Cloned (Comment) | Organism |
---|---|
expression in Escherichia coli | Candida parapsilosis |
Crystallization (Comment) | Organism |
---|---|
apo-enzyme, diffraction to 2.7 A. Enzyme forms a homotetramer with broken 2-2-2 symmetry. In the apo-form, the entrance of the NADPH pocket is blocked by a surface loop | Candida parapsilosis |
Protein Variants | Comment | Organism |
---|---|---|
additional information | site-directed mutagenesis, deletion of the N-terminal 31 residues, catalytic parameters similar to wild-type, decrase in melting temperature | Candida parapsilosis |
S172A | site-directed mutagenesis, decrease of melting temperature compared to wild-type. No catalytic activity | Candida parapsilosis |
S172T | site-directed mutagenesis, decrease of melting temperature compared to wild-type. Catalytic activity similar to wild-type | Candida parapsilosis |
S67D/H68D | site-directed mutagenesis, about 10fold increase and 20fold decrease for NADH and NADPH kcat/KM-value, respectively | Candida parapsilosis |
V270D | site-directed mutagenesis, catalytic acitivity similar to wild-type, mutant forms a dimer | Candida parapsilosis |
Y187A | site-directed mutagenesis, decrease of melting temperature compared to wild-type. No catalytic activity | Candida parapsilosis |
Y187F | site-directed mutagenesis, decrease of melting temperature compared to wild-type. No catalytic activity | Candida parapsilosis |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.59 | - |
NADPH | wild-type, pH 7.5, 35°C | Candida parapsilosis | |
0.63 | - |
NADPH | deletion mutant lacking 31 N-terminal amino acids, pH 7.5, 35°C | Candida parapsilosis | |
0.68 | - |
NADPH | mutant S172T, pH 7.5, 35°C | Candida parapsilosis | |
0.71 | - |
NADPH | mutant V270D, pH 7.5, 35°C | Candida parapsilosis | |
0.76 | - |
NADH | mutant S67D/H68D, pH 7.5, 35°C | Candida parapsilosis | |
7.89 | - |
NADH | wild-type, pH 7.5, 35°C | Candida parapsilosis | |
8.03 | - |
NADH | deletion mutant lacking 31 N-terminal amino acids, pH 7.5, 35°C | Candida parapsilosis | |
8.45 | - |
NADH | mutant V270D, pH 7.5, 35°C | Candida parapsilosis | |
8.67 | - |
NADH | mutant S172T, pH 7.5, 35°C | Candida parapsilosis | |
10.27 | - |
NADPH | mutant S67D/H68D, pH 7.5, 35°C | Candida parapsilosis |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Candida parapsilosis | B2KJ46 | fragment | - |
Purification (Comment) | Organism |
---|---|
recombinant protein | Candida parapsilosis |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
2-hydroxyacetophenone + NADH + H+ | - |
Candida parapsilosis | (S)-1-phenyl-1,2-ethanediol + NAD+ | - |
? | |
2-hydroxyacetophenone + NADPH + H+ | - |
Candida parapsilosis | (S)-1-phenyl-1,2-ethanediol + NADP+ | - |
? |
Subunits | Comment | Organism |
---|---|---|
tetramer | crystallization data | Candida parapsilosis |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
45 | - |
melting temperature, mutant Y187A | Candida parapsilosis |
46 | - |
melting temperature, mutant S172A | Candida parapsilosis |
46 | - |
melting temperature, mutant Y187F | Candida parapsilosis |
48 | - |
melting temperature, mutant S172T and N-terminal deletion mutant lacking 31 amino acids | Candida parapsilosis |
52 | - |
melting temperature, wild-type | Candida parapsilosis |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
1.13 | - |
NADH | mutant S172T, pH 7.5, 35°C | Candida parapsilosis | |
1.21 | - |
NADPH | mutant S67D/H68D, pH 7.5, 35°C | Candida parapsilosis | |
1.22 | - |
NADH | mutant V270D, pH 7.5, 35°C | Candida parapsilosis | |
1.24 | - |
NADH | deletion mutant lacking 31 N-terminal amino acids, pH 7.5, 35°C | Candida parapsilosis | |
1.26 | - |
NADH | mutant S67D/H68D, pH 7.5, 35°C | Candida parapsilosis | |
1.32 | - |
NADH | wild-type, pH 7.5, 35°C | Candida parapsilosis | |
1.34 | - |
NADPH | mutant V270D, pH 7.5, 35°C | Candida parapsilosis | |
1.36 | - |
NADPH | mutant S172T, pH 7.5, 35°C | Candida parapsilosis | |
1.39 | - |
NADPH | deletion mutant lacking 31 N-terminal amino acids, pH 7.5, 35°C | Candida parapsilosis | |
1.48 | - |
NADPH | wild-type, pH 7.5, 35°C | Candida parapsilosis |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NADH | NADPH is preferred over NADH | Candida parapsilosis | |
NADP+ | - |
Candida parapsilosis | |
NADPH | preferred over NADH | Candida parapsilosis |