Cloned (Comment) | Organism |
---|---|
expression in Escherichia coli | Mycobacterium tuberculosis |
Crystallization (Comment) | Organism |
---|---|
hanging drop vapor-diffusion method at 18°C, the crystal structure of HsaA to 2.5 A resolution reveales that the enzyme has the same fold, flavin-binding site, and catalytic residues as p-hydroxyphenyl acetate hydroxylase. HsaA prepared aerobically from the Escherichia coli host exists as a mixture of dimers and octamers in solution while HsaA prepared anaerobically from the rhodococcal host exists as a tetramer | Mycobacterium tuberculosis |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
riboflavin | 0.004 mM, 50% inhibition | Mycobacterium tuberculosis |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.097 | - |
3-hydroxy-9,10-seconandrost-1,3,5(10)-triene-9,17-dione | flavin: FADH2, pH 7.4, 25°C | Mycobacterium tuberculosis | |
0.1 | - |
O2 | flavin: FMNH2, pH 7.4, 25°C | Mycobacterium tuberculosis | |
0.2 | - |
3-hydroxy-9,10-seconandrost-1,3,5(10)-triene-9,17-dione | flavin: FMNH2, pH 7.4, 25°C | Mycobacterium tuberculosis | |
0.35 | - |
3,17-dihydroxy-9,10-seconandrost-1,3,5(10)-triene-9-one | flavin: FMNH2, pH 7.4, 25°C | Mycobacterium tuberculosis | |
1.44 | - |
3-hydroxybiphenyl | flavin: FMNH2, pH 7.4, 25°C | Mycobacterium tuberculosis |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
3-hydroxy-9,10-seconandrost-1,3,5(10)-triene-9,17-dione + FADH2 + O2 | Mycobacterium tuberculosis | the enzyme is involved in cholesterol catabolic pathway. This pathway contributes to the pathogenicity of Mycobacterium tuberculosis | 3,4-dihydroxy-9,10-seconandrost-1,3,5(10)-triene-9,17-dione + FAD + H2O | - |
? | |
3-hydroxy-9,10-seconandrost-1,3,5(10)-triene-9,17-dione + FADH2 + O2 | Mycobacterium tuberculosis H37Rv | the enzyme is involved in cholesterol catabolic pathway. This pathway contributes to the pathogenicity of Mycobacterium tuberculosis | 3,4-dihydroxy-9,10-seconandrost-1,3,5(10)-triene-9,17-dione + FAD + H2O | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Mycobacterium tuberculosis | P9WJA1 | P96852: oxygenase subunit hsaA. The reductase subunit hsaB of the two-component enzyme utilizes NADH to reduce the flavin, which is then transferred to the oxygenase subunit | - |
Mycobacterium tuberculosis H37Rv | P9WJA1 | P96852: oxygenase subunit hsaA. The reductase subunit hsaB of the two-component enzyme utilizes NADH to reduce the flavin, which is then transferred to the oxygenase subunit | - |
Purification (Comment) | Organism |
---|---|
- |
Mycobacterium tuberculosis |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
3,17-dihydroxy-9,10-seconandrost-1,3,5(10)-triene-9-one + FMNH2 + O2 | - |
Mycobacterium tuberculosis | 3,4,17-trihydroxy-9,10-seconandrost-1,3,5(10)-triene-9-one + FMN + H2O | - |
? | |
3,17-dihydroxy-9,10-seconandrost-1,3,5(10)-triene-9-one + FMNH2 + O2 | - |
Mycobacterium tuberculosis H37Rv | 3,4,17-trihydroxy-9,10-seconandrost-1,3,5(10)-triene-9-one + FMN + H2O | - |
? | |
3-hydroxy-9,10-seconandrost-1,3,5(10)-triene-9,17-dione + FADH2 + O2 | - |
Mycobacterium tuberculosis | 3,4-dihydroxy-9,10-seconandrost-1,3,5(10)-triene-9,17-dione + FAD + H2O | - |
? | |
3-hydroxy-9,10-seconandrost-1,3,5(10)-triene-9,17-dione + FADH2 + O2 | the enzyme is involved in cholesterol catabolic pathway. This pathway contributes to the pathogenicity of Mycobacterium tuberculosis | Mycobacterium tuberculosis | 3,4-dihydroxy-9,10-seconandrost-1,3,5(10)-triene-9,17-dione + FAD + H2O | - |
? | |
3-hydroxy-9,10-seconandrost-1,3,5(10)-triene-9,17-dione + FADH2 + O2 | - |
Mycobacterium tuberculosis H37Rv | 3,4-dihydroxy-9,10-seconandrost-1,3,5(10)-triene-9,17-dione + FAD + H2O | - |
? | |
3-hydroxy-9,10-seconandrost-1,3,5(10)-triene-9,17-dione + FADH2 + O2 | the enzyme is involved in cholesterol catabolic pathway. This pathway contributes to the pathogenicity of Mycobacterium tuberculosis | Mycobacterium tuberculosis H37Rv | 3,4-dihydroxy-9,10-seconandrost-1,3,5(10)-triene-9,17-dione + FAD + H2O | - |
? | |
3-hydroxy-9,10-seconandrost-1,3,5(10)-triene-9,17-dione + FMNH2 + O2 | - |
Mycobacterium tuberculosis | 3,4-dihydroxy-9,10-seconandrost-1,3,5(10)-triene-9,17-dione + FMN + H2O | - |
? | |
3-hydroxy-9,10-seconandrost-1,3,5(10)-triene-9,17-dione + FMNH2 + O2 | - |
Mycobacterium tuberculosis H37Rv | 3,4-dihydroxy-9,10-seconandrost-1,3,5(10)-triene-9,17-dione + FMN + H2O | - |
? | |
3-hydroxybiphenyl + FMNH2 + O2 | - |
Mycobacterium tuberculosis | 2,3-dihydroxybiphenyl + FMN + H2O | - |
? | |
additional information | the enzyme is able to catalyze the o-hydroxylation of a range of p-substituted phenols to generate the corresponding catechols | Mycobacterium tuberculosis | ? | - |
? | |
additional information | the enzyme is able to catalyze the o-hydroxylation of a range of p-substituted phenols to generate the corresponding catechols | Mycobacterium tuberculosis H37Rv | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
3-hydroxy-9,10-seconandrost-1,3,5(10)-triene-9,17-dione 4-hydroxylase | the two-component enzyme HsaAB consists of an oxygenase subunit HsaA and a reductase subunit hsaB that utilizes NADH to reduce the flavin, which is then transferred to the oxygenase subunit | Mycobacterium tuberculosis |
HsaAB | the two-component enzyme HsaAB consists of an oxygenase subunit HsaA and a reductase subunit hsaB that utilizes NADH to reduce the flavin, which is then transferred to the oxygenase subunit | Mycobacterium tuberculosis |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Mycobacterium tuberculosis |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.031 | - |
O2 | flavin: FMNH2, pH 7.4, 25°C | Mycobacterium tuberculosis | |
0.075 | - |
3-hydroxy-9,10-seconandrost-1,3,5(10)-triene-9,17-dione | flavin: FADH2, pH 7.4, 25°C | Mycobacterium tuberculosis | |
0.11 | - |
3-hydroxybiphenyl | flavin: FMNH2, pH 7.4, 25°C | Mycobacterium tuberculosis | |
0.2 | - |
3-hydroxy-9,10-seconandrost-1,3,5(10)-triene-9,17-dione | flavin: FMNH2, pH 7.4, 25°C | Mycobacterium tuberculosis | |
0.24 | - |
3,17-dihydroxy-9,10-seconandrost-1,3,5(10)-triene-9-one | flavin: FMNH2, pH 7.4, 25°C | Mycobacterium tuberculosis |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7 | - |
assay at | Mycobacterium tuberculosis |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
FADH2 | the enzyme can use FADH or FMNH2 as flavin cofactor | Mycobacterium tuberculosis | |
FMNH2 | the enzyme can use FADH or FMNH2 as flavin cofactor | Mycobacterium tuberculosis |
General Information | Comment | Organism |
---|---|---|
malfunction | hsaA deletion mutant (mutant with a deletion in the oxygenase subunit) does not grow on cholesterol but grows normally on pyruvate. The hsaA deletion mutant (mutant with a deletion in the oxygenase subunit) accumulates 3-hydroxy-9,10-seconandrost-1,3,5(10)-triene-9,17-dione | Mycobacterium tuberculosis |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.08 | - |
3-hydroxybiphenyl | flavin: FMNH2, pH 7.4, 25°C | Mycobacterium tuberculosis | |
0.32 | - |
O2 | flavin: FMNH2, pH 7.4, 25°C | Mycobacterium tuberculosis | |
0.7 | - |
3,17-dihydroxy-9,10-seconandrost-1,3,5(10)-triene-9-one | flavin: FMNH2, pH 7.4, 25°C | Mycobacterium tuberculosis | |
0.77 | - |
3-hydroxy-9,10-seconandrost-1,3,5(10)-triene-9,17-dione | flavin: FADH2, pH 7.4, 25°C | Mycobacterium tuberculosis | |
1 | - |
3-hydroxy-9,10-seconandrost-1,3,5(10)-triene-9,17-dione | flavin: FMNH2, pH 7.4, 25°C | Mycobacterium tuberculosis |