Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary extracted from

  • Lerchner, A.; Jarasch, A.; Meining, W.; Schiefner, A.; Skerra, A.
    Crystallographic analysis and structure-guided engineering of NADPH-dependent Ralstonia sp. alcohol dehydrogenase toward NADH cosubstrate specificity (2013), Biotechnol. Bioeng., 110, 2803-2814.
    View publication on PubMed

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
1.1.1.91 crystal structure of the apoenzyme as well as its NADP+-bound state with resolutions down to 2.8 A. ADH displays a homotetrameric quaternary structure that can be described as a dimer of homodimers while in each subunit a seven-stranded parallel beta-sheet, flanked by three alpha-helices on each side, forms a Rossmann fold-type dinucleotide binding domain. NADP+ specificity is largely governed by the residues Asn15, Gly37, Arg38, and Arg39 Ralstonia sp.

Protein Variants

EC Number Protein Variants Comment Organism
1.1.1.91 G37D mutation in NADP+-binding site, strong decrease in activity with NADP+ Ralstonia sp.
1.1.1.91 N15G/G37D/R38V/R39S mutation in NADP+-binding site, strong decrease in activity with NADP+ Ralstonia sp.
1.1.1.91 N15G/G37D/R38V/R39S/A86N/S88A mutation in NADP+-binding site, strong decrease in activity with NADP+ Ralstonia sp.

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.1.1.91 0.051
-
 NADP+ wild-type, pH 8.3, 30°C Ralstonia sp.
1.1.1.91 0.28
-
 NADP+ mutant N15G/G37D/R38V/R39S, pH 8.3, 30°C Ralstonia sp.
1.1.1.91 0.8
-
 NAD+ mutant N15G/G37D/R38V/R39S/A86N/S88A, pH 8.3, 30°C Ralstonia sp.
1.1.1.91 1.1
-
 NAD+ mutant N15G/G37D/R38V/R39S, pH 8.3, 30°C Ralstonia sp.
1.1.1.91 1.2
-
 (S)-1-phenylethanol mutant G37D, pH 8.3, 30°C Ralstonia sp.
1.1.1.91 2.3
-
 NAD+ mutant G37D, pH 8.3, 30°C Ralstonia sp.
1.1.1.91 2.3
-
 NADP+ mutant G37D, pH 8.3, 30°C Ralstonia sp.
1.1.1.91 3.1
-
 NAD+ wild-type, pH 8.3, 30°C Ralstonia sp.
1.1.1.91 5.5
-
 (R)-1-phenylethanol wild-type, pH 8.3, 30°C Ralstonia sp.
1.1.1.91 8.3
-
 (S)-1-phenylethanol mutant N15G/G37D/R38V/R39S, pH 8.3, 30°C Ralstonia sp.
1.1.1.91 9.9
-
 (S)-1-phenylethanol wild-type, pH 8.3, 30°C Ralstonia sp.
1.1.1.91 17.6
-
 (S)-1-phenylethanol mutant N15G/G37D/R38V/R39S/A86N/S88A, pH 8.3, 30°C Ralstonia sp.

Organism

EC Number Organism UniProt Comment Textmining
1.1.1.91 Ralstonia sp. C0IR58
-
-
1.1.1.91 Ralstonia sp. DSM Z 6428 C0IR58
-
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.1.91 (R)-1-phenylethanol + NAD+
-
 Ralstonia sp. 1-phenylacetaldehyde + NADH + H+
-
 ?
1.1.1.91 (R)-1-phenylethanol + NAD+
-
 Ralstonia sp. DSM Z 6428 1-phenylacetaldehyde + NADH + H+
-
 ?
1.1.1.91 (S)-1-phenylethanol + NADP+
-
 Ralstonia sp. 1-phenylacetaldehyde + NADPH + H+
-
 ?
1.1.1.91 (S)-1-phenylethanol + NADP+
-
 Ralstonia sp. DSM Z 6428 1-phenylacetaldehyde + NADPH + H+
-
 ?

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.1.1.91 0.00003
-
 NADP+ mutant N15G/G37D/R38V/R39S, pH 8.3, 30°C Ralstonia sp.
1.1.1.91 0.0025
-
 NADP+ mutant G37D, pH 8.3, 30°C Ralstonia sp.
1.1.1.91 0.0047
-
 (R)-1-phenylethanol wild-type, pH 8.3, 30°C Ralstonia sp.
1.1.1.91 0.0115
-
 (S)-1-phenylethanol wild-type, pH 8.3, 30°C Ralstonia sp.
1.1.1.91 0.022
-
 NAD+ wild-type, pH 8.3, 30°C Ralstonia sp.
1.1.1.91 0.023
-
 (S)-1-phenylethanol mutant N15G/G37D/R38V/R39S/A86N/S88A, pH 8.3, 30°C Ralstonia sp.
1.1.1.91 0.03
-
 NAD+ mutant N15G/G37D/R38V/R39S/A86N/S88A, pH 8.3, 30°C Ralstonia sp.
1.1.1.91 0.038
-
 NAD+ mutant G37D, pH 8.3, 30°C Ralstonia sp.
1.1.1.91 0.042
-
 (S)-1-phenylethanol mutant N15G/G37D/R38V/R39S, pH 8.3, 30°C Ralstonia sp.
1.1.1.91 0.08
-
 NAD+ mutant N15G/G37D/R38V/R39S, pH 8.3, 30°C Ralstonia sp.
1.1.1.91 0.783
-
 (S)-1-phenylethanol mutant G37D, pH 8.3, 30°C Ralstonia sp.
1.1.1.91 1.65
-
 NADP+ wild-type, pH 8.3, 30°C Ralstonia sp.

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.1.1.91 0.0001
-
 NADP+ mutant N15G/G37D/R38V/R39S, pH 8.3, 30°C Ralstonia sp.
1.1.1.91 0.0011
-
 NADP+ mutant G37D, pH 8.3, 30°C Ralstonia sp.
1.1.1.91 0.00115
-
 (S)-1-phenylethanol wild-type, pH 8.3, 30°C Ralstonia sp.
1.1.1.91 0.0013
-
 (S)-1-phenylethanol mutant N15G/G37D/R38V/R39S/A86N/S88A, pH 8.3, 30°C Ralstonia sp.
1.1.1.91 0.0047
-
 (R)-1-phenylethanol wild-type, pH 8.3, 30°C Ralstonia sp.
1.1.1.91 0.0052
-
 (S)-1-phenylethanol mutant N15G/G37D/R38V/R39S, pH 8.3, 30°C Ralstonia sp.
1.1.1.91 0.007
-
 NAD+ wild-type, pH 8.3, 30°C Ralstonia sp.
1.1.1.91 0.0167
-
 NAD+ mutant G37D, pH 8.3, 30°C Ralstonia sp.
1.1.1.91 0.038
-
 NAD+ mutant N15G/G37D/R38V/R39S/A86N/S88A, pH 8.3, 30°C Ralstonia sp.
1.1.1.91 0.073
-
 NAD+ mutant N15G/G37D/R38V/R39S, pH 8.3, 30°C Ralstonia sp.
1.1.1.91 0.65
-
 (S)-1-phenylethanol mutant G37D, pH 8.3, 30°C Ralstonia sp.
1.1.1.91 32.33
-
 NADP+ wild-type, pH 8.3, 30°C Ralstonia sp.