DNA encoding hyaluronan synthase from Pasteurella multocida and methods of use

• US 20050266460A1
• Filed: 05/09/2005
• Published: 12/01/2005
• Est. Priority Date: 04/02/1998
• Status: Active Grant

• Current Assignee: Paul Deangelis
• Original Assignee: Paul Deangelis
• Inventors: DeAngelis, Paul
• Granted Patent: US 7,232,684 B2
• Time in Patent Office: Days
• US Class Current: 435/6
• CPC Class Codes: A61K 39/102 Pasteurellales, e.g. Actinobacillus, Pasteurella; Haemophilus
• CPC Class Codes: A61K 47/36 Polysaccharides; Derivatives thereof, e.g. gums, starch, alginate, dextrin, hyaluronic acid, chitosan, inulin, agar or pectin
• CPC Class Codes: A61L 24/08 Polysaccharides A61L24/043 takes precedence
• CPC Class Codes: A61L 29/085 Macromolecular materials
• CPC Class Codes: C08L 5/08 Chitin; Chondroitin sulfate; Hyaluronic acid; Derivatives thereof
• 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

Abstract

The present invention relates to a nucleic acid segment having a coding region segment encoding enzymatically active Pasteurella multocida hyaluronate synthase (PmHAS), and to the use of this nucleic acid segment in the preparation of recombinant cells which produce hyaluronate synthase and its hyaluronic acid product.

Claims

1. A purified and isolated nucleic acid sequence encoding enzymatically active hyaluronan synthase, the nucleic acid sequence selected from the group consisting of:

(a) the nucleic acid sequence as set forth in SEQ ID NO;

2; and

(b) a nucleic acid sequence encoding an enzymatically active hyaluronan synthase, wherein a complement of the nucleic acid sequence is capable of hybridizing to the nucleic acid sequence of SEQ ID NO;

2 under standard hybridization conditions.

2. A procaryotic or eucaryotic host cell transformed or transfected with a nucleic acid segment according to claim 1 in a manner allowing the host cell to express hyaluronic acid.

3. An isolated nucleic acid segment consisting essentially of a nucleic acid segment encoding enzymatically active hyaluronan synthase from Pasteurella, wherein a complement of the nucleic acid sequence is capable of hybridizing to the nucleic acid sequence of SEQ ID NO:

2 under standard hybridization conditions.

4. A cDNA sequence according to claim 3.

5. A procaryotic or eucaryotic host cell transformed or transfected with a nucleic acid segment according to claim 4 in a manner allowing the host cell to express hyaluronic acid.

6. An isolated nucleic acid segment consisting essentially of a nucleic acid segment encoding enzymatically active hyaluronan synthase from Pasteurella, wherein the nucleic acid segment is essentially as set forth in SEQ ID NO:

2.

7. A cDNA sequence according to claim 6.

8. A procaryotic or eucaryotic host cell transformed or transfected with a nucleic acid segment according to claim 6 in a manner allowing the host cell to express hyaluronic acid.

9. An isolated nucleic acid segment consisting essentially of a nucleic acid segment encoding enzymatically active hyaluronan synthase from Pasteurella, wherein the nucleic acid segment is at least 85% identical to SEQ ID NO:

2.

10. A cDNA sequence according to claim 9.

11. A procaryotic or eucaryotic host cell transformed or transfected with a nucleic acid segment according to claim 9 in a manner allowing the host cell to express hyaluronic acid.

12. An isolated nucleic acid segment consisting essentially of a nucleic acid segment encoding enzymatically active hyaluronan synthase from Pasteurella, wherein the nucleic acid segment is essentially as set forth in SEQ ID NO:

2 and encodes a hyaluronan synthase, wherein the hyaluronan synthase has SEQ ID NO;

18 and DSD motifs therein.

13. A cDNA sequence according to claim 12.

14. A procaryotic or eucaryotic host cell transformed or transfected with a nucleic acid segment according to claim 12 in a manner allowing the host cell to express hyaluronic acid.

15. An isolated nucleic acid segment consisting essentially of a nucleic acid segment encoding enzymatically active hyaluronan synthase from Pasteurella, wherein the hyaluronate synthase encoded by the nucleic acid segment is at least 89% identical to SEQ ID NO:

1.

16. A cDNA sequence according to claim 15.

17. A procaryotic or eucaryotic host cell transformed or transfected with a nucleic acid segment according to claim 15 in a manner allowing the host cell to express hyaluronic acid.