RU-2026103941-A - KERATIN CF5, A METHOD FOR ITS PRODUCTION, A PHARMACEUTICAL COMPOSITION CONTAINING IT, AND THE USE OF KERATIN CF5

Inventors

• ЮЙ, Шишань
• ЛЮ, Юньбао
• Ли, Юн
• ЦАЙ, Цзе
• ВАН, Вэйпин
• ЧЖАН, Ми
• Ван, Сяолян
• Ван, Сяоцзин
• МА, Шуанган
• Ван, Лин
• ЦЮЙ, Цзин
• ЛИ, Ми
• ФЭН, Нан
• СЮЙ, Шаофэн

Assignees

• ИНСТИТЬЮТ ОФ МАТЕРИА МЕДИКА, ЧАЙНИЗ АКЭДЕМИ ОФ МЕДИКАЛ САЙЕНСИЗ

Dates

Publication Date

20260506

Application Date

20240521

Priority Date

20230717

Claims (20)

1. 1. Keratin CF5, where the amino acid sequence of keratin CF5 is:
2. (1) the amino acid sequence as set forth in SEQ ID NO. 1 in the sequence listing, or
3. (2) an amino acid sequence obtained by substituting, deleting or adding 1-45 amino acids in the amino acid sequence as shown in SEQ ID NO. 1 in the sequence listing, and keratin CF5 substantially retains the same biological function.
4. 2. Keratin CF5 according to claim 1, wherein conventional modifications may be made to keratin CF5, or a label for detection or purification may also be attached to keratin CF5, or it is a homologous protein of keratin CF5.
5. 3. The CF5 keratin of claim 2, wherein the common modification is acetylation, amidation, cyclization, glycosylation, phosphorylation, alkylation, biotinylation, modification with a fluorescent group, modification with polyethylene glycol (PEG), immobilization modification, sulfation, oxidation, methylation, deamination, disulfide bond formation or disulfide bond cleavage, or the label is His6, GST, EGFP, MBP, Nus, HA, IgG, FLAG, c-Myc or Profinity eXact.
6. 4. A nucleic acid molecule encoding keratin CF5 according to any one of claims 1-3.
7. 5. A nucleic acid molecule according to claim 4, wherein the nucleotide sequence of the nucleic acid molecule is:
8. (1) the nucleotide sequence as presented in SEQ ID NO. 2 in the sequence listing,
9. (2) a nucleotide sequence obtained by sequence optimization based on the nucleotide sequence as shown in SEQ ID NO. 2, or
10. (3) a nucleotide sequence complementary to the nucleotide sequence according to (1) or (2) above.
11. 6. An expression vector, wherein the expression vector comprises a nucleic acid molecule according to any one of claims 4-5.
12. 7. A host cell, wherein the host cell comprises the expression vector of claim 6 or the nucleic acid molecule of any one of claims 4-5 integrated into the genome of the host cell.
13. 8. The host cell of claim 7, wherein the host cell is a bacterium, yeast, aspergillus, plant cell, or insect cell.
14. 9. The host cell of claim 8, wherein the bacterium is Escherichia coli.
15. 10. A method for producing CF5 keratin according to any one of paragraphs 1-3, comprising the following steps:
16. A. a nucleic acid molecule corresponding to keratin CF5 according to any one of paragraphs 1-3 is synthesized, the nucleic acid molecule is ligated into an appropriate expression vector, the expression vector is transformed into a host cell, the host cell carrying the expression vector is cultured under certain conditions in a fermentation device, and expression of keratin CF5 is induced to obtain a solution of crude protein containing keratin CF5, and
17. B. The crude protein solution expressed in step A is separated, purified and dried to obtain keratin CF5.
18. 11. The method of claim 10, wherein in step A the host cell is primarily Escherichia coli, the CF5 keratin is expressed in inclusion bodies of the Escherichia coli, and the fermentation device is a shake flask or a fermentation vat.
19. 12. The method according to claim 10, wherein in step A, after inducing the expression of CF5 keratin, the impurities can be washed using a clearing agent, dissolved using a solution to obtain a solution of crude protein.
20. 13. The method according to claim 10, wherein in step B the separation and purification method is a purification method based on membrane technology of ultrafiltration or microfiltration, a purification method using column chromatography, a salting out method and a dialysis method.