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Literature summary extracted from

  • Narmandakh, A.; Bearne, S.L.
    Purification of recombinant mandelate racemase: improved catalytic activity (2010), Protein Expr. Purif., 69, 39-46.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
5.1.2.2 cloning of fusions proteins in Escherichia coli Pseudomonas putida

General Stability

EC Number General Stability Organism
5.1.2.2 recombinant enzyme exhibits only a slight loss of activity when the enzyme is dialyzed without 200 mM NaCl at 4°C for 8 h, a loss of 20% of its activity when the enzyme is stored for 1 h on ice in the absence of BSA (but stable in the presence of 0.1% BSA), and a 30% reduction in Vmax after storage for 20 days at -70°C in the presence of 200 mM NaCl Pseudomonas putida

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
5.1.2.2 0.3
-
 (S)-Mandelate recombinant mandelate racemase, putative misfolded form with N-terminal hexahistidine tag, Na-HEPES buffer (0.1 M, pH 7.5), 3.3 mM MgCl2 Pseudomonas putida
5.1.2.2 0.6
-
 (S)-Mandelate recombinant mrIII fusion protein bearing an N-terminal StrepII-tag and a C-terminal decahistidine tag, Na-HEPES buffer (0.1 M, pH 7.5), 3.3 mM MgCl2 Pseudomonas putida
5.1.2.2 0.8
-
 (S)-Mandelate recombinant mandelate racemase represents correctly folded enzyme with N-terminal hexahistidine tag, in Na-HEPES buffer (0.1 M, pH 7.5),3.3 mM MgCl2 Pseudomonas putida
5.1.2.2 1.1
-
 (S)-Mandelate recombinant mandelate racemase, replacement of the N-terminal hexahistidine tag by a StrepII-tag appears to ameliorate the folding problem yielding a single enzyme species, Na-HEPES buffer (0.1 M, pH 7.5), 3.3 mM MgCl2 Pseudomonas putida

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
5.1.2.2 Mg2+ enzymatic reaction is Mg2+-dependent 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 purification of recombinant MR as a fusion protein with an N-terminal hexahistidine tag using immobilized-nickel ion affinity chromatography and elution with a linear gradient of EDTA 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
-
 ?

Synonyms

EC Number Synonyms Comment Organism
5.1.2.2 mandelate racemase
-
 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

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
5.1.2.2 190
-
 (S)-Mandelate recombinant mandelate racemase, putative misfolded form with N-terminal hexahistidine tag, Na-HEPES buffer (0.1 M, pH 7.5), 3.3 mM MgCl2 Pseudomonas putida
5.1.2.2 472
-
 (S)-Mandelate recombinant mandelate racemase fusion protein bearing an N-terminal StrepII-tag and a C-terminal decahistidine tag, Na-HEPES buffer (0.1 M, pH 7.5), 3.3 mM MgCl2 Pseudomonas putida
5.1.2.2 940
-
 (S)-Mandelate recombinant mandelate racemase represents correctly folded enzyme, with N-terminal hexahistidine tag, in Na-HEPES buffer (0.1 M, pH 7.5), 3.3 mM MgCl2 Pseudomonas putida
5.1.2.2 1124
-
 (S)-Mandelate recombinant enzyme replacement of the N-terminal hexahistidine tag by a StrepII-tag appears to ameliorate the folding problem yielding a single enzyme species, Na-HEPES buffer (0.1 M, pH 7.5), 3.3 mM MgCl2 Pseudomonas putida

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
5.1.2.2 7.5
-
 assay at Pseudomonas putida

General Information

EC Number General Information Comment Organism
5.1.2.2 physiological function enzyme catalyzes the Mg2+-dependent 1,1-proton transfer that interconverts the enantiomers of mandelate Pseudomonas putida

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
5.1.2.2 1100
-
 (S)-Mandelate recombinant mandelate racemase, replacement of the N-terminal hexahistidine tag by a StrepII-tag appears to ameliorate the folding problem yielding a single enzyme species, Na-HEPES buffer (0.1 M, pH 7.5), 3.3 mM MgCl2 Pseudomonas putida
5.1.2.2 1200
-
 (S)-Mandelate recombinant mandelate racemase, represents correctly folded enzyme with N-terminal hexahistidine tag, in Na-HEPES buffer (0.1 M, pH 7.5), 3.3 mM MgCl2 Pseudomonas putida
5.1.2.2 7300
-
 (S)-Mandelate recombinant mandelate racemase, putative misfolded form with N-terminal hexahistidine tag, Na-HEPES buffer (0.1 M, pH 7.5), 3.3 mM MgCl2 Pseudomonas putida
5.1.2.2 8000
-
 (S)-Mandelate recombinant mandelate racemase, fusion protein bearing an N-terminal StrepII-tag and a C-terminal decahistidine tag, Na-HEPES buffer (0.1 M, pH 7.5), 3.3 mM MgCl2 Pseudomonas putida