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Literature summary for 1.17.1.9 extracted from

  • Schirwitz, K.; Schmidt, A.; Lamzin, V.S.
    High-resolution structures of formate dehydrogenase from Candida boidinii (2007), Protein Sci., 16, 1146-1156.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 1.17.1.9

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli Rosetta(DE3)-pLysS cells [Candida] boidinii

Crystallization (Commentary)

Crystallization (Comment) Organism
sitting drop vapour diffusion method with 0.1 M sodium-cacodylate pH 6.5, 0.15 M ammonium sulfate, 30% (w/v) PEG 8K (mutant K47E), and 0.1 M sodium cacodylate pH 6.0, 0.2 M ammonium sulfate, and 32.5% (w/v) PEG 8K (mutant K328V) [Candida] boidinii

Protein Variants

Protein Variants Comment Organism
K328V the mutation improves the crystallizability of the protein and also the catalytic properties and the stability of the enzyme, the specific activity of freshly prepared mutant is similar to that of wild type and native protein [Candida] boidinii
K47E the mutation improves the crystallizability of the protein and also the catalytic properties and the stability of the enzyme, the specific activity of freshly prepared mutant is similar to that of wild type and native protein [Candida] boidinii

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
5
-
 formate mutant enzyme K328V, at 20°C, after 2 weeks or 4 months of enzyme storage [Candida] boidinii
16
-
 formate mutant enzyme K47E, at 20°C, after 2 weeks of enzyme storage [Candida] boidinii
20
-
 formate mutant enzyme K47E, at 20°C, after 4 months of enzyme storage [Candida] boidinii
20
-
 formate native wild type enzyme, at 20°C, after 2 weeks of enzyme storage [Candida] boidinii
25
-
 NAD+ mutant enzyme K328V, at 20°C, after 2 weeks of enzyme storage [Candida] boidinii
25
-
 NAD+ mutant enzyme K47E, at 20°C, after 2 weeks of enzyme storage [Candida] boidinii
30
-
 NAD+ mutant enzyme K328V, at 20°C, after 4 months of enzyme storage [Candida] boidinii
35
-
 formate native wild type enzyme, at 20°C, after 4 months of enzyme storage [Candida] boidinii
40
-
 NAD+ mutant enzyme K47E, at 20°C, after 4 months of enzyme storage [Candida] boidinii
50
-
 NAD+ native wild type enzyme, at 20°C, after 2 weeks of enzyme storage [Candida] boidinii
90
-
 NAD+ native wild type enzyme, at 20°C, after 4 months of enzyme storage [Candida] boidinii

Organism

Organism UniProt Comment Textmining
[Candida] boidinii O13437
-
-

Purification (Commentary)

Purification (Comment) Organism
AMP-Sepharose column chromatography and Superdex 200 gel filtration [Candida] boidinii

Storage Stability

Storage Stability Organism
4°C, native enzyme in 150 mM NaCl in 20 mM bis-Tris-propane pH 7.4, 1 day, no loss of activity [Candida] boidinii
4°C, native enzyme in 150 mM NaCl in 20 mM bis-Tris-propane pH 7.4, 2 weeks, 29% loss of activity [Candida] boidinii
4°C, native enzyme in 150 mM NaCl in 20 mM bis-Tris-propane pH 7.4, 4 months, 86% loss of activity [Candida] boidinii

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
formate + NAD+
-
 [Candida] boidinii CO2 + NADH + H+
-
 ?

Subunits

Subunits Comment Organism
homodimer x-ray crystallography [Candida] boidinii

Synonyms

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

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
55
-
 at 55°C, where native FDH loses 50% activity within 20 min incubation due to heat inactivation, the mutants preserve 84% (K47E) and 70% (K328V) residual activity [Candida] boidinii

Cofactor

Cofactor Comment Organism Structure
NAD+
-
 [Candida] boidinii