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Literature summary for 3.5.2.5 extracted from

  • Martinez-Gomez, A.I.; Soriano-Maldonado, P.; Andujar-Sanchez, M.; Clemente-Jimenez, J.M.; Rodriguez-Vico, F.; Neira, J.L.; Las Heras-Vazquez, F.J.; Martinez-Rodriguez, S.
    Biochemical and mutational studies of allantoinase from Bacillus licheniformis CECT 20T (2014), Biochimie, 99, 178-188.
    View publication on PubMed
Search for more literature for 3.5.2.5

Cloned(Commentary)

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

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

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 [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

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 [Da] Molecular Weight Maximum [Da] Comment Organism
170000 215000 gel filtration Bacillus licheniformis

Natural Substrates/ Products (Substrates)

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

Organism UniProt Comment Textmining
Bacillus licheniformis L0CR34 gene pucH
-
Bacillus licheniformis CECT 20T L0CR34 gene pucH
-

Purification (Commentary)

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 and Products (Substrate)

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

Subunits Comment Organism
homotetramer
-
 Bacillus licheniformis

Synonyms

Synonyms Comment Organism
allantoin amidohydrolase
-
 Bacillus licheniformis
AllBali
-
 Bacillus licheniformis

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
50
-
-
 Bacillus licheniformis

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
additional information
-
 apparent Tm of 62°C Bacillus licheniformis

Turnover Number [1/s]

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

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
-
 Bacillus licheniformis

General Information

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 [mM/s]

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