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Literature summary for 1.1.1.44 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 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
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
N32D/R33L/T34S the mutant has an increased Km value on NADP+ and NAD+ compared to the wild type enzyme Thermotoga maritima
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

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.005
-
NADP+ wild type enzyme, at pH 7.5 and 70°C Thermotoga maritima
2.3
-
NADP+ mutant enzyme N32D, at pH 7.5 and 70°C Thermotoga maritima
2.5
-
NAD+ mutant enzyme N32E/R33I/T34I, at pH 7.5 and 70°C Thermotoga maritima
3.9
-
NAD+ mutant enzyme N32D/R33I/T34I, at pH 7.5 and 70°C Thermotoga maritima
4
-
NAD+ mutant enzyme N32D, at pH 7.5 and 70°C Thermotoga maritima
4.3
-
NAD+ wild type enzyme at pH 7.5 and 70°C Thermotoga maritima
7.5
-
NAD+ mutant enzyme N32D/R33L/T34S, at pH 7.5 and 70°C Thermotoga maritima
70
-
NADP+ mutant enzyme N32D/R33I/T34I, at pH 7.5 and 70°C Thermotoga maritima
79
-
NADP+ mutant enzyme N32E/R33I/T34I, at pH 7.5 and 70°C Thermotoga maritima
100
-
NADP+ Km above 100 mM, mutant enzyme N32D/R33L/T34S, at pH 7.5 and 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 + NADP+ Thermotoga maritima
-
D-ribulose 5-phosphate + CO2 + NADPH + H+
-
?

Organism

Organism UniProt Comment Textmining
Thermotoga maritima Q9WYR9
-
-

Purification (Commentary)

Purification (Comment) Organism
HisPur 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+ low activity with NAD+ Thermotoga maritima D-ribulose 5-phosphate + CO2 + NADH + H+
-
?
6-phospho-D-gluconate + NADP+
-
Thermotoga maritima D-ribulose 5-phosphate + CO2 + NADPH + H+
-
?

Synonyms

Synonyms Comment Organism
6-phosphogluconate dehydrogenase
-
Thermotoga maritima
6PGDH
-
Thermotoga maritima

Turnover Number [1/s]

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

Cofactor

Cofactor Comment Organism Structure
NAD+ low activity with NAD+ Thermotoga maritima
NADP+ natural cofactor Thermotoga maritima

kcat/KM [mM/s]

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