Hyaluronan synthase gene and uses thereof

• US 7,087,413 B2
• Filed: 09/14/2005
• Issued: 08/08/2006
• Est. Priority Date: 07/01/1994
• Status: Expired due to Fees

• Current Assignee: The Board of Regents of the University of Colorado (University of Colorado System)
• Original Assignee: The Board of Regents of the University of Colorado (University of Colorado System)
• Inventors: Weigel, Paul H., Kumari, Kshama, DeAngelis, Paul
• Primary Examiner(s): Wax, Robert A.
• Application Number:
  US11/226,480
• Publication Number:
  US 20060013829A1
• Time in Patent Office: 328 Days
• Field of Search: None
• US Class Current: 435/84
• CPC Class Codes: A61K 39/102 Pasteurellales, e.g. Actinobacillus, Pasteurella; Haemophilus
• CPC Class Codes: A61P 31/00 Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C07H 21/04 with deoxyribosyl as saccharide radical
• CPC Class Codes: C08B 37/0072 Hyaluronic acid, i.e. HA or hyaluronan; Derivatives thereof, e.g. crosslinked hyaluronic acid (hylan) or hyaluronates
• CPC Class Codes: C12N 9/1051 Hexosyltransferases (2.4.1)
• CPC Class Codes: C12P 19/04 Polysaccharides, i.e. compounds containing more than five saccharide radicals attached to each other by glycosidic bonds
• CPC Class Codes: C12P 19/26 Preparation of nitrogen-containing carbohydrates
• CPC Class Codes: Y10S 435/836 Bacillus licheniformis
• CPC Class Codes: Y10S 435/839 Bacillus subtilis
• CPC Class Codes: Y10S 435/848 Escherichia

Abstract

The present invention relates to a nucleic acid segment having a coding region segment encoding enzymatically active Streptococcus equisimilis hyaluronate synthase (seHAS), and to the use of this nucleic acid segment in the preparation of recombinant cells which produce hyaluronate synthase and its hyaluronic acid product. Hyaluronate is also known as hyaluronic acid or hyaluronan.

Claims

1. A method of producing hyaluronic acid, comprising the steps of:

providing a host cell having at least one expression construct comprising a hyaluronan synthase gene, wherein the hyaluronan synthase gene encodes an enzymatically active hyaluronan synthase that includes at least two amino acid motifs selected from the group consisting of;

2. The method of claim 1, further comprising the step of separating the hyaluronic acid from the host cell.

3. The method of claim 1, wherein the hyaluronan synthase gene is isolated from a prokaryotic source.

4. The method of claim 1, wherein the hyaluronan synthase gene is isolated from a microbial source.

5. A method of producing hyaluronic acid, comprising the steps of:

providing a host cell having at least one expression construct comprising a hyaluronan synthase gene, wherein the hyaluronan synthase gene encodes an enzymatically active hyaluronan synthase that includes at least three amino acid motifs selected from the group consisting of;

6. The method of claim 5, further comprising the step of separating the hyaluronic acid from the host cell.

7. The method of claim 5, wherein the hyaluronan synthase gene is isolated from a prokaryotic source.

8. The method of claim 5, wherein the hyaluronan synthase gene is isolated from a microbial source.

9. A method of producing hyaluronic acid, comprising the steps of:

providing a host cell having at least one expression construct comprising a hyaluronan synthase gene, wherein the hyaluronan synthase gene encodes an enzymatically active hyaluronan synthase that includes the amino acid motifs;

10. The method of claim 9, further comprising the step of separating the hyaluronic acid from the host cell.

11. The method of claim 9, wherein the hyaluronan synthase gene is isolated from a prokaryotic source.

12. The method of claim 9, wherein the hyaluronan synthase gene is isolated from a microbial source.

13. A method of producing hyaluronic acid, comprising the steps of:

providing a host cell having at least one expression construct comprising a hyaluronan synthase gene, wherein the hyaluronan synthase gene encodes an enzymatically active hyaluronan synthase that includes at least two amino acid motifs selected from the group consisting of;

14. The method of claim 13, further comprising the step of separating the hyaluronic acid from the host cell.

15. The method of claim 13, wherein the hyaluronan synthase gene is isolated from a prokaryotic source.

16. The method of claim 13, wherein the hyaluronan synthase gene is isolated from a microbial source.

17. A method of producing hyaluronic acid, comprising the steps of:

providing a host cell having at least one expression construct comprising a hyaluronan synthase gene, wherein the hyaluronan synthase gene encodes an enzymatically active hyaluronan synthase that includes at least three amino acid motifs selected from the group consisting of;

18. The method of claim 17, further comprising the step of separating the hyaluronic acid from the host cell.

19. The method of claim 17, wherein the hyaluronan synthase gene is isolated from a prokaryotic source.

20. The method of claim 17, wherein the hyaluronan synthase gene is isolated from a microbial source.

21. A method of producing hyaluronic acid, comprising the steps of:

providing a host cell having at least one expression construct comprising a hyaluronan synthase gene, wherein the hyaluronan synthase gene encodes an enzymatically active hyaluronan synthase that includes the amino acid motifs;

22. The method of claim 21, further comprising the step of separating the hyaluronic acid from the host cell.

23. The method of claim 21, wherein the hyaluronan synthase gene is isolated from a prokaryotic source.

24. The method of claim 21, wherein the hyaluronan synthase gene is isolated from a microbial source.