CN-121554567-B - Recombinant human XV type collagen and preparation method and application thereof

Abstract

The invention relates to the technical field of bioengineering, and particularly discloses recombinant human XV-type collagen and a preparation method and application thereof. The amino acid sequence of the recombinant human XV-type collagen is shown as SEQ ID NO. 1. The recombinant human XV-type collagen is derived from human collagen, has no rejection and sensitization risks, has good effects of promoting cell proliferation, promoting cell migration and the like, has good biocompatibility, and can be used as a safe and effective biomedical material.

Inventors

• WANG PAN
• LIU JIA
• WANG RUYAO
• YAO TIANYU
• XU HUAN
• LI CONGCONG
• FAN DAIDI

Assignees

• 西北大学

Dates

Publication Date

20260508

Application Date

20251129

Claims (9)

1. 1. The recombinant human XV-type collagen is characterized in that the amino acid sequence of the recombinant human XV-type collagen is shown as SEQ ID NO. 1.
2. 2. A gene encoding recombinant human XV type collagen according to claim 1, wherein the nucleotide sequence of said gene is shown in SEQ ID No. 2.
3. 3. An expression vector comprising the gene of claim 2.
4. 4. A host cell comprising the expression vector of claim 3.
5. 5. The method for producing recombinant human XV-type collagen according to claim 1, comprising culturing a host cell comprising the gene encoding the recombinant human XV-type collagen so as to express the recombinant human XV-type collagen.
6. 6. The method of claim 5, wherein the host cell is induced to express the recombinant human XV-type collagen using an inducer.
7. 7. The method of claim 6, wherein the inducer is methanol.
8. 8. The method of claim 5, further comprising purifying.
9. 9. The method of claim 8, wherein the purifying is performed by one or more of salting out, chromatography, affinity chromatography, acid-base precipitation, and membrane separation.

Description

Recombinant human XV type collagen and preparation method and application thereof Technical Field The invention relates to the technical field of bioengineering, in particular to recombinant human XV-type collagen and a preparation method and application thereof. Background Collagen is one of the most abundant proteins in the human body, and is mainly present in tissues such as skin, bones, tendons, ligaments and blood vessels, and plays a key role in maintaining tissue structure and function, accounting for about 30% of the total protein of the human body. Because of their excellent biological function, biocompatibility, biodegradability and other characteristics, collagen has become one of protein materials with great application potential in the fields of biological materials, regenerative medicine and the like. At present, most of the collagen used in the market is obtained by animal tissue extraction, but the traditional extraction methods have a plurality of problems. On one hand, the collagen structure is often changed in the extraction process and is different from the natural state in the body, and on the other hand, the collagen material has the defects of large immune rejection reaction, nonuniform product batch quality, unstable biological efficacy and the like, and the factors greatly limit the application range of the collagen material. With the rapid development of modern biotechnology such as genetic engineering, protein engineering and synthetic biology, recombinant protein expression technology has become an important choice. As a substitute for natural animal tissue collagen, the recombinant collagen has the characteristics of excellent biocompatibility, low immunogenicity, processability, controllable biological efficacy and the like, and has great potential for wide application in the fields of biological materials and biomedicine. XV-type collagen is widely distributed in various tissues, and is most strongly expressed in the basement membrane region, and its main function is to adhere the basement membrane to the underlying connective tissue matrix. This property makes XV-type collagen play a critical role in the tissue repair process, and particularly, is prominent in the repair of injuries to tissues such as skin, muscle, blood vessels and the like. However, although there is some progress in the research on the structure and function of XV-type collagen, the XV-type collagen is low in the animal body, which results in difficulty in extracting a single component. Disclosure of Invention In order to solve the technical problems, the invention provides recombinant human XV-type collagen, and a preparation method and application thereof. The invention provides recombinant human XV-type collagen, and the amino acid sequence of the recombinant human XV-type collagen is shown as SEQ ID NO. 1. The recombinant human XV-type collagen is based on the original amino acid sequence of the human XV-type collagen, and a novel recombinant human XV-type collagen is obtained by screening and retaining key fragments with high bioactivity and strong structural stability. The recombinant human XV-type collagen removes the full-length chain sequences of the C-end and the N-end of the collagen coding region, effectively avoids a series of antigen immune reactions, has a theoretical molecular weight of 37.30kDa, has moderate molecular weight and is easy to prepare. The invention also carries out the small-scale expression of the fermentation tank, realizes the high-efficiency secretion expression of the recombinant human XV-type collagen, and the yield of the 5L fermentation tank can reach 2.36g/L. The invention also provides a gene for encoding the recombinant human XV-type collagen, and the nucleotide sequence of the gene is shown as SEQ ID NO. 2. An expression vector comprising the gene. A host cell comprising said expression vector. The host cell is Escherichia coli or Pichia pastoris. A method for preparing recombinant human XV-type collagen comprises culturing host cells containing recombinant human XV-type collagen encoding genes to express the recombinant human XV-type collagen. Preferably, the host cell is induced to express the recombinant human XV-type collagen using an inducer. Preferably, the inducer is methanol. Preferably, purification is also included. Preferably, the purification employs one or more of salting out, chromatography, affinity chromatography, acid-base precipitation and membrane separation. The use of said recombinant human XV-type collagen in the preparation of a tissue repair product, said tissue comprising any of skin, muscle and vascular species. The material comprises any one of skin care products, health care products, tissue engineering materials and medical appliances. The recombinant human XV-type collagen is used for promoting cell proliferation. The recombinant human XV-type collagen of the present invention is used to promote cell migration. Compared with the prior