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 | 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 | Comment | Organism | Structure |
---|---|---|---|
NAD(P)+ | - |
Haloferax mediterranei | |
NAD+ | - |
Haloferax mediterranei | |
NADP+ | - |
Haloferax mediterranei |
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 | UniProt | Comment | Textmining |
---|---|---|---|
Haloferax mediterranei | - |
- |
- |
Haloferax mediterranei | Q977U7 | - |
- |
Purification (Comment) | Organism |
---|---|
- |
Haloferax mediterranei |
Renatured (Comment) | Organism |
---|---|
- |
Haloferax mediterranei |
Storage Stability | Organism |
---|---|
4°C, stable for weeks | Haloferax mediterranei |
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 | 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 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 | 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 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 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 |