Application | Comment | Organism |
---|---|---|
synthesis | because of its capacity to hydroxylate steroids, CYP106A2 is an interesting candidate for industrial production of steroids or their intermediates. CYP106A2 is known as a 15beta-hydroxylase, but also shows minor 11alpha-hydroxylase activity for progesterone. 11alpha-Hydroxyprogesterone is an important pharmaceutical compound with anti-androgenic and blood-pressure-regulating activity | Priestia megaterium |
Protein Variants | Comment | Organism |
---|---|---|
A106T/A395I | site-directed mutagenesis, the mutant shows 14.3fold higher regioselectivity for 11alpha-hydroxylation and 39.3fold increased catalytic efficiency compared to the wild-type enzyme | Priestia megaterium |
A106T/A395I/R409L | site-directed mutagenesis, the mutant shows 12.6fold higher regioselectivity for 11alpha-hydroxylation and 108fold increased catalytic efficiency compared to the wild-type enzyme | Priestia megaterium |
A106T/A395W/G379K/R409L | site-directed mutagenesis, the mutant shows higher regioselectivity for 11alpha-hydroxylation and increased catalytic efficiency compared to the wild-type enzyme | Priestia megaterium |
A243V/A395I | site-directed mutagenesis, the mutant shows higher regioselectivity for 11alpha-hydroxylation and increased catalytic efficiency compared to the wild-type enzyme | Priestia megaterium |
A243V/A395W/G397K | site-directed mutagenesis, the mutant shows higher regioselectivity for 11alpha-hydroxylation and increased catalytic efficiency compared to the wild-type enzyme | Priestia megaterium |
A395I | site-directed mutagenesis, the mutant shows 8.9fold higher 11alpha-hydroxylation activity compared to the wild-type enzyme | Priestia megaterium |
A395W/G397K | site-directed mutagenesis, the mutant shows 11.5fold higher 11alpha-hydroxylation activity compared to the wild-type enzyme | Priestia megaterium |
D217V/A395I | site-directed mutagenesis, the mutant shows higher regioselectivity for 11alpha-hydroxylation and increased catalytic efficiency compared to the wild-type enzyme | Priestia megaterium |
M155I/F165L/A395I | site-directed mutagenesis, the mutant shows higher regioselectivity for 11alpha-hydroxylation and increased catalytic efficiency compared to the wild-type enzyme | Priestia megaterium |
additional information | direction of the regioselectivity of the enzyme towards hydroxylation at position 11 in the C ring of the steroid through a combination of saturation mutagenesis and rational site-directed mutagenesis, with aid of data from a homology model of CYP106A2 containing docked progesterone, together with site-directed mutagenesis of active site residues. Distributions of amounts of monohydroxylated progesterone products (15beta-,11alpha-, 9alpha-, and 6beta-hydroxyprogesterone) in vivo of wild-type and mutant enzymes, overview | Priestia megaterium |
T247V/A395I | site-directed mutagenesis, the mutant shows higher regioselectivity for 11alpha-hydroxylation and increased catalytic efficiency compared to the wild-type enzyme | Priestia megaterium |
T247W/A395W/G379K | site-directed mutagenesis, the mutant shows higher regioselectivity for 11alpha-hydroxylation and increased catalytic efficiency compared to the wild-type enzyme | Priestia megaterium |
T248V/A395I | site-directed mutagenesis, the mutant shows higher regioselectivity for 11alpha-hydroxylation and increased catalytic efficiency compared to the wild-type enzyme | Priestia megaterium |
T89N/A106T/A395I/R409L | site-directed mutagenesis, the mutant shows higher regioselectivity for 11alpha-hydroxylation and increased catalytic efficiency compared to the wild-type enzyme | Priestia megaterium |
T89N/A395I | site-directed mutagenesis, the mutant shows 11.8fold higher regioselectivity for 11alpha-hydroxylation and 24.4fold increased catalytic efficiency compared to the wild-type enzyme. The production of 15beta-hydroxyprogesterone decreases from 50.4% of the wild-type to 4.8% of mutant T89N/A395I enzyme, whereas that of 11alpha-hydroxyprogesterone increases from 27.7% to 80.9% | Priestia megaterium |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.02 | - |
progesterone | pH not specified in the publication, 30°C, mutant A243V/A395I | Priestia megaterium | |
0.026 | - |
progesterone | pH not specified in the publication, 30°C, mutant A243V/A395W/G397K | Priestia megaterium | |
0.028 | - |
progesterone | pH not specified in the publication, 30°C, mutant D217V/A395I | Priestia megaterium | |
0.032 | - |
progesterone | pH not specified in the publication, 30°C, mutants A395I and M155I/F165L/A395I | Priestia megaterium | |
0.038 | - |
progesterone | pH not specified in the publication, 30°C, mutant T247V/A395I | Priestia megaterium | |
0.039 | - |
progesterone | pH not specified in the publication, 30°C, mutant T247W/A395W/G379K | Priestia megaterium | |
0.046 | - |
progesterone | pH not specified in the publication, 30°C, mutant A106T/A395I/R409L | Priestia megaterium | |
0.048 | - |
progesterone | pH not specified in the publication, 30°C, mutant A106T/A395W/G379K/R409L | Priestia megaterium | |
0.052 | - |
progesterone | pH not specified in the publication, 30°C, mutant T248V/A395I | Priestia megaterium | |
0.055 | - |
progesterone | pH not specified in the publication, 30°C, mutant A395W/G397K | Priestia megaterium | |
0.084 | - |
progesterone | pH not specified in the publication, 30°C, mutant T89N/A395I | Priestia megaterium | |
0.091 | - |
progesterone | pH not specified in the publication, 30°C, mutant A106T/A395I | Priestia megaterium | |
0.109 | - |
progesterone | pH not specified in the publication, 30°C, mutant T89N/A106T/A395I/R409L | Priestia megaterium | |
0.203 | - |
progesterone | pH not specified in the publication, 30°C, wild-type | Priestia megaterium |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
cytosol | - |
Priestia megaterium | 5829 | - |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
47500 | - |
x * 47500, SDS-PAGE | Priestia megaterium |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | Priestia megaterium | the enzyme is capable of converting useful pharmaceutical targets such as 3-oxo-DELTA4-steroids (e.g., testosterone, 11-deoxycorticosterone, androstenedione, and progesterone) mainly into their 15beta-hydroxy homologues. In addition, the enzyme hydroxylates progesterone to a minor extent at the 11alpha-, 9alpha-, and 6beta-positions | ? | - |
? | |
progesterone + AH2 + O2 | Priestia megaterium | - |
11alpha-hydroxyprogesterone + A + H2O | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Priestia megaterium | Q06069 | - |
- |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | the enzyme is capable of converting useful pharmaceutical targets such as 3-oxo-DELTA4-steroids (e.g., testosterone, 11-deoxycorticosterone, androstenedione, and progesterone) mainly into their 15beta-hydroxy homologues. In addition, the enzyme hydroxylates progesterone to a minor extent at the 11alpha-, 9alpha-, and 6beta-positions | Priestia megaterium | ? | - |
? | |
additional information | CYP106A2-catalyzed hydroxylations of progesterone at the 15beta-, 11alpha-, 9alpha-, and 6beta-positions | Priestia megaterium | ? | - |
? | |
progesterone + AH2 + O2 | - |
Priestia megaterium | 11alpha-hydroxyprogesterone + A + H2O | - |
? |
Subunits | Comment | Organism |
---|---|---|
? | x * 47500, SDS-PAGE | Priestia megaterium |
Synonyms | Comment | Organism |
---|---|---|
CYP106A2 | - |
Priestia megaterium |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
assay at | Priestia megaterium |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.3 | - |
progesterone | pH not specified in the publication, 30°C, wild-type | Priestia megaterium | |
0.9 | - |
progesterone | pH not specified in the publication, 30°C, mutant A243V/A395W/G397K | Priestia megaterium | |
1.1 | - |
progesterone | pH not specified in the publication, 30°C, mutant A243V/A395I | Priestia megaterium | |
1.2 | - |
progesterone | pH not specified in the publication, 30°C, mutant A395W/G397K | Priestia megaterium | |
1.3 | - |
progesterone | pH not specified in the publication, 30°C, mutant A395I | Priestia megaterium | |
1.9 | - |
progesterone | pH not specified in the publication, 30°C, mutant T247V/A395I | Priestia megaterium | |
2.5 | - |
progesterone | pH not specified in the publication, 30°C, mutant T89N/A106T/A395I/R409L | Priestia megaterium | |
2.7 | - |
progesterone | pH not specified in the publication, 30°C, mutant T247W/A395W/G379K | Priestia megaterium | |
2.9 | - |
progesterone | pH not specified in the publication, 30°C, mutant T89N/A395I | Priestia megaterium | |
3.5 | - |
progesterone | pH not specified in the publication, 30°C, mutant T248V/A395I | Priestia megaterium | |
4.3 | - |
progesterone | pH not specified in the publication, 30°C, mutant D217V/A395I | Priestia megaterium | |
4.5 | - |
progesterone | pH not specified in the publication, 30°C, mutant A106T/A395W/G379K/R409L | Priestia megaterium | |
5 | - |
progesterone | pH not specified in the publication, 30°C, mutant A106T/A395I | Priestia megaterium | |
6.4 | - |
progesterone | pH not specified in the publication, 30°C, mutant M155I/F165L/A395I | Priestia megaterium | |
6.9 | - |
progesterone | pH not specified in the publication, 30°C, mutant A106T/A395I/R409L | Priestia megaterium |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
1.4 | - |
progesterone | pH not specified in the publication, 30°C, wild-type | Priestia megaterium | |
21.7 | - |
progesterone | pH not specified in the publication, 30°C, mutant A395W/G397K | Priestia megaterium | |
23.2 | - |
progesterone | pH not specified in the publication, 30°C, mutant T89N/A106T/A395I/R409L | Priestia megaterium | |
34.2 | - |
progesterone | pH not specified in the publication, 30°C, mutant T89N/A395I | Priestia megaterium | |
34.5 | - |
progesterone | pH not specified in the publication, 30°C, mutant A243V/A395W/G397K | Priestia megaterium | |
42 | - |
progesterone | pH not specified in the publication, 30°C, mutants A395II | Priestia megaterium | |
49.9 | - |
progesterone | pH not specified in the publication, 30°C, mutant T247V/A395I | Priestia megaterium | |
53.1 | - |
progesterone | pH not specified in the publication, 30°C, mutant A243V/A395I | Priestia megaterium | |
55.1 | - |
progesterone | pH not specified in the publication, 30°C, mutant A106T/A395I | Priestia megaterium | |
66.8 | - |
progesterone | pH not specified in the publication, 30°C, mutant T248V/A395I | Priestia megaterium | |
70.6 | - |
progesterone | pH not specified in the publication, 30°C, mutant T247W/A395W/G379K | Priestia megaterium | |
93.4 | - |
progesterone | pH not specified in the publication, 30°C, mutant A106T/A395W/G379K/R409L | Priestia megaterium | |
151.4 | - |
progesterone | pH not specified in the publication, 30°C, mutant A106T/A395I/R409L | Priestia megaterium | |
151.5 | - |
progesterone | pH not specified in the publication, 30°C, mutant D217V/A395I | Priestia megaterium | |
201.8 | - |
progesterone | pH not specified in the publication, 30°C, mutant M155I/F165L/A395I | Priestia megaterium |