Application | Comment | Organism |
---|---|---|
synthesis | the enzyme might be a promising biocatalyst for producing chiral aromatic beta-hydroxy-alpha-amino acids | Delftia sp. |
Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli BL21 (DE3) | Delftia sp. |
General Stability | Organism |
---|---|
existence of the metal ions (Mn2+) enhances the stability of the enzyme | Delftia sp. |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.35 | - |
D-allo-threonine | pH 6.0, 50°C | Delftia sp. | |
0.83 | - |
D-threonine | pH 6.0, 50°C | Delftia sp. |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
MnCl2 | metal ion-free enzyme solution shows no activity. Metal ions are required for the reaction. The highest activity is recovered with Mn2+ | Delftia sp. |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
47000 | - |
gel filtration | Delftia sp. |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Delftia sp. | A0A031HCH9 | - |
- |
Delftia sp. RIT313 | A0A031HCH9 | - |
- |
Purification (Comment) | Organism |
---|---|
- |
Delftia sp. |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
21.3 | - |
pH 6.0, 50°C | Delftia sp. |
Storage Stability | Organism |
---|---|
-80°C, recombinant His-tagged enzyme remains stable for 6 months | Delftia sp. |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
D-allo-threonine | - |
Delftia sp. | glycine + acetaldehyde | - |
? | |
D-allo-threonine | - |
Delftia sp. RIT313 | glycine + acetaldehyde | - |
? | |
D-threonine | - |
Delftia sp. | glycine + acetaldehyde | - |
r | |
D-threonine | - |
Delftia sp. RIT313 | glycine + acetaldehyde | - |
r | |
glycine + 2-chlorobenzaldehyde | the reaction conversion and the diastereomeric excess of the 2-substituted substrates decrease in the order of F, H, Cl, Br | Delftia sp. | D-3-(2-chlorophenyl)serine | - |
r | |
glycine + 2-chlorobenzaldehyde | the reaction conversion and the diastereomeric excess of the 2-substituted substrates decrease in the order of F, H, Cl, Br | Delftia sp. RIT313 | D-3-(2-chlorophenyl)serine | - |
r | |
glycine + 2-fluorobenzaldehyde | the reaction conversion and the diastereomeric excess of the 2-substituted substrates decrease in the order of F, H, Cl, Br | Delftia sp. | D-3-(2-fluorophenyl)serine | - |
r | |
glycine + 2-nitrobenzaldehyde | the reaction conversion and the diastereomeric excess of the 2-substituted substrates decrease in the order of F, H, Cl, Br | Delftia sp. | D-3-(2-nitrophenyl)serine | - |
r | |
glycine + 3-chlorobenzaldehyde | - |
Delftia sp. | D-3-(3-chlorophenyl)serine | - |
r | |
glycine + 3-fluorobenzaldehyde | - |
Delftia sp. | D-3-(3-fluorophenyl)serine | - |
r | |
glycine + 3-nitrobenzaldehyde | - |
Delftia sp. | D-3-(3-nitrophenyl)serine | - |
r | |
glycine + 4-(methylsulfonyl)benzaldehyde | - |
Delftia sp. | D-3-(4-methylsulfonylphenyl)serine | - |
r | |
glycine + 4-fluorobenzaldehyde | - |
Delftia sp. | D-3-(4-fluorophenyl)serine | - |
r | |
glycine + 4-nitrobenzaldehyde | - |
Delftia sp. | D-3-(4-nitrophenyl)serine | - |
r | |
glycine + acetaldehyde | - |
Delftia sp. | D-allothreonine | - |
r | |
glycine + acetaldehyde | - |
Delftia sp. | D-threonine | - |
r | |
glycine + benzaldehyde | the erythro isomer is obtained as the major product when the temperature is above 15°C, and lowering the temperature result in an increase in threo isomer.Water-miscible organic solvents exert limited effect on the conversion, but greatly enhance the diastereomeric excess of the product, with 10% CH3CN giving the highest diastereomeric excess. For other organic solvents (THF, ethyl acetate, 2-methoxy-2-methylpropan, 1,4-dioxane, dichloromethane and toluene), lower stereoselectivity at the beta-carbon is observed | Delftia sp. | (2R,3S)-2-amino-3-hydroxy-3-phenylpropanoic acid + (2R,3R)-2-amino-3-hydroxy-3-phenylpropanoic acid | - |
r | |
glycine + benzaldehyde | the erythro isomer is obtained as the major product when the temperature is above 15°C, and lowering the temperature result in an increase in threo isomer.Water-miscible organic solvents exert limited effect on the conversion, but greatly enhance the diastereomeric excess of the product, with 10% CH3CN giving the highest diastereomeric excess. For other organic solvents (THF, ethyl acetate, 2-methoxy-2-methylpropan, 1,4-dioxane, dichloromethane and toluene), lower stereoselectivity at the beta-carbon is observed | Delftia sp. RIT313 | (2R,3S)-2-amino-3-hydroxy-3-phenylpropanoic acid + (2R,3R)-2-amino-3-hydroxy-3-phenylpropanoic acid | - |
r | |
additional information | the enzyme shows high activity toward aromatic aldehydes with electron-withdrawing substituents. The substrate profiling indicates that the enzyme accepts a wider range of acceptor substrates and is more active toward aromatic aldehydes bearing electron-withdrawing groups than those with electron-donating substituents. Molecular docking studies suggest that the substituent on the benzene ring of the substrate is critical in determining the enzyme activity and stereoselectivity by affecting the interaction between the beta-OH-group of the substrate and the manganese ion | Delftia sp. | ? | - |
? | |
additional information | the enzyme shows high activity toward aromatic aldehydes with electron-withdrawing substituents. The substrate profiling indicates that the enzyme accepts a wider range of acceptor substrates and is more active toward aromatic aldehydes bearing electron-withdrawing groups than those with electron-donating substituents. Molecular docking studies suggest that the substituent on the benzene ring of the substrate is critical in determining the enzyme activity and stereoselectivity by affecting the interaction between the beta-OH-group of the substrate and the manganese ion | Delftia sp. RIT313 | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
monomer | 1 * 47000, SDS-PAGE | Delftia sp. |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Delftia sp. |
50 | 60 | aldol addition | Delftia sp. |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
additional information | - |
existence of the metal ions (Mn2+) enhances the thermal stability of the enzyme | Delftia sp. |
25 | 60 | the enzyme retains about 70% activity from 25°C to 60°C after it is heated for 30 min | Delftia sp. |
70 | - |
30 min, 50% loss of activity | Delftia sp. |
80 | - |
30 min, 70% loss of activity | Delftia sp. |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.25 | - |
D-allo-threonine | pH 6.0, 50°C | Delftia sp. | |
0.39 | - |
D-threonine | pH 6.0, 50°C | Delftia sp. |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
6 | - |
aldol addition | Delftia sp. |
8 | - |
retro-aldol addition | Delftia sp. |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
5 | 10 | pH 5.0: about 55% of maximal activity, pH 10.0: about 45% of maximal activity, aldol addition | Delftia sp. |
6 | 9.5 | pH 6.0: about 65% of maximal activity, pH 9.5: about 75% of maximal activity, retro-aldol addition | Delftia sp. |
pH Stability | pH Stability Maximum | Comment | Organism |
---|---|---|---|
6 | - |
after being incubated at various pH values for 12 h, the enzyme retains the highest activity at pH 6.0 for both retro-aldol and aldol addition reactions | Delftia sp. |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
pyridoxal 5'-phosphate | dependent on | Delftia sp. |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.47 | - |
D-threonine | pH 6.0, 50°C | Delftia sp. | |
0.71 | - |
D-allo-threonine | pH 6.0, 50°C | Delftia sp. |