200000U/g, neutral proteinase enzyme activity are >=200000U/g, bromelain
Enzyme enzyme activity >=500000U/g, papain enzyme activity >=400000U/g.
7. the preparation method of inoxidizability coconut polypeptide according to claim 1 or 2, it is characterised in that:
The condition living of enzyme deactivation described in step (3) is 5~10min of enzyme deactivation at 90~95 DEG C；
The condition of centrifugation described in step (3) is that 5~10min is centrifugated under 8000~10000rpm.
8. the preparation method of inoxidizability coconut polypeptide according to claim 1 or 2, it is characterised in that:
In step (3), use molecule interception for the ultrafiltration membrane ultra-filtration and separation of 4000Da, polypeptide yield is smart coconut dregs of rice silty amount
9.6~25.8%, polypeptide molecular weight be 2547~3126Da；
The DPPH Scavenging activity of prepared coconut polypeptide be 57.3~84.5%, hydroxyl radical free radical clearance rate be 53.5~
74.2%.
9. a kind of inoxidizability coconut polypeptide, it is characterised in that: pass through preparation method system according to any one of claims 1 to 8
It is standby to obtain.
10. application of the inoxidizability coconut polypeptide as claimed in claim 9 as food or the antioxidant of cosmetics.
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| CN112280625A (en) * | 2020-11-04 | 2021-01-29 | 齐鲁工业大学 | Coconut flavor Lag beer and preparation method thereof |
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Application publication date: 20190426 |