Cloned (Comment) | Organism |
---|---|
expression oof wild-type and mutant enzyme in Escherichia coli strain Rosetta 2 (DE3) | Dictyostelium discoideum |
Protein Variants | Comment | Organism |
---|---|---|
H84A | site-directed mutagenesis, the mutant shows reduced racemase and dehydrase activities compared to the wild-type enzyme | Dictyostelium discoideum |
K111A | site-directed mutagenesis, the mutant shows reduced racemase and dehydrase activities compared to the wild-type enzyme | Dictyostelium discoideum |
K56A | site-directed mutagenesis, catalytically inactive mutant | Dictyostelium discoideum |
additional information | circular dichroism spectral analysis of wild-type and mutant enzymes, overview | Dictyostelium discoideum |
P150S | site-directed mutagenesis, the mutant shows reduced racemase and dehydrase activities compared to the wild-type enzyme | Dictyostelium discoideum |
R132A | site-directed mutagenesis, the mutant shows reduced racemase and dehydrase activities compared to the wild-type enzyme | Dictyostelium discoideum |
S80A | site-directed mutagenesis, catalytically inactive mutant | Dictyostelium discoideum |
S80C | site-directed mutagenesis, the mutant shows reduced racemase and dehydrase activities compared to the wild-type enzyme | Dictyostelium discoideum |
S81A | site-directed mutagenesis, Ser81 is located on the opposite side of K56, the mutation converts the enzyme from serine racemase to L-serine dehydrase, the mutant shows no racemase activity and has significantly reduced D-serine dehydrase activity, but it completely retains its L-serine dehydrase activity | Dictyostelium discoideum |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
L-serine | Dictyostelium discoideum | - |
D-serine | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Dictyostelium discoideum | - |
- |
- |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
L-serine = D-serine | the abstraction and addition of alpha-hydrogen to L- and D-serine are conducted by residues K56 and S81 at the si- and re-sides, respectively, of pyridoxal 5'-phosphate, K56 functions as a residue that abstracts the alpha-hydrogen from the Schiff base intermediate, two-base catalytic mechanism, overview | Dictyostelium discoideum |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
L-serine | - |
Dictyostelium discoideum | D-serine | - |
r | |
additional information | the enzyme catalyzes racemization and dehydration of both isomers of serine | Dictyostelium discoideum | ? | - |
? |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
assay at | Dictyostelium discoideum |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8.5 | - |
assay at | Dictyostelium discoideum |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
pyridoxal 5'-phosphate | dependent on, binding residue is Lys56 | Dictyostelium discoideum |
General Information | Comment | Organism |
---|---|---|
evolution | the eukaryotic serine racemase from Dictyostelium discoideum is a fold-type II pyridoxal 5'-phosphate-dependent enzyme | Dictyostelium discoideum |
additional information | circular dichroism spectral analysis of wild-type and mutant enzymes, overview | Dictyostelium discoideum |