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Literature summary for 1.1.1.343 extracted from

  • Chen, H.; Zhu, Z.; Huang, R.; Zhang, Y.P.
    Coenzyme engineering of a hyperthermophilic 6-phosphogluconate dehydrogenase from NADP+ to NAD+ with its application to biobatteries (2016), Sci. Rep., 6, 36311.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21(DE3) cells Thermotoga maritima

Protein Variants

Protein Variants Comment Organism
N32D the mutant exhibits an about 1.3fold increase of catalytic efficiency with NAD+ as cofactor compared to the wild type enzyme Thermotoga maritima
N32D/R33I/T34I the mutant exhibits an about 1.5fold increase of catalytic efficiency with NAD+ as cofactor compared to the wild type enzyme Thermotoga maritima
N32D/R33L/T34S the mutant exhibits an about 0.6fold decrease of catalytic efficiency with NAD+ as cofactor compared to the wild type enzyme Thermotoga maritima
N32E/R33I/T34I the mutant exhibits an about 3.5fold increase of catalytic efficiency with NAD+ as cofactor, with about 64000fold reversal of the coenzyme selectivity from NADP+ to NAD+ compared to the wild type enzyme Thermotoga maritima

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.5
-
NAD+ mutant enzyme N32E/R33I7T34I, in 100 mM HEPES (pH 7.5), at 70°C Thermotoga maritima
3.9
-
NAD+ mutant enzyme N32D/R33I/T34I, in 100 mM HEPES (pH 7.5), at 70°C Thermotoga maritima
4
-
NAD+ mutant enzyme N32D, in 100 mM HEPES (pH 7.5), at 70°C Thermotoga maritima
4.3
-
NAD+ wild type enzyme, in 100 mM HEPES (pH 7.5), at 70°C Thermotoga maritima
7.5
-
NAD+ mutant enzyme N32D/R33L/T34S, in 100 mM HEPES (pH 7.5), at 70°C Thermotoga maritima

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
6-phospho-D-gluconate + NAD+ Thermotoga maritima
-
D-ribulose 5-phosphate + CO2 + NADH + H+
-
?

Organism

Organism UniProt Comment Textmining
Thermotoga maritima Q9WYR9 cf EC 1.1.1.44
-

Purification (Commentary)

Purification (Comment) Organism
Ni-NTA resin column chromatography Thermotoga maritima

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
6-phospho-D-gluconate + NAD+
-
Thermotoga maritima D-ribulose 5-phosphate + CO2 + NADH + H+
-
?

Synonyms

Synonyms Comment Organism
6PGDH
-
Thermotoga maritima

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
23.3
-
NAD+ wild type enzyme, in 100 mM HEPES (pH 7.5), at 70°C Thermotoga maritima
25.4
-
NAD+ mutant enzyme N32D/R33L/T34S, in 100 mM HEPES (pH 7.5), at 70°C Thermotoga maritima
29.3
-
NAD+ mutant enzyme N32D, in 100 mM HEPES (pH 7.5), at 70°C Thermotoga maritima
32.2
-
NAD+ mutant enzyme N32D/R33I/T34I, in 100 mM HEPES (pH 7.5), at 70°C Thermotoga maritima
47.9
-
NAD+ mutant enzyme N32E/R33I7T34I, in 100 mM HEPES (pH 7.5), at 70°C Thermotoga maritima

Cofactor

Cofactor Comment Organism Structure
NAD+
-
Thermotoga maritima

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
3.4
-
NAD+ mutant enzyme N32D/R33L/T34S, in 100 mM HEPES (pH 7.5), at 70°C Thermotoga maritima
5.4
-
NAD+ wild type enzyme, in 100 mM HEPES (pH 7.5), at 70°C Thermotoga maritima
7.3
-
NAD+ mutant enzyme N32D, in 100 mM HEPES (pH 7.5), at 70°C Thermotoga maritima
8.3
-
NAD+ mutant enzyme N32D/R33I/T34I, in 100 mM HEPES (pH 7.5), at 70°C Thermotoga maritima
19.2
-
NAD+ mutant enzyme N32E/R33I7T34I, in 100 mM HEPES (pH 7.5), at 70°C Thermotoga maritima