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US 8,999,140 B2Granted
Glucose oxidase mutants, compositions, devices, kits and uses thereof
AssigneesRoche Diagnostics Operations, Inc., Ultizyme International Ltd.
InventorsKatsuhiro Kojima, Kazushige Mori, Sode Koji
Life of this patent
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Abstract
Compositions, devices, kits and methods are disclosed for assaying glucose with a glucose oxidase mutant that has been modified at an amino acid residue involved in the active site. The glucose oxidase mutant has reduced oxidase activity while substantially maintaining its dehydrogenase activity.
Claims17
A glucose oxidase mutant modified at one or more amino acid positions selected from:
- (a). a position corresponding to position 53 of the amino acid sequence set forth in SEQ ID NO:
- 1 by substituting the amino acid residue Thr with another amino acid residue;
- (b). a position corresponding to position 116 of the amino acid sequence set forth in SEQ ID NO:
- 1 by substituting the amino acid residue Ile with another amino acid residue;
- (c). a position corresponding to position 132 of the amino acid sequence set forth in SEQ ID NO:
- 1 by substituting the amino acid residue Ser or Thr with another amino acid residue;
- (d). a position corresponding to position 134 of the amino acid sequence set forth in SEQ ID NO:
- 1 by substituting the amino acid residue Thr with another amino acid residue;
- (e). a position corresponding to position 237 of the amino acid sequence set forth in SEQ ID NO:
- 1 by substituting the amino acid residue Ile or Phe with another amino acid residue;
- (f). a position corresponding to position 371 of the amino acid sequence set forth in SEQ ID NO:
- 1 by substituting the amino acid residue Val or Ala with another amino acid residue;
- (g). a position corresponding to position 373 of the amino acid sequence set forth in SEQ ID NO:
- 1 by substituting the amino acid residue Phe with another amino acid residue;
- (h). a position corresponding to position 434 of the amino acid sequence set forth in SEQ ID NO:
- 1 by substituting the amino acid residue Glu with another amino acid residue;
- (i). a position corresponding to position 436 of the amino acid sequence set forth in SEQ ID NO:
- 1 by substituting the amino acid residue Phe with another amino acid residue;
- (j). a position corresponding to position 448 of the amino acid sequence set forth in SEQ ID NO:
- 1 by substituting the amino acid residue Trp with another amino acid residue;
- and
- (k). a position corresponding to position 537 of the amino acid sequence set forth in SEQ ID NO:
- 1 by substituting the amino acid residue Trp with another amino acid residue.
20 more elements
A glucose oxidase mutant comprising an amino acid sequence selected from the group consisting of SEQ ID NOs:
- 1-21 modified at one or more positions corresponding to position 53, 116, 132, 134, 237, 371, 373, 434, 436, 448 or 537 of the amino acid sequence set forth in SEQ ID NO:
- 1, wherein the glucose oxidase mutant has a reduced oxidase activity when compared to a wild-type glucose oxidase.
A method of assaying glucose in a sample, the method comprising the steps of:
- contacting the sample with a glucose oxidase mutant modified at one or more amino acid positions selected from:
- (a). a position corresponding to position 53 of the amino acid sequence set forth in SEQ ID NO:
- 1 by substituting the amino acid residue Thr with another amino acid residue,
- (b). a position corresponding to position 116 of the amino acid sequence set forth in SEQ ID NO:
- 1 by substituting the amino acid residue Ile with another amino acid residue,
- (c). a position corresponding to position 132 of the amino acid sequence set forth in SEQ ID NO:
- 1 by substituting the amino acid residue Ser or Thr with another amino acid residue,
- (d). a position corresponding to position 134 of the amino acid sequence set forth in SEQ ID NO:
- 1 by substituting the amino acid residue Thr with another amino acid residue,
- (e). a position corresponding to position 237 of the amino acid sequence set forth in SEQ ID NO:
- 1 by substituting the amino acid residue Ile or Phe with another amino acid residue,
- (f). a position corresponding to position 371 of the amino acid sequence set forth in SEQ ID NO:
- 1 by substituting the amino acid residue Val or Ala with another amino acid residue,
- (g). a position corresponding to position 373 of the amino acid sequence set forth in SEQ ID NO:
- 1 by substituting the amino acid residue Phe with another amino acid residue,
- (h). a position corresponding to position 434 of the amino acid sequence set forth in SEQ ID NO:
- 1 by substituting the amino acid residue Glu with another amino acid residue,
- (i). a position corresponding to position 436 of the amino acid sequence set forth in SEQ ID NO:
- 1 by substituting the amino acid residue Phe with another amino acid residue,
- (j). a position corresponding to position 448 of the amino acid sequence set forth in SEQ ID NO:
- 1 by substituting the amino acid residue Trp with another amino acid residue, and
- (k). a position corresponding to position 537 of the amino acid sequence set forth in SEQ ID NO:
- 1 by substituting the amino acid residue Trp with another amino acid residue;
- and
- measuring an amount of glucose oxidized by the glucose oxidase.
22 more elements
Dependencies are inferred from the claim text.