Crystallization (Comment) | Organism |
---|---|
molecular modeling of structure containing its natural substrate. Residues H181, Y238 and T240 appear to stabilize the alpha NH3+ group, whereas R242 appears to interact with the COO- of the substrate. Residues V172, F173 and V130 create a hindered hydrophobic environment | Geobacillus stearothermophilus |
Protein Variants | Comment | Organism |
---|---|---|
F173A | mutation creates an additional large hydrophobic binding pocket in the active site, which favours the binding of aromatic substrates | Geobacillus stearothermophilus |
F173G | very low level of activity towards the tested substrates | Geobacillus stearothermophilus |
R242M | residue R242 appears to be involved in the binding of the alpha-carboxylic group of L-lysine and is located at the entrance of the active site | Geobacillus stearothermophilus |
R242M/F173A | mutant converts substrates 1-phenylethan-1-one (6%) and 1-phenylpropan-1-one (19%, ee > 99% R) | Geobacillus stearothermophilus |
R242M/F173V | mutant converts substrates 1-phenylethan-1-one, 1-phenylpropan-1-one 1-phenylbutan-1-one and 2,3-dihydro-4H-1-benzopyran-4-one (38%, 41%, 8%, 15% conversions, ee > 99% R) | Geobacillus stearothermophilus |
V130A/F173A | mutant exhibits a significant level of activity towards 1-phenylethan-1-one (74% conversion) and 1-phenylpropan-1-one (65% conversion), and all amine products are obtained in enantiomerically pure form (ee > 99% R) | Geobacillus stearothermophilus |
V130A/F173V | inactive | Geobacillus stearothermophilus |
V130G/F173A | mutant is stereospecific (ee > 99% R) for the amination of 1-phenylpropan-1-one (78% conversion) and other ketones | Geobacillus stearothermophilus |
V130G/F173V | inactive | Geobacillus stearothermophilus |
V172A/F173A | dramatic reduction of catalytic activity | Geobacillus stearothermophilus |
V172A/F173G | very low level of activity towards the tested substrates | Geobacillus stearothermophilus |
V172G/F173A | dramatic reduction of catalytic activity | Geobacillus stearothermophilus |
V172G/F173G | very low level of activity towards the tested substrates | Geobacillus stearothermophilus |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Geobacillus stearothermophilus | Q9AJC6 | - |
- |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
1-phenylbutan-1-one + NADH + H+ + NH3 | mutant F173A, 10 mM, 87% conversion, product in R-configuration, 99.8% ee | Geobacillus stearothermophilus | (1R)-1-phenylbutan-1-amine + NAD+ + H2O | - |
? | |
1-phenylethan-1-one + NADH + H+ + NH3 | mutant F173A, 10 mM, 99% conversion, product in R-configuration, 99.9% ee | Geobacillus stearothermophilus | (1R)-1-phenylethan-1-amine + NAD+ + H2O | - |
? | |
1-phenylpentan-1-one + NADH + H+ + NH3 | mutant F173A, 10 mM, 36% conversion | Geobacillus stearothermophilus | 1-phenylpentan-1-amine + NAD+ + H2O | - |
? | |
1-phenylpropan-1-one + NADH + H+ + NH3 | mutant F173A, 10 mM, 98% conversion, product in R-configuration, 99.9% ee | Geobacillus stearothermophilus | (1R)-1-phenylpropan-1-amine + NAD+ + H2O | - |
? | |
2,3-dihydro-4H-1-benzopyran-4-one + NADH + H+ + NH3 | mutant F173A, 10 mM, 82% conversion, product in R-configuration, 99.4% ee | Geobacillus stearothermophilus | (4R)-3,4-dihydro-2H-1-benzopyran-4-amine + NAD+ + H2O | - |
? | |
3,4-dihydronaphthalen-1(2H)-one + NADH + H+ + NH3 | mutant F173A, 10 mM, 79% conversion, product in R-configuration, 99% ee | Geobacillus stearothermophilus | (1R)-1,2,3,4-tetrahydronaphthalen-1-amine + NAD+ + H2O | - |
? | |
4-methylpentan-2-one + NADH + H+ + NH3 | mutant F173A, 10 mM, 76% conversion, product in R-configuration, 97% ee | Geobacillus stearothermophilus | (2R)-4-methylpentan-2-amine + NAD+ + H2O | - |
? | |
6-oxoheptanoic acid + NADH + H+ + NH3 | mutant F173A, 10 mM, 99% conversion, product in S-configuration, 99% ee | Geobacillus stearothermophilus | (6S)-6-aminoheptanoic acid + NAD+ + H2O | - |
? | |
6-oxohexanoic acid + NADH + H+ + NH3 | mutant F173A, 10 mM, 99% conversion | Geobacillus stearothermophilus | L-lysine + NAD+ + H2O | - |
? | |
cyclohexanone + NADH + H+ + NH3 | mutant F173A, 10 mM, 86% conversion | Geobacillus stearothermophilus | cyclohexanamine + NAD+ + H2O | - |
? | |
hexan-2-one + NADH + H+ + NH3 | mutant F173A, 10 mM, 87% conversion, product in R-configuration, 99% ee | Geobacillus stearothermophilus | (2R)-hexan-2-amine + NAD+ + H2O | - |
? | |
additional information | the alpha-carboxyl group is crucial for productive substrate binding and amination. No substrates: hexanal, heptanal, 2-heptanone, 2-octanone, acetophenone and alpha-chromanone | Geobacillus stearothermophilus | ? | - |
- |
|
pentan-2-one + NADH + H+ + NH3 | mutant F173A, 10 mM, 86% conversion, product in R-configuration, 89% ee | Geobacillus stearothermophilus | (2R)-pentan-2-amine + NAD+ + H2O | - |
? |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
50 | - |
- |
Geobacillus stearothermophilus |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
50 | - |
stable for everal days | Geobacillus stearothermophilus |
69 | - |
melting temperature | Geobacillus stearothermophilus |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
9 | - |
- |
Geobacillus stearothermophilus |