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Literature summary extracted from
- Structure and mechanism of an inverting alkylsulfatase from Pseudomonas sp. DSM6611 specific for secondary alkyl sulfates (2012), FEBS J., 279, 4374-4384 .
Cloned(Commentary)
| EC Number | Cloned (Comment) | Organism |
|---|---|---|
| 3.1.6.19 | expression in Escherichia coli | Pseudomonas sp. DSM 6611 |
Crystallization (Commentary)
| EC Number | Crystallization (Comment) | Organism |
|---|---|---|
| 3.1.6.19 | Pisa1 forms a dimer. Each protomer consists of three domains: the N-terminal, catalytic alphabetabetaalpha sandwich domain, an alpha-helical dimerization domain and an alpha,beta-mixed C-terminal domain. Comparison with the structure of alkyl sulfatase SdsA1 | Pseudomonas sp. DSM 6611 |
Protein Variants
| EC Number | Protein Variants | Comment | Organism |
|---|---|---|---|
| 3.1.6.19 | F250G | 190fold reduction in catalytic activity | Pseudomonas sp. DSM 6611 |
| 3.1.6.19 | H317A | 1300fold decrease in kcat, whereas Km is only slightly increased | Pseudomonas sp. DSM 6611 |
| 3.1.6.19 | S233F/F250G | three-fold to four-fold times lower kcat value and a 10fold higer Km value, variant displays activity towardstowards (S)-2-octyl sulfate | Pseudomonas sp. DSM 6611 |
| 3.1.6.19 | S233F/F250S | about 30% of wild-type activity | Pseudomonas sp. DSM 6611 |
| 3.1.6.19 | S233Y | 2.5fold reduction in catalytic activity | Pseudomonas sp. DSM 6611 |
| 3.1.6.19 | S233Y/F250G | three-fold to four-fold times lower kcat value | Pseudomonas sp. DSM 6611 |
| 3.1.6.19 | Y417D | inactive | Pseudomonas sp. DSM 6611 |
| 3.1.6.19 | Y417F | 114fold decrease in kcat value | Pseudomonas sp. DSM 6611 |
| 3.1.6.19 | Y417H | 238 fold decrease in kcat value | Pseudomonas sp. DSM 6611 |
KM Value [mM]
| EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 3.1.6.19 | 0.099 | - |
(2R)-octan-2-yl hydrogen sulfate | mutant S233Y, pH not specified in the publication, temperature not specified in the publication | Pseudomonas sp. DSM 6611 |  |
| 3.1.6.19 | 0.115 | - |
(2R)-octan-2-yl hydrogen sulfate | mutant S233F/F250S, pH not specified in the publication, temperature not specified in the publication | Pseudomonas sp. DSM 6611 | |
| 3.1.6.19 | 0.151 | - |
(2R)-octan-2-yl hydrogen sulfate | wild-type, pH not specified in the publication, temperature not specified in the publication | Pseudomonas sp. DSM 6611 | |
| 3.1.6.19 | 0.24 | - |
(2R)-octan-2-yl hydrogen sulfate | mutant H317A, pH not specified in the publication, temperature not specified in the publication | Pseudomonas sp. DSM 6611 | |
| 3.1.6.19 | 1.5 | 2 | (2R)-octan-2-yl hydrogen sulfate | mutant S233Y/F250G, pH not specified in the publication, temperature not specified in the publication | Pseudomonas sp. DSM 6611 | |
| 3.1.6.19 | 4.28 | - |
(2R)-octan-2-yl hydrogen sulfate | mutant F250G, pH not specified in the publication, temperature not specified in the publication | Pseudomonas sp. DSM 6611 | |
| 3.1.6.19 | 8.42 | - |
(2R)-octan-2-yl hydrogen sulfate | mutant S233F/F250G, pH not specified in the publication, temperature not specified in the publication | Pseudomonas sp. DSM 6611 | |
Metals/Ions
| EC Number | Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|---|
| 3.1.6.19 | Zn2+ | Pisa1 has a conserved binuclear zinc center | Pseudomonas sp. DSM 6611 | |
Organism
| EC Number | Organism | UniProt | Comment | Textmining |
|---|---|---|---|---|
| 3.1.6.19 | Pseudomonas sp. DSM 6611 | F8KAY7 | - |
- |
Substrates and Products (Substrate)
| EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 3.1.6.19 | (2R)-octan-2-yl hydrogen sulfate + H2O | - |
Pseudomonas sp. DSM 6611 | (S)-2-octanol + sulfate | - |
? | |
| 3.1.6.19 | octyl sulfate + H2O | - |
Pseudomonas sp. DSM 6611 | 1-octanol + sulfate | - |
? | |
Synonyms
| EC Number | Synonyms | Comment | Organism |
|---|---|---|---|
| 3.1.6.19 | Pisa1 | - |
Pseudomonas sp. DSM 6611 |
| 3.1.6.19 | sec-alkylsulfatase | - |
Pseudomonas sp. DSM 6611 |
Turnover Number [1/s]
| EC Number | Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 3.1.6.19 | 0.003 | - |
(2R)-octan-2-yl hydrogen sulfate | mutant H317A, pH not specified in the publication, temperature not specified in the publication | Pseudomonas sp. DSM 6611 | |
| 3.1.6.19 | 0.018 | - |
(2R)-octan-2-yl hydrogen sulfate | mutant Y417H, pH not specified in the publication, temperature not specified in the publication | Pseudomonas sp. DSM 6611 | |
| 3.1.6.19 | 0.02 | - |
(2R)-octan-2-yl hydrogen sulfate | mutant F250G, pH not specified in the publication, temperature not specified in the publication | Pseudomonas sp. DSM 6611 | |
| 3.1.6.19 | 0.038 | - |
(2R)-octan-2-yl hydrogen sulfate | mutant Y417F, pH not specified in the publication, temperature not specified in the publication | Pseudomonas sp. DSM 6611 | |
| 3.1.6.19 | 0.38 | - |
(2R)-octan-2-yl hydrogen sulfate | mutant S233F/F250G, pH not specified in the publication, temperature not specified in the publication | Pseudomonas sp. DSM 6611 | |
| 3.1.6.19 | 1.13 | - |
(2R)-octan-2-yl hydrogen sulfate | mutant S233F/F250S, pH not specified in the publication, temperature not specified in the publication | Pseudomonas sp. DSM 6611 | |
| 3.1.6.19 | 1.4 | - |
(2R)-octan-2-yl hydrogen sulfate | mutant S233Y/F250G, pH not specified in the publication, temperature not specified in the publication | Pseudomonas sp. DSM 6611 | |
| 3.1.6.19 | 1.73 | - |
(2R)-octan-2-yl hydrogen sulfate | mutant S233Y, pH not specified in the publication, temperature not specified in the publication | Pseudomonas sp. DSM 6611 | |
| 3.1.6.19 | 4.37 | - |
(2R)-octan-2-yl hydrogen sulfate | wild-type, pH not specified in the publication, temperature not specified in the publication | Pseudomonas sp. DSM 6611 | |
General Information
| EC Number | General Information | Comment | Organism |
|---|---|---|---|
| 3.1.6.19 | metabolism | Pisa1 shows a preference for secondary rather than primary alkyl sulfates, and enantioselectively hydrolyzes the (R)-enantiomer of rac-2-octyl sulfate, yielding (S)-2-octanol with inversion of absolute configuration as a result of C-O bond cleavage. An invariant histidine (His317) near the sulfate-binding site acts as general acid for crucial protonation of the sulfate leaving group | Pseudomonas sp. DSM 6611 |
kcat/KM [mM/s]
| EC Number | kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 3.1.6.19 | 0.005 | - |
(2R)-octan-2-yl hydrogen sulfate | mutant F250G, pH not specified in the publication, temperature not specified in the publication | Pseudomonas sp. DSM 6611 | |
| 3.1.6.19 | 0.014 | - |
(2R)-octan-2-yl hydrogen sulfate | mutant H317A, pH not specified in the publication, temperature not specified in the publication | Pseudomonas sp. DSM 6611 | |
| 3.1.6.19 | 0.046 | - |
(2R)-octan-2-yl hydrogen sulfate | mutant S233F/F250G, pH not specified in the publication, temperature not specified in the publication | Pseudomonas sp. DSM 6611 | |
| 3.1.6.19 | 0.919 | - |
(2R)-octan-2-yl hydrogen sulfate | mutant S233Y/F250G, pH not specified in the publication, temperature not specified in the publication | Pseudomonas sp. DSM 6611 | |
| 3.1.6.19 | 1.33 | - |
octyl sulfate | wild-type, pH not specified in the publication, temperature not specified in the publication | Pseudomonas sp. DSM 6611 | |
| 3.1.6.19 | 9.85 | - |
(2R)-octan-2-yl hydrogen sulfate | mutant S233F/F250S, pH not specified in the publication, temperature not specified in the publication | Pseudomonas sp. DSM 6611 | |
| 3.1.6.19 | 17.5 | - |
(2R)-octan-2-yl hydrogen sulfate | mutant S233Y, pH not specified in the publication, temperature not specified in the publication | Pseudomonas sp. DSM 6611 | |
| 3.1.6.19 | 28.9 | - |
(2R)-octan-2-yl hydrogen sulfate | wild-type, pH not specified in the publication, temperature not specified in the publication | Pseudomonas sp. DSM 6611 | |
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