Modified activin A

Patent metadata

Field	Value
Publication number	US-12291554-B2
Application number	US-202117321049-A
Country	US
Kind code	B2
Filing date	May 14, 2021
Priority date	Nov 15, 2018
Publication date	May 6, 2025
Grant date	May 6, 2025

Abstract

The purpose of the present invention is to provide a modified activin A.
The present invention provides activin A comprising a modified proregion.

First claim

The invention claimed is:

1. A recombinant activin A, comprising a modified proregion, wherein the modified proregion comprises an amino acid sequence selected from the group consisting of (a), (b) and (d):
(a) an amino acid sequence in which an amino acid sequence from position 182 to 199 of the amino acid sequence of SEQ ID NOs: 4, 5 or 6 is deleted;
(b) an amino acid sequence in which an amino acid sequence from position 182 to 199 of the amino acid sequence of SEQ ID NOs: 4, 5 or 6 is substituted with a spacer sequence consisting of 1 to 10 amino acids; and
(d) an amino acid sequence in which a least one of amino acids from position 180 to 201 of the amino acid sequence of SEQ ID NOs: 4, 5, or 6 has at least one amino acid inserted therein.

2. The recombinant activin A according to claim 1 , wherein the amino acid constituting the spacer sequence in the amino acid sequence (b) is at least one amino acid selected from the group consisting of glycine, alanine, and serine.

3. The recombinant activin A according to claim 1 , wherein the recombinant activin A has resistance to degradation by a protease other than Furin.

4. A recombinant activin A, comprising a modified proregion, wherein the modified proregion comprises an amino acid sequences selected from the group consisting of (e) to (g):
(e) an amino acid sequence selected from the group consisting of the amino acid sequence of SEQ ID NOs: 9 to 20;
(f) an amino acid sequence in which at least one amino acid is deleted, substituted or added in an amino acid sequence selected from the group consisting of the amino acid sequence of SEQ ID NOs: 9 to 20, wherein the recombinant activin A whose proregion is modified so that it has resistance to degradation by a protease other than Furin; and
(g) an amino acid sequence having at least 80% homology with an amino acid sequence selected from the group consisting of the amino acid sequence of SEQ ID NOs: 9 to 20, wherein the recombinant activin A comprising the amino acid sequence has resistance to degradation by a protease other than Furin.

5. A polynucleotide encoding the recombinant activin A according to claim 1 or 4 .

6. A vector comprising the polynucleotide according to claim 5 .

7. A transformant expressing the recombinant activin A according to claim 1 or 4 .

8. The transformant according to claim 7 , wherein the transformant is a plant transformant.

9. The transformant according to claim 8 , wherein the transformant expresses the recombinant activin A in apoplast.

10. A method for producing a recombinant activin A, the method comprising:
recovering the recombinant activin A expressed in the transformant according to claim 7 .

11. A method for producing a recombinant activin A, the method comprising:
(a) recovering the recombinant activin A expressed in the transformant according to claim 7 ; and
(b) treating the obtained recombinant activin A with Furin.

12. A mixture comprising:
(i) a recombinant activin A comprising a modified proregion and
(ii) a recombinant mature activin A,
wherein the modified proregion comprises an amino acid sequence selected from the group consisting of (a), (b), and (d):
(a) an amino acid sequence in which an amino acid sequence from position 182 to 199 of the amino acid sequence of SEQ ID NOs: 4, 5, or 6 is deleted;
(b) an amino acid sequence in which an amino acid sequence from position 182 to 199 of the amino acid sequence of SEQ ID NOs: 4, 5, or 6 is substituted with a spacer sequence consisting of 1 to 10 amino acids; and
(d) an amino acid sequence in which an amino acid sequence from position 180 to 201 of the amino acid sequence of SEQ ID NOs: 4, 5, or 6 has at least one amino acid inserted therein.

Assignees

• Mitsubishi Chem Corp
• Unibio Corp

Inventors

• Takayama Hidehito
• Watanabe Risa
• Takemoto Yutaka
• Tanaka Hiroyuki
• Kawabata Hiroshi
• Yoshinaka Kyoji

Classifications

• C12N15/8257

  for the production of primary gene products, e.g. pharmaceutical products, interferon · CPC title

• C12Y304/21075

  Furin (3.4.21.75) · CPC title

• C12P21/06

  produced by the hydrolysis of a peptide bond, e.g. hydrolysate products (preparing foodstuffs by protein hydrolysis A23J3/00) · CPC title

• C12N9/6454

  Dibasic site splicing serine proteases, e.g. kexin (3.4.21.61); furin (3.4.21.75) and other proprotein convertases · CPC title

• C07K2319/50

  containing protease site · CPC title