阅读说明：本技术 一种促进皮肤、骨骼、肌肉功能的胶原蛋白三肽结构 (Collagen tripeptide structure for promoting skin, bone and muscle functions ) 是由 林倩如 王颖 于 2021-08-03 设计创作，主要内容包括：本发明公开了一种促进皮肤、骨骼、肌肉功能的胶原蛋白三肽结构,胶原蛋白肽的氨基酸序列为Pro-Hyp-Gly。本发明采用上述的一种促进皮肤、骨骼、肌肉功能的胶原蛋白三肽结构,胶原三肽结构稳定,有效的促进皮肤和骨骼、肌肉的发育。(The invention discloses a collagen tripeptide structure for promoting skin, bone and muscle functions, wherein the amino acid sequence of a collagen peptide is Pro-Hyp-Gly. The collagen tripeptide structure for promoting the functions of skin, bones and muscles is stable, and the development of the skin, the bones and the muscles is effectively promoted.)

1. A collagen tripeptide structure for promoting skin, bone and muscle functions, characterized in that: the tripeptide amino acid sequence of the collagen peptide is Pro-Hyp-Gly.

2. A collagen tripeptide structure for promoting skin, bone and muscle function according to claim 1, wherein: the concentration of the collagen peptide is 1-5 mug/mL.

3. A collagen tripeptide structure for promoting skin, bone and muscle function according to claim 1, wherein: the derived sequence of collagen is expanded based on the basic amino acid sequence, the amino acid sequence is not more than 10, and the molecular weight is not more than 1200 Da.

Technical Field

The invention relates to the technical field of collagen, in particular to a collagen tripeptide structure for promoting skin, bone and muscle functions.

Background

Collagen is the most abundant protein in many vertebrates and invertebrates, belongs to structural proteins, is the main fibrous component of skin, bone, cartilage, blood vessels and teeth, and is present in all organs.

The high mechanical strength of collagen is closely related to the protein structure of collagen. In collagen, special amino groups constitute a stable triple helix structure, various acting forces between amino acids and peptide chains maintain the stability of collagen, and the performance characteristics of collagen are the external reflection of amino acid sequence arrangement.

The amino acid arrangement of collagen can trigger mutation of certain amino acid residues, loss of original stability, and cause lesions. The properties depend on the structure, and the stability of collagen is an outward manifestation of its particular structure. The collagen structure is of great significance to people with congenital and acquired connective tissue diseases.

Disclosure of Invention

The invention aims to provide a collagen peptide for promoting the functions of skin and bones, which has a stable collagen structure and effectively promotes the development of the skin and the bones.

In order to achieve the aim, the invention provides a collagen peptide for promoting skin and bone functions, wherein the basic amino acid sequence of the collagen peptide is Pro-Hyp-Gly.

Preferably, the concentration of the collagen peptide is 1-5. mu.g/mL.

Preferably, the collagen derived sequence is extended based on a base amino acid sequence, the amino acid sequence is no more than 10, and the molecular weight is no more than 1200 Da.

Therefore, the collagen tripeptide structure for promoting the functions of skin, bones and muscles is adopted, the tripeptide structure is stable, the oriented tripeptide structure of proline, hydroxyproline and glycine has a remarkable effect on the development of skin and bones, and the function of a body is promoted to be enhanced.

The stability of the collagen helix depends on the synergistic effects of various intermolecular and intramolecular interactions, the combined action of imino acids, hydrogen bonds, van der waals forces, and hydration in stabilizing the collagen molecular structure, while hydrophobic interactions and ionic interactions provide intermolecular forces. However, the amino acid sequence of collagen determines the stability and intermolecular interactions of the molecules. The collagen peptide of the present invention has improved stability of the helix by a high content of imino acids (proline, hydroxyproline). Glycine improves its stability by optimizing the range of concepts in polypeptide chain folding. Hydroxylation of proline improves stability in the helical structure. Proline and hydroxyproline are imino acids, and the stability of the triple helix structure is improved.

The technical solution of the present invention is further described in detail by the following examples.

Detailed Description

The technical solution of the present invention is further illustrated by the following examples.

A collagen peptide for promoting skin and bone functions has a basic amino acid sequence of Pro-Hyp-Gly. The concentration of the collagen peptide is 1-5 mug/mL.

The derived sequence of collagen is extended based on the basic amino acid sequence, the amino acid sequence is not more than 10, and the molecular weight is 1200 Da.

Test of

Effect of collagen peptide on fibroblast

Human skin fibroblasts (Hs27 cells (ATCC)) were cultured using complete medium consisting essentially of basal medium high-glucose DMEM, 10% fetal bovine serum (v/v), 1% diabase (penicillin and streptomycin).

Placing the fibroblasts at 37 deg.C and 5% CO₂The culture medium of (1) was changed every 2 days. Inoculating the cells to 96-well culture plate with 100 μ L/well and concentration of 5 × 10 when the cells are about 90% of the culture flask⁴cells/mL. After 24h of cell growth, the culture was aspirated and washed 1 time with 200. mu.LPBS. Adding 200 μ LPBS, and mixing at a concentration of 80mJ/cm²UVB irradiation, PBS was aspirated. Wherein 200 μ L of 2 μ g/mL tripeptide collagen peptide is added into each well of the test group, the same amount of PBS is added into the control group, the culture is continued for 72h, and then the culture solution is removed by suction, and the cell survival rate is detected.

As a result, the number of fibroblasts in the test group was found to be significantly greater than that in the control group.

(II) variation of collagen peptide content

Hs27 skin fibroblasts were spread in 24-well culture vessels with 1 x 10 each⁴After that, at 5% CO₂After incubation at 37 ℃ for 24 hours, 200. mu.L of 2. mu.g/mL tripeptide type collagen peptide was added to each well of the test group, and an equal amount of PBS was added to the control group, followed by further incubation for 72 hours. Cell proliferation was measured by crystal violet assay.

After removing the medium from each well again, 500ml of 0.1% crystal violet solution was added to 1 well and stained for 5 minutes, and then the crystal violet solution was removed, washed 3 times with distilled water and repeated 4 times until the wells became clear. Then, 1ml of 95% ethanol was added thereto and stirred for 20min to dissolve the crystal violet stained by the cells. The solution was dispensed into 96 well containers at 200ml per well and the relative cell proliferation was calculated for the test and control groups.

As a result, the number of fibroblasts in the test group was found to be significantly greater than that in the control group.

(III) Effect of collagen peptides on bone cells

After culturing mouse osteoblasts and osteoclasts in an osteoblast culture medium for 24 hours, the cells were divided into a control group and a test group, the test group was added with 1mg/mL tripeptide collagen peptide, the control group was added with bovine serum albumin, the cells were cultured for 14 days, and then the alkaline phosphatase activity was quantitatively measured with a fluorometer, and osteoblast pictures of the culture medium were observed with a microscope. As a result, it was found that the alkaline phosphatase activity of the test group (2.13. + -. 0.85 mU/10)⁵Cells) were significantly higher than the control group (0.42. + -. 0.16 mU/10)⁵Cells), collagen peptide composition bone cell character changed, and bovine serum albumin control group bone cell ablation.

Effect of (tetra) collagen peptides on muscle cells

After ether inhalation anesthesia of rats, ketamine (22mg/kg) was injected into the abdominal cavity, and a transverse incision was made at the level of the left 4 th rib, and the pectoralis major muscle was removed by about 1cm³And trimming it to 1mm³Small muscle granules of size. After blood cells were washed out with Hanks 'solution, myogranules were carefully attached to the bottom of a cell culture flask previously coated with gelatin, and an appropriate amount of myoblast proliferation medium (15% calf serum, penicillin 100U/ml, streptomycin 100U/ml in Ham's F-10 medium) was added. Placing the bottle bottom upwards at 37 deg.C with 5% CO₂And in an incubator with saturated humidity, after 4 hours, the culture bottle is turned slightly, and the muscle granules are soaked in the culture solution. And (3) replacing the solution once, and passaging after the cells grow to full bottom of the bottle.

10. mu.L of the second generation of subculture cells were placed in a new culture flask, and divided into a test group and a control group, and 200. mu.L of collagen peptide and 200. mu.L of LPBS were added, respectively, to culture the cells. Relative cell proliferation was calculated for the test group versus the control group.

As a result, the number of muscle cells in the test group was found to be significantly greater than that in the control group.

Therefore, the collagen tripeptide structure for promoting the functions of skin, bones and muscles is adopted, and the collagen tripeptide peptide chain structure is stable, so that the development of skin, bones and muscles is effectively promoted.

Finally, it should be noted that: the above embodiments are only for illustrating the technical solutions of the present invention and not for limiting the same, and although the present invention is described in detail with reference to the preferred embodiments, those of ordinary skill in the art should understand that: modifications and equivalents may be made to the invention without departing from the spirit and scope of the invention.