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Literature summary for 1.1.5.9 extracted from

  • Yamaoka, H.; Yamashita, Y.; Ferri, S.; Sode, K.
    Site directed mutagenesis studies of FAD-dependent glucose dehydrogenase catalytic subunit of Burkholderia cepacia (2008), Biotechnol. Lett., 30, 1967-1972.
    View publication on PubMed

Application

Application Comment Organism
diagnostics glucose sensor/glucose determination Burkholderia cepacia

Cloned(Commentary)

Cloned (Comment) Organism
expressed as recombinant protein in Escherichia coli strain DH5alpha Burkholderia cepacia
expression in Escherichia coli Burkholderia cepacia

Protein Variants

Protein Variants Comment Organism
A472C site directed mutagenesis Burkholderia cepacia
A472D site directed mutagenesis Burkholderia cepacia
A472E site directed mutagenesis Burkholderia cepacia
A472F site directed mutagenesis Burkholderia cepacia
A472F mutant with higher substrate specificity for glucose in relation to maltose, constructed by site-directed mutagenesis Burkholderia cepacia
A472G site directed mutagenesis Burkholderia cepacia
A472H site directed mutagenesis Burkholderia cepacia
A472I site directed mutagenesis Burkholderia cepacia
A472K site directed mutagenesis Burkholderia cepacia
A472L site directed mutagenesis Burkholderia cepacia
A472M site directed mutagenesis Burkholderia cepacia
A472N site directed mutagenesis Burkholderia cepacia
A472P site directed mutagenesis Burkholderia cepacia
A472Q site directed mutagenesis Burkholderia cepacia
A472R site directed mutagenesis Burkholderia cepacia
A472S site directed mutagenesis Burkholderia cepacia
A472T site directed mutagenesis Burkholderia cepacia
A472V site directed mutagenesis Burkholderia cepacia
A472W site directed mutagenesis Burkholderia cepacia
A472Y site directed mutagenesis Burkholderia cepacia
N475A site directed mutagenesis Burkholderia cepacia
N475C site directed mutagenesis Burkholderia cepacia
N475D site directed mutagenesis Burkholderia cepacia
N475D mutant with higher substrate specificity for glucose in relation to maltose, constructed by site-directed mutagenesis Burkholderia cepacia
N475E site directed mutagenesis Burkholderia cepacia
N475F site directed mutagenesis Burkholderia cepacia
N475G site directed mutagenesis Burkholderia cepacia
N475H site directed mutagenesis Burkholderia cepacia
N475I site directed mutagenesis Burkholderia cepacia
N475K site directed mutagenesis Burkholderia cepacia
N475L site directed mutagenesis Burkholderia cepacia
N475M site directed mutagenesis Burkholderia cepacia
N475P site directed mutagenesis Burkholderia cepacia
N475Q site directed mutagenesis Burkholderia cepacia
N475R site directed mutagenesis Burkholderia cepacia
N475S site directed mutagenesis Burkholderia cepacia
N475T site directed mutagenesis Burkholderia cepacia
N475V site directed mutagenesis Burkholderia cepacia
N475W site directed mutagenesis Burkholderia cepacia
N475Y site directed mutagenesis Burkholderia cepacia

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
17
-
D-glucose A472F mutant, higher specificity toward glucose compared to the wild type enzyme, no activity towards maltose Burkholderia cepacia
17
-
glucose FADGDH mutant A472F Burkholderia cepacia
28
-
D-glucose N475D mutant, higher specificity toward glucose compared to the wild type enzyme, no activity towards maltose Burkholderia cepacia
28
-
glucose FADGDH mutant N475D Burkholderia cepacia

Organism

Organism UniProt Comment Textmining
Burkholderia cepacia
-
-
-

Purification (Commentary)

Purification (Comment) Organism
-
Burkholderia cepacia
by DEAE-5PW and Superdex-200 10/300GL column chromatography Burkholderia cepacia

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
additional information
-
590 micromol/min/mg Vmax for the FADGDH mutant A472F Burkholderia cepacia
additional information
-
770 micromol/min/mg Vmax for the FADGDH mutant N475D Burkholderia cepacia
4
-
A472F mutant towards maltose Burkholderia cepacia
21
-
N475D mutant towards maltose Burkholderia cepacia
118
-
N475D mutant towards glucose Burkholderia cepacia
120
-
A472F mutant towards glucose Burkholderia cepacia
139
-
wild type towards maltose Burkholderia cepacia
430
-
enzymatic activity of wild-type FADGDH in an assay with 40 mM maltose Burkholderia cepacia
543
-
wild type towards glucose Burkholderia cepacia
950
-
enzymatic activity of wild-type FADGDH in an assay with 40 mM glucose Burkholderia cepacia

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
D-glucose + 2,6-dichlorophenolindophenol
-
Burkholderia cepacia D-glucono-1,5-lactone + reduced 2,6-dichlorophenolindophenol
-
?
D-glucose + acceptor
-
Burkholderia cepacia D-glucono-1,5-lactone + reduced acceptor
-
?
D-maltose + 2,6-dichlorophenolindophenol
-
Burkholderia cepacia ? + reduced 2,6-dichlorophenolindophenol
-
?
D-maltose + acceptor
-
Burkholderia cepacia ?
-
?

Synonyms

Synonyms Comment Organism
FAD-dependent glucose dehydrogenase
-
Burkholderia cepacia
FAD-dependent glucose dehydrogenase FADGDH Burkholderia cepacia
FADGDH
-
Burkholderia cepacia
glucose dehydrogenase
-
Burkholderia cepacia

Cofactor

Cofactor Comment Organism Structure
FAD
-
Burkholderia cepacia