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Literature summary for 1.1.1.2 extracted from

  • Solanki, K.; Abdallah, W.; Banta, S.
    Engineering the cofactor specificity of an alcohol dehydrogenase via single mutations or insertions distal to the 2'-phosphate group of NADP(H) (2017), Protein Eng. Des. Sel., 30, 373-380 .
    View publication on PubMed
Search for more literature for 1.1.1.2

Protein Variants

Protein Variants Comment Organism
G211C site-directed mutagenesis, cofactor binding and kinetic analysis compared to the wild-type enzyme Pyrococcus furiosus
G211S site-directed mutagenesis, cofactor binding and kinetic analysis compared to the wild-type enzyme Pyrococcus furiosus
additional information insertion mutants of Ala, Gly, Ser or Cys between positions 211 and 212 (termed G211InsA, G211InsG, G211InsS, and G211InsC, respectively) are created by site-directed mutagenesis of the wild-type AdhD gene in the pET-20b vector. For some mutants (G211S, G211C, G211InsA and G211InsS), with NAD+ as a cofactor, the KA or KB values are observed to be higher than the maximum concentration of substrate or cofactor utilized. So, for mutants G211S, G211C, G211InsA and G211InsS, kinetic data are also measured using assay reaction mixtures containing 1-450 mM 2,3-butanediol and 1-5500 microM NAD+. In the case of NAD+, the on-rate of cofactor (kss 1 ) is found to decrease in all of the mutants except for the G211S and insertion of Gly (G211InsG). Kinetic analysis, detailed overview Pyrococcus furiosus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
 additional information kinetic analysis of wild-type and mutant enzymes, fit to the ordered bi-bi rate equation, steady-state kinetics, detailed overview. For some mutants (G211S, G211C, G211InsA and G211InsS), with NAD+ as a cofactor, the KA or KB values are observed to be higher than the maximum concentration of substrate or cofactor utilized. So, for mutants G211S, G211C, G211InsA and G211InsS, kinetic data is also measured using assay reaction mixtures containing 1-450 mM 2,3-butanediol and 0.001-5.50 mM NAD+ Pyrococcus furiosus

Organism

Organism UniProt Comment Textmining
Pyrococcus furiosus
-
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2,3-butanediol + NADP+
-
 Pyrococcus furiosus acetoin + NADPH + H+
-
 r

Synonyms

Synonyms Comment Organism
AdhD
-
 Pyrococcus furiosus
alcohol dehydrogenase D
-
 Pyrococcus furiosus

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
45
-
 assay at Pyrococcus furiosus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8.8
-
 assay at Pyrococcus furiosus

Cofactor

Cofactor Comment Organism Structure
additional information cofactor binding energies and kinetics with wild-type and mutant enzymes, overview Pyrococcus furiosus
NADP+
-
 Pyrococcus furiosus
NADPH
-
 Pyrococcus furiosus

General Information

General Information Comment Organism
additional information structure homology model of AdhD and its mutants generated using the model of prostaglandin F synthase from Trypanosoma brucei (PDB ID 1VBJ, 40.26% identity). The homology models are aligned with the structure of 2,5-DKGR from Corynebacterium (PDB ID 1A80 with bound NADPH, and PDB ID 1M9H with bound NADH), overview Pyrococcus furiosus