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Literature summary for 1.4.99.6 extracted from

  • Akita, H.; Suzuki, H.; Doi, K.; Ohshima, T.
    Efficient synthesis of D-branched-chain amino acids and their labeled compounds with stable isotopes using D-amino acid dehydrogenase (2014), Appl. Microbiol. Biotechnol., 98, 1135-1143.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli Rosetta (DE3) cells Ureibacillus thermosphaericus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
32.3
-
2-oxo-4-methylvaleric acid at pH 10.5 and 50°C Ureibacillus thermosphaericus

Organism

Organism UniProt Comment Textmining
Ureibacillus thermosphaericus
-
-
-
Ureibacillus thermosphaericus A1 (NBRC 108682)
-
-
-

Purification (Commentary)

Purification (Comment) Organism
Sepharose column chromatography, and gel filtration Ureibacillus thermosphaericus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
(3S,3R)-2-oxo-3-methylvaleric acid + NH3 + NADPH + H+
-
Ureibacillus thermosphaericus D-isoleucine + H2O + NADP+ yield of 45% after incubation for 1 h at 50 °C ?
(3S,3R)-2-oxo-3-methylvaleric acid + NH3 + NADPH + H+
-
Ureibacillus thermosphaericus A1 (NBRC 108682) D-isoleucine + H2O + NADP+ yield of 45% after incubation for 1 h at 50 °C ?
2-oxo-3-methylbutyric acid + NH3 + NADPH + H+
-
Ureibacillus thermosphaericus D-valine + H2O + NADP+ yield of 83% after incubation for 1 h at 50 °C ?
2-oxo-3-methylbutyric acid + NH3 + NADPH + H+
-
Ureibacillus thermosphaericus A1 (NBRC 108682) D-valine + H2O + NADP+ yield of 83% after incubation for 1 h at 50 °C ?
2-oxo-4-methylvaleric acid + NH3 + NADPH + H+
-
Ureibacillus thermosphaericus D-leucine + H2O + NADP+ yield of 91% after incubation for 1 h at 50 °C ?
2-oxo-4-methylvaleric acid + NH3 + NADPH + H+
-
Ureibacillus thermosphaericus A1 (NBRC 108682) D-leucine + H2O + NADP+ yield of 91% after incubation for 1 h at 50 °C ?

Synonyms

Synonyms Comment Organism
D-amino acid dehydrogenase
-
Ureibacillus thermosphaericus
DAADH
-
Ureibacillus thermosphaericus
NADP+-dependent D-amino acid dehydrogenase
-
Ureibacillus thermosphaericus

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
65
-
temperature optimum for D-leucine synthesis Ureibacillus thermosphaericus

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
5700
-
2-oxo-4-methylvaleric acid at pH 10.5 and 50°C Ureibacillus thermosphaericus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
10.5
-
pH-optimum for D-leucine synthesisin glycine-KOH buffer Ureibacillus thermosphaericus

Cofactor

Cofactor Comment Organism Structure
NADP+ the yield is highest in the presence of 0.5 mM NADP+ Ureibacillus thermosphaericus
NADPH dependent on Ureibacillus thermosphaericus

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
180
-
2-oxo-4-methylvaleric acid at pH 10.5 and 50°C Ureibacillus thermosphaericus