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Literature summary for 1.17.1.9 extracted from

  • Andreadeli, A.; Platis, D.; Tishkov, V.; Popov, V.; Labrou, N.E.
    Structure-guided alteration of coenzyme specificity of formate dehydrogenase by saturation mutagenesis to enable efficient utilization of NADP(+) (2008), FEBS J., 275, 3859-3869.
    View publication on PubMed
Search for more literature for 1.17.1.9

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli [Candida] boidinii
expressed in Escherichia coli BL21(DE3) cells [Candida] boidinii

Protein Variants

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 [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

Organism UniProt Comment Textmining
[Candida] boidinii
-
-
-
[Candida] boidinii O13437
-
-

Purification (Commentary)

Purification (Comment) Organism
Cibacron Blue 3GA-Sepharose affinity column chromatography [Candida] boidinii

Substrates and Products (Substrate)

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

Synonyms Comment Organism
FDH
-
 [Candida] boidinii
NAD+-dependent formate dehydrogenase
-
 [Candida] boidinii

Turnover Number [1/s]

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

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
 mutant enzyme D195Q/Y196H [Candida] boidinii

pH Range

pH Minimum pH Maximum Comment Organism
5.5 8.5 mutant enzyme D195Q/Y196H [Candida] boidinii

Cofactor

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