EC Number | Cloned (Comment) | Organism |
---|---|---|
1.1.1.184 | SCR overexpressed from pETSCR plasmid in Escherichia coli BL21 (DE3) as His6-tagged proteins | Candida parapsilosis |
EC Number | Crystallization (Comment) | Organism |
---|---|---|
1.1.1.184 | by the hanging-drop vapor diffusion method, to 2.7 A resolution, crystal form belongs to space group P212121 with cell dimensions of a=104.7 A, b=142.8 A, and c=151.8 A. It forms a homotetramer with a broken 2-2-2 symmetry. SCR contains an extended N-terminal peptide (i.e., residues 1-25) and a short helix (residues 26-30) projecting out from the core domain that may stabilize the oligomer. In the apo-SCR structure, the entrance of the NADPH pocket is blocked by a surface loop | Candida parapsilosis |
EC Number | Protein Variants | Comment | Organism |
---|---|---|---|
1.1.1.184 | DELTA31 | deletion of the N-terminal 31 residues, kinetic parameters KM and kcat are essentially the same as those of the wild-type. It forms a tetramer in solution, which is similar to the wild-type but is less stable. Melting temperature is 48°C, which is lower than that of the wild-type (52°C) | Candida parapsilosis |
1.1.1.184 | S172A | melting temperature is 46°C, which is lower than that of the wild-type (52°C). Shows no activity | Candida parapsilosis |
1.1.1.184 | S172T | melting temperature is 48°C, which is lower than that of the wild-type (52°C). Activity is essentially the same as the wild-type | Candida parapsilosis |
1.1.1.184 | S67D/H68D | double-point mutation inside the coenzyme-binding pocket results in a nearly 10fold increase and a 20fold decrease in the kcat/KM value when NADH and NADPH are used as cofactors, respectively, with kcat remaining essentially the same. Shows similar thermal stability to wild-type | Candida parapsilosis |
1.1.1.184 | V270D | renders the SCR as a homodimer, rather than a tetramer, without affecting the enzymatic activity. Melting temperature is 45°C, which is lower than that of the wild-type (52°C) | Candida parapsilosis |
1.1.1.184 | Y187A | melting temperature is 45°C, which is lower than that of the wild-type (52°C). Shows no activity | Candida parapsilosis |
1.1.1.184 | Y187F | melting temperature is 46°C, which is lower than that of the wild-type (52°C). Shows no activity | Candida parapsilosis |
EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
1.1.1.184 | 0.59 | - |
2-hydroxyacetophenone | wild-type, with NADPH as cofactor, at 35°C in 100 mM acetate buffer (pH 5.0) | Candida parapsilosis | |
1.1.1.184 | 0.63 | - |
2-hydroxyacetophenone | mutant DELTA31, with NADPH as cofactor, at 35°C in 100 mM acetate buffer (pH 5.0) | Candida parapsilosis | |
1.1.1.184 | 0.68 | - |
2-hydroxyacetophenone | mutant S172T, with NADPH as cofactor, at 35°C in 100 mM acetate buffer (pH 5.0) | Candida parapsilosis | |
1.1.1.184 | 0.71 | - |
2-hydroxyacetophenone | mutant V270D, with NADPH as cofactor, at 35°C in 100 mM acetate buffer (pH 5.0) | Candida parapsilosis | |
1.1.1.184 | 0.76 | - |
2-hydroxyacetophenone | mutant S67D/H68D, with NADH as cofactor, at 35°C in 100 mM acetate buffer (pH 5.0) | Candida parapsilosis | |
1.1.1.184 | 7.89 | - |
2-hydroxyacetophenone | wild-type, with NADH as cofactor, at 35°C in 100 mM acetate buffer (pH 5.0) | Candida parapsilosis | |
1.1.1.184 | 8.03 | - |
2-hydroxyacetophenone | mutant DELTA31, with NADH as cofactor, at 35°C in 100 mM acetate buffer (pH 5.0) | Candida parapsilosis | |
1.1.1.184 | 8.45 | - |
2-hydroxyacetophenone | mutant V270D, with NADH as cofactor, at 35°C in 100 mM acetate buffer (pH 5.0) | Candida parapsilosis | |
1.1.1.184 | 8.67 | - |
2-hydroxyacetophenone | mutant S172T, with NADH as cofactor, at 35°C in 100 mM acetate buffer (pH 5.0) | Candida parapsilosis | |
1.1.1.184 | 10.27 | - |
2-hydroxyacetophenone | mutant S67D/H68D, with NADPH as cofactor, at 35°C in 100 mM acetate buffer (pH 5.0) | Candida parapsilosis |
EC Number | Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|---|
1.1.1.184 | 58000 | - |
gel filtration, mutant V270D | Candida parapsilosis |
1.1.1.184 | 100000 | - |
analytic ultracentrifugation, mutant DELTA31 | Candida parapsilosis |
1.1.1.184 | 112000 | - |
analytic ultracentrifugation, wild-type | Candida parapsilosis |
EC Number | Organism | UniProt | Comment | Textmining |
---|---|---|---|---|
1.1.1.184 | Candida parapsilosis | B2KJ46 | - |
- |
EC Number | Purification (Comment) | Organism |
---|---|---|
1.1.1.184 | by affinity chromatography on a Ni2+ column and by gel filtration | Candida parapsilosis |
EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
1.1.1.184 | 2-hydroxyacetophenone + NADPH + H+ | - |
Candida parapsilosis | (S)-1-phenyl-1,2-ethanediol + NADP+ | - |
? |
EC Number | Subunits | Comment | Organism |
---|---|---|---|
1.1.1.184 | homodimer | analytic ultracentrifugation analysis, mutant V270D | Candida parapsilosis |
1.1.1.184 | homotetramer | crystallography, wild-type | Candida parapsilosis |
EC Number | Synonyms | Comment | Organism |
---|---|---|---|
1.1.1.184 | SCR | - |
Candida parapsilosis |
1.1.1.184 | short-chain (S)-1-phenyl-1,2-ethanediol dehydrogenase | - |
Candida parapsilosis |
EC Number | Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
1.1.1.184 | 1.13 | - |
2-hydroxyacetophenone | mutant S172T, with NADH as cofactor, at 35°C in 100 mM acetate buffer (pH 5.0) | Candida parapsilosis | |
1.1.1.184 | 1.21 | - |
2-hydroxyacetophenone | mutant S67D/H68D, with NADPH as cofactor, at 35°C in 100 mM acetate buffer (pH 5.0) | Candida parapsilosis | |
1.1.1.184 | 1.22 | - |
2-hydroxyacetophenone | mutant V270D, with NADH as cofactor, at 35°C in 100 mM acetate buffer (pH 5.0) | Candida parapsilosis | |
1.1.1.184 | 1.24 | - |
2-hydroxyacetophenone | mutant DELTA31, with NADH as cofactor, at 35°C in 100 mM acetate buffer (pH 5.0) | Candida parapsilosis | |
1.1.1.184 | 1.26 | - |
2-hydroxyacetophenone | mutant S67D/H68D, with NADH as cofactor, at 35°C in 100 mM acetate buffer (pH 5.0) | Candida parapsilosis | |
1.1.1.184 | 1.32 | - |
2-hydroxyacetophenone | wild-type, with NADH as cofactor, at 35°C in 100 mM acetate buffer (pH 5.0) | Candida parapsilosis | |
1.1.1.184 | 1.34 | - |
2-hydroxyacetophenone | mutant V270D, with NADPH as cofactor, at 35°C in 100 mM acetate buffer (pH 5.0) | Candida parapsilosis | |
1.1.1.184 | 1.36 | - |
2-hydroxyacetophenone | mutant S172T, with NADPH as cofactor, at 35°C in 100 mM acetate buffer (pH 5.0) | Candida parapsilosis | |
1.1.1.184 | 1.39 | - |
2-hydroxyacetophenone | mutant DELTA31, with NADPH as cofactor, at 35°C in 100 mM acetate buffer (pH 5.0) | Candida parapsilosis | |
1.1.1.184 | 1.48 | - |
2-hydroxyacetophenone | wild-type, with NADPH as cofactor, at 35°C in 100 mM acetate buffer (pH 5.0) | Candida parapsilosis |
EC Number | Cofactor | Comment | Organism | Structure |
---|---|---|---|---|
1.1.1.184 | NAD(P)H | the wild-type exhibits coenzyme specificity for NADPH over NADH, whereas NADH has stronger inducing ability than NADPH for the S67D/H68D mutant | Candida parapsilosis |