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Literature summary for 5.1.2.2 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.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 5.1.2.2

Cloned(Commentary)

Cloned (Comment) Organism
overexpression of Strep-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) Pseudomonas putida

Crystallization (Commentary)

Crystallization (Comment) Organism
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

Protein Variants Comment Organism
A25V site-directed mutagenesis Pseudomonas putida
T24S site-directed mutagenesis Pseudomonas putida
V22A site-directed mutagenesis Pseudomonas putida
V22F site-directed mutagenesis Pseudomonas putida
V22I site-directed mutagenesis Pseudomonas putida
V26A site-directed mutagenesis Pseudomonas putida
V26A/V29L site-directed mutagenesis Pseudomonas putida
V26F site-directed mutagenesis Pseudomonas putida
V26L site-directed mutagenesis Pseudomonas putida
V29A site-directed mutagenesis Pseudomonas putida
V29F site-directed mutagenesis Pseudomonas putida
V29L site-directed mutagenesis Pseudomonas putida
Y54F site-directed mutagenesis Pseudomonas putida
Y54L site-directed mutagenesis Pseudomonas putida

Inhibitors

Inhibitors Comment Organism Structure
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
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

Metals/Ions Comment Organism Structure
Mg2+ dependent on Pseudomonas putida

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
(S)-mandelate Pseudomonas putida
-
 (R)-mandelate
-
 ?

Organism

Organism UniProt Comment Textmining
Pseudomonas putida P11444
-
-

Purification (Commentary)

Purification (Comment) Organism
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)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
(S)-mandelate
-
 Pseudomonas putida (R)-mandelate
-
 ?

Subunits

Subunits Comment Organism
homooctamer generated from a tetramer of dimers Pseudomonas putida
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]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
25
-
 assay at Pseudomonas putida

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
 assay at Pseudomonas putida