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Literature summary for 3.5.1.84 extracted from

  • Esquirol, L.; Peat, T.S.; Wilding, M.; Lucent, D.; French, N.G.; Hartley, C.J.; Newman, J.; Scott, C.
    Structural and biochemical characterization of the biuret hydrolase (BiuH) from the cyanuric acid catabolism pathway of Rhizobium leguminasorum bv. viciae 3841 (2018), PLoS One, 13, e0192736 .
    View publication on PubMed
Search for more literature for 3.5.1.84

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli Rhizobium leguminosarum bv. viciae

Crystallization (Commentary)

Crystallization (Comment) Organism
crystals suitable for X-ray analysis are grown using a combination of seeding, in situ proteolysis, formulation variation and additive screening. Structures of the enzyme, the enzyme with bound inhibitor and variants of the enzyme Rhizobium leguminosarum bv. viciae

Protein Variants

Protein Variants Comment Organism
C175S the mutant enzyme binds biuret but is non-catalytic. The structure of the inactive C175S mutant enzyme with substrate bound in the active site reveals that an active site cysteine (Cys175), aspartic acid (Asp36) and lysine (Lys142) form a catalytic triad, which is consistent with biochemical studies of enzyme variants Rhizobium leguminosarum bv. viciae
F41A kcat/Km for buiret is 540fold lower than the wild-type value Rhizobium leguminosarum bv. viciae
F41L kcat/Km for buiret is 713fold lower than the wild-type value Rhizobium leguminosarum bv. viciae
F41W kcat/Km for buiret is 73fold lower than the wild-type value Rhizobium leguminosarum bv. viciae
F41Y kcat/Km for buiret is 139fold lower than the wild-type value Rhizobium leguminosarum bv. viciae
K142A kcat/Km for buiret is 297fold lower than the wild-type value Rhizobium leguminosarum bv. viciae
Q215A kcat/Km for buiret is 419fold lower than the wild-type value Rhizobium leguminosarum bv. viciae
Q215E kcat/Km for buiret is 1302fold lower than the wild-type value Rhizobium leguminosarum bv. viciae
Q215N kcat/Km for buiret is 38fold lower than the wild-type value Rhizobium leguminosarum bv. viciae

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.048
-
 Biuret pH 8.5, 28°C, mutant enzyme Q215N Rhizobium leguminosarum bv. viciae
0.079
-
 Biuret pH 8.5, 28°C, wild-type enzyme Rhizobium leguminosarum bv. viciae
0.127
-
 Biuret pH 8.5, 28°C, mutant enzyme F41W Rhizobium leguminosarum bv. viciae
0.139
-
 Biuret pH 8.5, 28°C, mutant enzyme K142A Rhizobium leguminosarum bv. viciae
0.333
-
 Biuret pH 8.5, 28°C, mutant enzyme F41L Rhizobium leguminosarum bv. viciae
0.505
-
 Biuret pH 8.5, 28°C, mutant enzyme F41A Rhizobium leguminosarum bv. viciae
0.749
-
 Biuret pH 8.5, 28°C, mutant enzyme F41Y Rhizobium leguminosarum bv. viciae
2.073
-
 Biuret pH 8.5, 28°C, mutant enzyme Q215A Rhizobium leguminosarum bv. viciae
2.528
-
 Biuret pH 8.5, 28°C, mutant enzyme Q215E Rhizobium leguminosarum bv. viciae

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
biuret + H2O Rhizobium leguminosarum bv. viciae the enzyme is involved in cyanuric acid mineralization urea-1-carboxylate + NH3
-
 ?

Organism

Organism UniProt Comment Textmining
Rhizobium leguminosarum bv. viciae Q1M7F4
-
-

Purification (Commentary)

Purification (Comment) Organism
-
 Rhizobium leguminosarum bv. viciae

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
biuret + H2O
-
 Rhizobium leguminosarum bv. viciae urea-1-carboxylate + NH3
-
 ?
biuret + H2O the enzyme is involved in cyanuric acid mineralization Rhizobium leguminosarum bv. viciae urea-1-carboxylate + NH3
-
 ?

Synonyms

Synonyms Comment Organism
BiuH
-
 Rhizobium leguminosarum bv. viciae

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.07
-
 Biuret pH 8.5, 28°C, mutant enzyme F41L Rhizobium leguminosarum bv. viciae
0.07
-
 Biuret pH 8.5, 28°C, mutant enzyme K142A Rhizobium leguminosarum bv. viciae
0.14
-
 Biuret pH 8.5, 28°C, mutant enzyme F41A Rhizobium leguminosarum bv. viciae
0.19
-
 Biuret pH 8.5, 28°C, mutant enzyme Q215N Rhizobium leguminosarum bv. viciae
0.26
-
 Biuret pH 8.5, 28°C, mutant enzyme F41W Rhizobium leguminosarum bv. viciae
0.29
-
 Biuret pH 8.5, 28°C, mutant enzyme Q215E Rhizobium leguminosarum bv. viciae
0.74
-
 Biuret pH 8.5, 28°C, mutant enzyme Q215A Rhizobium leguminosarum bv. viciae
0.81
-
 Biuret pH 8.5, 28°C, mutant enzyme F41Y Rhizobium leguminosarum bv. viciae
11.9
-
 Biuret pH 8.5, 28°C, wild-type enzyme Rhizobium leguminosarum bv. viciae

General Information

General Information Comment Organism
metabolism the enzyme is involved in cyanuric acid mineralization Rhizobium leguminosarum bv. viciae

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.115
-
 Biuret pH 8.5, 28°C, mutant enzyme Q215E Rhizobium leguminosarum bv. viciae
0.21
-
 Biuret pH 8.5, 28°C, mutant enzyme F41L Rhizobium leguminosarum bv. viciae
0.277
-
 Biuret pH 8.5, 28°C, mutant enzyme F41A Rhizobium leguminosarum bv. viciae
0.357
-
 Biuret pH 8.5, 28°C, mutant enzyme Q215A Rhizobium leguminosarum bv. viciae
0.504
-
 Biuret pH 8.5, 28°C, mutant enzyme K142A Rhizobium leguminosarum bv. viciae
1.081
-
 Biuret pH 8.5, 28°C, mutant enzyme F41Y Rhizobium leguminosarum bv. viciae
2.047
-
 Biuret pH 8.5, 28°C, mutant enzyme F41W Rhizobium leguminosarum bv. viciae
3.958
-
 Biuret pH 8.5, 28°C, mutant enzyme Q215N Rhizobium leguminosarum bv. viciae
149.742
-
 Biuret pH 8.5, 28°C, wild-type enzyme Rhizobium leguminosarum bv. viciae