Literature summary extracted from
Lietzan, A.D.; Nagar, M.; Pellmann, E.A.; Bourque, J.R.; Bearne, S.L.; St Maurice, M.
Structure of mandelate racemase with bound intermediate analogues benzohydroxamate and cupferron (2012), Biochemistry, 51, 1160-1170.
Cloned(Commentary)
EC Number |
Cloned (Comment) |
Organism |
---|
5.1.2.2 |
overexpression of Strep-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) |
Pseudomonas putida |
Crystallization (Commentary)
EC Number |
Crystallization (Comment) |
Organism |
---|
5.1.2.2 |
purified recombinant homooctameric wild-type enzyme complexed with two analogues of the putative aci-carboxylate intermediate, benzohydroxamate and Cupferron, X-ray diffraction structure at 2.2 A resolution |
Pseudomonas putida |
Protein Variants
EC Number |
Protein Variants |
Comment |
Organism |
---|
5.1.2.2 |
A25V |
site-directed mutagenesis |
Pseudomonas putida |
5.1.2.2 |
T24S |
site-directed mutagenesis |
Pseudomonas putida |
5.1.2.2 |
V22A |
site-directed mutagenesis |
Pseudomonas putida |
5.1.2.2 |
V22F |
site-directed mutagenesis |
Pseudomonas putida |
5.1.2.2 |
V22I |
site-directed mutagenesis |
Pseudomonas putida |
5.1.2.2 |
V26A |
site-directed mutagenesis |
Pseudomonas putida |
5.1.2.2 |
V26A/V29L |
site-directed mutagenesis |
Pseudomonas putida |
5.1.2.2 |
V26F |
site-directed mutagenesis |
Pseudomonas putida |
5.1.2.2 |
V26L |
site-directed mutagenesis |
Pseudomonas putida |
5.1.2.2 |
V29A |
site-directed mutagenesis |
Pseudomonas putida |
5.1.2.2 |
V29F |
site-directed mutagenesis |
Pseudomonas putida |
5.1.2.2 |
V29L |
site-directed mutagenesis |
Pseudomonas putida |
5.1.2.2 |
Y54F |
site-directed mutagenesis |
Pseudomonas putida |
5.1.2.2 |
Y54L |
site-directed mutagenesis |
Pseudomonas putida |
Inhibitors
EC Number |
Inhibitors |
Comment |
Organism |
Structure |
---|
5.1.2.2 |
benzohydroxamate |
a reasonable mimic of the transition state and/or intermediate that chelates the active site divalent metal ion and is bound in a conformation with the phenyl ring coplanar with the hydroxamate moiety, active site binding, structure comparison with bound Cupferron, overview |
Pseudomonas putida |
|
5.1.2.2 |
Cupferron |
a reasonable mimic of the transition state and/or intermediate that chelates the active site divalent metal ion and is bound in a conformation with the phenyl ring coplanar with the diazeniumdiolate moiety, active site binding, structure comparison with bound benzohydroxamate, overview |
Pseudomonas putida |
|
Metals/Ions
EC Number |
Metals/Ions |
Comment |
Organism |
Structure |
---|
5.1.2.2 |
Mg2+ |
dependent on |
Pseudomonas putida |
|
Natural Substrates/ Products (Substrates)
EC Number |
Natural Substrates |
Organism |
Comment (Nat. Sub.) |
Natural Products |
Comment (Nat. Pro.) |
Rev. |
Reac. |
---|
5.1.2.2 |
(S)-mandelate |
Pseudomonas putida |
- |
(R)-mandelate |
- |
? |
|
Organism
EC Number |
Organism |
UniProt |
Comment |
Textmining |
---|
5.1.2.2 |
Pseudomonas putida |
P11444 |
- |
- |
Purification (Commentary)
EC Number |
Purification (Comment) |
Organism |
---|
5.1.2.2 |
recombinant Strep-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by affinity chromatography and dialysis |
Pseudomonas putida |
Substrates and Products (Substrate)
EC Number |
Substrates |
Comment Substrates |
Organism |
Products |
Comment (Products) |
Rev. |
Reac. |
---|
5.1.2.2 |
(S)-mandelate |
- |
Pseudomonas putida |
(R)-mandelate |
- |
? |
|
Subunits
EC Number |
Subunits |
Comment |
Organism |
---|
5.1.2.2 |
homooctamer |
generated from a tetramer of dimers |
Pseudomonas putida |
5.1.2.2 |
More |
the enzyme is composed of three distinct structural domains: an N-terminal capping domain consisting of a three-stranded beta-sheet with an antiparallel four-alpha-helix bundle, a central domain consisting of a (beta/alpha)7 beta-barrel, and a short C-terminal domain composed of external beta-strands |
Pseudomonas putida |
Temperature Optimum [°C]
EC Number |
Temperature Optimum [°C] |
Temperature Optimum Maximum [°C] |
Comment |
Organism |
---|
5.1.2.2 |
25 |
- |
assay at |
Pseudomonas putida |
pH Optimum
EC Number |
pH Optimum Minimum |
pH Optimum Maximum |
Comment |
Organism |
---|
5.1.2.2 |
7.5 |
- |
assay at |
Pseudomonas putida |