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Literature summary for 1.17.1.9 extracted from

  • Alekseeva, A.; Serenko, A.; Kargov, I.; Savin, S.; Kleymenov, S.; Tishkov, V.
    Engineering catalytic properties and thermal stability of plant formate dehydrogenase by single-point mutations (2012), Protein Eng., 25, 781-788.
    View publication on PubMed
Search for more literature for 1.17.1.9

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21(DE3)CodonPlus/pLysS cells Glycine max

Protein Variants

Protein Variants Comment Organism
F290D the Km for formate is increased while KM for NAD+ does not change. The mutation increases the Tm values by 4.3°C. The catalytic efficiency for NAD+ is about 2fold higher compared to the wild type enzyme Glycine max
F290N the Km for formate is increased while KM for NAD+ does not change. The mutation increases the Tm values by 2.9°C. The catalytic efficiency for NAD+ is about 2fold higher compared to the wild type enzyme Glycine max
F290S the Km for formate is increased while Km for NAD+ slightly increases. The mutation increases the Tm values by 7.8°C. The catalytic efficiency for NAD+ is slightly decreased compared to the wild type enzyme Glycine max

Inhibitors

Inhibitors Comment Organism Structure
azide
-
 Glycine max

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.5
-
 formate wild type enzyme, in 0.1 M sodium phosphate buffer, pH 7.0, at 30°C Glycine max
4.1
-
 formate mutant enzyme F290S, in 0.1 M sodium phosphate buffer, pH 7.0, at 30°C Glycine max
4.5
-
 formate mutant enzyme F290N, in 0.1 M sodium phosphate buffer, pH 7.0, at 30°C Glycine max
5
-
 formate mutant enzyme F290D, in 0.1 M sodium phosphate buffer, pH 7.0, at 30°C Glycine max
9.1
-
 NAD+ mutant enzyme F290S, in 0.1 M sodium phosphate buffer, pH 7.0, at 30°C Glycine max
12.8
-
 NAD+ mutant enzyme F290D, in 0.1 M sodium phosphate buffer, pH 7.0, at 30°C Glycine max
13.3
-
 NAD+ wild type enzyme, in 0.1 M sodium phosphate buffer, pH 7.0, at 30°C Glycine max
14
-
 NAD+ mutant enzyme F290N, in 0.1 M sodium phosphate buffer, pH 7.0, at 30°C Glycine max

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
formate + NAD+ Glycine max
-
 CO2 + NADH + H+
-
 ?

Organism

Organism UniProt Comment Textmining
Glycine max
-
-
-

Purification (Commentary)

Purification (Comment) Organism
ammonium sulfate precipitation, phenyl Sepharose column chromatography, and Sephadex G-25 gel filtration Glycine max

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
formate + NAD+
-
 Glycine max CO2 + NADH + H+
-
 ?

Synonyms

Synonyms Comment Organism
FDH
-
 Glycine max

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.8
-
 NAD+ mutant enzyme F290N, in 0.1 M sodium phosphate buffer, pH 7.0, at 30°C Glycine max
2.8
-
 formate mutant enzyme F290N, in 0.1 M sodium phosphate buffer, pH 7.0, at 30°C Glycine max
2.9
-
 NAD+ wild type enzyme, in 0.1 M sodium phosphate buffer, pH 7.0, at 30°C Glycine max
2.9
-
 formate wild type enzyme, in 0.1 M sodium phosphate buffer, pH 7.0, at 30°C Glycine max
4.1
-
 NAD+ mutant enzyme F290S, in 0.1 M sodium phosphate buffer, pH 7.0, at 30°C Glycine max
4.1
-
 formate mutant enzyme F290S, in 0.1 M sodium phosphate buffer, pH 7.0, at 30°C Glycine max
5.1
-
 NAD+ mutant enzyme F290D, in 0.1 M sodium phosphate buffer, pH 7.0, at 30°C Glycine max
5.1
-
 formate mutant enzyme F290D, in 0.1 M sodium phosphate buffer, pH 7.0, at 30°C Glycine max

Cofactor

Cofactor Comment Organism Structure
NAD+
-
 Glycine max

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.62
-
 formate mutant enzyme F290N, in 0.1 M sodium phosphate buffer, pH 7.0, at 30°C Glycine max
1
-
 formate mutant enzyme F290S, in 0.1 M sodium phosphate buffer, pH 7.0, at 30°C Glycine max
1.02
-
 formate mutant enzyme F290D, in 0.1 M sodium phosphate buffer, pH 7.0, at 30°C Glycine max
1.93
-
 formate wild type enzyme, in 0.1 M sodium phosphate buffer, pH 7.0, at 30°C Glycine max
200
-
 NAD+ mutant enzyme F290N, in 0.1 M sodium phosphate buffer, pH 7.0, at 30°C Glycine max
220
-
 NAD+ wild type enzyme, in 0.1 M sodium phosphate buffer, pH 7.0, at 30°C Glycine max
400
-
 NAD+ mutant enzyme F290D, in 0.1 M sodium phosphate buffer, pH 7.0, at 30°C Glycine max
450
-
 NAD+ mutant enzyme F290S, in 0.1 M sodium phosphate buffer, pH 7.0, at 30°C Glycine max