Application | Comment | Organism |
---|---|---|
synthesis | the enzyme catalyzes the Baeyer-Villiger oxidation of 2-butanone, yielding ethyl acetate and methyl propanoate as products. Methyl propanoate is of industrial interest as a precursor of acrylic plastic. Various residues near the substrate and NADP+ binding sites are subjected to saturation mutagenesis to enhance both the activity on 2-butanone and the regioselectivity toward methyl propanoate | Acinetobacter calcoaceticus |
Cloned (Comment) | Organism |
---|---|
expression in Escherichia coli | Acinetobacter calcoaceticus |
Protein Variants | Comment | Organism |
---|---|---|
F281H | cultures expressing the F281H variant produce 30 to 40% methyl propanoate, while the wild-type enzyme produce 26% methyl propanoate | Acinetobacter calcoaceticus |
I491A | cultures expressing the I491A variant produce 30 to 40% methyl propanoate, while the wild-type enzyme produce 26% methyl propanoate | Acinetobacter calcoaceticus |
I491A | mutant exhibits a significant improvement over the wild type enzyme in the desired regioselectivity using 2-butanone as a substrate (40% vs 26% methyl propanoate, respectively) | Acinetobacter calcoaceticus |
L244R | cultures expressing the L244R variant produce 30 to 40% methyl propanoate, while the wild-type enzyme produce 26% methyl propanoate | Acinetobacter calcoaceticus |
L435N | cultures expressing the L435N variant produce 30 to 40% methyl propanoate, while the wild-type enzyme produce 26% methyl propanoate | Acinetobacter calcoaceticus |
T56S | mutant, which exhibits a higher conversion yield (92%) and kcat (0.5/s) than wild type AcCHMO (52% and 0.3/s, respectively). The uncoupling rate for the T56S mutant is also significantly lower than that for the wild type enzyme | Acinetobacter calcoaceticus |
T56S/I491A | mutant with higher conversion and improved regioselectivity | Acinetobacter calcoaceticus |
T56S/L435N/I491A | mutant produces less total product than wild-type enzyme | Acinetobacter calcoaceticus |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Acinetobacter calcoaceticus | - |
- |
- |
Purification (Comment) | Organism |
---|---|
- |
Acinetobacter calcoaceticus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
2-butanone + NADPH + H+ + O2 | - |
Acinetobacter calcoaceticus | methyl propanoate + NADP+ + H2O | - |
? |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.16 | - |
2-butanone | pH 7.0, 25°C, mutant enzyme L435N | Acinetobacter calcoaceticus | |
0.19 | - |
2-butanone | pH 7.0, 25°C, mutant enzyme I491A | Acinetobacter calcoaceticus | |
0.2 | - |
2-butanone | pH 7.0, 25°C, mutant enzyme F281H | Acinetobacter calcoaceticus | |
0.24 | - |
2-butanone | pH 7.0, 25°C, mutant enzyme T56S/L244R/I491 | Acinetobacter calcoaceticus | |
0.26 | - |
2-butanone | pH 7.0, 25°C, mutant enzyme T56S/I491A | Acinetobacter calcoaceticus | |
0.28 | - |
2-butanone | pH 7.0, 25°C, wild-type enzyme | Acinetobacter calcoaceticus | |
0.31 | - |
2-butanone | pH 7.0, 25°C, mutant enzyme L244R | Acinetobacter calcoaceticus | |
0.33 | - |
2-butanone | pH 7.0, 25°C, mutant enzyme T56S/L435N/I491 | Acinetobacter calcoaceticus | |
0.41 | - |
2-butanone | pH 7.0, 25°C, mutant enzyme T56S/L244R | Acinetobacter calcoaceticus | |
0.47 | - |
2-butanone | pH 7.0, 25°C, mutant enzyme T56S | Acinetobacter calcoaceticus |