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Literature summary extracted from

  • Huang, R.; Chen, H.; Zhong, C.; Kim, J.E.; Zhang, Y.H.
    High-throughput screening of coenzyme preference change of thermophilic 6-phosphogluconate dehydrogenase from NADP+ to NAD+ (2016), Sci. Rep., 6, 32644.
    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 Moorella thermoacetica
1.1.1.343 expressed in Escherichia coli TOP10 and BL21(DE3) cells Moorella thermoacetica

Protein Variants

EC Number Protein Variants Comment Organism
1.1.1.44 A30C/R31I/T32K the mutant exhibits a 118.5fold reversal of coenzyme selectivity from NADP+ to NAD+ Moorella thermoacetica
1.1.1.44 A30D/R31I/T32I the mutant exhibits a 4278fold reversal of coenzyme selectivity from NADP+ to NAD+ Moorella thermoacetica
1.1.1.44 A30E/R31I/T32D the mutant exhibits a 1293fold reversal of coenzyme selectivity from NADP+ to NAD+ Moorella thermoacetica
1.1.1.44 R31I the mutant exhibits a 6.9fold reversal of coenzyme selectivity from NADP+ to NAD+ Moorella thermoacetica
1.1.1.44 R31I/T32G the mutant exhibits a 22.1fold reversal of coenzyme selectivity from NADP+ to NAD+ Moorella thermoacetica
1.1.1.44 R31T the mutant exhibits a 3.5fold reversal of coenzyme selectivity from NADP+ to NAD+ Moorella thermoacetica
1.1.1.343 A30C/R31I/T32K the catalytic efficiency of the mutant for NAD+ is increased by about 1.6fold compared to the wild type enzyme Moorella thermoacetica
1.1.1.343 A30D/R31I/T32I the catalytic efficiency of the mutant for NAD+ is increased by 54fold, with a 4278fold reversal of coenzyme selectivity from NADP+ to NAD+ Moorella thermoacetica
1.1.1.343 A30E/R31I/T32D the catalytic efficiency of the mutant for NAD+ is increased by about 9.5fold compared to the wild type enzyme Moorella thermoacetica
1.1.1.343 R31I the catalytic efficiency of the mutant for NAD+ is increased by about 4.7fold compared to the wild type enzyme Moorella thermoacetica
1.1.1.343 R31I/T32G the catalytic efficiency of the mutant for NAD+ is increased by about 4fold compared to the wild type enzyme Moorella thermoacetica
1.1.1.343 R31T the catalytic efficiency of the mutant for NAD+ is increased by about 1.7fold compared to the wild type enzyme Moorella thermoacetica

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.1.1.44 0.01187
-
 NAD+ mutant enzyme A30D/R31I/T32I, at pH 7.5 and 50°C Moorella thermoacetica
1.1.1.44 0.0139
-
 NADP+ wild type enzyme, at pH 7.5 and 50°C Moorella thermoacetica
1.1.1.44 0.0265
-
 NADP+ mutant enzyme R31I, at pH 7.5 and 50°C Moorella thermoacetica
1.1.1.44 0.0355
-
 NADP+ mutant enzyme R31T, at pH 7.5 and 50°C Moorella thermoacetica
1.1.1.44 0.1044
-
 NADP+ mutant enzyme R31I/T32G, at pH 7.5 and 50°C Moorella thermoacetica
1.1.1.44 0.127
-
 NAD+ mutant enzyme A30E/R31I/T32D, at pH 7.5 and 50°C Moorella thermoacetica
1.1.1.44 0.228
-
 NADP+ mutant enzyme A30D/R31I/T32I, at pH 7.5 and 50°C Moorella thermoacetica
1.1.1.44 0.354
-
 NAD+ mutant enzyme R31I, at pH 7.5 and 50°C Moorella thermoacetica
1.1.1.44 0.362
-
 NAD+ mutant enzyme R31I/T32G, at pH 7.5 and 50°C Moorella thermoacetica
1.1.1.44 0.404
-
 NAD+ mutant enzyme A30C/R31I/T32K, at pH 7.5 and 50°C Moorella thermoacetica
1.1.1.44 0.605
-
 NAD+ mutant enzyme R31T, at pH 7.5 and 50°C Moorella thermoacetica
1.1.1.44 0.66
-
 NADP+ mutant enzyme A30E/R31I/T32D, at pH 7.5 and 50°C Moorella thermoacetica
1.1.1.44 0.698
-
 NADP+ mutant enzyme A30C/R31I/T32K, at pH 7.5 and 50°C Moorella thermoacetica
1.1.1.44 1.397
-
 NAD+ wild type enzyme, at pH 7.5 and 50°C Moorella thermoacetica
1.1.1.343 0.01187
-
 NAD+ mutant enzyme A30D/R31I/T32I, in 100 mM HEPES (pH 7.5), at 50°C Moorella thermoacetica
1.1.1.343 0.0404
-
 NAD+ mutant enzyme A30C/R31I/T32K, in 100 mM HEPES (pH 7.5), at 50°C Moorella thermoacetica
1.1.1.343 0.127
-
 NAD+ mutant enzyme A30E/R31I/T32D, in 100 mM HEPES (pH 7.5), at 50°C Moorella thermoacetica
1.1.1.343 0.354
-
 NAD+ mutant enzyme R31I, in 100 mM HEPES (pH 7.5), at 50°C Moorella thermoacetica
1.1.1.343 0.362
-
 NAD+ mutant enzyme R31I/T32K, in 100 mM HEPES (pH 7.5), at 50°C Moorella thermoacetica
1.1.1.343 0.605
-
 NAD+ mutant enzyme R31T, in 100 mM HEPES (pH 7.5), at 50°C Moorella thermoacetica
1.1.1.343 1.397
-
 NAD+ wild type enzyme, in 100 mM HEPES (pH 7.5), at 50°C Moorella thermoacetica

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+ Moorella thermoacetica
-
 D-ribulose 5-phosphate + CO2 + NADPH + H+
-
 ?
1.1.1.343 6-phospho-D-gluconate + NAD+ Moorella thermoacetica the wild type enzyme shows very low activity with NAD+, mutations at A30, R31, and T32 increase the catalytic activity with NAD+ D-ribulose 5-phosphate + CO2 + NADH + H+
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
1.1.1.44 Moorella thermoacetica A0A1J5NX54
-
-
1.1.1.343 Moorella thermoacetica A0A1J5NX54
-
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.1.44 6-phospho-D-gluconate + NAD+ very low activity with NAD+ Moorella thermoacetica D-ribulose 5-phosphate + CO2 + NADH + H+
-
 ?
1.1.1.44 6-phospho-D-gluconate + NADP+
-
 Moorella thermoacetica D-ribulose 5-phosphate + CO2 + NADPH + H+
-
 ?
1.1.1.343 6-phospho-D-gluconate + NAD+ the wild type enzyme shows very low activity with NAD+, mutations at A30, R31, and T32 increase the catalytic activity with NAD+ Moorella thermoacetica D-ribulose 5-phosphate + CO2 + NADH + H+
-
 ?

Synonyms

EC Number Synonyms Comment Organism
1.1.1.44 6-phosphogluconate dehydrogenase
-
 Moorella thermoacetica
1.1.1.44 6PGDH
-
 Moorella thermoacetica
1.1.1.343 6-phosphogluconate dehydrogenase
-
 Moorella thermoacetica
1.1.1.343 6PGDH
-
 Moorella thermoacetica

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.1.1.44 1.81
-
 NADP+ mutant enzyme A30D/R31I/T32I, at pH 7.5 and 50°C Moorella thermoacetica
1.1.1.44 3.1
-
 NADP+ mutant enzyme A30E/R31I/T32D, at pH 7.5 and 50°C Moorella thermoacetica
1.1.1.44 5.75
-
 NAD+ mutant enzyme A30D/R31I/T32I, at pH 7.5 and 50°C Moorella thermoacetica
1.1.1.44 6
-
 NAD+ mutant enzyme A30C/R31I/T32K, at pH 7.5 and 50°C Moorella thermoacetica
1.1.1.44 6.23
-
 NADP+ mutant enzyme A30C/R31I/T32K, at pH 7.5 and 50°C Moorella thermoacetica
1.1.1.44 8.73
-
 NADP+ wild type enzyme, at pH 7.5 and 50°C Moorella thermoacetica
1.1.1.44 9.12
-
 NAD+ mutant enzyme R31T, at pH 7.5 and 50°C Moorella thermoacetica
1.1.1.44 10.8
-
 NAD+ mutant enzyme A30E/R31I/T32D, at pH 7.5 and 50°C Moorella thermoacetica
1.1.1.44 10.83
-
 NADP+ mutant enzyme R31T, at pH 7.5 and 50°C Moorella thermoacetica
1.1.1.44 11.53
-
 NADP+ mutant enzyme R31I, at pH 7.5 and 50°C Moorella thermoacetica
1.1.1.44 11.9
-
 NADP+ mutant enzyme R31I/T32G, at pH 7.5 and 50°C Moorella thermoacetica
1.1.1.44 12.6
-
 NAD+ wild type enzyme, at pH 7.5 and 50°C Moorella thermoacetica
1.1.1.44 12.94
-
 NAD+ mutant enzyme R31I/T32G, at pH 7.5 and 50°C Moorella thermoacetica
1.1.1.44 15.01
-
 NAD+ mutant enzyme R31I, at pH 7.5 and 50°C Moorella thermoacetica
1.1.1.343 5.75
-
 NAD+ mutant enzyme A30D/R31I/T32I, in 100 mM HEPES (pH 7.5), at 50°C Moorella thermoacetica
1.1.1.343 6
-
 NAD+ mutant enzyme A30C/R31I/T32K, in 100 mM HEPES (pH 7.5), at 50°C Moorella thermoacetica
1.1.1.343 9.12
-
 NAD+ mutant enzyme R31T, in 100 mM HEPES (pH 7.5), at 50°C Moorella thermoacetica
1.1.1.343 10.8
-
 NAD+ mutant enzyme A30E/R31I/T32D, in 100 mM HEPES (pH 7.5), at 50°C Moorella thermoacetica
1.1.1.343 12.6
-
 NAD+ wild type enzyme, in 100 mM HEPES (pH 7.5), at 50°C Moorella thermoacetica
1.1.1.343 12.94
-
 NAD+ mutant enzyme R31I/T32K, in 100 mM HEPES (pH 7.5), at 50°C Moorella thermoacetica
1.1.1.343 15.01
-
 NAD+ mutant enzyme R31I, in 100 mM HEPES (pH 7.5), at 50°C Moorella thermoacetica

Cofactor

EC Number Cofactor Comment Organism Structure
1.1.1.44 NAD+ the catalytic efficiency for NADP+ is far higher than that for NAD+ Moorella thermoacetica
1.1.1.44 NADP+ the catalytic efficiency for NADP+ is far higher than that for NAD+ Moorella thermoacetica
1.1.1.343 NAD+
-
 Moorella thermoacetica

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 4.7
-
 NADP+ mutant enzyme A30E/R31I/T32D, at pH 7.5 and 50°C Moorella thermoacetica
1.1.1.44 7.9
-
 NADP+ mutant enzyme A30D/R31I/T32I, at pH 7.5 and 50°C Moorella thermoacetica
1.1.1.44 8.9
-
 NADP+ mutant enzyme A30C/R31I/T32K, at pH 7.5 and 50°C Moorella thermoacetica
1.1.1.44 9
-
 NAD+ wild type enzyme, at pH 7.5 and 50°C Moorella thermoacetica
1.1.1.44 14.8
-
 NAD+ mutant enzyme A30C/R31I/T32K, at pH 7.5 and 50°C Moorella thermoacetica
1.1.1.44 15
-
 NAD+ mutant enzyme R31T, at pH 7.5 and 50°C Moorella thermoacetica
1.1.1.44 35.8
-
 NAD+ mutant enzyme R31I/T32G, at pH 7.5 and 50°C Moorella thermoacetica
1.1.1.44 42.4
-
 NAD+ mutant enzyme R31I, at pH 7.5 and 50°C Moorella thermoacetica
1.1.1.44 85.1
-
 NAD+ mutant enzyme A30E/R31I/T32D, at pH 7.5 and 50°C Moorella thermoacetica
1.1.1.44 114
-
 NADP+ mutant enzyme R31I/T32G, at pH 7.5 and 50°C Moorella thermoacetica
1.1.1.44 305.6
-
 NADP+ mutant enzyme R31T, at pH 7.5 and 50°C Moorella thermoacetica
1.1.1.44 435
-
 NADP+ mutant enzyme R31I, at pH 7.5 and 50°C Moorella thermoacetica
1.1.1.44 484.2
-
 NAD+ mutant enzyme A30D/R31I/T32I, at pH 7.5 and 50°C Moorella thermoacetica
1.1.1.44 628.8
-
 NADP+ wild type enzyme, at pH 7.5 and 50°C Moorella thermoacetica
1.1.1.343 9
-
 NAD+ wild type enzyme, in 100 mM HEPES (pH 7.5), at 50°C Moorella thermoacetica
1.1.1.343 14.8
-
 NAD+ mutant enzyme A30C/R31I/T32K, in 100 mM HEPES (pH 7.5), at 50°C Moorella thermoacetica
1.1.1.343 15
-
 NAD+ mutant enzyme R31T, in 100 mM HEPES (pH 7.5), at 50°C Moorella thermoacetica
1.1.1.343 35.8
-
 NAD+ mutant enzyme R31I/T32K, in 100 mM HEPES (pH 7.5), at 50°C Moorella thermoacetica
1.1.1.343 42.4
-
 NAD+ mutant enzyme R31I, in 100 mM HEPES (pH 7.5), at 50°C Moorella thermoacetica
1.1.1.343 85.1
-
 NAD+ mutant enzyme A30E/R31I/T32D, in 100 mM HEPES (pH 7.5), at 50°C Moorella thermoacetica
1.1.1.343 484.2
-
 NAD+ mutant enzyme A30D/R31I/T32I, in 100 mM HEPES (pH 7.5), at 50°C Moorella thermoacetica