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Literature summary extracted from

  • Fogal, S.; Beneventi, E.; Cendron, L.; Bergantino, E.
    Structural basis for double cofactor specificity in a new formate dehydrogenase from the acidobacterium Granulicella mallensis MP5ACTX8 (2015), Appl. Microbiol. Biotechnol., 99, 9541-9554.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.17.1.10 expressed in Escherichia coli BL21(DE3) cells Granulicella mallensis

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
1.17.1.10 apoprotein and holoprotein, either in complex with NAD+ or NADP+, hanging drop vapor diffusion method, using 0.1 M MES pH 6.5, 12% (w/v) PEG 2000 MME or 0. 1 M Na acetate pH 4.6, 2.0 M Na formate Granulicella mallensis

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.17.1.10 DMSO 1-33% residual activity at 40% (v/v) Granulicella mallensis
1.17.1.10 ethanol complete inhibition at 40% (v/v) Granulicella mallensis
1.17.1.10 formaldehyde 79.8% residual activity at 0.1 M Granulicella mallensis
1.17.1.10 Isopropanol 33-66% residual activity at 20 and 40% (v/v) Granulicella mallensis
1.17.1.10 methanol complete inhibition at 40% (v/v) Granulicella mallensis
1.17.1.10 Sodium azide complete inhibition at 1 mM Granulicella mallensis
1.17.1.10 Sodium nitrate 44.3% residual activity at 0.1 M Granulicella mallensis
1.17.1.10 tert-butanol 33-66% residual activity at 10, 20 and 40% (v/v) Granulicella mallensis
1.17.1.10 Triton X-100 33-66% residual activity at 5 and 10% (v/v) Granulicella mallensis

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.17.1.10 0.85
-
 NADP+ at pH 7.0 and 30°C Granulicella mallensis
1.17.1.10 6.5
-
 NAD+ at pH 7.0 and 30°C Granulicella mallensis
1.17.1.10 80
-
 formate with NAD+ as cosubstrate, at pH 7.0 and 30°C Granulicella mallensis
1.17.1.10 200
-
 formate with NADP+ as cosubstrate, at pH 7.0 and 30°C Granulicella mallensis

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.17.1.10 45200
-
 2 * 45200, SDS-PAGE Granulicella mallensis
1.17.1.10 79000
-
 gel filtration Granulicella mallensis

Organism

EC Number Organism UniProt Comment Textmining
1.17.1.10 Granulicella mallensis G8NVB5
-
-
1.17.1.10 Granulicella mallensis MP5ACTX8 G8NVB5
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.17.1.10 Ni-NTA column chromatography, and Superdex 200 gel filtration Granulicella mallensis

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.17.1.10 formate + NAD+
-
 Granulicella mallensis CO2 + NADH + H+
-
 ?
1.17.1.10 formate + NAD+
-
 Granulicella mallensis MP5ACTX8 CO2 + NADH + H+
-
 ?
1.17.1.10 formate + NADP+ NADP+ is the preferred cofactor Granulicella mallensis CO2 + NADPH
-
 ?
1.17.1.10 formate + NADP+ NADP+ is the preferred cofactor Granulicella mallensis MP5ACTX8 CO2 + NADPH
-
 ?
1.17.1.10 additional information no activity is detected with malic acid, sodium citrate, sodium nitrate, succinic acid, sodium bicarbonate, sodium phosphite, ethanol, methanol, formaldehyde, sodium acetate, or sodium lactate Granulicella mallensis ?
-
 ?
1.17.1.10 additional information no activity is detected with malic acid, sodium citrate, sodium nitrate, succinic acid, sodium bicarbonate, sodium phosphite, ethanol, methanol, formaldehyde, sodium acetate, or sodium lactate Granulicella mallensis MP5ACTX8 ?
-
 ?

Subunits

EC Number Subunits Comment Organism
1.17.1.10 homodimer 2 * 45200, SDS-PAGE Granulicella mallensis

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
1.17.1.10 25 50 the enzyme remains stable after incubation for 25 h at 25°C and 37°C, while its activity is reduced to about 35% after 3 min at 50°C and is completely lost after 15 min at 50°C Granulicella mallensis

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.17.1.10 3.96
-
 NADP+ at pH 7.0 and 30°C Granulicella mallensis
1.17.1.10 5.77
-
 NAD+ at pH 7.0 and 30°C Granulicella mallensis

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.17.1.10 6
-
 with NADP+ as cofactor Granulicella mallensis
1.17.1.10 7.5
-
 with NADP as cofactor Granulicella mallensis

Cofactor

EC Number Cofactor Comment Organism Structure
1.17.1.10 NAD+ the enzyme shows double cofactor specificity, with NADP+ preferred over NAD+ at acidic pH values Granulicella mallensis
1.17.1.10 NADP+ the enzyme shows double cofactor specificity, with NADP+ preferred over NAD+ at acidic pH values Granulicella mallensis