Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli BL21(DE3) cells | Microbacterium oxydans |
Protein Variants | Comment | Organism |
---|---|---|
L199A | the mutant shows generally lower activity (decrease of 15-97%) towards most substrates compared to wild type enzyme with the exception of the larger substrates, such as cyclooctanamine and bicyclic (S)-1-aminotetraline | Microbacterium oxydans |
L353M | the mutant shows 7-445% higher activity towards primary aliphatic amines with cycloalkane or aromatic moieties | Microbacterium oxydans |
M226A | the mutant displays an enhanced activity (5-400%) towards most substrates | Microbacterium oxydans |
Y321A | the mutant shows higher catalytic efficiency towards cyclooctanamine compared to the wild type enzyme | Microbacterium oxydans |
Y321F | the mutant shows higher catalytic efficiency towards cyclooctanamine compared to the wild type enzyme | Microbacterium oxydans |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.07 | - |
(S)-1-aminotetraline | mutant enzyme Y321F, at pH 6.5 and 30°C | Microbacterium oxydans | |
0.08 | - |
(S)-1-aminotetraline | mutant enzyme Y321A, at pH 6.5 and 30°C | Microbacterium oxydans | |
0.08 | - |
(S)-1-aminotetraline | wild type enzyme, at pH 6.5 and 30°C | Microbacterium oxydans | |
0.21 | - |
(S)-1-aminotetraline | mutant enzyme L353M, at pH 6.5 and 30°C | Microbacterium oxydans | |
0.22 | - |
cyclooctanamine | mutant enzyme Y321F, at pH 6.5 and 30°C | Microbacterium oxydans | |
0.24 | - |
cyclooctanamine | mutant enzyme Y321A, at pH 6.5 and 30°C | Microbacterium oxydans | |
0.26 | - |
(S)-1-aminotetraline | mutant enzyme M226A, at pH 6.5 and 30°C | Microbacterium oxydans | |
0.32 | - |
(S)-1-aminotetraline | mutant enzyme L199A, at pH 6.5 and 30°C | Microbacterium oxydans | |
0.59 | - |
cyclooctanamine | wild type enzyme, at pH 6.5 and 30°C | Microbacterium oxydans | |
1.08 | - |
Cyclohexylamine | wild type enzyme, at pH 6.5 and 30°C | Microbacterium oxydans | |
1.89 | - |
cyclooctanamine | mutant enzyme M226A, at pH 6.5 and 30°C | Microbacterium oxydans | |
1.91 | - |
cyclooctanamine | mutant enzyme L199A, at pH 6.5 and 30°C | Microbacterium oxydans | |
2.04 | - |
cyclooctanamine | mutant enzyme L353M, at pH 6.5 and 30°C | Microbacterium oxydans | |
2.28 | - |
Cyclohexylamine | mutant enzyme Y321F, at pH 6.5 and 30°C | Microbacterium oxydans | |
3.39 | - |
Cyclohexylamine | mutant enzyme L353M, at pH 6.5 and 30°C | Microbacterium oxydans | |
4.76 | - |
Cyclohexylamine | mutant enzyme M226A, at pH 6.5 and 30°C | Microbacterium oxydans | |
5.94 | - |
Cyclohexylamine | mutant enzyme Y321A, at pH 6.5 and 30°C | Microbacterium oxydans | |
7.71 | - |
Cyclohexylamine | mutant enzyme L199A, at pH 6.5 and 30°C | Microbacterium oxydans |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
50000 | - |
x * 50000, SDS-PAGE | Microbacterium oxydans |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
cyclohexylamine + O2 + H2O | Microbacterium oxydans | best substrate | cyclohexanone + NH3 + H2O2 | - |
? | |
cyclohexylamine + O2 + H2O | Microbacterium oxydans IH-35A | best substrate | cyclohexanone + NH3 + H2O2 | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Microbacterium oxydans | - |
- |
- |
Microbacterium oxydans IH-35A | - |
- |
- |
Purification (Comment) | Organism |
---|---|
DEAE column chromatography, and gel filtration | Microbacterium oxydans |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
0.013 | - |
with 2-ethyl-1-hexylamine as substrate, wild type enzyme, at pH 6.5 and 30°C | Microbacterium oxydans |
0.018 | - |
with (R)-1-(4-chlorophenyl)ethylamine as substrate, wild type enzyme, at pH 6.5 and 30°C | Microbacterium oxydans |
0.08 | - |
with N-methylcyclohexanamine as substrate, wild type enzyme, at pH 6.5 and 30°C | Microbacterium oxydans |
0.084 | - |
with (R)-1-(4-fluorophenyl)ethanamine as substrate, wild type enzyme, at pH 6.5 and 30°C | Microbacterium oxydans |
0.11 | - |
with 4-methylbenzylamine as substrate, wild type enzyme, at pH 6.5 and 30°C | Microbacterium oxydans |
0.27 | - |
with (S)-1-p-tolylethanamine as substrate, wild type enzyme, at pH 6.5 and 30°C | Microbacterium oxydans |
0.33 | - |
with cyclopentylamine as substrate, wild type enzyme, at pH 6.5 and 30°C | Microbacterium oxydans |
0.34 | - |
with benzylamine as substrate, wild type enzyme, at pH 6.5 and 30°C | Microbacterium oxydans |
0.68 | - |
with 2-amino-4-phenylbutane as substrate, wild type enzyme, at pH 6.5 and 30°C | Microbacterium oxydans |
0.97 | - |
with 2-aminocyclohexanol as substrate, wild type enzyme, at pH 6.5 and 30°C | Microbacterium oxydans |
1.033 | - |
with cyclooctanamine as substrate, wild type enzyme, at pH 6.5 and 30°C | Microbacterium oxydans |
1.13 | - |
with 2-aminopentane as substrate, wild type enzyme, at pH 6.5 and 30°C | Microbacterium oxydans |
1.2 | - |
with 2-aminoheptane as substrate, wild type enzyme, at pH 6.5 and 30°C | Microbacterium oxydans |
1.49 | - |
with 2-aminonorbornane hydrochloride as substrate, wild type enzyme, at pH 6.5 and 30°C | Microbacterium oxydans |
1.66 | - |
with (S)-1-phenylpropropan-1-amine as substrate, wild type enzyme, at pH 6.5 and 30°C | Microbacterium oxydans |
2.35 | - |
with (S)-1-aminoindan as substrate, wild type enzyme, at pH 6.5 and 30°C | Microbacterium oxydans |
2.36 | - |
with 1-phenylethanamine as substrate, wild type enzyme, at pH 6.5 and 30°C | Microbacterium oxydans |
2.62 | - |
with (S)-1-(4-chlorophenyl)ethylamine as substrate, wild type enzyme, at pH 6.5 and 30°C | Microbacterium oxydans |
2.79 | - |
with (S)-2-aminohexane as substrate, wild type enzyme, at pH 6.5 and 30°C | Microbacterium oxydans |
3.058 | - |
with 2-methylcyclohexanamine as substrate, wild type enzyme, at pH 6.5 and 30°C | Microbacterium oxydans |
3.37 | - |
with 1-(4-fluorophenyl)ethanamine as substrate, wild type enzyme, at pH 6.5 and 30°C | Microbacterium oxydans |
3.51 | - |
with (S)-(-)-1-phenylethylamine as substrate, wild type enzyme, at pH 6.5 and 30°C | Microbacterium oxydans |
3.6 | - |
with 2-amino-5-methylhexane as substrate, wild type enzyme, at pH 6.5 and 30°C | Microbacterium oxydans |
4.56 | - |
with (S)-1-(4-fluorophenyl)ethanamine as substrate, wild type enzyme, at pH 6.5 and 30°C | Microbacterium oxydans |
5.6 | - |
with cyclohexylamine as substrate, wild type enzyme, at pH 6.5 and 30°C | Microbacterium oxydans |
5.69 | - |
with (S)-1-aminotetraline as substrate, wild type enzyme, at pH 6.5 and 30°C | Microbacterium oxydans |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
(R)-1-(4-chlorophenyl)ethylamine + O2 + H2O | - |
Microbacterium oxydans | 1-(4-chlorophenyl)ethanone + NH3 + H2O2 | - |
? | |
(R)-1-(4-fluorophenyl)ethanamine + O2 + H2O | - |
Microbacterium oxydans | 1-(4-fluorophenyl)ethanone + NH3 + H2O2 | - |
? | |
(R)-1-aminotetraline + O2 + H2O | - |
Microbacterium oxydans | ? | - |
? | |
(S)-1-(4-chlorophenyl)ethylamine + O2 + H2O | - |
Microbacterium oxydans | 1-(4-chlorophenyl)ethanone + NH3 + H2O2 | - |
? | |
(S)-1-(4-fluorophenyl)ethanamine + O2 + H2O | - |
Microbacterium oxydans | 1-(4-fluorophenyl)ethanone + NH3 + H2O2 | - |
? | |
(S)-1-aminoindan + O2 + H2O | - |
Microbacterium oxydans | ? | - |
? | |
(S)-1-aminotetraline + O2 + H2O | - |
Microbacterium oxydans | ? | - |
? | |
(S)-1-aminotetraline + O2 + H2O | - |
Microbacterium oxydans | tetralone + NH3 + H2O2 | - |
? | |
(S)-1-p-tolylethanamine + O2 + H2O | - |
Microbacterium oxydans | 1-(4-methylphenyl)ethanone + NH3 + H2O2 | - |
? | |
(S)-1-phenylethylamine + O2 + H2O | - |
Microbacterium oxydans | ? | - |
? | |
(S)-1-phenylpropan-1-amine + O2 + H2O | - |
Microbacterium oxydans | 1-phenylpropionaldehyde + NH3 + H2O2 | - |
? | |
(S)-2-aminohexane + O2 + H2O | - |
Microbacterium oxydans | hexan-2-one + NH3 + H2O2 | - |
? | |
1-(4-fluorophenyl)ethanamine + O2 + H2O | - |
Microbacterium oxydans | 1-(4-fluorophenyl)ethanone + NH3 + H2O2 | - |
? | |
1-phenylethanamine + O2 + H2O | - |
Microbacterium oxydans | 1-phenylacetaldehyde + NH3 + H2O2 | - |
? | |
2-amino-4-phenylbutane + O2 + H2O | - |
Microbacterium oxydans | 4-phenylbutan-2-one + NH3 + H2O2 | - |
? | |
2-amino-5-methylhexane + O2 + H2O | - |
Microbacterium oxydans | 5-methylhexane-2-one + NH3 + H2O2 | - |
? | |
2-aminocyclohexanol + O2 + H2O | - |
Microbacterium oxydans | ? | - |
? | |
2-aminoheptane + O2 + H2O | - |
Microbacterium oxydans | heptan-2-one + NH3 + H2O2 | - |
? | |
2-aminonorbornane + O2 + H2O | - |
Microbacterium oxydans | ? | - |
? | |
2-aminopentane + O2 + H2O | - |
Microbacterium oxydans | pentan-2-one + NH3 + H2O2 | - |
? | |
2-ethyl-1-hexylamine + O2 + H2O | - |
Microbacterium oxydans | 2-ethylhexanaldehyde + NH3 + H2O2 | - |
? | |
2-methylcyclohexanamine + O2 + H2O | - |
Microbacterium oxydans | 2-methylcyclohexanone + NH3 + H2O2 | - |
? | |
4-methylbenzylamine + O2 + H2O | - |
Microbacterium oxydans | 4-methylbenzaldehyde + NH3 + H2O2 | - |
? | |
benzylamine + O2 + H2O | - |
Microbacterium oxydans | benzaldehyde + NH3 + H2O2 | - |
? | |
benzylamine + O2 + H2O | - |
Microbacterium oxydans IH-35A | benzaldehyde + NH3 + H2O2 | - |
? | |
cyclohexylamine + O2 + H2O | - |
Microbacterium oxydans | cyclohexanone + NH3 + H2O2 | - |
? | |
cyclohexylamine + O2 + H2O | best substrate | Microbacterium oxydans | cyclohexanone + NH3 + H2O2 | - |
? | |
cyclohexylamine + O2 + H2O | - |
Microbacterium oxydans IH-35A | cyclohexanone + NH3 + H2O2 | - |
? | |
cyclohexylamine + O2 + H2O | best substrate | Microbacterium oxydans IH-35A | cyclohexanone + NH3 + H2O2 | - |
? | |
cyclooctanamine + O2 + H2O | - |
Microbacterium oxydans | cyclooctanone + NH3 + H2O2 | - |
? | |
cyclopentylamine + O2 + H2O | best substrate | Microbacterium oxydans | cyclopentanone + NH3 + H2O2 | - |
? | |
additional information | deracemization of 1-aminotetraline by coupling wild type enzyme with a nonselective reducing agent NH3·BH3 is a route to norsertraline | Microbacterium oxydans | ? | - |
? | |
additional information | trace activity with (R)-2-aminohexane, 1-hexylamine, (R)-(-)-1-phenylethylamine, (R)-1-p-tolylethanamine, (R)-1-aminoindan, (S)-(-)-Nalpha-dimethylbenzylamine, 2-methylpiperidine, 4-methylpiperidine, 2-methyl-1,2,3,4-tetrahydroquinoline, and N,N-dimethylcyclohexylamine. No activity with 1-naphthalen-1-ylethanamine | Microbacterium oxydans | ? | - |
? | |
additional information | deracemization of 1-aminotetraline by coupling wild type enzyme with a nonselective reducing agent NH3·BH3 is a route to norsertraline | Microbacterium oxydans IH-35A | ? | - |
? | |
additional information | trace activity with (R)-2-aminohexane, 1-hexylamine, (R)-(-)-1-phenylethylamine, (R)-1-p-tolylethanamine, (R)-1-aminoindan, (S)-(-)-Nalpha-dimethylbenzylamine, 2-methylpiperidine, 4-methylpiperidine, 2-methyl-1,2,3,4-tetrahydroquinoline, and N,N-dimethylcyclohexylamine. No activity with 1-naphthalen-1-ylethanamine | Microbacterium oxydans IH-35A | ? | - |
? | |
N-methylcyclohexanamine + O2 + H2O | - |
Microbacterium oxydans | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
? | x * 50000, SDS-PAGE | Microbacterium oxydans |
Synonyms | Comment | Organism |
---|---|---|
CHAO | - |
Microbacterium oxydans |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
40 | - |
the wild type enzyme is stable at 40°C for at least 30 min, but decreases rapidly above 40°C | Microbacterium oxydans |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
2.96 | - |
(S)-1-aminotetraline | mutant enzyme Y321A, at pH 6.5 and 30°C | Microbacterium oxydans | |
3.03 | - |
cyclooctanamine | wild type enzyme, at pH 6.5 and 30°C | Microbacterium oxydans | |
3.32 | - |
(S)-1-aminotetraline | mutant enzyme Y321F, at pH 6.5 and 30°C | Microbacterium oxydans | |
3.44 | - |
cyclooctanamine | mutant enzyme Y321A, at pH 6.5 and 30°C | Microbacterium oxydans | |
3.79 | - |
cyclooctanamine | mutant enzyme Y321F, at pH 6.5 and 30°C | Microbacterium oxydans | |
4.77 | - |
Cyclohexylamine | mutant enzyme L199A, at pH 6.5 and 30°C | Microbacterium oxydans | |
5.72 | - |
(S)-1-aminotetraline | wild type enzyme, at pH 6.5 and 30°C | Microbacterium oxydans | |
6.62 | - |
cyclooctanamine | mutant enzyme L199A, at pH 6.5 and 30°C | Microbacterium oxydans | |
7.06 | - |
cyclooctanamine | mutant enzyme M226A, at pH 6.5 and 30°C | Microbacterium oxydans | |
7.14 | - |
(S)-1-aminotetraline | mutant enzyme L199A, at pH 6.5 and 30°C | Microbacterium oxydans | |
7.68 | - |
cyclooctanamine | mutant enzyme L353M, at pH 6.5 and 30°C | Microbacterium oxydans | |
8.52 | - |
Cyclohexylamine | mutant enzyme Y321A, at pH 6.5 and 30°C | Microbacterium oxydans | |
8.54 | - |
(S)-1-aminotetraline | mutant enzyme M226A, at pH 6.5 and 30°C | Microbacterium oxydans | |
10.62 | - |
(S)-1-aminotetraline | mutant enzyme L353M, at pH 6.5 and 30°C | Microbacterium oxydans | |
10.93 | - |
Cyclohexylamine | mutant enzyme Y321F, at pH 6.5 and 30°C | Microbacterium oxydans | |
11.48 | - |
Cyclohexylamine | wild type enzyme, at pH 6.5 and 30°C | Microbacterium oxydans | |
15.25 | - |
Cyclohexylamine | mutant enzyme L353M, at pH 6.5 and 30°C | Microbacterium oxydans | |
16.33 | - |
Cyclohexylamine | mutant enzyme M226A, at pH 6.5 and 30°C | Microbacterium oxydans |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
6.5 | - |
- |
Microbacterium oxydans |
pH Stability | pH Stability Maximum | Comment | Organism |
---|---|---|---|
6 | 9 | the enzyme retains more than 70% of its original activity over a broad range of pH 6.09.0 at 4°C for 24 h. However, stability decreases quickly in more acidic condition | Microbacterium oxydans |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
FAD | - |
Microbacterium oxydans |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.62 | - |
Cyclohexylamine | mutant enzyme L199A, at pH 6.5 and 30°C | Microbacterium oxydans | |
1.43 | - |
Cyclohexylamine | mutant enzyme Y321A, at pH 6.5 and 30°C | Microbacterium oxydans | |
3.43 | - |
Cyclohexylamine | mutant enzyme M226A, at pH 6.5 and 30°C | Microbacterium oxydans | |
3.47 | - |
cyclooctanamine | mutant enzyme L199A, at pH 6.5 and 30°C | Microbacterium oxydans | |
3.74 | - |
cyclooctanamine | mutant enzyme M226A, at pH 6.5 and 30°C | Microbacterium oxydans | |
3.76 | - |
cyclooctanamine | mutant enzyme L353M, at pH 6.5 and 30°C | Microbacterium oxydans | |
4.49 | - |
Cyclohexylamine | mutant enzyme L353M, at pH 6.5 and 30°C | Microbacterium oxydans | |
4.79 | - |
Cyclohexylamine | mutant enzyme Y321F, at pH 6.5 and 30°C | Microbacterium oxydans | |
5.14 | - |
cyclooctanamine | wild type enzyme, at pH 6.5 and 30°C | Microbacterium oxydans | |
10.63 | - |
Cyclohexylamine | wild type enzyme, at pH 6.5 and 30°C | Microbacterium oxydans | |
14.33 | - |
cyclooctanamine | mutant enzyme Y321A, at pH 6.5 and 30°C | Microbacterium oxydans | |
17.23 | - |
cyclooctanamine | mutant enzyme Y321F, at pH 6.5 and 30°C | Microbacterium oxydans | |
22.31 | - |
(S)-1-aminotetraline | mutant enzyme L199A, at pH 6.5 and 30°C | Microbacterium oxydans | |
32.85 | - |
(S)-1-aminotetraline | mutant enzyme M226A, at pH 6.5 and 30°C | Microbacterium oxydans | |
37 | - |
(S)-1-aminotetraline | mutant enzyme Y321A, at pH 6.5 and 30°C | Microbacterium oxydans | |
47.43 | - |
(S)-1-aminotetraline | mutant enzyme Y321F, at pH 6.5 and 30°C | Microbacterium oxydans | |
50.57 | - |
(S)-1-aminotetraline | mutant enzyme L353M, at pH 6.5 and 30°C | Microbacterium oxydans | |
71.5 | - |
(S)-1-aminotetraline | wild type enzyme, at pH 6.5 and 30°C | Microbacterium oxydans |