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Literature summary for 1.1.1.119 extracted from

  • Pire, C.; Esclapez, J.; Diaz, S.; Perez-Pomares, F.; Ferrer, J.; Bonete, M.
    Alteration of coenzyme specificity in halophilic NAD(P)+ glucose dehydrogenase by site-directed mutagenesis (2009), J. Mol. Catal. B, 59, 261-265.
No PubMed abstract available

Cloned(Commentary)

Cloned (Comment) Organism
expressed as recombinant protein in Escherichia coli strain BL21(DE3) Haloferax mediterranei
expressed in Escherichia coli BL21 (DE3) cells Haloferax mediterranei
expression as inclusion bodies in Escherichia coli Haloferax mediterranei

Protein Variants

Protein Variants Comment Organism
G206D less efficient with NADP+ than the wild type, 1.6fold increase in ratio kcat/Km for NAD+ Haloferax mediterranei
G206D prefers NAD+ over NADP+ Haloferax mediterranei
G206D the mutant is less efficient with NADP+ than the wild type enzyme, the relation kcat/KNAD+ is 1.6times higher than in the wild type, resulting in an enzyme that prefers NAD+ over NADP+ Haloferax mediterranei
G206D/R207I active only with NAD+ Haloferax mediterranei
G206D/R207I no activity with NADP+, highest reaction rate with cofactor NAD+ of all mutants tested Haloferax mediterranei
G206D/R207I the mutant shows no activity with NADP+, when the coenzyme NAD+ is incubated with this double mutant, it reaches the highest kcat value, between 1.5 and 2times higher than the kcat of the wild type enzyme with NADP+, and between 3 and 4times higher than the kcat of the wild type with NAD+ Haloferax mediterranei
G206D/R207I/R208N active only with NAD+ Haloferax mediterranei
G206D/R207I/R208N no activity with NADP+ Haloferax mediterranei
G206D/R207I/R208N the mutant shows no activity with NADP+ Haloferax mediterranei
R207I less efficient with NAD+ or NADP+ than the wild-type enzyme Haloferax mediterranei
R207I less efficient with NADP+ than the wild type Haloferax mediterranei
R207I the mutant is less efficient with NADP+ than the wild type enzyme, shows an increase of 48times in Km value with NADP+ when compared with the wild type accompanied by a decrease in kcat, which clearly makes the R207I mutant less efficient in catalysis with NADP+, the R207I substitution also makes the enzyme less efficient with NAD+, with a decrease of 4 times in kcat/Km, this substitution also increases the Km for glucose Haloferax mediterranei

Inhibitors

Inhibitors Comment Organism Structure
NAD(P)+
-
Haloferax mediterranei
NAD+
-
Haloferax mediterranei
NADP+
-
Haloferax mediterranei

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.035
-
NADP+ wild type enzyme Haloferax mediterranei
0.035
-
NAD(P)+ with NADP+ as coenzyme Haloferax mediterranei
0.035
-
NADP+ wild-type, pH 8.8, 40°C Haloferax mediterranei
0.4
-
NAD+ wild type enzyme Haloferax mediterranei
0.4
-
NAD(P)+ with NAD+ as coenzyme Haloferax mediterranei
0.4
-
NAD+ wild-type, pH 8.8, 40°C Haloferax mediterranei
0.49
-
NAD+ mutant enzyme G206D Haloferax mediterranei
0.49
-
NAD+ mutant G206D, pH 8.8, 40°C Haloferax mediterranei
1.3
-
NAD+ mutant enzyme G206D/R207I/R208N Haloferax mediterranei
1.3
-
NAD+ mutant enzyme R207I Haloferax mediterranei
1.3
-
NAD+ mutant G206D/R207I/R208N, pH 8.8, 40°C Haloferax mediterranei
1.3
-
NAD+ mutant R207I, pH 8.8, 40°C Haloferax mediterranei
1.7
-
NADP+ mutant enzyme R207I Haloferax mediterranei
1.7
-
NADP+ mutant R207I, pH 8.8, 40°C Haloferax mediterranei
2.6
-
NADP+ mutant enzyme G206D Haloferax mediterranei
2.6
-
NADP+ mutant G206D, pH 8.8, 40°C Haloferax mediterranei
2.7
-
NAD+ mutant enzyme G206D/R207I Haloferax mediterranei
2.7
-
NAD+ mutant G206D/R207I, pH 8.8, 40°C Haloferax mediterranei
2.8
-
D-glucose with NADP+ as coenzyme Haloferax mediterranei
2.8
-
D-glucose wild-type, pH 8.8, 40°C Haloferax mediterranei
12.9
-
D-glucose with NAD+ as coenzyme Haloferax mediterranei
12.9
-
D-glucose cosubstrate NAD+, wild-type, pH 8.8, 40°C Haloferax mediterranei
12.9
-
beta-D-glucose wild type enzyme, using NAD+ as cosubstrate Haloferax mediterranei
15
-
D-glucose cosubstrate NAD+, mutant G206D, pH 8.8, 40°C Haloferax mediterranei
15
-
beta-D-glucose mutant enzyme G206D, using NAD+ as cosubstrate Haloferax mediterranei
24
-
beta-D-glucose mutant enzyme R207I, using NADP+ as cosubstrate Haloferax mediterranei
24
-
D-glucose mutant R207I, pH 8.8, 40°C Haloferax mediterranei
31.1
-
beta-D-glucose wild type enzyme, using NADP+ as cosubstrate Haloferax mediterranei
32
-
D-glucose cosubstrate NAD+, mutant R207I, pH 8.8, 40°C Haloferax mediterranei
32
-
beta-D-glucose mutant enzyme R207I, using NAD+ as cosubstrate Haloferax mediterranei
52
-
beta-D-glucose mutant enzyme G206D, using NADP+ as cosubstrate Haloferax mediterranei
52
-
D-glucose mutant G206D, pH 8.8, 40°C Haloferax mediterranei
57
-
D-glucose cosubstrate NAD+, mutant G206D/R207I/R208N, pH 8.8, 40°C Haloferax mediterranei
57
-
beta-D-glucose mutant enzyme G206D/R207I/R208N, using NAD+ as cosubstrate Haloferax mediterranei
80
-
D-glucose cosubstrate NAD+, mutant G206D/R207I, pH 8.8, 40°C Haloferax mediterranei
80
-
beta-D-glucose mutant enzyme G206D/R207I, using NAD+ as cosubstrate Haloferax mediterranei

Organism

Organism UniProt Comment Textmining
Haloferax mediterranei
-
-
-
Haloferax mediterranei Q977U7
-
-

Purification (Commentary)

Purification (Comment) Organism
-
Haloferax mediterranei

Renatured (Commentary)

Renatured (Comment) Organism
-
Haloferax mediterranei

Storage Stability

Storage Stability Organism
4°C, stable for weeks Haloferax mediterranei

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
beta-D-glucose + NAD+ the enzyme has a strong preference for NADP+ over NAD+ Haloferax mediterranei D-glucono-1,5-lactone + NADH + H+
-
?
beta-D-glucose + NADP+ the enzyme has a strong preference for NADP+ over NAD+ Haloferax mediterranei D-glucono-1,5-lactone + NADPH + H+
-
?
D-glucose + NAD(P)+
-
Haloferax mediterranei D-glucono-1,5-lactone + NAD(P)H + H+
-
?
D-glucose + NAD+
-
Haloferax mediterranei D-glucono-1,5-lactone + NADH + H+
-
?
D-glucose + NAD+
-
Haloferax mediterranei D-glucono-1,5-lactone + NADH
-
?
D-glucose + NADP+
-
Haloferax mediterranei D-glucono-1,5-lactone + NADPH
-
?

Synonyms

Synonyms Comment Organism
beta-D-glucose: NAD(P) 1-oxidoreductase
-
Haloferax mediterranei
GDH
-
Haloferax mediterranei
halophilic glucose dehydrogenase
-
Haloferax mediterranei
Hm GDH
-
Haloferax mediterranei
NAD(P)+ glucose dehydrogenase
-
Haloferax mediterranei

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.047
-
D-glucose cosubstrate NAD+, mutant G206D/R207I/R208N, pH 8.8, 40°C Haloferax mediterranei
0.072
-
D-glucose cosubstrate NAD+, mutant R207I, pH 8.8, 40°C Haloferax mediterranei
0.2
-
D-glucose mutant R207I, pH 8.8, 40°C Haloferax mediterranei
0.26
-
D-glucose cosubstrate NAD+, wild-type, pH 8.8, 40°C Haloferax mediterranei
0.26
-
D-glucose mutant G206D, pH 8.8, 40°C Haloferax mediterranei
0.5
-
D-glucose wild-type, pH 8.8, 40°C Haloferax mediterranei
0.57
-
D-glucose cosubstrate NAD+, mutant G206D, pH 8.8, 40°C Haloferax mediterranei
0.9
-
D-glucose cosubstrate NAD+, mutant G206D/R207I, pH 8.8, 40°C Haloferax mediterranei

Cofactor

Cofactor Comment Organism Structure
NAD+ poor substrate for wild-type Haloferax mediterranei
NADP+
-
Haloferax mediterranei
NADP+ the Km value of the wild type enzyme is 11fold lower for NADP+ than for NAD+, indicating that the enzyme has a strong preference for NADP+ Haloferax mediterranei

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
0.09
-
NADP+ wild-type, pH 8.8, 40°C Haloferax mediterranei
0.09
-
NAD(P)+ with NADP+ as coenzyme Haloferax mediterranei
0.69
-
NADP+ mutant G206D, pH 8.8, 40°C Haloferax mediterranei
1
-
NAD+ mutant G206D, pH 8.8, 40°C Haloferax mediterranei
1.17
-
NAD+ mutant G206D/R207I, pH 8.8, 40°C Haloferax mediterranei
1.3
-
NADP+ mutant R207I, pH 8.8, 40°C Haloferax mediterranei
1.7
-
NAD+ mutant G206D/R207I/R208N, pH 8.8, 40°C Haloferax mediterranei
2
-
NAD+ wild-type, pH 8.8, 40°C Haloferax mediterranei
2
-
NAD(P)+ with NAD+ as coenzyme Haloferax mediterranei
2.1
-
NAD+ mutant R207I, pH 8.8, 40°C Haloferax mediterranei

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.0000008
-
D-glucose cosubstrate NAD+, mutant G206D/R207I/R208N, pH 8.8, 40°C Haloferax mediterranei
0.000002
-
D-glucose cosubstrate NAD+, mutant R207I, pH 8.8, 40°C Haloferax mediterranei
0.000005
-
D-glucose mutant G206D, pH 8.8, 40°C Haloferax mediterranei
0.000008
-
D-glucose mutant R207I, pH 8.8, 40°C Haloferax mediterranei
0.000012
-
D-glucose cosubstrate NAD+, mutant G206D/R207I, pH 8.8, 40°C Haloferax mediterranei
0.00002
-
D-glucose cosubstrate NAD+, wild-type, pH 8.8, 40°C Haloferax mediterranei
0.000035
-
NAD+ mutant G206D/R207I/R208N, pH 8.8, 40°C Haloferax mediterranei
0.000038
-
D-glucose cosubstrate NAD+, mutant G206D, pH 8.8, 40°C Haloferax mediterranei
0.000055
-
NAD+ mutant R207I, pH 8.8, 40°C Haloferax mediterranei
0.0001
-
NADP+ mutant G206D, pH 8.8, 40°C Haloferax mediterranei
0.00012
-
NADP+ mutant R207I, pH 8.8, 40°C Haloferax mediterranei
0.00019
-
D-glucose wild-type, pH 8.8, 40°C Haloferax mediterranei
0.00033
-
NAD+ mutant G206D/R207I, pH 8.8, 40°C Haloferax mediterranei
0.00073
-
NAD+ wild-type, pH 8.8, 40°C Haloferax mediterranei
0.00112
-
NAD+ mutant G206D, pH 8.8, 40°C Haloferax mediterranei
0.015
-
NADP+ wild-type, pH 8.8, 40°C Haloferax mediterranei