CN116602910B - Oligopeptide composition and application thereof in preparation of skin repair and anti-aging products - Google Patents
Oligopeptide composition and application thereof in preparation of skin repair and anti-aging products Download PDFInfo
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- WSGCRSMLXFHGRM-DEVHWETNSA-N (2s)-2-[[(2s)-6-amino-2-[[(2s,3r)-2-[[(2s,3r)-2-[[(2s)-6-amino-2-(hexadecanoylamino)hexanoyl]amino]-3-hydroxybutanoyl]amino]-3-hydroxybutanoyl]amino]hexanoyl]amino]-3-hydroxypropanoic acid Chemical compound CCCCCCCCCCCCCCCC(=O)N[C@@H](CCCCN)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CO)C(O)=O WSGCRSMLXFHGRM-DEVHWETNSA-N 0.000 claims abstract description 23
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- 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
- C12P1/00—Preparation of compounds or compositions, not provided for in groups C12P3/00 - C12P39/00, by using microorganisms or enzymes
- C12P1/04—Preparation of compounds or compositions, not provided for in groups C12P3/00 - C12P39/00, by using microorganisms or enzymes by using bacteria
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- 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
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- 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
- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/99—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from microorganisms other than algae or fungi, e.g. protozoa or bacteria
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
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- 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
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
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- A61K2800/40—Chemical, physico-chemical or functional or structural properties of particular ingredients
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- A61K2800/592—Mixtures of compounds complementing their respective functions
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- 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
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- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
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Abstract
The invention discloses an oligopeptide composition and application thereof in preparing skin repair and anti-aging products. The oligopeptide composition comprises 20-300 parts of lactobacillus fermentation product filtrate, 10-100 parts of oligopeptide-1, 10-100 parts of palmitoyl pentapeptide-4 and 10-100 parts of acetyl tetrapeptide-9; the lactobacillus fermentation product filtrate is prepared by activating lactobacillus, inoculating the lactobacillus into a seed culture medium for culture to obtain seed liquid, and then inoculating the seed liquid into a fermentation culture medium for culture; the seed culture medium comprises 1.35-6.25 g/L of acanthopanax polysaccharide; the fermentation medium comprises 1.35-6.25 g/L of acanthopanax polysaccharide and 1.35-5.55 g/L of lycium barbarum polysaccharide. According to the embodiment of the application, the culture medium containing acanthopanax polysaccharide and wolfberry polysaccharide is used as a raw material, lactobacillus paracasei is used for fermenting the culture medium, and the fermentation product is compounded with oligopeptide-1, palmitoyl pentapeptide-4 and acetyl tetrapeptide-9 to obtain the oligopeptide composition, so that the oligopeptide composition has the remarkable effects of resisting oxidation, inhibiting tyrosine, promoting collagen regeneration and the like due to the synergistic effect of the components.
Description
Technical Field
The invention relates to the field of daily chemicals and skin care products, in particular to an oligopeptide composition and application thereof in preparing skin repair and anti-aging products.
Background
Skin aging generally has two manifestations of natural aging and photoaging. The former mainly refers to aging caused by nonresistant factors such as inheritance, gravity, endocrine, immune functions and the like existing in a organism with the increase of age; the latter is mainly due to the direct effect of a series of environmental exposures such as ultraviolet light, climate change and environmental pollution on the photoaging of the skin. Based on the aging mechanism of skin, the anti-aging pathway is discussed, mainly including: protecting the skin from external environmental stimuli; removing redundant free radicals in cells; repairing and supplementing skin cells. The anti-aging cosmetic is one of cosmetics for realizing anti-aging effect
The company finds the break from the formulation by combining years of research and development experience of cosmetic formulations, particularly the advantages in the anti-aging field of cosmetics, finds the application of polypeptides such as oligopeptides in the aspects of preventing aging, slowing down aging, smoothing wrinkles and the like, and has a special magic effect; meanwhile, the development of the raw materials of the fermentation product with the effects of resisting aging and repairing skin is researched and developed by experts in the fields of biological engineering of company tissues and the like, and the raw materials are compounded, so that the oligopeptide composition with the effects of promoting skin repair and resisting aging is finally developed.
Disclosure of Invention
The invention aims to overcome the defects of the prior art and provide an oligopeptide composition and application thereof in preparing skin repair and anti-aging products so as to solve the problems in the technical background.
In order to achieve the above object, the present invention is realized by the following technical scheme:
in a first aspect, the invention provides an oligopeptide composition, which comprises 20-300 parts of lactobacillus fermentation product filtrate, 10-100 parts of oligopeptide-1, 10-100 parts of palmitoyl pentapeptide-4 and 10-100 parts of acetyl tetrapeptide-9;
the lactobacillus fermentation product filtrate is prepared by activating lactobacillus, inoculating the lactobacillus into a seed culture medium for culture to obtain seed liquid, and then inoculating the seed liquid into a fermentation culture medium for culture.
As a further embodiment, the oligopeptide composition comprises 150 parts lactobacillus fermentation product filtrate, 60 parts oligopeptide-1, 60 parts palmitoyl pentapeptide-4, 60 parts acetyl tetrapeptide-9.
As a further scheme, the seed culture medium comprises 1.35-6.25 g/L of acanthopanax polysaccharide, 10g/L of peptone, 10g/L of beef extract and 5g/L, KH of yeast extract 2 PO 4 2g/L, trisodium citrate 2g/L, sodium acetate 2g/L, glucose 20g/L, tween, 1mL/L, mgSO 4 ·7H 2 O 0.58g/L、MnSO 4 ·4H 2 O 0.25g/L。
As a further scheme, the fermentation medium comprises 1.35-6.25 g/L of acanthopanax polysaccharide, 1.35-5.55 g/L of lycium polysaccharide, 10g/L of peptone, 10g/L of beef extract and 5g/L, KH of yeast extract 2 PO 4 2g/L, trisodium citrate 2g/L, sodium acetate 2g/L, glucose 20g/L, tween, 1mL/L, mgSO 4 ·7H 2 O 0.58g/L、MnSO 4 ·4H 2 O 0.25g/L。
As a further scheme, the seed culture medium comprises 3.55g/L of acanthopanax polysaccharide, 10g/L of peptone, 10g/L of beef extract and 5g/L, KH of yeast extract 2 PO 4 2g/L, trisodium citrate 2g/L, sodium acetate 2g/L, glucose 20g/L, tween, 1mL/L, mgSO 4 ·7H 2 O 0.58g/L、MnSO 4 ·4H 2 O0.25 g/L; the fermentation medium comprises 3.55g/L of acanthopanax polysaccharide, 2.75g/L of wolfberry polysaccharide, 10g/L of peptone, 10g/L of beef extract and 5g/L, KH of yeast extract 2 PO 4 2g/L, trisodium citrate 2g/L, sodium acetate 2g/L, glucose 20g/L, tween, 1mL/L, mgSO 4 ·7H 2 O 0.58g/L、MnSO 4 ·4H 2 O 0.25g/L。
In a second aspect, the invention provides the use of the oligopeptide composition for preparing a skin repair and anti-aging product.
Compared with the prior art, the invention has the beneficial effects that:
1. according to the embodiment of the application, the culture medium containing acanthopanax polysaccharide and wolfberry polysaccharide is used as a raw material, lactobacillus paracasei is used for fermenting the culture medium, and the fermentation product is compounded with oligopeptide-1, palmitoyl pentapeptide-4 and acetyl tetrapeptide-9 to obtain the oligopeptide composition, so that the oligopeptide composition has the remarkable effects of resisting oxidation, inhibiting tyrosine, promoting collagen regeneration and the like due to the synergistic effect of the components.
2. The oligopeptide composition disclosed by the application has excellent effects of resisting oxidation, inhibiting tyrosine, promoting collagen regeneration and the like in-vitro experiments, has very outstanding anti-aging skin repair effect, improves skin luster, lightens skin color and has good market application prospect.
Drawings
FIG. 1 is a bar graph of the effect of an oligopeptide composition of the present invention on tyrosinase inhibition rate and DPPH radical scavenging rate;
FIG. 2 is a bar graph of the effect of an oligopeptide composition according to the invention on the cell proliferation rate of human skin fibroblasts;
FIG. 3 is a bar graph showing the effect of oligopeptide composition according to the present invention on the amount of secreted type I collagen in human skin fibroblasts.
Detailed Description
Other advantages and effects of the present invention will become apparent to those skilled in the art from the following disclosure, which describes the embodiments of the present invention with reference to specific examples. The invention may be practiced or carried out in other embodiments that depart from the specific details, and the details of the present description may be modified or varied from the spirit and scope of the present invention. It should be noted that the following embodiments and features in the embodiments may be combined with each other without conflict.
Part of raw materials in the application:
the acanthopanax polysaccharide has the content of 90 percent and is purchased from Shaanxi Huidae plant development Co., ltd;
the wolfberry polysaccharide content is 90%, and is purchased from Shaanxi Huidae plant development Co.
Example 1: the embodiment provides an oligopeptide composition, which comprises 20 parts of lactobacillus fermentation product filtrate, 100 parts of oligopeptide-1, 100 parts of palmitoyl pentapeptide-4 and 100 parts of acetyl tetrapeptide-9 in parts by weight;
the preparation method of the lactobacillus fermentation product filtrate comprises the following steps:
(1) Taking 1 count of lactobacillus paracasei (product number NA-Jh487880, purchased from NodeA Gene technology (Wuhan) Co., ltd.) for freeze-drying, dissolving with 0.5mL of MRS liquid culture solution, inoculating to 1 MRS flat plate to grow single colony, diluting the single colony with 0.5mL of sterile water, inoculating to seed culture medium at a volume ratio of 3v/v%, and culturing at 37 ℃ for 18h to obtain seed solution; wherein, MRS liquid medium: peptone 10g/L, beef extract 10g/L, yeast extract 5g/L、KH 2 PO 4 2g/L, trisodium citrate 2g/L, sodium acetate 2g/L, glucose 20g/L, tween, 1mL/L, mgSO 4 ·7H 2 O 0.58g/L、MnSO 4 ·4H 2 O0.25 g/L; adjusting pH to 6.7+ -0.1, sterilizing at 121deg.C for 20 min. The seed culture medium comprises acanthopanax polysaccharide 6.25g/L, peptone 10g/L, beef extract 10g/L, yeast extract 5g/L, KH 2 PO 4 2g/L, trisodium citrate 2g/L, sodium acetate 2g/L, glucose 20g/L, tween, 1mL/L, mgSO 4 ·7H 2 O 0.58g/L、MnSO 4 ·4H 2 O0.25 g/L. Adjusting pH to 6.7+ -0.1, sterilizing at 121deg.C for 20 min.
(2) Inoculating the seed solution into a fermentation medium according to the volume ratio of 5v/v%, fermenting and culturing for 36 hours at 37 ℃, filtering, and performing pasteurization to obtain lactobacillus fermentation product filtrate. The fermentation medium comprises acanthopanax polysaccharide 1.35g/L, matrimony vine polysaccharide 5.55g/L, peptone 10g/L, beef extract 10g/L, yeast extract 5g/L, KH 2 PO 4 2g/L, trisodium citrate 2g/L, sodium acetate 2g/L, glucose 20g/L, tween, 1mL/L, mgSO 4 ·7H 2 O 0.58g/L、MnSO 4 ·4H 2 O0.25 g/L. Adjusting pH to 6.7+ -0.1, sterilizing at 121deg.C for 20 min.
Example 2: the embodiment provides an oligopeptide composition, which comprises 150 parts of lactobacillus fermentation product filtrate, 60 parts of oligopeptide-1, 60 parts of palmitoyl pentapeptide-4 and 60 parts of acetyl tetrapeptide-9 in parts by weight; the preparation method of the lactobacillus fermentation product filtrate is similar to that of the example 1, and the seed culture medium and the fermentation culture medium are slightly different; wherein the seed culture medium in the step (1) comprises 3.55g/L of acanthopanax polysaccharide, 10g/L of peptone, 10g/L of beef extract and 5g/L, KH of yeast extract 2 PO 4 2g/L, trisodium citrate 2g/L, sodium acetate 2g/L, glucose 20g/L, tween, 1mL/L, mgSO 4 ·7H 2 O 0.58g/L、MnSO 4 ·4H 2 O0.25 g/L; the fermentation medium in the step (2) comprises 3.55g/L of acanthopanax polysaccharide, 2.75g/L of lycium polysaccharide, 10g/L of peptone, 10g/L of beef extract and 5g/L, KH of yeast extract 2 PO 4 2g/L, trisodium citrate 2g/L, sodium acetate 2g/L, and glucose20g/L、Tween 80 1mL/L、MgSO 4 ·7H 2 O 0.58g/L、MnSO 4 ·4H 2 O 0.25g/L。
Example 3: the embodiment provides an oligopeptide composition, which comprises 300 parts of lactobacillus fermentation product filtrate, 10 parts of oligopeptide-1, 10 parts of palmitoyl pentapeptide-4 and 10 parts of acetyl tetrapeptide-9 in parts by weight; the preparation method of the lactobacillus fermentation product filtrate is similar to that of the embodiment 1, the seed culture medium and the fermentation culture medium are slightly different, wherein the seed culture medium in the step (1) comprises 1.35g/L of acanthopanax polysaccharide, 10g/L of peptone, 10g/L of beef extract and 5g/L, KH of yeast extract 2 PO 4 2g/L, trisodium citrate 2g/L, sodium acetate 2g/L, glucose 20g/L, tween, 1mL/L, mgSO 4 ·7H 2 O 0.58g/L、MnSO 4 ·4H 2 O0.25 g/L; the fermentation medium in the step (2) comprises acanthopanax polysaccharide 6.25g/L, matrimony vine polysaccharide 1.35g/L, peptone 10g/L, beef extract 10g/L, yeast extract 5g/L, KH 2 PO 4 2g/L, trisodium citrate 2g/L, sodium acetate 2g/L, glucose 20g/L, tween, 1mL/L, mgSO 4 ·7H 2 O 0.58g/L、MnSO 4 ·4H 2 O 0.25g/L。
Comparative example 1
This embodiment is similar to embodiment 2, except that: removing lactobacillus fermentation product filtrate.
Comparative example 2
This embodiment is similar to embodiment 2, except that: oligopeptide-1 was removed.
Comparative example 3
This embodiment is similar to embodiment 2, except that: palmitoyl pentapeptide-4 was removed.
Comparative example 4
This embodiment is similar to embodiment 2, except that: removing acetyl tetrapeptide-9.
Comparative example 5
This example is identical to the formulation of example 2, except that the preparation method of the lactobacillus fermentation product filtrate in the formulation is different, and in the preparation step of the lactobacillus fermentation product filtrate, the seed culture medium used in step (1) comprises: peptone 10g/L, beefPaste 10g/L, yeast paste 5g/L, KH 2 PO 4 2g/L, trisodium citrate 2g/L, sodium acetate 2g/L, glucose 20g/L, tween, 1mL/L, mgSO 4 ·7H 2 O 0.58g/L、MnSO 4 ·4H 2 O0.25 g/L; adjusting pH to 6.7+ -0.1, sterilizing at 121deg.C for 20 min.
The fermentation medium used in step (2) comprises: peptone 10g/L, beef extract 10g/L, yeast extract 5g/L, KH 2 PO 4 2g/L, trisodium citrate 2g/L, sodium acetate 2g/L, glucose 20g/L, tween, 1mL/L, mgSO 4 ·7H 2 O 0.58g/L、MnSO 4 ·4H 2 O0.25 g/L; adjusting pH to 6.7+ -0.1, sterilizing at 121deg.C for 20 min.
Comparative example 6
This example is identical to the formulation of example 2, except that the preparation method of the lactobacillus fermentation product filtrate in the formulation is different, and in the preparation step of the lactobacillus fermentation product filtrate, step (2) is identical to example 2; the seed medium used in step (1) comprises: peptone 10g/L, beef extract 10g/L, yeast extract 5g/L, KH 2 PO 4 2g/L, trisodium citrate 2g/L, sodium acetate 2g/L, glucose 20g/L, tween, 1mL/L, mgSO 4 ·7H 2 O 0.58g/L、MnSO 4 ·4H 2 O0.25 g/L; adjusting pH to 6.7+ -0.1, sterilizing at 121deg.C for 20 min.
Comparative example 7
This example is identical to the formulation of example 2, except that the preparation method of the lactobacillus fermentation product filtrate in the formulation is different, and in the preparation step of the lactobacillus fermentation product filtrate, step (1) is identical to example 2; the fermentation medium used in step (2) comprises: lycium barbarum polysaccharide 2.75g/L, peptone 10g/L, beef extract 10g/L, yeast extract 5g/L, KH 2 PO 4 2g/L, trisodium citrate 2g/L, sodium acetate 2g/L, glucose 20g/L, tween, 1mL/L, mgSO 4 ·7H 2 O 0.58g/L、MnSO 4 ·4H 2 O0.25 g/L; adjusting pH to 6.7+ -0.1, sterilizing at 121deg.C for 20 min.
Comparative example 8
This example is identical to example 2 in formulationExcept that the preparation method of the lactobacillus fermentation product filtrate in the formulation is different, in the preparation step of the lactobacillus fermentation product filtrate, the step (1) is the same as in example 2; the fermentation medium used in step (2) comprises: 3.55g/L of acanthopanax polysaccharide, 10g/L of peptone, 10g/L of beef extract and 5g/L, KH of yeast extract 2 PO 4 2g/L, trisodium citrate 2g/L, sodium acetate 2g/L, glucose 20g/L, tween, 1mL/L, mgSO 4 ·7H 2 O 0.58g/L、MnSO 4 ·4H 2 O0.25 g/L; adjusting pH to 6.7+ -0.1, sterilizing at 121deg.C for 20 min.
1. To further elucidate the efficacy of the oligopeptide compositions provided in the examples of the present application, the following performance tests were also performed in the examples of the present application:
1. inhibition tyrosinase assay
Sample to be measured: the oligopeptide compositions, lactobacillus fermentation product filtrate, oligopeptide-1, palmitoyl pentapeptide-4 and acetyl tetrapeptide-9 provided in examples 1-3 and comparative examples 1-8, respectively; wherein the oligopeptide compositions provided in examples 1 to 3 and comparative examples 1 to 8 were respectively prepared into 3wt% solutions (prepared by 3% of the total mass of the raw materials of the formulation after weighing according to the corresponding formulation) with 0.2M PBS buffer solution and pH6.80, the lactobacillus fermentation product filtrate was prepared into a solution containing 1.36% of lactobacillus fermentation product filtrate with PBS buffer solution and pH6.80, the oligopeptide-1 was prepared into a solution containing 0.55% of oligopeptide-1 with PBS buffer solution and the palmitoyl pentapeptide-4 was prepared into a solution containing 0.55% of palmitoyl pentapeptide-4 with PBS buffer solution and the acetyl tetrapeptide-9 was prepared into a solution containing 0.55% of acetyl tetrapeptide-9 with PBS buffer solution and pH 6.80.
The test method comprises the following steps: tyrosinase catalytic reaction systems (table 1) were constructed to evaluate the inhibition effect of the oligopeptide compositions provided in examples 1 to 3 and comparative examples 1 to 8 on tyrosinase. Carrying out an experiment on the tyrosinase inhibition test of each sample to be tested according to a tyrosinase catalytic reaction system, and carrying out 3 repetitions on each sample; the tyrosinase catalytic reaction system is divided into a, b, c, d four reaction tubes for sample addition, after the sample addition is finished, the reaction is carried out for 10min at a constant temperature in a water bath at 37 ℃, and the a, b, c, d light absorption value is measured at 475 nm. Tyrosinase inhibition rate (%) = [1- (c-d)/(a-b) ]. Times.100%, wherein a, b, c, d is the absorbance at 475nm after completion of the reaction of the corresponding reaction tube, and the tyrosinase inhibition rate of each experimental group was calculated, and the test results are shown in Table 3 and FIG. 1.
TABLE 1
| Reaction system | a | b | c | d |
| Sample to be measured (ul) | 0 | 0 | 200 | 200 |
| PBS（ul） | 1000 | 1200 | 800 | 1000 |
| 500U/mL tyrosinase solution (ul) | 200 | 0 | 200 | 0 |
| 0.5wt% tyrosine solution (ul) | 300 | 300 | 300 | 300 |
2. DPPH radical scavenging test
Sample to be measured: the oligopeptide compositions, lactobacillus fermentation product filtrate, oligopeptide-1, palmitoyl pentapeptide-4 and acetyl tetrapeptide-9 provided in examples 1-3 and comparative examples 1-8, respectively; wherein the oligopeptide compositions provided in examples 1 to 3 and comparative examples 1 to 8 were respectively prepared into 3wt% solutions with 0.2M PBS buffer solution and pH6.80 PBS buffer solution, the lactobacillus fermentation product filtrate was prepared into a solution containing 1.36% lactobacillus fermentation product filtrate with pH6.80 PBS buffer solution, the oligopeptide-1 was prepared into a solution containing 0.55% oligopeptide-1 with pH6.80 PBS buffer solution, the palmitoyl pentapeptide-4 was prepared into a solution containing 0.55% palmitoyl pentapeptide-4 with pH6.80 PBS buffer solution, and the acetyl tetrapeptide-9 was prepared into a solution containing 0.55% acetyl tetrapeptide-9 with pH6.80 PBS buffer solution.
The test method comprises the following steps: DPPH radical scavenging reaction system (Table 2) was constructed to evaluate the antioxidant activity of the oligopeptide compositions provided in examples 1 to 3 and comparative examples 1 to 8. Each sample to be tested is subjected to an experiment according to a tyrosinase catalytic reaction system, and each sample is subjected to 3 repetitions; the DPPH free radical scavenging reaction system is divided into three reaction tubes A1, A2 and A3 for sample addition, after sample addition, the reaction system is kept stand for 30min at a constant temperature and a dark place in a water bath at 25 ℃ and the absorbance values of A0, A1 and A2 are measured at 517 nm. DPPH clearance rate= [1- (A1-A2)/A0 ]. Times.100%, wherein A0, A1 and A2 are absorbance values at 517nm after the reaction of the corresponding reaction tube is finished, DPPH radical clearance rate of each experimental group is calculated, and test results are shown in Table 3 and FIG. 1.
TABLE 2
| Reaction system | A0 | A1 | A2 |
| Sample to be measured (ml) | － | 0.5 | 0.5 |
| 0.2mM DPPH methanol solution (ml) | 0.5 | 0.5 | － |
| Deionized water (ml) | 0.5 | － | 0.5 |
TABLE 3 Table 3
| Group of | DPPH radical scavenging% | Tyrosinase inhibition rate% |
| Example 1 | 92.73±0.54 a | 85.28±0.71 a |
| Example 2 | 95.87±1.13 a | 86.71±1.12 a |
| Example 3 | 95.24±0.82 a | 84.17±1.10 a |
| Comparative example 1 | 57.96±1.38 b | 24.90±0.78 c |
| Comparative example 2 | 61.93±0.63 b | 42.82±0.61 b |
| Comparative example 3 | 63.21±0.45 b | 44.69±0.78 b |
| Comparative example 4 | 65.41±0.58 b | 46.14±0.13 b |
| Comparative example 5 | 63.59±0.53 b | 19.45±0.56 c |
| Comparative example 6 | 66.17±0.81 b | 21.02±0.40 c |
| Comparative example 7 | 66.39±0.43 b | 25.11±0.19 c |
| Comparative example 8 | 64.79±0.87 b | 23.75±0.38 c |
| Lactobacillus fermentation product filtrate | 43.39±0.53 c | 32.79±0.35 bc |
| Oligopeptide-1 | 37.89±0.51 c | 14.22±0.30 c |
| Palmitoyl pentapeptide-4 | 36.69±0.56 c | 18.67±0.44 c |
| Acetyl tetrapeptide-9 | 40.56±0.70 c | 16.09±0.17 c |
As can be seen from Table 3 and FIG. 1, the tyrosinase inhibition rate and DPPH free radical removal rate of the oligopeptide composition provided in examples 1-3 are significantly higher than those of the oligopeptide composition provided in comparative examples 1-8, the lactobacillus fermentation product filtrate, oligopeptide-1, palmitoyl pentapeptide-4 and acetyl tetrapeptide-9, which indicates that the oligopeptide composition provided in the examples of the present application has better antioxidant capacity and stronger tyrosinase inhibition rate; tyrosinase is the most critical in the formation process of melanin in skin, and can inhibit tyrosinase activity to play a role in whitening, and free radicals or oxidants can influence the metabolic capacity of the body to generate health problems, so that the body can produce skin aging due to oxidation, and the anti-oxidation can delay skin aging. Example 2 shows that the tyrosinase inhibition rate and DPPH free radical removal rate of the oligopeptide composition are both significantly improved compared with comparative examples 1-8, indicating the synergistic effect of the lactobacillus fermentation product filtrate, oligopeptide-1, palmitoyl pentapeptide-4 and acetyl tetrapeptide-9 in the oligopeptide composition; compared with comparative examples 1 and 5-8, the comparative examples 2-4 have significantly improved tyrosinase inhibition rate, which indicates that lactobacillus paracasei fermentation product filtrate obtained by fermenting lactobacillus paracasei with a culture medium containing acanthopanax polysaccharide and wolfberry polysaccharide has more remarkable whitening effect with oligopeptide-1, palmitoyl pentapeptide-4 and acetyl tetrapeptide-9.
2. To further elucidate the efficacy of the oligopeptide compositions provided in the examples of the present application, the following cell experiments were also performed:
and (3) cells: human skin fibroblast HSF, cat No. BFN608007204, purchased from the Shanghai cell bank.
3. Human fibroblast in vitro proliferation Activity
Sample and group: 9 test groups and 1 negative control group were set, and the test groups were each prepared by using the oligopeptide compositions of example 2 and comparative examples 1 to 8 of the present application, and after the oligopeptide compositions were prepared into mother solutions by using deionized water, the respective mother solutions were prepared by using DMEM media, which were each prepared to be DMEM media containing 3wt% of the oligopeptide composition. The negative control group used DMEM medium.
The test method comprises the following steps: taking human skin fibroblasts cultured to the 4 th-6 th generation logarithmic growth phase, preparing the cells into suspension, inoculating 100ul of cells per well and 10000 cells per well into a 96-well plate, arranging 3 compound wells per group, and adding 100ul of DMEM culture medium into the other group as a blank control. The well plate was spread with 5% CO at 37 ℃ 2 The culture was carried out overnight under the conditions.
After the cells are adhered, the culture medium is discarded, the administration is carried out respectively, the culture medium corresponding to the experimental group is added (10% fetal bovine serum is also added to the culture medium corresponding to the experimental group and the negative control group during the administration culture, PBS is used for cleaning and then serum-free DMEM is added again during the test by using the MTT detection kit), the culture is continued for 24 hours in a cell culture box, the culture medium in the holes is discarded, 10 mu L of mixed solution of MTT and 90 mu L of serum-free DMEM is added to each hole, after the incubation is carried out for 4 hours in the cell culture box, the operation is carried out according to the MTT detection kit (MTT cell proliferation and cytotoxicity detection kit, solarbio), the absorbance values of the experimental group, the negative control group and the blank control group at 490nm of the detection wavelength of an enzyme-labeled instrument are calculated, and the cell proliferation rate is shown in the table 4 and the table 2;
test group: HSF cell, DMEM culture solution and oligopeptide composition
Negative control group: HSF cell+DMEM culture solution
Blank control group: DMEM culture solution
Cell proliferation rate% (cell viability) = (test group-blank control group)/(negative control group-blank control group) ×100%,
TABLE 4 Table 4
| Group of | Cell proliferation Rate (%) |
| Example 2 | 158.15±1.07 a |
| Comparative example 1 | 108.21±1.10 abc |
| Comparative example 2 | 124.92±0.59 ab |
| Comparative example 3 | 126.31±0.82 ab |
| Comparative example 4 | 124.80±0.91 ab |
| Comparative example 5 | 110.36±1.35 abc |
| Comparative example 6 | 111.31±0.96 abc |
| Comparative example 7 | 112.83±0.69 abc |
| Comparative example 8 | 112.31±0.80 abc |
| Negative control group | 100 |
As can be seen from table 4 and fig. 2, the oligopeptide composition provided in example 2 has significantly higher cell proliferation rate of cultured HSF cells than that of comparative examples 1-8, which indicates that the oligopeptide composition provided in example 2 can promote cell proliferation, and the lactobacillus fermentation product filtrate, oligopeptide-1, palmitoyl pentapeptide-4 and acetyl tetrapeptide-9 in the oligopeptide composition act synergistically; the comparative examples 2 to 4 showed a remarkable increase in cell proliferation rate as compared with comparative examples 1 and 5 to 8, showing that the lactobacillus fermentation product filtrate in the oligopeptide composition, which was obtained by fermenting lactobacillus paracasei with a medium containing acanthopanax polysaccharide and lycium barbarum polysaccharide, had a more remarkable effect in promoting HSF cell proliferation.
4. Cell secretion collagen I content
Sample and group: 9 test groups and 1 negative control group were set, the test groups were each corresponding to the oligopeptide compositions of example 2 and comparative examples 1 to 8 of the present application, and after the oligopeptide compositions were each prepared into mother solutions with deionized water, DMEM medium was used to prepare DMEM medium containing 10% fetal bovine serum in a volume fraction of 3% oligopeptide composition. The negative control group used DMEM medium containing 10% fetal bovine serum by volume fraction.
The test method comprises the following steps: taking human skin fibroblast cultured to 4 th-6 th generation logarithmic growth phase, inoculating 5000 cells per well and 100ul per well into 96-well plate, arranging 3 multiple wells per group, placing into cell incubator at 37deg.C and 5% CO 2 Culturing overnight under the condition;
after the cells are attached, the culture medium is discarded, PBS is cleaned, the administration is respectively carried out, the culture medium corresponding to the experimental group is added, after the incubation for 36 hours in a 37 ℃ incubator, a cell supernatant sample is taken, the content of collagen is measured by a human type I collagen (COL-I) ELISA kit (Shanghai Sitang Biotech Co., ltd.), and the type I collagen synthesis promotion effect on human fibroblasts is evaluated. The results of the type I collagen content test are shown in table 5 and fig. 3.
TABLE 5
| Group of | Type I collagen (ng/mL) |
| Example 2 | 66.56±0.59 a |
| Comparative example 1 | 29.80±0.29 abc |
| Comparative example 2 | 45.01±0.67 ab |
| Comparative example 3 | 46.78±0.73 ab |
| Comparative example 4 | 44.68±1.20 ab |
| Comparative example 5 | 30.84±0.31 abc |
| Comparative example 6 | 31.75±0.35 abc |
| Comparative example 7 | 33.39±0.41 abc |
| Comparative example 8 | 32.04±0.18 abc |
| Negative control group | 18.85±0.28 |
From table 5 and fig. 3, the oligopeptide composition provided in example 2 has a significantly higher effect of promoting HSF cells to secrete type i collagen than that of comparative examples 1-8, which indicates that the oligopeptide composition provided in example 2 can promote collagen production, so that skin is smooth and elastic, has the potential of developing cosmetics with anti-wrinkle repair and anti-aging effects on skin, and the synergistic effect of lactobacillus fermentation product filtrate, oligopeptide-1, palmitoyl pentapeptide-4 and acetyl tetrapeptide-9 in the oligopeptide composition promotes HSF to produce collagen; compared with comparative examples 1 and 5-8, comparative examples 2-4 have significantly improved secretion levels of collagen, showing that the lactobacillus fermentation product filtrate in the oligopeptide composition, which is obtained by fermenting lactobacillus paracasei with a culture medium containing acanthopanax polysaccharide and lycium barbarum polysaccharide, has a good synergistic effect in promoting secretion of type I collagen by HSF cells.
The oligopeptide composition provided by the application can be applied to the preparation of skin care products, wherein the skin care products have the effects of resisting aging, repairing skin, whitening and the like.
The foregoing examples merely illustrate specific embodiments of the invention, which are described in greater detail and are not to be construed as limiting the scope of the invention. It should be noted that it will be apparent to those skilled in the art that several variations and modifications can be made without departing from the spirit of the invention, which are all within the scope of the invention.
Claims (4)
1. An oligopeptide composition, which is characterized by comprising 20-300 parts of lactobacillus fermentation product filtrate, 10-100 parts of oligopeptide-1, 10-100 parts of palmitoyl pentapeptide-4 and 10-100 parts of acetyl tetrapeptide-9;
the lactobacillus fermentation product filtrate is prepared by activating lactobacillus, inoculating the lactobacillus into a seed culture medium for culture to obtain seed liquid, and then inoculating the seed liquid into a fermentation culture medium for culture;
the seed culture medium comprises 1.35-6.25 g/L of acanthopanax polysaccharide, 10g/L of peptone, 10g/L of beef extract and 5g/L, KH of yeast extract 2 PO 4 2g/L, trisodium citrate 2g/L, sodium acetate 2g/L, glucose 20g/L, tween, 1mL/L, mgSO 4 ·7H 2 O 0.58g/L、MnSO 4 ·4H 2 O 0.25g/L；
The fermentation culture medium comprises 1.35-6.25 g/L of acanthopanax polysaccharide, 1.35-5.55 g/L of medlar polysaccharide, 10g/L of peptone, 10g/L of beef extract and 5g/L, KH of yeast extract 2 PO 4 2g/L, trisodium citrate 2g/L, sodium acetate 2g/L, glucose 20g/L, tween, 1mL/L, mgSO 4 ·7H 2 O 0.58g/ L、MnSO 4 ·4H 2 O0.25g/L。
2. An oligopeptide composition according to claim 1, comprising 150 parts lactobacillus fermentation product filtrate, 60 parts oligopeptide-1, 60 parts palmitoyl pentapeptide-4, 60 parts acetyl tetrapeptide-9.
3. The oligopeptide composition according to claim 1, wherein the seed medium comprises acanthopanax polysaccharide 3.55g/L, peptone 10g/L, beef extract 10g/L, yeast extract 5g/L, KH 2 PO 4 2g/L, trisodium citrate 2g/L, sodium acetate 2g/L, glucose 20g/L, tween, 1mL/L, mgSO 4 ·7H 2 O 0.58g/L、MnSO 4 ·4H 2 O0.25 g/L; the fermentation medium comprises 3.55g/L of acanthopanax polysaccharide, 2.75g/L of wolfberry polysaccharide, 10g/L of peptone, 10g/L of beef extract and 5g/L, KH of yeast extract 2 PO 4 2g/L, trisodium citrate 2g/L, sodium acetate 2g/L, glucose 20g/L, tween, 1mL/L, mgSO 4 ·7H 2 O 0.58g/L、MnSO 4 ·4H 2 O 0.25g/L。
4. Use of the oligopeptide composition of any one of claims 1-3 for the preparation of a skin repair and anti-aging product.
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| CN111686029A (en) * | 2020-07-28 | 2020-09-22 | 广州市柏亚化妆品有限公司 | Anti-aging wrinkle-removing composition |
| CN114085271A (en) * | 2021-11-29 | 2022-02-25 | 北京远胜达生物科技发展有限公司 | A medicine or antioxidant whitening cosmetic prepared from microorganism fermentation liquid |
| CN115554220A (en) * | 2022-10-24 | 2023-01-03 | 广州瑞隽生物科技有限公司 | Microbial fermentation stock solution with skin care effect and preparation method and application thereof |
| CN116019185A (en) * | 2022-12-14 | 2023-04-28 | 江南大学 | Application of lactobacillus paracasei in preparation of fermented medlar |
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