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Literature summary 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)

EC Number Cloned (Comment) Organism
1.1.1.44 expressed in Escherichia coli BL21(DE3) cells Thermotoga maritima
1.1.1.343 expressed in Escherichia coli BL21(DE3) cells Thermotoga maritima

Protein Variants

EC Number Protein Variants Comment Organism
1.1.1.44 N32D the mutant exhibits an 460times higher Km value on NADP+ and a slightly decreased Km value of 4.0 mM on NAD+ compared with the wild type enzyme Thermotoga maritima
1.1.1.44 N32D/R33I/T34I the mutant exhibits a far higher Km value of on NADP+ and a slightly decreased Km value on NAD+ compared with the wild type enzyme Thermotoga maritima
1.1.1.44 N32D/R33L/T34S the mutant has an increased Km value on NADP+ and NAD+ compared to the wild type enzyme Thermotoga maritima
1.1.1.44 N32E/R33I/T34I the mutant exhibits an about 6400fold reversal of the coenzyme selectivity from NADP+ to NAD+ compared to the wild type enzyme Thermotoga maritima
1.1.1.343 N32D the mutant exhibits an about 1.3fold increase of catalytic efficiency with NAD+ as cofactor compared to the wild type enzyme Thermotoga maritima
1.1.1.343 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
1.1.1.343 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
1.1.1.343 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]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.1.1.44 0.005
-
 NADP+ wild type enzyme, at pH 7.5 and 70°C Thermotoga maritima
1.1.1.44 2.3
-
 NADP+ mutant enzyme N32D, at pH 7.5 and 70°C Thermotoga maritima
1.1.1.44 2.5
-
 NAD+ mutant enzyme N32E/R33I/T34I, at pH 7.5 and 70°C Thermotoga maritima
1.1.1.44 3.9
-
 NAD+ mutant enzyme N32D/R33I/T34I, at pH 7.5 and 70°C Thermotoga maritima
1.1.1.44 4
-
 NAD+ mutant enzyme N32D, at pH 7.5 and 70°C Thermotoga maritima
1.1.1.44 4.3
-
 NAD+ wild type enzyme at pH 7.5 and 70°C Thermotoga maritima
1.1.1.44 7.5
-
 NAD+ mutant enzyme N32D/R33L/T34S, at pH 7.5 and 70°C Thermotoga maritima
1.1.1.44 70
-
 NADP+ mutant enzyme N32D/R33I/T34I, at pH 7.5 and 70°C Thermotoga maritima
1.1.1.44 79
-
 NADP+ mutant enzyme N32E/R33I/T34I, at pH 7.5 and 70°C Thermotoga maritima
1.1.1.44 100
-
 NADP+ Km above 100 mM, mutant enzyme N32D/R33L/T34S, at pH 7.5 and 70°C Thermotoga maritima
1.1.1.343 2.5
-
 NAD+ mutant enzyme N32E/R33I7T34I, in 100 mM HEPES (pH 7.5), at 70°C Thermotoga maritima
1.1.1.343 3.9
-
 NAD+ mutant enzyme N32D/R33I/T34I, in 100 mM HEPES (pH 7.5), at 70°C Thermotoga maritima
1.1.1.343 4
-
 NAD+ mutant enzyme N32D, in 100 mM HEPES (pH 7.5), at 70°C Thermotoga maritima
1.1.1.343 4.3
-
 NAD+ wild type enzyme, in 100 mM HEPES (pH 7.5), at 70°C Thermotoga maritima
1.1.1.343 7.5
-
 NAD+ mutant enzyme N32D/R33L/T34S, in 100 mM HEPES (pH 7.5), at 70°C Thermotoga maritima

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.1.1.44 6-phospho-D-gluconate + NADP+ Thermotoga maritima
-
 D-ribulose 5-phosphate + CO2 + NADPH + H+
-
 ?
1.1.1.343 6-phospho-D-gluconate + NAD+ Thermotoga maritima
-
 D-ribulose 5-phosphate + CO2 + NADH + H+
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
1.1.1.44 Thermotoga maritima Q9WYR9
-
-
1.1.1.343 Thermotoga maritima Q9WYR9 cf EC 1.1.1.44
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.1.1.44 HisPur Ni-NTA resin column chromatography Thermotoga maritima
1.1.1.343 Ni-NTA resin column chromatography Thermotoga maritima

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.1.44 6-phospho-D-gluconate + NAD+ low activity with NAD+ Thermotoga maritima D-ribulose 5-phosphate + CO2 + NADH + H+
-
 ?
1.1.1.44 6-phospho-D-gluconate + NADP+
-
 Thermotoga maritima D-ribulose 5-phosphate + CO2 + NADPH + H+
-
 ?
1.1.1.343 6-phospho-D-gluconate + NAD+
-
 Thermotoga maritima D-ribulose 5-phosphate + CO2 + NADH + H+
-
 ?

Synonyms

EC Number Synonyms Comment Organism
1.1.1.44 6-phosphogluconate dehydrogenase
-
 Thermotoga maritima
1.1.1.44 6PGDH
-
 Thermotoga maritima
1.1.1.343 6PGDH
-
 Thermotoga maritima

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.1.1.44 15.9
-
 NADP+ mutant enzyme N32D/R33I/T34I, at pH 7.5 and 70°C Thermotoga maritima
1.1.1.44 15.9
-
 NADP+ mutant enzyme N32E/R33I/T34I, at pH 7.5 and 70°C Thermotoga maritima
1.1.1.44 16.7
-
 NADP+ mutant enzyme N32D, at pH 7.5 and 70°C Thermotoga maritima
1.1.1.44 17.6
-
 NADP+ wild type enzyme, at pH 7.5 and 70°C Thermotoga maritima
1.1.1.44 23.3
-
 NAD+ wild type enzyme at pH 7.5 and 70°C Thermotoga maritima
1.1.1.44 25.4
-
 NAD+ mutant enzyme N32D/R33L/T34S, at pH 7.5 and 70°C Thermotoga maritima
1.1.1.44 29.3
-
 NAD+ mutant enzyme N32D, at pH 7.5 and 70°C Thermotoga maritima
1.1.1.44 32.2
-
 NAD+ mutant enzyme N32D/R33I/T34I, at pH 7.5 and 70°C Thermotoga maritima
1.1.1.44 47.9
-
 NAD+ mutant enzyme N32E/R33I/T34I, at pH 7.5 and 70°C Thermotoga maritima
1.1.1.343 23.3
-
 NAD+ wild type enzyme, in 100 mM HEPES (pH 7.5), at 70°C Thermotoga maritima
1.1.1.343 25.4
-
 NAD+ mutant enzyme N32D/R33L/T34S, in 100 mM HEPES (pH 7.5), at 70°C Thermotoga maritima
1.1.1.343 29.3
-
 NAD+ mutant enzyme N32D, in 100 mM HEPES (pH 7.5), at 70°C Thermotoga maritima
1.1.1.343 32.2
-
 NAD+ mutant enzyme N32D/R33I/T34I, in 100 mM HEPES (pH 7.5), at 70°C Thermotoga maritima
1.1.1.343 47.9
-
 NAD+ mutant enzyme N32E/R33I7T34I, in 100 mM HEPES (pH 7.5), at 70°C Thermotoga maritima

Cofactor

EC Number Cofactor Comment Organism Structure
1.1.1.44 NAD+ low activity with NAD+ Thermotoga maritima
1.1.1.44 NADP+ natural cofactor Thermotoga maritima
1.1.1.343 NAD+
-
 Thermotoga maritima

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.1.1.44 0.2
-
 NADP+ mutant enzyme N32D/R33I/T34I, at pH 7.5 and 70°C Thermotoga maritima
1.1.1.44 0.2
-
 NADP+ mutant enzyme N32E/R33I/T34I, at pH 7.5 and 70°C Thermotoga maritima
1.1.1.44 3.4
-
 NAD+ mutant enzyme N32D/R33L/T34S, at pH 7.5 and 70°C Thermotoga maritima
1.1.1.44 5.4
-
 NAD+ wild type enzyme at pH 7.5 and 70°C Thermotoga maritima
1.1.1.44 7.3
-
 NAD+ mutant enzyme N32D, at pH 7.5 and 70°C Thermotoga maritima
1.1.1.44 7.3
-
 NADP+ mutant enzyme N32D, at pH 7.5 and 70°C Thermotoga maritima
1.1.1.44 8.3
-
 NAD+ mutant enzyme N32D/R33I/T34I, at pH 7.5 and 70°C Thermotoga maritima
1.1.1.44 19.2
-
 NAD+ mutant enzyme N32E/R33I/T34I, at pH 7.5 and 70°C Thermotoga maritima
1.1.1.44 3520
-
 NADP+ wild type enzyme, at pH 7.5 and 70°C Thermotoga maritima
1.1.1.343 3.4
-
 NAD+ mutant enzyme N32D/R33L/T34S, in 100 mM HEPES (pH 7.5), at 70°C Thermotoga maritima
1.1.1.343 5.4
-
 NAD+ wild type enzyme, in 100 mM HEPES (pH 7.5), at 70°C Thermotoga maritima
1.1.1.343 7.3
-
 NAD+ mutant enzyme N32D, in 100 mM HEPES (pH 7.5), at 70°C Thermotoga maritima
1.1.1.343 8.3
-
 NAD+ mutant enzyme N32D/R33I/T34I, in 100 mM HEPES (pH 7.5), at 70°C Thermotoga maritima
1.1.1.343 19.2
-
 NAD+ mutant enzyme N32E/R33I7T34I, in 100 mM HEPES (pH 7.5), at 70°C Thermotoga maritima