Cloned (Comment) | Organism |
---|---|
gene pucH, DNA and amino acid sequence determination and analysis, recombinant expression of wild-type and mutant His-tagged enzymes in Escherichia coli strain BL21(DE3) | Bacillus licheniformis |
Protein Variants | Comment | Organism |
---|---|---|
D319A | site-directed mutagenesis, inactive mutant, altered thermal denaturation compared to the wild-type enzyme | Bacillus licheniformis |
D319N | site-directed mutagenesis, inactive mutant, altered thermal denaturation compared to the wild-type enzyme | Bacillus licheniformis |
S292A | site-directed mutagenesis, the mutant shows reduced activity and altered thermal denaturation compared to the wild-type enzyme | Bacillus licheniformis |
T155A | site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme | Bacillus licheniformis |
T155Y | site-directed mutagenesis, the mutant shows reduced activity and altered thermal denaturation compared to the wild-type enzyme | Bacillus licheniformis |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
8-Hydroxyquinoline-5-sulfonic acid | complete inhibition | Bacillus licheniformis | |
Acetohydroxamic acid | - |
Bacillus licheniformis | |
DTT | complete inhibition | Bacillus licheniformis | |
H2O2 | allosteric effect of H2O2 toward allantoinase | Bacillus licheniformis | |
L-cysteine | complete inhibition | Bacillus licheniformis |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
17.7 | - |
(S)-allantoin | pH 7.5, 50°C, recombinant wild-type enzyme | Bacillus licheniformis | |
20.3 | - |
(S)-allantoin | pH 7.5, 50°C, recombinant mutant T155Y | Bacillus licheniformis | |
20.9 | - |
(S)-allantoin | pH 7.5, 50°C, recombinant mutant T155A | Bacillus licheniformis | |
35.7 | - |
(S)-allantoin | pH 7.5, 50°C, recombinant mutant S292A | Bacillus licheniformis |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Co2+ | activates, most effective metal cofactor, inverting the enantioselectivity of AllBali | Bacillus licheniformis | |
Mn2+ | activates | Bacillus licheniformis | |
Zn2+ | activates slightly | Bacillus licheniformis |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
170000 | 215000 | gel filtration | Bacillus licheniformis |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
(S)-allantoin + H2O | Bacillus licheniformis | - |
allantoate | - |
? | |
(S)-allantoin + H2O | Bacillus licheniformis CECT 20T | - |
allantoate | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Bacillus licheniformis | L0CR34 | gene pucH | - |
Bacillus licheniformis CECT 20T | L0CR34 | gene pucH | - |
Purification (Comment) | Organism |
---|---|
recombinant wild-type and mutant His-tagged enzymes from Escherichia coli strain BL21(DE3) by cobalt affinity chromatography, gel fltration, and ultrafiltration | Bacillus licheniformis |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
(S)-allantoin + H2O | - |
Bacillus licheniformis | allantoate | - |
? | |
(S)-allantoin + H2O | best substrate, enantioselectivity of Co2+-enzyme toward allantoin enantiomers | Bacillus licheniformis | allantoate | - |
? | |
(S)-allantoin + H2O | - |
Bacillus licheniformis CECT 20T | allantoate | - |
? | |
(S)-allantoin + H2O | best substrate, enantioselectivity of Co2+-enzyme toward allantoin enantiomers | Bacillus licheniformis CECT 20T | allantoate | - |
? | |
5-ethyl-hydantoin + H2O | low activity | Bacillus licheniformis | ? | - |
? | |
5-ethyl-hydantoin + H2O | low activity | Bacillus licheniformis CECT 20T | ? | - |
? | |
dihydrouracil + H2O | low activity | Bacillus licheniformis | ? | - |
? | |
dihydrouracil + H2O | low activity | Bacillus licheniformis CECT 20T | ? | - |
? | |
hydantoin + H2O | low activity | Bacillus licheniformis | ? | - |
? | |
hydantoin + H2O | low activity | Bacillus licheniformis CECT 20T | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
homotetramer | - |
Bacillus licheniformis |
Synonyms | Comment | Organism |
---|---|---|
allantoin amidohydrolase | - |
Bacillus licheniformis |
AllBali | - |
Bacillus licheniformis |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
50 | - |
- |
Bacillus licheniformis |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
additional information | - |
apparent Tm of 62°C | Bacillus licheniformis |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
5.2 | - |
(S)-allantoin | pH 7.5, 50°C, recombinant mutant T155Y | Bacillus licheniformis | |
10.1 | - |
(S)-allantoin | pH 7.5, 50°C, recombinant mutant T155A | Bacillus licheniformis | |
24.4 | - |
(S)-allantoin | pH 7.5, 50°C, recombinant wild-type enzyme and mutant S292A | Bacillus licheniformis |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
- |
Bacillus licheniformis |
General Information | Comment | Organism |
---|---|---|
additional information | residue Ser292 is likely implicated in the binding of the allantoin ring through the carbonyl group of the polypeptide main chain, which is the common mechanism observed in other members of the amidohydrolase family. Modeling and docking studies | Bacillus licheniformis |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.3 | - |
(S)-allantoin | pH 7.5, 50°C, recombinant mutant T155Y | Bacillus licheniformis | |
0.5 | - |
(S)-allantoin | pH 7.5, 50°C, recombinant mutant T155A | Bacillus licheniformis | |
0.7 | - |
(S)-allantoin | pH 7.5, 50°C, recombinant mutant S292A | Bacillus licheniformis | |
1.4 | - |
(S)-allantoin | pH 7.5, 50°C, recombinant wild-type enzyme | Bacillus licheniformis |