Genetically engineered pyrroloquinoline quinone dependent glucose dehydrogenase comprising an amino acid insertion
AssigneeRoche Diagnostics Operations, Inc.
InventorsMara Boenitz-Dulat, Jessica Laggerbauer, Rainer Schmuck, Peter Kratzsch, Wolfgang-Reinhold Knappe
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Abstract
The present invention relates to improved variants of soluble pyrroloquinoline quinone (PQQ)-dependent glucose dehydrogenases (s-GDH) comprising an amino acid insertion between positions 428 and 429 as corresponding to the amino acid sequence known from Acinetobacter calcoaceticus, to genes encoding such variant s-GDH, to proteins of such s-GDH variants with improved substrate specificity for glucose, and to different applications of these s-GDH variants, particularly for determining concentrations of sugars, especially of glucose in a sample.
Claims13
A variant of PQQ-dependent soluble glucose dehydrogenase (s-GDH), said variant comprising the sequence of SEQ ID NO:
- 2 modified by the insertion of one amino acid residue between the amino acid positions corresponding to positions 428 and 429 of the s-GDH wild-type sequence known from A. calcoaceticus(SEQ ID NO:
- 2), a substitution of threonine at position 348 of SEQ ID NO:
- 2 with an amino acid residue selected from the group consisting of alanine, glycine and serine, and optionally, one or more additional amino acid substitutions, wherein said variant is at least 90% identical to SEQ ID NO:
- 2 , and has s-GDH activity.
Dependencies are inferred from the claim text.