Cloned (Comment) | Organism |
---|---|
overexpression of wild-type and mutant enzymes in strain XL1-Blue | Escherichia coli |
Crystallization (Comment) | Organism |
---|---|
purified recombinant wild-type and mutant enzymes, hanging or sitting drop vapour diffusion method, at 4°C and 21°C, 0.006 ml protein solutions: about 10 mg/ml protein, 1.8 M K2PO4, pH 10.0, N-octyl-beta-R-glucopyranoside 6% w/v, + 2 ml reservoir solution: 1.8 M K2PO4, pH 10.0, 3-4 days, X-ray diffraction structure determination and analysis at 2.35-2.5 A resolution | Escherichia coli |
Protein Variants | Comment | Organism |
---|---|---|
additional information | detailed structural properties of the mutant enzymes based on the crystal structure models | Escherichia coli |
T44V | site-directed mutagenesis, reduced activity, structure is similar to the wild-type enzyme | Escherichia coli |
Y107F | site-directed mutagenesis, reduced activity, structure is similar to the wild-type enzyme | Escherichia coli |
Y133F | site-directed mutagenesis, reduced activity, structure is similar to the wild-type enzyme | Escherichia coli |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.08 | - |
pyruvate | mutant T44V, pH 8.0, 30°C | Escherichia coli | |
0.09 | - |
(S)-aspartate 4-semialdehyde | mutant T44V, pH 8.0, 30°C | Escherichia coli | |
0.11 | - |
(S)-aspartate 4-semialdehyde | wild-type enzyme, pH 8.0, 30°C | Escherichia coli | |
0.16 | - |
pyruvate | mutant Y107F, pH 8.0, 30°C | Escherichia coli | |
0.26 | - |
pyruvate | wild-type enzyme, pH 8.0, 30°C | Escherichia coli | |
0.58 | - |
(S)-aspartate 4-semialdehyde | mutant Y107F, pH 8.0, 30°C | Escherichia coli | |
2.7 | - |
(S)-aspartate 4-semialdehyde | mutant Y133F, pH 8.0, 30°C | Escherichia coli | |
35 | - |
pyruvate | mutant Y133F, pH 8.0, 30°C | Escherichia coli |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
(S)-aspartate 4-semialdehyde + pyruvate | Escherichia coli | branch point reaction in the biosynthesis of lysine | dihydrodipicolinate + H2O | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | P0A6L2 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant wild-type 5.8fold, recombinant mutant Y107F 17.6fold, recombinant mutant T44V 15.8fold, and recombinant mutant Y133F 18fold | Escherichia coli |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
pyruvate + L-aspartate-4-semialdehyde = (2S,4S)-4-hydroxy-2,3,4,5-tetrahydrodipicolinate + H2O | ping-pong kinetic and catalytic mechanism involves a catalytic triad which consists of Tyr133, Thr44, and Tyr107, overview | Escherichia coli |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
0.0034 | - |
purified mutant T44V | Escherichia coli |
0.008 | - |
purified mutant Y133F | Escherichia coli |
0.27 | - |
purified mutant Y107F | Escherichia coli |
1.81 | - |
purified wild-type enzyme | Escherichia coli |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
(S)-aspartate 4-semialdehyde + pyruvate | - |
Escherichia coli | dihydrodipicolinate + H2O | - |
? | |
(S)-aspartate 4-semialdehyde + pyruvate | branch point reaction in the biosynthesis of lysine | Escherichia coli | dihydrodipicolinate + H2O | - |
? |
Subunits | Comment | Organism |
---|---|---|
tetramer | homotetramer of (alpha/beta)8 barrels, each containing one active site, crystal structure | Escherichia coli |
Synonyms | Comment | Organism |
---|---|---|
DHDPS | - |
Escherichia coli |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.12 | - |
(S)-aspartate 4-semialdehyde | mutant T44V, pH 8.0, 30°C | Escherichia coli | |
0.7 | - |
(S)-aspartate 4-semialdehyde | mutant Y133F, pH 8.0, 30°C | Escherichia coli | |
10.8 | - |
(S)-aspartate 4-semialdehyde | mutant Y107F, pH 8.0, 30°C | Escherichia coli | |
124 | - |
(S)-aspartate 4-semialdehyde | wild-type enzyme, pH 8.0, 30°C | Escherichia coli |