Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli | [Candida] boidinii |
expressed in Escherichia coli BL21(DE3) cells | [Candida] boidinii |
Protein Variants | Comment | Organism |
---|---|---|
D195/Y196P | the mutant shows an improvement in overall catalytic efficiency with NADP+ and a decrease in the efficiency with NAD+ as cofactors compared to the wild type enzyme | [Candida] boidinii |
D195/Y196S | the mutant shows an improvement in overall catalytic efficiency with NADP+ and a decrease in the efficiency with NAD+ as cofactors compared to the wild type enzyme | [Candida] boidinii |
D195A | reduced activity | [Candida] boidinii |
D195A | increased activity with NADP+, decreased activity with NAD+ compared to the wild type enzyme | [Candida] boidinii |
D195A | mutant shows a noticeable decrease of Km for NADP+ (approximately 10fold) with concomitant increase of kcat (approximately 10000fold) for NADP+ compared to the wild type enzyme, in addition the Km for NAD+ is dramatically increased | [Candida] boidinii |
D195N | reduced activity | [Candida] boidinii |
D195N | increased activity with NADP+, decreased activity with NAD+ compared to the wild type enzyme | [Candida] boidinii |
D195N | mutant shows a noticeable decrease of Km for NADP+ (approximately 10fold) with concomitant increase of kcat (approximately 10000fold) for NADP+ compared to the wild type enzyme, in addition the Km for NAD+ is dramatically increased | [Candida] boidinii |
D195Q | reduced activity | [Candida] boidinii |
D195Q | increased activity with NADP+, decreased activity with NAD+ compared to the wild type enzyme | [Candida] boidinii |
D195Q | mutant shows a noticeable decrease of Km for NADP+ (approximately 10fold) with concomitant increase of kcat (approximately 10000fold) for NADP+ compared to the wild type enzyme, in addition the Km for NAD+ is dramatically increased | [Candida] boidinii |
D195Q/Y196H | the double mutant shows a more than 20000000fold improvement in overall catalytic efficiency with NADP+ and a more than 900fold decrease in the efficiency with NAD+ as cofactors | [Candida] boidinii |
D195Q/Y196H | the mutant shows a more than 20000000fold improvement in overall catalytic efficiency with NADP+ and a more than 900fold decrease in the efficiency with NAD+ as cofactors compared to the wild type enzyme | [Candida] boidinii |
D195Q/Y196H | the double mutant shows a more than 20000000fold improvement in overall catalytic efficiency with NADP+ and a more than 900fold decrease in the efficiency with NAD+ as cofactors, exhibits high catalytic activity with NADP+ as coenzyme (14% of the catalytic activity of the wild type for NAD+) | [Candida] boidinii |
D195Q/Y196P | the double mutant shows increased activity with NADP+, decreased activity with NAD+ compared to the wild type enzyme | [Candida] boidinii |
D195Q/Y196P | the double mutant shows an improvement in overall catalytic efficiency with NADP+ and a decrease in the efficiency with NAD+ as cofactors | [Candida] boidinii |
D195Q/Y196S | the double mutant shows decreased activity with NADP+ and NAD+ compared to the wild type enzyme | [Candida] boidinii |
D195Q/Y196S | the double mutant shows an improvement in overall catalytic efficiency with NADP+ and a decrease in the efficiency with NAD+ as cofactors | [Candida] boidinii |
D195S | reduced activity | [Candida] boidinii |
D195S | increased activity with NADP+, decreased activity with NAD+ compared to the wild type enzyme | [Candida] boidinii |
D195S | mutant shows a noticeable decrease of Km for NADP+ (approximately 10fold) with concomitant increase of kcat (approximately 10000fold) for NADP+ compared to the wild type enzyme, in addition the Km for NAD+ is dramatically increased | [Candida] boidinii |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.015 | - |
NAD+ | wild type enzyme, in 0.1 M potassium phosphate buffer (pH 7.5) | [Candida] boidinii | |
0.015 | - |
NAD+ | wild type enzyme, in 0.1 M potassium phosphate buffer (pH 7.5), at 37°C | [Candida] boidinii | |
0.13 | - |
NAD+ | mutant enzyme D195Q/Y196P, in 0.1 M potassium phosphate buffer (pH 7.5) | [Candida] boidinii | |
0.13 | - |
NAD+ | mutant enzyme D195Q/Y196P, in 0.1 M potassium phosphate buffer (pH 7.5), at 37°C | [Candida] boidinii | |
0.96 | - |
NAD+ | mutant enzyme D195Q, in 0.1 M potassium phosphate buffer (pH 7.5) | [Candida] boidinii | |
0.96 | - |
NAD+ | mutant enzyme D195Q, in 0.1 M potassium phosphate buffer (pH 7.5), at 37°C | [Candida] boidinii | |
1.5 | - |
NAD+ | mutant enzyme D195S, in 0.1 M potassium phosphate buffer (pH 7.5) | [Candida] boidinii | |
1.5 | - |
NAD+ | mutant enzyme D195S, in 0.1 M potassium phosphate buffer (pH 7.5), at 37°C | [Candida] boidinii | |
1.7 | - |
NADP+ | mutant enzyme D195Q/Y196H, in 0.1 M potassium phosphate buffer (pH 7.5) | [Candida] boidinii | |
1.7 | - |
NADP+ | mutant enzyme D195Q/Y196H, in 0.1 M potassium phosphate buffer (pH 7.5), at 37°C | [Candida] boidinii | |
1.8 | - |
NAD+ | mutant enzyme D195Q/Y196H, in 0.1 M potassium phosphate buffer (pH 7.5) | [Candida] boidinii | |
1.8 | - |
NAD+ | mutant enzyme D195Q/Y196H, in 0.1 M potassium phosphate buffer (pH 7.5), at 37°C | [Candida] boidinii | |
3.3 | - |
NADP+ | mutant enzyme D195A, in 0.1 M potassium phosphate buffer (pH 7.5) | [Candida] boidinii | |
3.3 | - |
NADP+ | mutant enzyme D195A, in 0.1 M potassium phosphate buffer (pH 7.5), at 37°C | [Candida] boidinii | |
3.7 | - |
NADP+ | mutant enzyme D195Q/Y196P, in 0.1 M potassium phosphate buffer (pH 7.5) | [Candida] boidinii | |
3.7 | - |
NADP+ | mutant enzyme D195Q/Y196P, in 0.1 M potassium phosphate buffer (pH 7.5), at 37°C | [Candida] boidinii | |
4.5 | - |
NADP+ | mutant enzyme D195Q, in 0.1 M potassium phosphate buffer (pH 7.5) | [Candida] boidinii | |
4.5 | - |
NADP+ | mutant enzyme D195Q, in 0.1 M potassium phosphate buffer (pH 7.5), at 37°C | [Candida] boidinii | |
4.8 | - |
NAD+ | mutant enzyme D195A, in 0.1 M potassium phosphate buffer (pH 7.5) | [Candida] boidinii | |
4.8 | - |
NAD+ | mutant enzyme D195A, in 0.1 M potassium phosphate buffer (pH 7.5), at 37°C | [Candida] boidinii | |
5.01 | - |
NAD+ | mutant enzyme D195N, in 0.1 M potassium phosphate buffer (pH 7.5) | [Candida] boidinii | |
5.01 | - |
NAD+ | mutant enzyme D195N, in 0.1 M potassium phosphate buffer (pH 7.5), at 37°C | [Candida] boidinii | |
5.1 | - |
NAD+ | mutant enzyme D195Q/Y196S, in 0.1 M potassium phosphate buffer (pH 7.5) | [Candida] boidinii | |
5.1 | - |
NAD+ | mutant enzyme D195Q/Y196S, in 0.1 M potassium phosphate buffer (pH 7.5), at 37°C | [Candida] boidinii | |
6.2 | - |
NADP+ | mutant enzyme D195Q/Y196S, in 0.1 M potassium phosphate buffer (pH 7.5) | [Candida] boidinii | |
6.2 | - |
NADP+ | mutant enzyme D195S, in 0.1 M potassium phosphate buffer (pH 7.5) | [Candida] boidinii | |
6.2 | - |
NADP+ | mutant enzyme D195Q/Y196S, in 0.1 M potassium phosphate buffer (pH 7.5), at 37°C | [Candida] boidinii | |
6.2 | - |
NADP+ | mutant enzyme D195S, in 0.1 M potassium phosphate buffer (pH 7.5), at 37°C | [Candida] boidinii | |
13.2 | - |
NADP+ | mutant enzyme D195N, in 0.1 M potassium phosphate buffer (pH 7.5) | [Candida] boidinii | |
13.2 | - |
NADP+ | mutant enzyme D195N, in 0.1 M potassium phosphate buffer (pH 7.5), at 37°C | [Candida] boidinii | |
38 | - |
NADP+ | Km above 38 mM, wild type enzyme, in 0.1 M potassium phosphate buffer (pH 7.5) | [Candida] boidinii | |
38 | - |
NADP+ | Km above 38 mM, wild type enzyme, in 0.1 M potassium phosphate buffer (pH 7.5), at 37°C | [Candida] boidinii |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
[Candida] boidinii | - |
- |
- |
[Candida] boidinii | O13437 | - |
- |
Purification (Comment) | Organism |
---|---|
Cibacron Blue 3GA-Sepharose affinity column chromatography | [Candida] boidinii |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
formate + NAD+ | - |
[Candida] boidinii | CO2 + NADH + H+ | - |
? | |
formate + NAD+ | high activity | [Candida] boidinii | CO2 + NADH + H+ | - |
? | |
formate + NAD+ | FDH is highly specific to NAD+ and virtually fails to catalyze the reaction with NADP+ | [Candida] boidinii | CO2 + NADH + H+ | - |
? | |
formate + NADP+ | FDH is highly specific to NAD+ and virtually fails to catalyze the reaction with NADP+ | [Candida] boidinii | CO2 + NADPH | - |
? | |
formate + NADP+ | low activity | [Candida] boidinii | CO2 + NADPH + H+ | - |
? | |
formate + NADP+ | the wild type enzyme virtually fails to catalyze the reaction with NADP+ | [Candida] boidinii | CO2 + NADPH + H+ | - |
? |
Synonyms | Comment | Organism |
---|---|---|
FDH | - |
[Candida] boidinii |
NAD+-dependent formate dehydrogenase | - |
[Candida] boidinii |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.00004 | - |
NADP+ | wild type enzyme, in 0.1 M potassium phosphate buffer (pH 7.5) | [Candida] boidinii | |
0.00004 | - |
NADP+ | wild type enzyme, in 0.1 M potassium phosphate buffer (pH 7.5), at 37°C | [Candida] boidinii | |
0.052 | - |
NADP+ | mutant enzyme D195A, in 0.1 M potassium phosphate buffer (pH 7.5) | [Candida] boidinii | |
0.052 | - |
NADP+ | mutant enzyme D195A, in 0.1 M potassium phosphate buffer (pH 7.5), at 37°C | [Candida] boidinii | |
0.21 | - |
NAD+ | mutant enzyme D195N, in 0.1 M potassium phosphate buffer (pH 7.5) | [Candida] boidinii | |
0.21 | - |
NAD+ | mutant enzyme D195N, in 0.1 M potassium phosphate buffer (pH 7.5), at 37°C | [Candida] boidinii | |
0.26 | - |
NADP+ | mutant enzyme D195N, in 0.1 M potassium phosphate buffer (pH 7.5) | [Candida] boidinii | |
0.26 | - |
NAD+ | mutant enzyme D195Q, in 0.1 M potassium phosphate buffer (pH 7.5) | [Candida] boidinii | |
0.26 | - |
NADP+ | mutant enzyme D195Q, in 0.1 M potassium phosphate buffer (pH 7.5) | [Candida] boidinii | |
0.26 | - |
NADP+ | mutant enzyme D195N, in 0.1 M potassium phosphate buffer (pH 7.5), at 37°C | [Candida] boidinii | |
0.26 | - |
NAD+ | mutant enzyme D195Q, in 0.1 M potassium phosphate buffer (pH 7.5), at 37°C | [Candida] boidinii | |
0.26 | - |
NADP+ | mutant enzyme D195Q, in 0.1 M potassium phosphate buffer (pH 7.5), at 37°C | [Candida] boidinii | |
0.34 | - |
NADP+ | mutant enzyme D195Q/Y196P, in 0.1 M potassium phosphate buffer (pH 7.5) | [Candida] boidinii | |
0.34 | - |
NADP+ | mutant enzyme D195Q/Y196S, in 0.1 M potassium phosphate buffer (pH 7.5) | [Candida] boidinii | |
0.34 | - |
NAD+ | mutant enzyme D195S, in 0.1 M potassium phosphate buffer (pH 7.5) | [Candida] boidinii | |
0.34 | - |
NADP+ | mutant enzyme D195S, in 0.1 M potassium phosphate buffer (pH 7.5) | [Candida] boidinii | |
0.34 | - |
NADP+ | mutant enzyme D195Q/Y196P, in 0.1 M potassium phosphate buffer (pH 7.5), at 37°C | [Candida] boidinii | |
0.34 | - |
NADP+ | mutant enzyme D195Q/Y196S, in 0.1 M potassium phosphate buffer (pH 7.5), at 37°C | [Candida] boidinii | |
0.34 | - |
NAD+ | mutant enzyme D195S, in 0.1 M potassium phosphate buffer (pH 7.5), at 37°C | [Candida] boidinii | |
0.34 | - |
NADP+ | mutant enzyme D195S, in 0.1 M potassium phosphate buffer (pH 7.5), at 37°C | [Candida] boidinii | |
0.4 | - |
NAD+ | mutant enzyme D195Q/Y196S, in 0.1 M potassium phosphate buffer (pH 7.5) | [Candida] boidinii | |
0.4 | - |
NAD+ | mutant enzyme D195Q/Y196S, in 0.1 M potassium phosphate buffer (pH 7.5), at 37°C | [Candida] boidinii | |
0.44 | - |
NADP+ | mutant enzyme D195Q/Y196H, in 0.1 M potassium phosphate buffer (pH 7.5) | [Candida] boidinii | |
0.44 | - |
NADP+ | mutant enzyme D195Q/Y196H, in 0.1 M potassium phosphate buffer (pH 7.5), at 37°C | [Candida] boidinii | |
0.49 | - |
NAD+ | mutant enzyme D195Q/Y196H, in 0.1 M potassium phosphate buffer (pH 7.5) | [Candida] boidinii | |
0.49 | - |
NAD+ | mutant enzyme D195Q/Y196H, in 0.1 M potassium phosphate buffer (pH 7.5), at 37°C | [Candida] boidinii | |
0.76 | - |
NAD+ | mutant enzyme D195A, in 0.1 M potassium phosphate buffer (pH 7.5) | [Candida] boidinii | |
0.76 | - |
NAD+ | mutant enzyme D195A, in 0.1 M potassium phosphate buffer (pH 7.5), at 37°C | [Candida] boidinii | |
0.87 | - |
NAD+ | mutant enzyme D195Q/Y196P, in 0.1 M potassium phosphate buffer (pH 7.5) | [Candida] boidinii | |
0.87 | - |
NAD+ | mutant enzyme D195Q/Y196P, in 0.1 M potassium phosphate buffer (pH 7.5), at 37°C | [Candida] boidinii | |
3.7 | - |
NAD+ | wild type enzyme, in 0.1 M potassium phosphate buffer (pH 7.5) | [Candida] boidinii | |
3.7 | - |
NAD+ | wild type enzyme, in 0.1 M potassium phosphate buffer (pH 7.5), at 37°C | [Candida] boidinii |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
mutant enzyme D195Q/Y196H | [Candida] boidinii |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
5.5 | 8.5 | mutant enzyme D195Q/Y196H | [Candida] boidinii |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NAD+ | highly specific to NAD+ | [Candida] boidinii | |
NAD+ | most efficient cofactor | [Candida] boidinii | |
NADP+ | low activity compared to NADP+ | [Candida] boidinii | |
NADP+ | the wild type enzyme virtually fails to use NADP+ as a cofacor | [Candida] boidinii |