Improved substrate specificity of water-soluble pyrroloquinoline quinone glucose dehydrogenase by a peptide ligand.
A new approach in altering the substrate specificity of enzyme is proposed using glucose dehydrogenase, with pyrroroquinoine quinone (PQQGDH) as co-factor, as the model. This approach is based on the selection of random peptide phage displayed library. Using an M13 phage-display random peptide library, we have selected peptide ligands. Among the peptide ligands, a 7-mer peptide, composed of Thr-Thr-Ala-Thr-Glu-Tyr-Ser, caused PQQGDH substrate specificity to decrease significantly toward disaccharides, such as maltose and lactose, while a smaller effect was observed toward glucose. Consequently, this peptide narrowed the substrate specificity of PQQGDH, without a significant loss of the enzyme activity.
Yoshida
Hiromi
H
Department of Biotechnology, Faculty of Technology, Tokyo University of Agriculture and Technology, 2-24-16 Nakamachi, Koganei, Tokyo 184-8588, Japan.
Yagi
Yukiko
Y
Ikebukuro
Kazunori
K
Sode
Koji
K
D000595:Amino Acid Sequence; D002241:Carbohydrates / Q000737:chemistry* / Q000145:classification; D003067:Coenzymes / Q000737:chemistry; D004789:Enzyme Activation; D005948:Glucose Dehydrogenases / Q000737:chemistry*; D008024:Ligands; D046911:Macromolecular Substances; D008969:Molecular Sequence Data; D019151:Peptide Library; D010455:Peptides / Q000737:chemistry*; D015202:Protein Engineering / Q000379:methods*; D021122:Protein Subunits / Q000737:chemistry; D011786:Quality Control; D020539:Sequence Analysis, Protein / Q000379:methods; D012995:Solubility; D013379:Substrate Specificity; D014867:Water / Q000737:chemistry
D002241:Carbohydrates; D003067:Coenzymes; D008024:Ligands; D046911:Macromolecular Substances; D019151:Peptide Library; D010455:Peptides; D021122:Protein Subunits; D014867:Water; D005948:Glucose Dehydrogenases; C033955:glucose dehydrogenase (pyrroloquinoline-quinone)