Improved mutants of pyrroloquinoline quinone dependent...

C - Chemistry – Metallurgy – 12 – N

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C12N 9/04 (2006.01)

Patent

CA 2649135

The present invention discloses a mutant of PQQ-dependent soluble glucose dehydrogenase (s-GDH; EC 1.1.5.2) with improved specificity for glucose as compared to maltose, having a substitution of threonine at position 348 by either glycine, alanine or serine, wherein said mutant additionally comprises, at least one mutation for improving the stability of the mutant and one or more mutation(s) for improving the affinity of the mutant to glucose, and/or one or more mutation(s) for further improving the specificity of the mutant for glucose as compared to maltose, and wherein position 348 correspond to the amino acid positions known from the A. calcoaceticuss-GDH wild-type sequence. Also disclosed are genes encoding such mutant s-GDH, and different applications of these s-GDH mutants, particularly for determining the concentration of glucose in a sample.

La présente invention concerne un mutant de la glucose déshydrogénase soluble dépendante de la PQQ (s-GDH; EC 1.1.5.2) ayant une meilleure spécificité vis-à-vis du glucose par rapport au maltose, ayant une substitution de la thréonine au niveau de la position 348 par soit une glycine, soit une alanine, soit une sérine, ledit mutant comprenant en plus au moins une mutation servant à améliorer la stabilité du mutant et une ou plusieurs mutations servant à améliorer l'affinité du mutant vis-à-vis du glucose et/ou une ou plusieurs mutations servant à améliorer davantage la spécificité du mutant vis-à-vis du glucose par rapport au maltose et ladite position 348 correspondant aux positions des acides aminés connues d'après la séquence de type sauvage de s-GDH de A. calcoaceticus. L'invention concerne également des gènes codant pour de telles s-GDH mutantes et différentes applications de ces mutants de la s-GDH, en particulier pour déterminer la concentration de glucose dans un échantillon.

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