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Literature summary for 5.1.99.3 extracted from

  • Cendron, L.; Ramazzina, I.; Puggioni, V.; Maccacaro, E.; Liuzzi, A.; Secchi, A.; Zanotti, G.; Percudani, R.
    The structure and function of a microbial allantoin racemase reveal the origin and conservation of a catalytic mechanism (2016), Biochemistry, 55, 6421-6432 .
    View publication on PubMed
Search for more literature for 5.1.99.3

Crystallization (Commentary)

Crystallization (Comment) Organism
in cubic and orthorhombic crystal forms, at 2.10 and 2.15 A resolution, respectively Pseudomonas fluorescens

Protein Variants

Protein Variants Comment Organism
E78D mutant shows diminished catalytic activity affecting kcat and KM for both enantiomers Pseudomonas fluorescens
E78Q loss of catalytic activity Pseudomonas fluorescens
additional information The Pseudomonas protein lacks a cysteine residue (C79 in the Klebsiella pneumoniae AllR sequence) that is essential for the two-base catalytic mechanism of KpAllR. The residue is substituted with glycine Pseudomonas fluorescens

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.1
-
 (R)-allantoin wild-type, pH 7.6, temperature not specified in the publication Pseudomonas fluorescens
2.3
-
 (R)-allantoin mutant E78D, pH 7.6, temperature not specified in the publication Pseudomonas fluorescens
2.5
-
 (S)-allantoin wild-type, pH 7.6, temperature not specified in the publication Pseudomonas fluorescens

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
55000
-
 gel filtration Pseudomonas fluorescens

Organism

Organism UniProt Comment Textmining
Pseudomonas fluorescens E3SAZ9
-
-
Pseudomonas fluorescens DSM 50090 E3SAZ9
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
(R)-allantoin
-
 Pseudomonas fluorescens (S)-allantoin
-
 r
(R)-allantoin
-
 Pseudomonas fluorescens DSM 50090 (S)-allantoin
-
 r
(S)-allantoin
-
 Pseudomonas fluorescens (R)-allantoin
-
 r
(S)-allantoin
-
 Pseudomonas fluorescens DSM 50090 (R)-allantoin
-
 r
additional information Allantoin can undergo racemization through formation of a bicyclic intermediate (faster) or proton exchange at the chiral center (slower). The velocity of the reaction via a bicyclic intermediate is unaffected by the enzyme, whereas the velocity of the reaction via the chiral center is increased by 7 orders of magnitude Pseudomonas fluorescens ?
-
 ?
additional information Allantoin can undergo racemization through formation of a bicyclic intermediate (faster) or proton exchange at the chiral center (slower). The velocity of the reaction via a bicyclic intermediate is unaffected by the enzyme, whereas the velocity of the reaction via the chiral center is increased by 7 orders of magnitude Pseudomonas fluorescens DSM 50090 ?
-
 ?

Synonyms

Synonyms Comment Organism
AllR
-
 Pseudomonas fluorescens
BFW91_04965
-
 Pseudomonas fluorescens

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
43
-
 (R)-allantoin mutant E78D, pH 7.6, temperature not specified in the publication Pseudomonas fluorescens
57
-
 (R)-allantoin wild-type, pH 7.6, temperature not specified in the publication Pseudomonas fluorescens
129
-
 (S)-allantoin wild-type, pH 7.6, temperature not specified in the publication Pseudomonas fluorescens

General Information

General Information Comment Organism
physiological function mechanism of racemization involves residue Glu78 acting as a base for the abstraction of the C5 proton of (S)-allantoin and Cys 180 transferring a proton on the opposite side of the allantoin ring. In the stereoinversion of (R)-allantoin, proton abstraction is performed by Cys 180 and reprotonation by Glu 78 Pseudomonas fluorescens

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
16
-
 (S)-allantoin mutant E78D, pH 7.6, temperature not specified in the publication Pseudomonas fluorescens
19
-
 (R)-allantoin mutant E78D, pH 7.6, temperature not specified in the publication Pseudomonas fluorescens
52
-
 (S)-allantoin wild-type, pH 7.6, temperature not specified in the publication Pseudomonas fluorescens
52
-
 (R)-allantoin wild-type, pH 7.6, temperature not specified in the publication Pseudomonas fluorescens