Cloned (Comment) | Organism |
---|---|
expression in Escherichia coli | Paenarthrobacter ureafaciens |
Crystallization (Comment) | Organism |
---|---|
single anonymous dispersion phasing of mercury derivative to 2.2 A, and construction of models of the native enzyme and mutant S174A to 1.9 A resolution. The overall structure of the molecule is a compact mixed alpha/beta fold containing a central irregular-sheet composed of 11 beta-strands surrounded by 20 alpha-helices | Paenarthrobacter ureafaciens |
Protein Variants | Comment | Organism |
---|---|---|
C316A | 18% of wild-type hydrolytic activity, decrease in the Km to approximately one-quarter the level of wild-type NylA, CD spectra similar to wild-type | Paenarthrobacter ureafaciens |
C316D | 1.5% of wild-type activity | Paenarthrobacter ureafaciens |
C316E | 1%% of wild-type activity | Paenarthrobacter ureafaciens |
C316G | 185% of wild-type activity | Paenarthrobacter ureafaciens |
C316K | 3.6% of wild-type activity | Paenarthrobacter ureafaciens |
C316N | 4.3% of wild-type activity | Paenarthrobacter ureafaciens |
C316S | 107% of wild-type activity | Paenarthrobacter ureafaciens |
K72A | decreases the hydrolytic and esterolytic activities to undetectable levels | Paenarthrobacter ureafaciens |
N125A | 14% of wild-type hydrolytic activity, CD spectra similar to wild-type | Paenarthrobacter ureafaciens |
S150A | decreases the hydrolytic and esterolytic activities to undetectable levels | Paenarthrobacter ureafaciens |
S174A | decreases the hydrolytic and esterolytic activities to undetectable levels | Paenarthrobacter ureafaciens |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.85 | - |
6-amino-hexanoate cyclic dimer | mutant C316A, pH 7.0, 30°C | Paenarthrobacter ureafaciens | |
1.8 | - |
6-amino-hexanoate cyclic dimer | mutant C316S, pH 7.0, 30°C | Paenarthrobacter ureafaciens | |
3.51 | - |
6-amino-hexanoate cyclic dimer | wild-type, pH 7.0, 30°C | Paenarthrobacter ureafaciens | |
3.66 | - |
6-amino-hexanoate cyclic dimer | mutant C316G, pH 7.0, 30°C | Paenarthrobacter ureafaciens | |
13.2 | - |
6-amino-hexanoate cyclic dimer | mutant N125A, pH 7.0, 30°C | Paenarthrobacter ureafaciens | |
19 | - |
6-amino-hexanoate cyclic dimer | mutant C316D, pH 7.0, 30°C | Paenarthrobacter ureafaciens |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Paenarthrobacter ureafaciens | - |
- |
- |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
1,8-diazacyclotetradecane-2,9-dione + H2O = N-(6-aminohexanoyl)-6-aminohexanoate | a Ser174-cis-Ser150-Lys72 triad constitutes the catalytic center, the backbone N in Ala171 and Ala172 are involved in oxyanion stabilization, Cys316-S forms a hydrogen bond with nitrogen N7 of 6-amino-hexanoate cyclic dimer at the uncleaved amide bond in two equivalent amide bonds of 6-amino-hexanoate cyclic dimer | Paenarthrobacter ureafaciens |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
0.04 | - |
mutant C316E, pH 7.0, 30°C | Paenarthrobacter ureafaciens |
0.063 | - |
mutant C316D, pH 7.0, 30°C | Paenarthrobacter ureafaciens |
0.15 | - |
mutant C316K, pH 7.0, 30°C | Paenarthrobacter ureafaciens |
0.18 | - |
mutant C316N, pH 7.0, 30°C | Paenarthrobacter ureafaciens |
0.57 | - |
mutant N125A, pH 7.0, 30°C | Paenarthrobacter ureafaciens |
0.73 | - |
mutant C316A, pH 7.0, 30°C | Paenarthrobacter ureafaciens |
4.1 | - |
wild-type, pH 7.0, 30°C | Paenarthrobacter ureafaciens |
4.46 | - |
mutant C316S, pH 7.0, 30°C | Paenarthrobacter ureafaciens |
7.7 | - |
mutant C316G, pH 7.0, 30°C | Paenarthrobacter ureafaciens |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
6-amino-hexanoate cyclic dimer + H2O | - |
Paenarthrobacter ureafaciens | 6-aminohexanoate linear dimer | - |
? | |
additional information | enzyme additionally catalyzes the hydrolysis of amide compounds and esters such as glutamine, malonamide, indoleacetamide, 4-nitrophenylacetate, with a specific activity beolw 0.5% of the activity with 6-amino-hexanoate cyclic dimer | Paenarthrobacter ureafaciens | ? | - |
? |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.14 | - |
6-amino-hexanoate cyclic dimer | mutant C316D, pH 7.0, 30°C | Paenarthrobacter ureafaciens | |
0.71 | - |
6-amino-hexanoate cyclic dimer | mutant C316A, pH 7.0, 30°C | Paenarthrobacter ureafaciens | |
1.12 | - |
6-amino-hexanoate cyclic dimer | mutant N125A, pH 7.0, 30°C | Paenarthrobacter ureafaciens | |
4.83 | - |
6-amino-hexanoate cyclic dimer | mutant C316S, pH 7.0, 30°C | Paenarthrobacter ureafaciens | |
4.97 | - |
6-amino-hexanoate cyclic dimer | wild-type, pH 7.0, 30°C | Paenarthrobacter ureafaciens | |
9.15 | - |
6-amino-hexanoate cyclic dimer | mutant C316G, pH 7.0, 30°C | Paenarthrobacter ureafaciens |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.007 | - |
6-amino-hexanoate cyclic dimer | mutant C316D, pH 7.0, 30°C | Paenarthrobacter ureafaciens | |
0.085 | - |
6-amino-hexanoate cyclic dimer | mutant N125A, pH 7.0, 30°C | Paenarthrobacter ureafaciens | |
0.83 | - |
6-amino-hexanoate cyclic dimer | mutant C316A, pH 7.0, 30°C | Paenarthrobacter ureafaciens | |
1.42 | - |
6-amino-hexanoate cyclic dimer | wild-type, pH 7.0, 30°C | Paenarthrobacter ureafaciens | |
2.5 | - |
6-amino-hexanoate cyclic dimer | mutant C316G, pH 7.0, 30°C | Paenarthrobacter ureafaciens | |
2.68 | - |
6-amino-hexanoate cyclic dimer | mutant C316S, pH 7.0, 30°C | Paenarthrobacter ureafaciens |