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Literature summary for 5.1.2.2 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
Search for more literature for 5.1.2.2

Cloned(Commentary)

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

General Stability

General Stability Organism
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]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
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
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
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
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

Metals/Ions Comment Organism Structure
Mg2+ enzymatic reaction is Mg2+-dependent 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
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)

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

Synonyms

Synonyms Comment Organism
mandelate racemase
-
 Pseudomonas putida

Temperature Optimum [°C]

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

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
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
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
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
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

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

General Information

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

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
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
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
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
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