Method for preparing ultra-low-molecular-weight keratin peptide, and use thereof

A technology of keratin peptide and keratin, which is applied in the field of preparation and utilization of ultra-low molecular keratin peptide, can solve the problems of low economy and productivity, achieve excellent skin keratin permeability, prevent skin aging or skin The effect of wrinkles

Active Publication Date: 2019-03-01
KYUNGPOOK NAT UNIV IND ACADEMIC COOP FOUND
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In particular, in order to develop functional cosmetic materials that are not used in medicine, the separation method using the above-mentioned expensive equipment is very low in terms of economy and productivity, so there is a limit to its application in actual industry

Method used

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  • Method for preparing ultra-low-molecular-weight keratin peptide, and use thereof
  • Method for preparing ultra-low-molecular-weight keratin peptide, and use thereof
  • Method for preparing ultra-low-molecular-weight keratin peptide, and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0076] Example 1 . Preparation of Keratin Peptides

[0077] 1.1 Culture and preparation of keratin hydrolyzate

[0078] In order to prepare the keratin peptide mixture, as shown in the following table 1, using a nitrogen source, under the temperature condition of 70 ℃, in the growth medium supplemented with chicken feathers, Fervidobacterium islandicum (Fervidobacterium islandicum) AW-1 (Fervidobacterium islandicum) AW-1 ( KCTC4680) were anaerobically cultured to obtain keratin hydrolyzate.

[0079] Table 1

[0080] .

[0081] After that, the above-mentioned keratin hydrolyzate was subjected to the following operation: After primary filtration of the decomposed chicken feather residue with filter paper (5 μm, No. 20, Hyundai Micro, Korea), it was centrifuged at 10,000×g at 4°C 20 minutes, and the supernatant was recovered. The recovered supernatant is used as a sample in the separation and purification process of functional ultra-low molecular weight keratin peptides...

Embodiment 2

[0088] Example 2 . Cytotoxicity assay of keratin peptides in human skin fibroblasts and gene expression induced by ultraviolet B Inhibitory effect of metalloproteinase-1 expression

[0089] In order to examine the cytotoxicity of the keratin peptide prepared by the method of Example 1 above, MTT assay was performed using human dermal fibroblast.

[0090] First, human fibroblasts were dispensed into 96-well plates such that 5×10 3 cell / well, using Dulbecco's Modified Eagle's Medium (Dulbecco's Modified Eagle's Medium) mixed with 10% fetal bovine serum (FBS) and penicillin-streptomycin (GIBCO Invitrogen, Auckland, NZ) sMedium, DMEM), at 37°C, 5% CO 2 Under the condition of , cultured for 6 hours. 200 mg of 3-(4,5-dimethylthiazol-2)-2,5-diphenyltetrazolium bromide (MTT, 3-(4,5-dimethylthiazol-2-yl)-2,5 -diphenyl-tetrazolium bromide) powder was dissolved in 40 ml of phosphate-buffered saline (PBS), and then filtered to prepare a 5 mg / mL MTT solution. Six hours after the ...

Embodiment 3

[0098] Embodiment 3. Amino Acid Sequence Identification and Synthesis of Keratin Peptides

[0099] As mentioned above, in order to clarify the amino acid sequence of the keratin peptide exhibiting this activity in the No. 2 and KP7 samples whose anti-aging ability and wrinkle improvement ability were confirmed, peptide identification by LC-MS / MS method was carried out .

[0100] Specifically, the chicken (Gallus gallus ver. 5.0) feather keratin sequence (feather keratin sequence) was obtained from the Genbank (Genbank) in order to be used for the database construction of keratin peptide mapping (mapping), and through AW- Transcript analysis of strain 1 confirmed the selection of proteases involved in chicken feather decomposition and peptide cleavage sites, and a database for LC-MS / MS results analysis was constructed on this basis. Electrospray quadrupole time-of-flight (ESI-Q-TOF, produced by Thermo (Dionex) of the United States, model: UHPLC Ultimate 3000, operating system...

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Abstract

The present invention relates to a method for preparing an ultra-low-molecular-weight keratin peptide and a use thereof. In particular, the present invention relates to: an ultra-low-molecular-weightkeratin peptide preparation method using the cultivation of microorganisms having keratin decomposition activity in a culture medium including keratins, ultrafiltration, ion exchange chromatography, and gel filtration chromatography; a peptide prepared according to said method; and cosmetic and food compositions for preventing or reducing skin aging or skin wrinkles, comprising said peptide. The keratin peptide preparation method according to the present invention enables environmentally friendly biological treatment of waste resources and effective refinement and recovery of an antiaging, functional, ultra-low-molecular-weight keratin peptide. In addition, the keratin peptide according to the present invention has inhibitory activity against the expression and activity of MMP-1, which isan enzyme that causes skin aging by decomposing collagen, thus has excellent effects of preventing skin aging and reducing skin wrinkles, is appropriate for use as cosmetic, pharmaceutical, and food compositions for preventing, reducing, or treating skin aging and skin wrinkles by having no toxicity to skin cells, and thus can be usefully used for efficient and quick production and development ofhigh value-added functional cosmetic materials.

Description

technical field [0001] The present invention relates to the preparation method and utilization of ultra-low molecular weight keratin peptide. Specifically, the present invention relates to a method for producing ultra-low-molecular keratin peptides using culture of microorganisms having keratinolytic activity in a medium containing keratin, ultrafiltration, ion exchange chromatography, and gel filtration chromatography, and to The prepared peptide and the cosmetic and food composition for preventing or improving skin aging or skin wrinkles containing the peptide. Background technique [0002] As interest in elastic and beautiful skin increases among men, women and children, in the cosmetics market, there is an increasing trend from existing health-oriented cosmetics to functions that can satisfy the desire for beauty and contribute to wrinkle improvement Excavation of sexual material. In particular, keratin peptides are basic natural amino acid polymers (less than 50 amino...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/47C07K1/20C07K1/30C07K1/36B01D15/32A61K8/65A23L33/17
CPCC12P21/06A61K8/65A23L33/18B01D15/34B01D61/145B01D15/361A23V2002/00A61Q19/08C07K14/4741A23V2200/318A23V2250/542A23L33/17B01D15/327C07K1/20C07K1/303C07K1/36C07K1/18C07K1/16C07K1/34
Inventor 李东佑姜南朱李勇直陈炫秀吕寅赫宋京燮李在恩
Owner KYUNGPOOK NAT UNIV IND ACADEMIC COOP FOUND
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