Cloned (Comment) | Organism |
---|---|
gene ddh, expression of wild-type enzyme and mutant Q154L/D158G/T173I/R199M/H249N in Escherichia coli strain Rosetta (DE3) | Ureibacillus thermosphaericus |
Protein Variants | Comment | Organism |
---|---|---|
Q154L/D158G/T173I/R199M/H249N | construction of a thermostable, NADP+-dependent D-amino acid dehydrogenase (DAADH) from the meso-diaminopimelate dehydrogenase of strain A1 by introducing five point mutations into amino acid residues located in the active site. In the presence of NADP+, the mutant enzyme catalyzes the oxidative deamination of several D-amino acids, including D-cyclohexylalanine, D-isoleucine, and D-2-aminooctanoate, but not of meso-diaminopimelate. The corresponding 2-oxo acids are aminated in the presence of NADPH and ammonia in the reverse reaction, mutant substrate specificity, overview. The mutant enzyme is also more thermostable than its parental meso-diaminopimelate dehydrogenase | Ureibacillus thermosphaericus |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
2-mercaptoethanol | complete inhibition at 5 mM | Ureibacillus thermosphaericus | |
4-chloromercuribenzoate | complete inhibition at 0.1 mM | Ureibacillus thermosphaericus | |
HgCl2 | complete inhibition at 0.1 mM | Ureibacillus thermosphaericus | |
additional information | no inhibition by EDTA, NaN3, Na2S and iodoacetate at 1 mM | Ureibacillus thermosphaericus | |
N-ethylmaleimide | strong inhibition at 1 mM | Ureibacillus thermosphaericus | |
Ni2+ | strong inhibition at 1 mM | Ureibacillus thermosphaericus | |
thioglycolate | complete inhibition at 5 mM | Ureibacillus thermosphaericus | |
Zn2+ | strong inhibition at 1 mM | Ureibacillus thermosphaericus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | Michaelis-Menten kinetics | Ureibacillus thermosphaericus |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
MnSO4 | slight activation, mutant enzyme | Ureibacillus thermosphaericus |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
meso-2,6-diaminoheptanedioate + H2O + NADP+ | Ureibacillus thermosphaericus | - |
L-2-amino-6-oxoheptanedioate + NH3 + NADPH + H+ | - |
r | |
meso-2,6-diaminoheptanedioate + H2O + NADP+ | Ureibacillus thermosphaericus A1 | - |
L-2-amino-6-oxoheptanedioate + NH3 + NADPH + H+ | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Ureibacillus thermosphaericus | G1UII1 | gene ddh | - |
Ureibacillus thermosphaericus A1 | G1UII1 | gene ddh | - |
Purification (Comment) | Organism |
---|---|
recombinant wild-type enzyme and mutant Q154L/D158G/T173I/R199M/H249N from Escherichia coli strain Rosetta (DE3) by heat treatment and chelating affinity chromatograhy | Ureibacillus thermosphaericus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
meso-2,6-diaminoheptanedioate + H2O + NADP+ | - |
Ureibacillus thermosphaericus | L-2-amino-6-oxoheptanedioate + NH3 + NADPH + H+ | - |
r | |
meso-2,6-diaminoheptanedioate + H2O + NADP+ | - |
Ureibacillus thermosphaericus A1 | L-2-amino-6-oxoheptanedioate + NH3 + NADPH + H+ | - |
r |
Synonyms | Comment | Organism |
---|---|---|
meso-diaminopimelate dehydrogenase | - |
Ureibacillus thermosphaericus |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
50 | - |
assay at | Ureibacillus thermosphaericus |
60 | - |
mutant enzyme | Ureibacillus thermosphaericus |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
65 | - |
purified mutant enzyme, 30 min, pH 7.2, fully stable, half-liife is 80 min | Ureibacillus thermosphaericus |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
10.5 | - |
reductive amination | Ureibacillus thermosphaericus |
11.5 | - |
oxidative deamination | Ureibacillus thermosphaericus |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NADP+ | - |
Ureibacillus thermosphaericus | |
NADPH | - |
Ureibacillus thermosphaericus |
General Information | Comment | Organism |
---|---|---|
malfunction | substrate specificity of mut-DAADH Q154L/D158G/T173I/R199M/H249N , overview | Ureibacillus thermosphaericus |