Hyaluronan synthase gene and uses therof

• US 6,833,264 B1
• Filed: 12/21/1999
• Issued: 12/21/2004
• Est. Priority Date: 10/31/1997
• Status: Expired due to Term

• 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:
  US09/469,200
• Time in Patent Office: 1,827 Days
• Field of Search: 435/320.1, 435/252.3, 435/252.31, 435/252.33, 435/69.1, 435/193, 435/836, 435/839, 435/848, 435/254.11, 435/325, 536/23.2
• US Class Current: 435/252.31
• 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. An isolated and purified nucleic acid segment encoding the hyaluronate synthase of SEQ ID NO:

2.

3. A recombinant expression vector comprising the nucleic acid segment of claim 1.

4. The recombinant expression vector of claim 3, wherein the recombinant expression vector is a plasmid.

5. The recombinant expression vector of claim 3, wherein the recombinant expression vector is an expression vector having a promoter operatively linked to the nucleic acid segment of claim 1.

6. The recombinant expression vector of claim 3, 4 or 5, placed in a recombinant strain of Bacillus or Escherichia or Enterococcus.

7. The recombinant expression vector of claim 6 wherein the recombinant strain of Bacillus is selected from the group consisting of Bacillus subtilis, Bacillus licheniformis, or Bacillus thuringiensis.

12. The recombinant expression vector of claim 5, wherein the promoter is a heterologous promoter.

13. The recombinant host cell of claim 6 wherein said host cell produces hyaluronic acid.

2. An isolated and purified nucleic acid segment encoding a hyaluronate synthase, wherein the isolated and purified nucleic acid segment comprises the nucleic acid segment of SEQ ID NO:

1.

8. A recombinant expression vector comprising the nucleic acid segment of claim 2.

9. The recombinant expression vector of claim 8, wherein the recombinant expression vector is a plasmid.

10. A recombinant host cell comprising the recombinant expression vector of claim 8 or 9, which is a strain of Bacillus or Escherichia or Enterococcus.

11. The recombinant host cell of claim 10 wherein the recombinant strain of Bacillus is selected from the group consisting of Bacillus subtilis, Bacillus licheniformis, or Bacillus thuringiensis.

14. The recombinant host cell of claim 10 wherein said host cell produces hyaluronic acid.