CN109055466A - A kind of preparation method of pea peptide and its application in whitening field - Google Patents
A kind of preparation method of pea peptide and its application in whitening field Download PDFInfo
- Publication number
- CN109055466A CN109055466A CN201810852566.5A CN201810852566A CN109055466A CN 109055466 A CN109055466 A CN 109055466A CN 201810852566 A CN201810852566 A CN 201810852566A CN 109055466 A CN109055466 A CN 109055466A
- Authority
- CN
- China
- Prior art keywords
- pea
- peptide
- whitening
- preparation
- pea peptide
- 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
- 108090000765 processed proteins & peptides Proteins 0.000 title claims abstract description 32
- 230000002087 whitening effect Effects 0.000 title claims abstract description 20
- 238000002360 preparation method Methods 0.000 title claims abstract description 11
- 230000000694 effects Effects 0.000 claims abstract description 16
- 238000006243 chemical reaction Methods 0.000 claims abstract description 15
- 102000004169 proteins and genes Human genes 0.000 claims abstract description 14
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 14
- 239000000843 powder Substances 0.000 claims abstract description 10
- 239000002537 cosmetic Substances 0.000 claims abstract description 8
- 238000001976 enzyme digestion Methods 0.000 claims abstract description 8
- 230000002401 inhibitory effect Effects 0.000 claims abstract description 8
- 150000001875 compounds Chemical class 0.000 claims abstract description 6
- 235000013376 functional food Nutrition 0.000 claims abstract description 6
- 235000013305 food Nutrition 0.000 claims abstract description 5
- 108091005804 Peptidases Proteins 0.000 claims abstract description 4
- 239000004365 Protease Substances 0.000 claims abstract description 4
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 claims abstract description 4
- 102000004190 Enzymes Human genes 0.000 claims description 19
- 108090000790 Enzymes Proteins 0.000 claims description 19
- WTDRDQBEARUVNC-LURJTMIESA-N L-DOPA Chemical compound OC(=O)[C@@H](N)CC1=CC=C(O)C(O)=C1 WTDRDQBEARUVNC-LURJTMIESA-N 0.000 claims description 17
- 108010084695 Pea Proteins Proteins 0.000 claims description 16
- 235000019702 pea protein Nutrition 0.000 claims description 16
- WTDRDQBEARUVNC-UHFFFAOYSA-N L-Dopa Natural products OC(=O)C(N)CC1=CC=C(O)C(O)=C1 WTDRDQBEARUVNC-UHFFFAOYSA-N 0.000 claims description 14
- 239000007788 liquid Substances 0.000 claims description 14
- 235000018102 proteins Nutrition 0.000 claims description 13
- 102000003425 Tyrosinase Human genes 0.000 claims description 12
- 108060008724 Tyrosinase Proteins 0.000 claims description 12
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 8
- 238000001816 cooling Methods 0.000 claims description 4
- 238000001125 extrusion Methods 0.000 claims description 4
- 238000001035 drying Methods 0.000 claims description 3
- 206010040829 Skin discolouration Diseases 0.000 claims description 2
- 235000005072 Vigna sesquipedalis Nutrition 0.000 claims description 2
- 244000090207 Vigna sesquipedalis Species 0.000 claims description 2
- 238000002835 absorbance Methods 0.000 claims description 2
- 235000021278 navy bean Nutrition 0.000 claims description 2
- 239000000758 substrate Substances 0.000 claims description 2
- 230000007062 hydrolysis Effects 0.000 claims 1
- 238000006460 hydrolysis reaction Methods 0.000 claims 1
- 239000003112 inhibitor Substances 0.000 description 13
- 239000000047 product Substances 0.000 description 10
- 230000007071 enzymatic hydrolysis Effects 0.000 description 9
- 238000006047 enzymatic hydrolysis reaction Methods 0.000 description 9
- 238000012360 testing method Methods 0.000 description 8
- 238000006911 enzymatic reaction Methods 0.000 description 6
- 238000005259 measurement Methods 0.000 description 5
- 102000004196 processed proteins & peptides Human genes 0.000 description 5
- AHMIDUVKSGCHAU-UHFFFAOYSA-N Dopaquinone Natural products OC(=O)C(N)CC1=CC(=O)C(=O)C=C1 AHMIDUVKSGCHAU-UHFFFAOYSA-N 0.000 description 4
- AHMIDUVKSGCHAU-LURJTMIESA-N L-dopaquinone Chemical compound [O-]C(=O)[C@@H]([NH3+])CC1=CC(=O)C(=O)C=C1 AHMIDUVKSGCHAU-LURJTMIESA-N 0.000 description 4
- 230000008859 change Effects 0.000 description 4
- MVORZMQFXBLMHM-QWRGUYRKSA-N Gly-His-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)CN)CC1=CN=CN1 MVORZMQFXBLMHM-QWRGUYRKSA-N 0.000 description 3
- 239000012141 concentrate Substances 0.000 description 3
- 238000000502 dialysis Methods 0.000 description 3
- 239000012153 distilled water Substances 0.000 description 3
- 108010038983 glycyl-histidyl-lysine Proteins 0.000 description 3
- 238000000034 method Methods 0.000 description 3
- XKZQKPRCPNGNFR-UHFFFAOYSA-N 2-(3-hydroxyphenyl)phenol Chemical compound OC1=CC=CC(C=2C(=CC=CC=2)O)=C1 XKZQKPRCPNGNFR-UHFFFAOYSA-N 0.000 description 2
- XUMBMVFBXHLACL-UHFFFAOYSA-N Melanin Chemical compound O=C1C(=O)C(C2=CNC3=C(C(C(=O)C4=C32)=O)C)=C2C4=CNC2=C1C XUMBMVFBXHLACL-UHFFFAOYSA-N 0.000 description 2
- 229920002472 Starch Polymers 0.000 description 2
- 238000010521 absorption reaction Methods 0.000 description 2
- 239000003963 antioxidant agent Substances 0.000 description 2
- 230000003078 antioxidant effect Effects 0.000 description 2
- 235000006708 antioxidants Nutrition 0.000 description 2
- 230000009849 deactivation Effects 0.000 description 2
- 238000007254 oxidation reaction Methods 0.000 description 2
- 239000008363 phosphate buffer Substances 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 235000019698 starch Nutrition 0.000 description 2
- 239000008107 starch Substances 0.000 description 2
- 239000013589 supplement Substances 0.000 description 2
- 230000001629 suppression Effects 0.000 description 2
- 241000208340 Araliaceae Species 0.000 description 1
- 208000024172 Cardiovascular disease Diseases 0.000 description 1
- 206010061218 Inflammation Diseases 0.000 description 1
- 235000005035 Panax pseudoginseng ssp. pseudoginseng Nutrition 0.000 description 1
- 235000003140 Panax quinquefolius Nutrition 0.000 description 1
- 235000010627 Phaseolus vulgaris Nutrition 0.000 description 1
- 244000046052 Phaseolus vulgaris Species 0.000 description 1
- 230000002292 Radical scavenging effect Effects 0.000 description 1
- 230000001153 anti-wrinkle effect Effects 0.000 description 1
- 230000036772 blood pressure Effects 0.000 description 1
- 238000006555 catalytic reaction Methods 0.000 description 1
- 125000002091 cationic group Chemical group 0.000 description 1
- 235000010980 cellulose Nutrition 0.000 description 1
- 229920002678 cellulose Polymers 0.000 description 1
- 238000004891 communication Methods 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- MHUWZNTUIIFHAS-CLFAGFIQSA-N dioleoyl phosphatidic acid Chemical compound CCCCCCCC\C=C/CCCCCCCC(=O)OCC(COP(O)(O)=O)OC(=O)CCCCCCC\C=C/CCCCCCCC MHUWZNTUIIFHAS-CLFAGFIQSA-N 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 230000002500 effect on skin Effects 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- 230000005714 functional activity Effects 0.000 description 1
- 235000008434 ginseng Nutrition 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 230000004957 immunoregulator effect Effects 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 230000004054 inflammatory process Effects 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 229960004502 levodopa Drugs 0.000 description 1
- 230000031700 light absorption Effects 0.000 description 1
- 238000013507 mapping Methods 0.000 description 1
- 239000008055 phosphate buffer solution Substances 0.000 description 1
- 229920001184 polypeptide Polymers 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 230000036559 skin health Effects 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 230000036967 uncompetitive effect Effects 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
- 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
- A23L33/185—Vegetable proteins
-
- 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
- A61Q17/00—Barrier preparations; Preparations brought into direct contact with the skin for affording protection against external influences, e.g. sunlight, X-rays or other harmful rays, corrosive materials, bacteria or insect stings
- A61Q17/04—Topical preparations for affording protection against sunlight or other radiation; Topical sun tanning preparations
-
- 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
- A61Q19/02—Preparations for care of the skin for chemically bleaching or whitening the skin
-
- 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
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/74—Biological properties of particular ingredients
- A61K2800/78—Enzyme modulators, e.g. Enzyme agonists
- A61K2800/782—Enzyme inhibitors; Enzyme antagonists
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/80—Process related aspects concerning the preparation of the cosmetic composition or the storage or application thereof
- A61K2800/805—Corresponding aspects not provided for by any of codes A61K2800/81 - A61K2800/95
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/80—Process related aspects concerning the preparation of the cosmetic composition or the storage or application thereof
- A61K2800/84—Products or compounds obtained by lyophilisation, freeze-drying
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/80—Process related aspects concerning the preparation of the cosmetic composition or the storage or application thereof
- A61K2800/92—Oral administration
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Animal Behavior & Ethology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Dermatology (AREA)
- Molecular Biology (AREA)
- Nutrition Science (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Genetics & Genomics (AREA)
- Polymers & Plastics (AREA)
- Biochemistry (AREA)
- Food Science & Technology (AREA)
- Mycology (AREA)
- General Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Birds (AREA)
- Water Supply & Treatment (AREA)
- Biotechnology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Biophysics (AREA)
- Microbiology (AREA)
- Epidemiology (AREA)
- Medicinal Chemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- Gerontology & Geriatric Medicine (AREA)
- Cosmetics (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention discloses a kind of preparation method of pea peptide and its applications in whitening field, are related to functional food and cosmetic field.The present invention is by squeezing Fed Protein Powder of Pea Insteal, compound protease carries out enzyme digestion reaction to prepare pea peptide, shortens enzymolysis time, substantially increases enzymolysis efficiency.Our experiments show that pea peptide has good inhibiting effect, half-inhibitory concentration IC for tyrosinase50For 21.3 mg/mL.The pea peptide can be applied in whitening product and functional food field, such as is added in food or in cosmetics by a certain percentage, can reach the effect of whitening.
Description
Technical field
The invention belongs to the Related products such as cosmetics and functional food field, disclose a kind of pea peptide preparation method and
Its application in whitening field.
Background technique
With the continuous improvement of people's living standards, skin health problem obtains pay attention to day by day.Ultraviolet light in sunlight
It (UVB) is to cause dermal melanin calmness factor the most significant, skin darkening can be caused by being exposed under ultraviolet light irradiation for a long time,
So that influencing physically and mentally healthy people and human communication and relationship.Therefore, find a kind of high efficiency, low side effects of whitening medicaments at
Divide and has become an important and urgent task.
Pea (Pea) has very extensive plantation all over the world, is the main edible beans of people.As in the world most
One of big pea country of origin, China possess vast pea cultivated area, and total output also is located at forefront.In terms of ingredient, pea
Beans are rich in substances such as starch, protein, celluloses.Wherein in ripe seed respectively containing protein 21% ~ 28% and starch 48 ~
52%.It is international studies have shown that pea protein is by digesting pea protein peptide obtained has certain functional activity.Than
Such as, Ndiaye et al. research discovery pea protein enzymolysis liquid has anti-oxidant, reduction inflammation and immunoregulatory effect.And Huan
The research of Li et al. people then shows that the product after pea protein enzymatic hydrolysis is cationic polypeptide, which can effectively adjust Ca MK
The activity of II, thus can be used as the prevention aspect that health care product is applied to cardiovascular disease.Samson et al. then has found pea
Peptide can inhibit the activity of ACE enzyme, to play the role of blood pressure lowering.
Have no the pea peptide in the application report in whitening field through retrieval.
Summary of the invention
Application of the pea peptide in whitening product and functional food with whitening function that the present invention provides a kind of.
To solve the deficiencies in the prior art, there is whitening, sun-proof, anti-oxidant, anti-wrinkle, depth simultaneously the present invention provides a kind of
The skin-lightening cosmetic and whitening function food of the peptide containing pea of layer moisture-keeping efficacy.
The preparation method of pea peptide of the present invention:
The Fed Protein Powder of Pea Insteal that water content is 25% is squeezed in single-screw extrusion machine, extruding condition: 60 DEG C of barrel temperature,
170 r/min of screw speed, 12 mm of die hole aperture.Fed Protein Powder of Pea Insteal after extruding is crushed to 60 mesh through cooling.
Enzymolysis process are as follows: smashed Fed Protein Powder of Pea Insteal will be squeezed and be dissolved in the phosphate buffer solution that pH value is 6.5,
It is digested with compound protease again, obtains enzymolysis liquid.
Wherein:
Compound protease digests optimum condition: enzymolysis temperature is 40 ~ 42 DEG C, and the pH value of enzyme digestion reaction is 6.5 ~ 7.0, is added
Enzyme amount is 4.5 ~ 4.9%(E/S), and pea protein concentration is 12.5 ~ 14 %(g/mL in enzyme digestion reaction), enzymolysis time is 10 ~ 11
h。
After enzymatic hydrolysis, enzyme-removal temperature is 100 DEG C, and the enzyme deactivation time is 15 min.
Pea protein enzymolysis liquid is separated using bag filter: using the bag filter of 7 KDa, 3.5 KDa, 1 KDa, to enzymolysis liquid
Successively separate.Bag filter is pre-processed, about 2/3rds enzymolysis liquid, two end seals are added into the bag filter handled well
It is good, it is placed in distilled water and is stirred, change a water at regular intervals, there is no color changes up to peritoneal effluent, will appear
After liquid is concentrated with Rotary Evaporators, concentrate is freeze-dried, and obtains pea Gly-His-Lys.
Wherein, enzymolysis product obtains four fractions through dialysis: isolated less than 1 KDa, 1~3.5 KDa, 3.5~7
KDa and enzymolysis liquid greater than 74 different components of KDa.Measure its O2 .-Free radical scavenging activity is respectively 81.3%, 35.2%,
20.3%,12.4%.It was found that molecular weight is most strong less than the enzymolysis liquid activity of 1 KDa, molecular weight is had focused largely in 200~800 Da
Left and right illustrates that the peptide obtained after enzymatic hydrolysis is 2~8 peptides mostly.
The application of pea peptide of the present invention:
When the mass concentration of pea peptide is 25 ~ 30 mg/mL, inhibiting rate can reach 62.9 ~ 73.8%, half-inhibitory concentration IC50
For 21.3 mg/mL.
Pea peptide can be used in the application of whitening product functional food field, such as is added in food or changes by a certain percentage
In cosmetic, the effect of whitening can reach.
Specific embodiment
The present invention is further described by the following examples, but the protection that these embodiments are not intended to restrict the invention
Range and the scope of application.
Embodiment 1
2 kg Fed Protein Powder of Pea Insteal are weighed, moisture content 25% is squeezed in single-screw extrusion machine, and extruding condition is as follows: machine barrel temperature
60 DEG C, 170 r/min of screw speed, 12 mm of die hole aperture of degree.Fed Protein Powder of Pea Insteal after extruding is through cooling, drying, crushing, and 60
Mesh choosing, pack storage, referred to as pea protein extrudate.The pea protein extrudate sample of 500 g is weighed, compound protein is added
Enzyme enzymatic hydrolysis, enzymatic hydrolysis condition are as follows: enzymolysis temperature is 40 DEG C, and the pH value of enzyme digestion reaction is 6.5, and enzyme concentration is 4.5 %(E/S),
Pea protein concentration is 12.5%(g/mL in enzyme digestion reaction), enzymolysis time is 10 h.After enzymatic hydrolysis, enzyme-removal temperature is 100 DEG C,
The enzyme deactivation time is 15 min.
Pea protein enzymolysis liquid is separated using the bag filter of 1 KDa, bag filter is pre-processed, to the dialysis handled well
About 2/3rds enzymolysis liquid is added in bag, both ends seal, are placed in distilled water and are stirred, and change at regular intervals primary
Water, up to peritoneal effluent, there is no color changes, and after peritoneal effluent is concentrated with Rotary Evaporators, concentrate is freeze-dried, and obtain
To pea Gly-His-Lys.
Tyrosinase vigor is measured with being catalyzed the diphenol enzyme activity of L-3,4 dihydroxyphenylalanine oxidation reaction generation DOPA quinone.Add in test tube
Enter different amounts of L-3,4 dihydroxyphenylalanine (1.0 mg/mL), respectively 0.4,1,2 ml, the supplement phosphate buffer of pH=6.8 to 2.8 mL, 30
After DEG C water-bath keeps the temperature 10 min, 0.2 mL of tyrosinase (270 U/mL) is added and is uniformly mixed.The measurement of tyrosinase activity is former
Reason is to convert red product DOPA quinone for L-3,4 dihydroxyphenylalanine based on enzymatic reaction, and the product has absorption maximum at 475 nm.With
A per minute475Increasing by 0.001 is 1 enzyme activity unit, the speed of enzymatic reaction A per minute475Value added indicates.From mixed
Conjunction uniformly starts to measure, and measurement in every 30 seconds is primary, surveys until 15 min.Enzyme activity is calculated by following two formula and enzyme inhibits
Rate.In triplicate, mean value is calculated.
In formula: A1There is A when L-3,4 dihydroxyphenylalanine for no inhibitor475；A2A when for no inhibitor without L-3,4 dihydroxyphenylalanine475；A3To there is suppression
A when preparation and L-3,4 dihydroxyphenylalanine475；A4To there is A of the inhibitor without L-3,4 dihydroxyphenylalanine when475。
According to above-mentioned tyrosinase vigour-testing method, different amounts of pea peptide, respectively 5,10,15,20,25 are added
Mg/mL measures the absorbance change of this reaction, measures since being uniformly mixed, and measurement in every 30 seconds is primary, measures 10 min, counts
Calculate its inhibiting rate.
When the mass concentration of pea peptide is 25 mg/mL, inhibiting rate can reach 62.9%, half-inhibitory concentration IC50For
21.3 mg/mL。
Embodiment 2
2 kg Fed Protein Powder of Pea Insteal are weighed, moisture content 25% is squeezed in single-screw extrusion machine, and extruding condition is as follows: machine barrel temperature
60 DEG C, 170 r/min of screw speed, 12 mm of die hole aperture of degree.Fed Protein Powder of Pea Insteal after extruding is through cooling, drying, crushing, and 60
Mesh choosing, pack storage, referred to as pea protein extrudate.The pea protein extrudate sample of 500 g is weighed, compound protein is added
Enzyme enzymatic hydrolysis, enzymatic hydrolysis condition are as follows: enzymolysis temperature is 42 DEG C, and the pH value of enzyme digestion reaction is 7.0, and enzyme concentration is 4.9 %(E/S),
Pea protein concentration is 14%(g/mL in enzyme digestion reaction), enzymolysis time is 11 h.After enzymatic hydrolysis, enzyme-removal temperature is 100 DEG C, is gone out
The enzyme time is 15 min.
Pea protein enzymolysis liquid is separated using the bag filter of 1 KDa, bag filter is pre-processed, to the dialysis handled well
About 2/3rds enzymolysis liquid is added in bag, both ends seal, are placed in distilled water and are stirred, and change at regular intervals primary
Water, up to peritoneal effluent, there is no color changes, and after peritoneal effluent is concentrated with Rotary Evaporators, concentrate is freeze-dried, and obtain
To pea Gly-His-Lys.
Tyrosinase vigor is measured with being catalyzed the diphenol enzyme activity of L-3,4 dihydroxyphenylalanine oxidation reaction generation DOPA quinone.Add in test tube
Enter different amounts of L-3,4 dihydroxyphenylalanine (1.0 mg/mL), respectively 0.4,1,2 mL, the supplement phosphate buffer of pH=6.8 to 2.8 mL, 30
After DEG C water-bath keeps the temperature 10 min, 0.2 mL of tyrosinase (270 U/mL) is added and is uniformly mixed.The measurement of tyrosinase activity is former
Reason is to convert red product DOPA quinone for L-3,4 dihydroxyphenylalanine based on enzymatic reaction, and the product has absorption maximum at 475 nm.With
A per minute475Increasing by 0.001 is 1 enzyme activity unit, the speed of enzymatic reaction A per minute475Value added indicates.From mixed
Conjunction uniformly starts to measure, and measurement in every 30 seconds is primary, surveys until 15 min.Enzyme activity is calculated by following two formula and enzyme inhibits
Rate.In triplicate, mean value is calculated.
In formula: A1There is A when L-3,4 dihydroxyphenylalanine for no inhibitor475；A2A when for no inhibitor without L-3,4 dihydroxyphenylalanine475；A3To there is suppression
A when preparation and L-3,4 dihydroxyphenylalanine475；A4To there is A of the inhibitor without L-3,4 dihydroxyphenylalanine when475。
According to above-mentioned tyrosinase vigour-testing method, the concentration (0,15,25 mg/mL) of fixed inhibitor measures L-
The light absorption value of enzyme reaction, i.e. enzymatic when DOPA concentration of substrate (0.133,0.200,0.267,0.333,0.400 mg/mL) is different
The remnant enzyme activity of reaction finds out corresponding reaction rateA/min calculates mean value in triplicate.By Lineweaver-Burk
Double-reciprocal plot method mapping, with L-3,4 dihydroxyphenylalanine concentration inverse for abscissa, determine the type of enzymatic reaction inhibiting effect, obtain
Michaelis constant KmWith maximum reaction velocity Vmax。
Tyrosinase catalysis L-DOPA is to follow Michaelis-Menten dynamics, passes through analytic dynamics curve and ginseng
Number, including apparent K_m (Km) and maximum reaction velocity (Vm) variation judge the inhibiting effect type of pea peptide.This two
Straight line is parallel to each other, wherein not plus the test group kinetic parameter K of pea inhibitor peptidesm=0.095 mg/mL, Vmax=0.036A/min.The test group kinetic parameter K of pea inhibitor peptides (15 mg/mL) is addedm=0.107 mg/mL, Vmax =0.040.The test group kinetic parameter K of pea inhibitor peptides (25 mg/mL)m=0.115 mg/mL, Vmax =0.044.The results show that the test group K of inhibitor is added compared to control groupmValue reduces, VmaxReduce, KmAnd VmaxRatio
It is constant, show that pea inhibitor peptides are uncompetitive inhibitor to the inhibiting effect type of tyrosinase.
Claims (3)
1. a kind of preparation method of pea peptide and its application in whitening field, it is characterised in that: pea peptide is added 2.5%
It is mixed evenly into food or cosmetics, functional food or cosmetics with whitening function can be obtained.
2. a kind of preparation method of pea peptide as described in claim 1 and its application in whitening field, which is characterized in that
Pea peptide the preparation method comprises the following steps:
(1) Fed Protein Powder of Pea Insteal that water content is 25% is squeezed in single-screw extrusion machine, extruding condition: barrel temperature 60
DEG C, 170 r/min of screw speed, 12 mm of die hole aperture；
(2) Fed Protein Powder of Pea Insteal after extruding is through cooling, drying, crushing, and the choosing of 60 meshes, pack storage is pea protein extrudate；
Using compound protease in pH value 6.5 ~ 7.0,40 ~ 42 DEG C of temperature, 10 ~ 11 h of hydrolysis time, concentration of substrate 12.5 ~
14%, enzyme concentration is 4.5 ~ 4.9% progress enzyme digestion reactions to prepare pea protein enzymolysis liquid；
(3) bag filter for using 1 KDa, separates enzymolysis liquid, peritoneal effluent is concentrated with Rotary Evaporators, is lyophilized, obtains pea
Beans peptide；
(4) tyrosinase and L-3,4 dihydroxyphenylalanine are reacted in the presence of various concentration pea peptide, after 30 DEG C of water-baths keep the temperature 10 min,
And measure the absorbance of product after the reaction of the two；
(5) inhibiting rate is calculated by the two reaction rate.
3. a kind of preparation method of pea peptide as described in claim 1 and its application in whitening field, it is characterised in that:
The whitening function food or skin-lightening cosmetic are to carry out whitening as approach to reduce tyrosinase activity.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810852566.5A CN109055466A (en) | 2018-07-30 | 2018-07-30 | A kind of preparation method of pea peptide and its application in whitening field |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810852566.5A CN109055466A (en) | 2018-07-30 | 2018-07-30 | A kind of preparation method of pea peptide and its application in whitening field |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN109055466A true CN109055466A (en) | 2018-12-21 |
Family
ID=64831560
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201810852566.5A Pending CN109055466A (en) | 2018-07-30 | 2018-07-30 | A kind of preparation method of pea peptide and its application in whitening field |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN109055466A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112961892A (en) * | 2021-02-25 | 2021-06-15 | 厦门元之道生物科技有限公司 | Pea polypeptide and preparation method thereof |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101974593A (en) * | 2010-11-11 | 2011-02-16 | 湖北远成药业有限公司 | Method for producing pea peptide through enzyme method |
| CN103436580A (en) * | 2013-08-27 | 2013-12-11 | 山东天久生物技术有限公司 | Preparation method of pea polypeptide |
| CN106260498A (en) * | 2016-08-08 | 2017-01-04 | 山东理工大学 | The manufacture method of a kind of Semen Pisi sativi protein peptide and application |
| CN107668314A (en) * | 2017-11-02 | 2018-02-09 | 林峰 | A kind of industrialized production pea active peptide and preparation method |
| CN107890436A (en) * | 2017-12-04 | 2018-04-10 | 山东理工大学 | A kind of cosmetics for whitening antioxidation |
-
2018
- 2018-07-30 CN CN201810852566.5A patent/CN109055466A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101974593A (en) * | 2010-11-11 | 2011-02-16 | 湖北远成药业有限公司 | Method for producing pea peptide through enzyme method |
| CN103436580A (en) * | 2013-08-27 | 2013-12-11 | 山东天久生物技术有限公司 | Preparation method of pea polypeptide |
| CN106260498A (en) * | 2016-08-08 | 2017-01-04 | 山东理工大学 | The manufacture method of a kind of Semen Pisi sativi protein peptide and application |
| CN107668314A (en) * | 2017-11-02 | 2018-02-09 | 林峰 | A kind of industrialized production pea active peptide and preparation method |
| CN107890436A (en) * | 2017-12-04 | 2018-04-10 | 山东理工大学 | A kind of cosmetics for whitening antioxidation |
Non-Patent Citations (2)
| Title |
|---|
| 张欣: "甘草提取物与水解豌豆蛋白在乳化体系中的协同抗氧化性研究", 《中国博士学位论文全文数据库电子期刊 (工程科技Ⅰ辑)》 * |
| 张欣等: "加热和碱性处理豌豆蛋白对其不同酶水解物抗氧化性的影响", 《食品科学》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112961892A (en) * | 2021-02-25 | 2021-06-15 | 厦门元之道生物科技有限公司 | Pea polypeptide and preparation method thereof |
| CN112961892B (en) * | 2021-02-25 | 2022-12-02 | 厦门元之道生物科技有限公司 | Pea polypeptide and preparation method thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| JP4658348B2 (en) | Collagen production promoter, collagenase inhibitor, fibroblast proliferation agent, elastase inhibitor, estrogen-like agent, and skin cosmetics | |
| KR101936294B1 (en) | Composition comprising for skin-whitening and anti-wrinkling extract of Rumex acetosella L. or extract of Hydrangea serrata | |
| CN106399437B (en) | Preparation method of small water turtle polypeptide and small water turtle polypeptide mask | |
| CN105219586A (en) | A kind of fermented type gas water beverage and preparation method thereof | |
| KR102314654B1 (en) | Composition for skin wrinkle improvement or anti-inflammation comprising extract of fermented Centella asiatica with improved antioxidant activity as effective component | |
| CN111000770A (en) | Whitening cream containing pearl peptide | |
| CN110236981A (en) | A kind of whitening plant polyose composition and its application | |
| KR20160116800A (en) | Composition for moisturizing skin | |
| CN114134088B (en) | Choerospondias axillaris fermentation product and preparation method and application thereof | |
| CN111529474A (en) | Multi-effect composition for moisturizing, whitening and antioxidant repair and preparation method and application thereof | |
| KR101458274B1 (en) | Cosmetic Composition Comprising Fermented Ginseng Extracts | |
| CN109055466A (en) | A kind of preparation method of pea peptide and its application in whitening field | |
| KR102544792B1 (en) | Composition Comprising Complex Extract of seaweeds Fermented by Lactobacillus Brevis with Anti-Wrinkling and Elasticity, Antioxidant, and Skin Hydration Property as Active Ingredient | |
| JP2011055837A (en) | Food and drink for beautification | |
| LU501500B1 (en) | Preparation method of pea peptide and application thereof in field of whitening | |
| JP2007269659A (en) | Antioxidant composition | |
| KR101063351B1 (en) | The extracts and fractions of Hippophae rhamnoides L. | |
| KR102599579B1 (en) | Nanorization-treated fermented Hibiscus manihot with enhanced collagen, amino acid, polyphenol or flavonoid content and manufacturing method thereof | |
| CN113116757B (en) | Collagenase inhibitor, moisturizing mask containing collagenase inhibitor and preparation method of moisturizing mask | |
| CN108404108A (en) | A kind of compound maize oligopeptide capsule and preparation method thereof | |
| KR20090085876A (en) | The extracts and fractions of hippophae rhamnoides l | |
| JP5479754B2 (en) | Powdered composition and tablet, and method for preventing browning of γ-aminobutyric acid and browning inhibitor | |
| KR102331119B1 (en) | Composition for preventing skin anti-aging comprising fermented black rice extract and fermented peanut sprout extract | |
| JP4703829B2 (en) | Prophylactic / therapeutic agent for inflammatory diseases | |
| KR20130136048A (en) | Compositions for improving skin aging comprising fucosterol |
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 | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20181221 |