Activating Compound | Comment | Organism | Structure |
---|---|---|---|
2-mercaptoethanol | 10 mM, 108% of initial activity | Sinorhizobium meliloti | |
dithiothreitol | 10 mM, 111% of initial activity | Sinorhizobium meliloti |
Cloned (Comment) | Organism |
---|---|
- |
Sinorhizobium meliloti |
Crystallization (Comment) | Organism |
---|---|
to 1.85 A resolution. Enzyme consists of two domains, an (alpha/beta)8 TIM-like barrel domain, where the catalytic centre is located, and a smaller beta-sheet sandwich domain of unknown function. The C-terminal tails of each subunit extend toward another monomer in a swapping-like manner, creating a hydrogen bond network which suggests its implication in protein oligomerisation. Mutational and structural evidence suggest the involvement of a conserved tyrosine in the reaction mechanism of the enzyme | Sinorhizobium meliloti |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
EDTA | 10 mM, 90% residual activity | Sinorhizobium meliloti | |
Hg2+ | 2 mM, 0% residual activity | Sinorhizobium meliloti | |
additional information | not inhibitory: iodoacetamide | Sinorhizobium meliloti |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
1 | - |
DL-5-methyldihydrouracil | pH 8.0, 40°C | Sinorhizobium meliloti | |
3 | - |
dihydrouracil | pH 8.0, 40°C | Sinorhizobium meliloti | |
5 | - |
D-5-benzylhydantoin | pH 8.0, 40°C | Sinorhizobium meliloti | |
10 | - |
D-5-isobutylhydantoin | pH 8.0, 40°C | Sinorhizobium meliloti | |
14 | - |
D-5-butylhydantoin | pH 8.0, 40°C | Sinorhizobium meliloti | |
18 | - |
D-5-ethylhydantoin | pH 8.0, 40°C | Sinorhizobium meliloti | |
20 | - |
DL-5-(p-hydroxyphenyl)hydantoin | pH 8.0, 40°C | Sinorhizobium meliloti | |
26 | - |
D-5-propylhydantoin | pH 8.0, 40°C | Sinorhizobium meliloti | |
44 | - |
D-5-isopropylhydantoin | pH 8.0, 40°C | Sinorhizobium meliloti | |
66 | - |
D-5-methyl-thio-ethylhydantoin | pH 8.0, 40°C | Sinorhizobium meliloti | |
72 | - |
D-5-methylhydantoin | pH 8.0, 40°C | Sinorhizobium meliloti | |
81 | - |
DL-6-isopropyldihydrouracil | pH 8.0, 40°C | Sinorhizobium meliloti | |
111 | - |
DL-6-methyldihydrouracil | pH 8.0, 40°C | Sinorhizobium meliloti | |
125 | - |
hydantoin | pH 8.0, 40°C | Sinorhizobium meliloti |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Co2+ | 2 mM, 331% of initial activity | Sinorhizobium meliloti | |
Cu2+ | 2 mM, 150% of initial activity | Sinorhizobium meliloti | |
Mg2+ | 2 mM, 121% of initial activity | Sinorhizobium meliloti | |
Mn2+ | 2 mM, 407% of initial activity | Sinorhizobium meliloti | |
Ni2+ | 2 mM, 346% of initial activity | Sinorhizobium meliloti | |
Zn2+ | 2 mM, 432% of initial activity | Sinorhizobium meliloti |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Sinorhizobium meliloti | - |
strain CECT4114 | - |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
D-5-benzylhydantoin + H2O | - |
Sinorhizobium meliloti | N-carbamoyl-D-phenylalanine | - |
? | |
D-5-butylhydantoin + H2O | - |
Sinorhizobium meliloti | ? | - |
? | |
D-5-ethylhydantoin + H2O | - |
Sinorhizobium meliloti | ? | - |
? | |
D-5-isobutylhydantoin + H2O | - |
Sinorhizobium meliloti | ? | - |
? | |
D-5-isopropylhydantoin + H2O | - |
Sinorhizobium meliloti | ? | - |
? | |
D-5-methyl-thio-ethylhydantoin + H2O | - |
Sinorhizobium meliloti | ? | - |
? | |
D-5-methylhydantoin + H2O | - |
Sinorhizobium meliloti | N-carbamoyl-D-alanine | - |
? | |
D-5-propylhydantoin + H2O | - |
Sinorhizobium meliloti | ? | - |
? | |
dihydrouracil + H2O | - |
Sinorhizobium meliloti | ureidopropionate | - |
? | |
DL-5-(p-hydroxyphenyl)hydantoin + H2O | - |
Sinorhizobium meliloti | N-carbamoyl-p-hydroxyphenylglycine | - |
? | |
DL-5-methyldihydrouracil + H2O | - |
Sinorhizobium meliloti | ? | - |
? | |
DL-6-isopropyldihydrouracil + H2O | - |
Sinorhizobium meliloti | ? | - |
? | |
DL-6-methyldihydrouracil + H2O | - |
Sinorhizobium meliloti | ? | - |
? | |
hydantoin + H2O | - |
Sinorhizobium meliloti | N-carbamoyl glycine | - |
? | |
additional information | enzyme presents both hydantoinase and dihydropyrimidinase activities, with higher affinity for the natural six-membered ring substrates. For the five-membered ring substrates, affinity is greater for those with aliphatic and apolar groups in the 5th carbon atom, with the highest rates of hydrolysis for D-5-methyl and D-5-ethyl hydantoin. Enzyme is enantioselective for the D-isomer of monosubstituted 5-hydantoins. Activity with substrate analogues such as allantoin, 5,5-dimethylhydantoin, 5-benzyl,5-methyl-hydantoin, succinimide, rodamine or tiazoledione is not detected under the conditions assayed | Sinorhizobium meliloti | ? | - |
? |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
60 | - |
- |
Sinorhizobium meliloti |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
60 | - |
6 h, 95% residual activity | Sinorhizobium meliloti |
70 | - |
30 min, 25% residual activity | Sinorhizobium meliloti |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
1 | - |
DL-5-(p-hydroxyphenyl)hydantoin | pH 8.0, 40°C | Sinorhizobium meliloti | |
1 | - |
DL-5-methyldihydrouracil | pH 8.0, 40°C | Sinorhizobium meliloti | |
1 | - |
DL-6-isopropyldihydrouracil | pH 8.0, 40°C | Sinorhizobium meliloti | |
2 | - |
dihydrouracil | pH 8.0, 40°C | Sinorhizobium meliloti | |
2 | - |
D-5-isobutylhydantoin | pH 8.0, 40°C | Sinorhizobium meliloti | |
2 | - |
D-5-benzylhydantoin | pH 8.0, 40°C | Sinorhizobium meliloti | |
3 | - |
D-5-butylhydantoin | pH 8.0, 40°C | Sinorhizobium meliloti | |
5 | - |
D-5-isopropylhydantoin | pH 8.0, 40°C | Sinorhizobium meliloti | |
6 | - |
D-5-propylhydantoin | pH 8.0, 40°C | Sinorhizobium meliloti | |
8 | - |
DL-6-methyldihydrouracil | pH 8.0, 40°C | Sinorhizobium meliloti | |
12 | - |
hydantoin | pH 8.0, 40°C | Sinorhizobium meliloti | |
17 | - |
D-5-ethylhydantoin | pH 8.0, 40°C | Sinorhizobium meliloti | |
18 | - |
D-5-methyl-thio-ethylhydantoin | pH 8.0, 40°C | Sinorhizobium meliloti | |
197 | - |
D-5-methylhydantoin | pH 8.0, 40°C | Sinorhizobium meliloti |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8 | - |
- |
Sinorhizobium meliloti |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.012 | - |
DL-6-isopropyldihydrouracil | pH 8.0, 40°C | Sinorhizobium meliloti | |
0.05 | - |
DL-5-(p-hydroxyphenyl)hydantoin | pH 8.0, 40°C | Sinorhizobium meliloti | |
0.072 | - |
DL-6-methyldihydrouracil | pH 8.0, 40°C | Sinorhizobium meliloti | |
0.096 | - |
hydantoin | pH 8.0, 40°C | Sinorhizobium meliloti | |
0.114 | - |
D-5-isopropylhydantoin | pH 8.0, 40°C | Sinorhizobium meliloti | |
0.2 | - |
D-5-isobutylhydantoin | pH 8.0, 40°C | Sinorhizobium meliloti | |
0.214 | - |
D-5-butylhydantoin | pH 8.0, 40°C | Sinorhizobium meliloti | |
0.231 | - |
D-5-propylhydantoin | pH 8.0, 40°C | Sinorhizobium meliloti | |
0.273 | - |
D-5-methyl-thio-ethylhydantoin | pH 8.0, 40°C | Sinorhizobium meliloti | |
0.4 | - |
D-5-benzylhydantoin | pH 8.0, 40°C | Sinorhizobium meliloti | |
0.667 | - |
dihydrouracil | pH 8.0, 40°C | Sinorhizobium meliloti | |
0.944 | - |
D-5-ethylhydantoin | pH 8.0, 40°C | Sinorhizobium meliloti | |
1 | - |
DL-5-methyldihydrouracil | pH 8.0, 40°C | Sinorhizobium meliloti | |
2.736 | - |
D-5-methylhydantoin | pH 8.0, 40°C | Sinorhizobium meliloti |