Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli BL21(DE3) CodonPlus cells | Rhodococcus rhodochrous |
Crystallization (Comment) | Organism |
---|---|
sitting drop vapor diffusion method, using 1.8-2.2 M MgSO4, 0.1 M MES/HCl, pH 6.2-6.5 | Rhodococcus rhodochrous |
Protein Variants | Comment | Organism |
---|---|---|
K295A | the mutant is 3times more active than the wild type enzyme | Rhodococcus rhodochrous |
L500Y | the mutant is about 2.5times more active than the wild type enzyme | Rhodococcus rhodochrous |
P157Q | the mutant shows wild type activity | Rhodococcus rhodochrous |
T345L | the mutation turns the enzyme inactive against progesterone without altering the catalytic efficiency for phenylacetone | Rhodococcus rhodochrous |
V291A | the mutant is less active than the wild type enzyme | Rhodococcus rhodochrous |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.2 | - |
phenylacetone | mutant enzyme L500Y, in 50 mM Tris/HCl, pH 7.5, temperature not specified in the publication | Rhodococcus rhodochrous | |
0.2 | - |
phenylacetone | mutant enzyme V291A, in 50 mM Tris/HCl, pH 7.5, temperature not specified in the publication | Rhodococcus rhodochrous | |
0.6 | - |
phenylacetone | mutant enzyme P157Q, in 50 mM Tris/HCl, pH 7.5, temperature not specified in the publication | Rhodococcus rhodochrous | |
0.8 | - |
phenylacetone | mutant enzyme K295A, in 50 mM Tris/HCl, pH 7.5, temperature not specified in the publication | Rhodococcus rhodochrous | |
1 | - |
phenylacetone | wild type enzyme, in 50 mM Tris/HCl, pH 7.5, temperature not specified in the publication | Rhodococcus rhodochrous | |
2.6 | - |
phenylacetone | mutant enzyme T345L, in 50 mM Tris/HCl, pH 7.5, temperature not specified in the publication | Rhodococcus rhodochrous |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
progesterone + NADPH + H+ + O2 | Rhodococcus rhodochrous | - |
Testosterone acetate + NADP+ + H2O | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Rhodococcus rhodochrous | O50641 | - |
- |
Purification (Comment) | Organism |
---|---|
Ni2+-affinity column chromatography and Superdex 75 gel filtration | Rhodococcus rhodochrous |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
phenylacetone + NADPH + H+ + O2 | - |
Rhodococcus rhodochrous | benzylacetate + NADP+ + H2O | - |
? | |
progesterone + NADPH + H+ + O2 | - |
Rhodococcus rhodochrous | Testosterone acetate + NADP+ + H2O | - |
? |
Synonyms | Comment | Organism |
---|---|---|
Baeyer-Villiger monooxygenase | - |
Rhodococcus rhodochrous |
Steroid monooxygenase | - |
Rhodococcus rhodochrous |
STMO | - |
Rhodococcus rhodochrous |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.2 | - |
phenylacetone | mutant enzyme T345L, in 50 mM Tris/HCl, pH 7.5, temperature not specified in the publication | Rhodococcus rhodochrous | |
0.6 | - |
phenylacetone | mutant enzyme P157Q, in 50 mM Tris/HCl, pH 7.5, temperature not specified in the publication | Rhodococcus rhodochrous | |
1 | - |
phenylacetone | mutant enzyme L500Y, in 50 mM Tris/HCl, pH 7.5, temperature not specified in the publication | Rhodococcus rhodochrous | |
1.5 | - |
phenylacetone | wild type enzyme, in 50 mM Tris/HCl, pH 7.5, temperature not specified in the publication | Rhodococcus rhodochrous | |
1.8 | - |
phenylacetone | mutant enzyme V291A, in 50 mM Tris/HCl, pH 7.5, temperature not specified in the publication | Rhodococcus rhodochrous | |
3 | - |
phenylacetone | mutant enzyme K295A, in 50 mM Tris/HCl, pH 7.5, temperature not specified in the publication | Rhodococcus rhodochrous |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
FAD | - |
Rhodococcus rhodochrous | |
NADPH | - |
Rhodococcus rhodochrous |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.1 | - |
progesterone | kcat_Km less than 0.1 1/sec*min, mutant enzyme L500Y, in 50 mM Tris/HCl, pH 7.5, temperature not specified in the publication | Rhodococcus rhodochrous | |
1.2 | - |
phenylacetone | mutant enzyme K295A, in 50 mM Tris/HCl, pH 7.5, temperature not specified in the publication | Rhodococcus rhodochrous | |
1.2 | - |
phenylacetone | mutant enzyme T345L, in 50 mM Tris/HCl, pH 7.5, temperature not specified in the publication | Rhodococcus rhodochrous | |
1.4 | - |
progesterone | mutant enzyme K295A, in 50 mM Tris/HCl, pH 7.5, temperature not specified in the publication | Rhodococcus rhodochrous | |
1.5 | - |
phenylacetone | wild type enzyme, in 50 mM Tris/HCl, pH 7.5, temperature not specified in the publication | Rhodococcus rhodochrous | |
1.7 | - |
progesterone | mutant enzyme V291A, in 50 mM Tris/HCl, pH 7.5, temperature not specified in the publication | Rhodococcus rhodochrous | |
1.8 | - |
progesterone | wild type enzyme, in 50 mM Tris/HCl, pH 7.5, temperature not specified in the publication | Rhodococcus rhodochrous | |
2.1 | - |
phenylacetone | mutant enzyme L500Y, in 50 mM Tris/HCl, pH 7.5, temperature not specified in the publication | Rhodococcus rhodochrous | |
2.2 | - |
phenylacetone | mutant enzyme V291A, in 50 mM Tris/HCl, pH 7.5, temperature not specified in the publication | Rhodococcus rhodochrous | |
2.6 | - |
phenylacetone | mutant enzyme P157Q, in 50 mM Tris/HCl, pH 7.5, temperature not specified in the publication | Rhodococcus rhodochrous | |
3.9 | - |
progesterone | mutant enzyme P157Q, in 50 mM Tris/HCl, pH 7.5, temperature not specified in the publication | Rhodococcus rhodochrous | |
5.2 | - |
progesterone | mutant enzyme T345L, in 50 mM Tris/HCl, pH 7.5, temperature not specified in the publication | Rhodococcus rhodochrous |