Activating Compound | Comment | Organism | Structure |
---|---|---|---|
pyridoxal 5'-phosphate | the addition to the reaction mixture immediately before carrying out the enzymatic assay has no influence on activity, but preincubation increases the activity about twofold | Pseudomonas putida | |
pyridoxal 5'-phosphate | the optimum of activation is obtained with a 0.05 mM concentration | Pseudomonas putida |
Application | Comment | Organism |
---|---|---|
synthesis | L-phenylalanine production as a building block of aspartame | Pseudomonas putida |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
CoCl2 | retains 35% activity at 10 mM | Pseudomonas putida | |
EDTA | complete inhibition at 10 mM | Pseudomonas putida | |
HgCl2 | irreversible and complete inhibition is observed at concentrations about 0.1 mM | Pseudomonas putida | |
KCN | irreversible inhibitor | Pseudomonas putida | |
MnCl2 | retains 65% activity at 10 mM | Pseudomonas putida | |
p-chloromercuribenzoate | irreversible and complete inhibition is observed at concentrations about 0.1 mM | Pseudomonas putida | |
phenyl 2-oxopropanoate | - |
Pseudomonas putida | |
phenylhydrazine | irreversible inhibitor; retains 56% activity at 0.01 mM, retains 18% activity at 0.1 mM | Pseudomonas putida | |
pyridoxal 5'-phosphate | at concentrations higher than 0.05 mM inhibitory effects are observed | Pseudomonas putida | |
Semicarbazide | irreversible inhibitor; retains 59% activity at 0.01 mM, retains 24% activity at 0.1 mM | Pseudomonas putida |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
36000 | - |
2 * 36000, SDS-PAGE | Pseudomonas putida |
72000 | - |
gel filtration | Pseudomonas putida |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
L-aspartate + 3-phenylpyruvate | Pseudomonas putida | - |
oxaloacetate + L-phenylalanine | oxaloacetate decomposition drives the reaction towards L-phenylalanine | r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Pseudomonas putida | - |
- |
- |
Purification (Comment) | Organism |
---|---|
centrifugation, streptomycin sulfate precipitation, several chromatographic steps and disc-gel electrophoresis | Pseudomonas putida |
Storage Stability | Organism |
---|---|
-18°C, 50% glycerol without loss of activity for over two years | Pseudomonas putida |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
2-oxoglutarate + L-aspartate | 34% activity with respect to L-phenylpyruvate | Pseudomonas putida | L-glutamate + oxaloaspartate | - |
? | |
2-oxoglutarate + L-phenylalanine | reverse reaction, 31% activity with respect to oxalacetate | Pseudomonas putida | L-glutamate + 3-phenylpyruvate | - |
r | |
2-oxoisovalerate + L-aspartate | 6% activity with respect to L-phenylpyruvate | Pseudomonas putida | L-norvaline + oxaloacetate | - |
? | |
2-oxoisovalerate + L-phenylalanine | reverse reaction, 8% activity with respect to oxalacetate | Pseudomonas putida | L-norvaline + 3-phenylpyruvate | - |
r | |
3-phenylpyruvate + L-histidine | 4% activity with respect to L-aspartate | Pseudomonas putida | 3-(1H-imidazol-4-yl)-2-oxopropanoate + L-phenylalanine | - |
? | |
3-phenylpyruvate + L-tyrosine | 27% activity with respect to aspartate | Pseudomonas putida | p-hydroxyphenylpyruvate + L-phenylalanine | - |
? | |
beta-methyl-DL-aspartate + 3-phenylpyruvate | 2% activity with respect to L-aspartate | Pseudomonas putida | 3-methyl-2-oxobutanedioate + L-phenylalanine | - |
? | |
DL-p-fluorophenylalanine + oxaloacetate | reverse reaction, 51% activity with respect to L-phenylalanine | Pseudomonas putida | p-fluorophenylpyruvate + L-aspartate | - |
r | |
indolepyruvate + L-aspartate | 10% activity with respect to L-phenylpyruvate | Pseudomonas putida | oxaloaspartate + L-tryptophan | - |
? | |
L-aspartate + 3-phenylpyruvate | - |
Pseudomonas putida | oxaloacetate + L-phenylalanine | oxaloacetate decomposition drives the reaction towards L-phenylalanine | r | |
L-tryptophan + 3-phenylpyruvate | 40% activity with respect to aspartate | Pseudomonas putida | 3-indole-2-oxopropanoate + L-phenylalanine | - |
? | |
L-tryptophan + oxaloacetate | reverse reaction, 77% activity with respect to L-phenylalanine | Pseudomonas putida | 3-indole-2-oxopropanoate + L-aspartate | - |
r | |
additional information | cysteine, glutamine, arginine, alanine, threonine, L-aspartate dibutylester, L-phenylglycine, isoleucine, valine, serine, lysine, ketobutyrate, alpha-keto-beta-n-valerate, beta-hydroxypyruvate, prephenate are inert as substrates | Pseudomonas putida | ? | - |
? | |
additional information | D amino acids are not utilized as substrates | Pseudomonas putida | ? | - |
? | |
additional information | modification of the carboxyl group of L-phenylalanine exhibits great influence on the enzyme substrate interaction as do substituents at C-2 or shortening the side chain | Pseudomonas putida | ? | - |
? | |
additional information | aspartate derivatives are used only with low activity if a modification at the position of the beta carbon is made | Pseudomonas putida | ? | - |
? | |
additional information | addition of substituents in the aromatic ring in the para position decreases the affinity of the derivative to the enzyme apparently with increasing bulk of the group, for a given halogen, position at the aromatic ring has only marginal influence on the reaction rate | Pseudomonas putida | ? | - |
? | |
oxaloacetate + L-histidine | reverse reaction, 4% activity with respect to L-phenylalanine | Pseudomonas putida | 3-(1H-imidazol-4-yl)-2-oxopropanoate + L-aspartate | - |
r | |
oxaloacetate + L-tyrosine | reverse reaction, 38% activity with respect to L-phenylalanine | Pseudomonas putida | p-hydroxyphenylpyruvate + L-aspartate | - |
r | |
p-hydroxyphenylpyruvate + L-aspartate | 54% activity with respect to L-phenylpyruvate | Pseudomonas putida | oxaloacetate + L-tyrosine | - |
? | |
p-hydroxyphenylpyruvate + L-phenylalanine | reverse reaction, 52% activity with respect to oxalacetate | Pseudomonas putida | L-tyrosine + 3-phenylpyruvate | - |
r |
Subunits | Comment | Organism |
---|---|---|
dimer | 2 * 36000, SDS-PAGE | Pseudomonas putida |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
assay at | Pseudomonas putida |
55 | - |
maximum reaction velocity is observed | Pseudomonas putida |
Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | 55 | - |
Pseudomonas putida |