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Literature summary for 1.17.1.9 extracted from

  • Yamamoto, H.; Mitsuhashi, K.; Kimoto, N.; Kobayashi, Y.; Esaki, N.
    Robust NADH-regenerator: Improved alpha-haloketone-resistant formate dehydrogenase (2005), Appl. Microbiol. Biotechnol., 67, 33-39.
    View publication on PubMed

Activating Compound

Activating Compound Comment Organism Structure
ethyl acetate wild-type, 21% increase of activity Mycolicibacterium vaccae

Application

Application Comment Organism
biotechnology use of enzyme for regeneration of NADH. Coexpression of alpha-haloketone resistant enzyme mutants and a carbonyl reductase from Kluyveromyces aestuarii for production of ethyl-(S)-4-chloro-3-hydroxybutanoate with optical purity of product of 99% enantiomeric excess, from ethyl-4-chloroacetoacetate Mycolicibacterium vaccae

Protein Variants

Protein Variants Comment Organism
C146S/C256V 85% increase of activity in presence of ethyl acetate, 104% residual activity in presence of 20 mM ethyl-4-chloroacetoacetate Mycolicibacterium vaccae
C256V 11% increase of activity in presence of ethyl acetate, 100% residual activity in presence of 20 mM ethyl-4-chloroacetoacetate Mycolicibacterium vaccae
C6A/C146A/C256V 119% increase of activity in presence of ethyl acetate, 106% residual activity in presence of 20 mM ethyl-4-chloroacetoacetate Mycolicibacterium vaccae
C6A/C146S/C256V 37% increase of activity in presence of ethyl acetate, 108% residual activity in presence of 20 mM ethyl-4-chloroacetoacetate Mycolicibacterium vaccae
C6A/C146V/C256V 15% decrease of activity in presence of ethyl acetate, 100% residual activity in presence of 20 mM ethyl-4-chloroacetoacetate Mycolicibacterium vaccae
C6A/C256S 9% increase of activity in presence of ethyl acetate, 97% residual activity in presence of 20 mM ethyl-4-chloroacetoacetate Mycolicibacterium vaccae
C6A/C256V 11% increase of activity in presence of ethyl acetate, 94.5% residual activity in presence of 20 mM ethyl-4-chloroacetoacetate Mycolicibacterium vaccae
C6S/C256A 25% increase of activity in presence of ethyl acetate, 120% residual activity in presence of 20 mM ethyl-4-chloroacetoacetate Mycolicibacterium vaccae
C6S/C256S 5% increase of activity in presence of ethyl acetate, 125% residual activity in presence of 20 mM ethyl-4-chloroacetoacetate Mycolicibacterium vaccae
C6S/C256V 10% increase of activity in presence of ethyl acetate, 94% residual activity in presence of 20 mM ethyl-4-chloroacetoacetate Mycolicibacterium vaccae
C6V/C256S 5% increase of activity in presence of ethyl acetate, 92% residual activity in presence of 20 mM ethyl-4-chloroacetoacetate Mycolicibacterium vaccae

Inhibitors

Inhibitors Comment Organism Structure
ethyl-4-chloroacetoacetate wild-type, 7.43% residual activity at 20 mM Mycolicibacterium vaccae

Organism

Organism UniProt Comment Textmining
Mycolicibacterium vaccae
-
-
-

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.285
-
mutant C6S/C256S, pH 6.5, 30°C Mycolicibacterium vaccae
0.504
-
mutant C6S/C256V, pH 6.5, 30°C Mycolicibacterium vaccae
0.635
-
mutant C6S/C256A, pH 6.5, 30°C Mycolicibacterium vaccae
1.15
-
mutant C6A/C146A/C256V, pH 6.5, 30°C Mycolicibacterium vaccae
1.49
-
mutant C6A/C146V/C256V, pH 6.5, 30°C Mycolicibacterium vaccae
1.62
-
mutant C6V/C256S, pH 6.5, 30°C Mycolicibacterium vaccae
1.83
-
mutant C6A/C256V, pH 6.5, 30°C Mycolicibacterium vaccae
1.98
-
mutant C6A/C256S, pH 6.5, 30°C Mycolicibacterium vaccae
2.29
-
mutant C146S/C256V, pH 6.5, 30°C Mycolicibacterium vaccae
2.58
-
mutant C256V, pH 6.5, 30°C Mycolicibacterium vaccae
2.65
-
mutant C6A/C146S/C256V, pH 6.5, 30°C Mycolicibacterium vaccae
3.24
-
wild-type, pH 6.5, 30°C Mycolicibacterium vaccae

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
formate + NAD+
-
Mycolicibacterium vaccae CO2 + NADH + H+
-
?

Cofactor

Cofactor Comment Organism Structure
NAD+
-
Mycolicibacterium vaccae
NADH
-
Mycolicibacterium vaccae