Polynucleotide sequence for production of glucose oxidase in recombinant systems

• US 5,266,688 A
• Filed: 03/06/1992
• Issued: 11/30/1993
• Est. Priority Date: 06/21/1988
• Status: Expired due to Term

• Current Assignee: Chiron Corporation (Bayer AG)
• Original Assignee: Chiron Corporation (Bayer AG)
• Inventors: Rosenberg, Steven
• Primary Examiner(s): Schwartz, Richard A.
• Assistant Examiner(s): BROWN, GARY
• Application Number:
  US07/848,406
• Time in Patent Office: 634 Days
• Field of Search: 536/27, 536/23.2
• US Class Current: 536/23.2
• CPC Class Codes: C07K 16/40 against enzymes
• CPC Class Codes: C12N 15/81 for yeasts
• CPC Class Codes: C12N 9/0006 acting on CH-OH groups as donors (1.1)

Abstract

The present invention provides recombinant polynucleotides which encode glucose oxidase (GO). It also provides recombinant expression systems which produce, and when desired, secrete active GO and GO analogs into the extracellular medium.

Claims

1. A recombinant molecule comprising a polynucleotide encoding a polypeptide which exhibits glucose oxidase activity, wherein said polynucleotide encodes the polypeptide shown in FIG. 5B.

2. The molecule of claim 1, further comprising expression control sequences operationally linked to said polynucleotide, wherein the expression control sequences control expression of the polypeptide.

3. The molecule of claim 2, further comprising a secretion control sequence which controls secretion of the polypeptide.

4. The molecule of claim 2, wherein the expression control sequences control expression of the polypeptide in prokaryotic cells.

5. The molecule of claim 2, wherein the expression control sequences control expression of the polypeptide in eukaryotic cells.

6. The molecule of claim 5, wherein the expression control sequences control expression of the polypeptide in yeast.

7. The molecule of claim 6, further comprising a secretion control sequence.

8. The molecule of claim 3, wherein the expression control sequences comprise the ADH2-GAP hybrid promoter and the GAP terminator, and the secretion control sequence is selected from the group consisting of the alpha factor sequence and the GO prepro sequence.

9. The molecule of claim 8, wherein the molecule is selected from the group consisting of pAB24AGS_GO GO, pAB24AG_alpha GO, pAB24AGSGO, and pAB24AG5/8GO.

10. A recombinant molecule comprising a polynucleotide encoding a polypeptide which exhibits glucose oxidase (GO) activity, wherein said polynucleotide has the sequence shown in FIG. 5B.

11. The molecule of claim 10, further comprising expression control sequences operationally linked to said polynucleotide, wherein the expression control sequences control expression of the polypeptide.

12. The molecule of claim 11, further comprising a secretion control sequence which controls secretion of the polypeptide.

13. The molecule of claim 11, wherein the expression control sequences control expression of the polypeptide in prokaryotic cells.

14. The molecule of claim 11, wherein the expression control sequences control expression of the polypeptide in eukaryotic cells.

15. The molecule of claim 14, wherein the expression control sequences control expression of the polypeptide in yeast.

16. The molecule of claim 15, further comprising a secretion control sequence which controls secretion of the polypeptide.

17. The molecule of claim 12, wherein the expression control sequences comprise the ADH2-GAP hybrid promoter and the GAP terminator, and the secretion control sequence is selected from the group consisting of the alpha factor and the GO prepro sequence.

18. The molecule of claim 17, wherein the molecule is selected from the group consisting of pAB24AGS_GO GO, pAB24AG_alpha GO, pAG24AGSGO, and pAB24AG@GO.