Flavin-bound glucose dehydrogenases, a method for producing a flavin-bound glucose dehydrogenase, and yeast transformant used for the same
A heat-resistant flavin-bound glucose dehydrogenase having a high substrate specificity for D-glucose, an method for producing the same, and a transformant used for the same. A flavin-bound glucose dehydrogenase gene encoding a flavin-bound glucose dehydrogenase derived from Mucor is introduced into yeast, Zygosaccharomyces , to obtain a transformant. Subsequently, the yeast transformant is cultured to obtain a flavin-bound glucose dehydrogenase from the culture. The heat-resistant flavin-bound glucose dehydrogenase is less susceptible to the effects of dissolved oxygen and allows accurate measurement of glucose even in the presence of sugar compounds other than glucose in a sample.
1. A flavin-bound glucose dehydrogenase having the amino acid sequence of SEQ ID NO: 3, and having the following properties (i) to (iv):
(i) action: glucose dehydrogenase activity in the presence of an electron acceptor;
(ii) molecular weight: the molecular weight of the polypeptide chain portion of the protein is approximately 80 kDa;
(iii) substrate specificity: lower reactivity to maltose, D-galactose, and D-xylose than that to D-glucose; and
(iv) 50% or above residual activity after heat treatment at 50° C. for 15 minutes, wherein the flavin-bound glucose dehydrogenase is produced by a yeast transformant.
2. A flavin-bound glucose dehydrogenase having an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 3, and having the following properties (i) to (iv):
(i) action: glucose dehydrogenase activity in the presence of an electron acceptor;
(ii) molecular weight: the molecular weight of the polypeptide chain portion of the protein is approximately 80 kDa;
(iii) substrate specificity: lower reactivity to maltose, D-galactose, and D-xylose than that to D-glucose; and
(iv) 50% or above residual activity after heat treatment at 50° C. for 15 minutes, wherein the flavin-bound glucose dehydrogenase is produced by a yeast transformant.
3. The flavin-bound glucose dehydrogenase of claim 1 , wherein said flavin-bound glucose dehydrogenase is obtained from a microorganism classified as Mucoromycotina.
4. The flavin-bound glucose dehydrogenase of claim 2 , wherein said flavin-bound glucose dehydrogenase is obtained from a microorganism classified as Mucoromycotina.
5. The flavin-bound glucose dehydrogenase of claim 1 , wherein said glucose dehydrogenase is obtained from a microorganism classified as Mucoromycetes.
6. The flavin-bound glucose dehydrogenase of claim 1 , wherein said glucose dehydrogenase is obtained from a microorganism classified as Mucorales.
7. The flavin-bound glucose dehydrogenase of claim 1 , wherein said glucose dehydrogenase is obtained from a microorganism classified as Mucoraceae.
8. The flavin-bound glucose dehydrogenase of claim 2 , wherein said glucose dehydrogenase is obtained from a microorganism classified as Mucoromycetes.
9. The flavin-bound glucose dehydrogenase of claim 2 , wherein said glucose dehydrogenase is obtained from a microorganism classified as Mucorales.
10. The flavin-bound glucose dehydrogenase of claim 2 , wherein said glucose dehydrogenase is obtained from a microorganism classified as Mucoraceae.