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Literature summary for 5.1.1.1 extracted from

  • Wu, D.; Hu, T.; Zhang, L.; Chen, J.; Du, J.; Ding, J.; Jiang, H.; Shen, X.
    Residues Asp164 and Glu165 at the substrate entryway function potently in substrate orientation of alanine racemase from Escherichia coli: Enzymatic characterization with crystal structure analysis (2008), Protein Sci., 17, 1066-1076.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 5.1.1.1

Cloned(Commentary)

Cloned (Comment) Organism
Escherichia coli strain BL21 (DE3) was transformed with pET28a–Alr (1 wild-type and 8 single point mutants) Escherichia coli

Protein Variants

Protein Variants Comment Organism
D164A alanine racemase Alr from Escherichia coli with single point mutation from D to A at position 164 Escherichia coli
D164K alanine racemase Alr from Escherichia coli with single point mutation from D to K at position 164 Escherichia coli
E165A alanine racemase Alr from Escherichia coli with single point mutation from E to A at position 165 Escherichia coli
E165K alanine racemase Alr from Escherichia coli with single point mutation from E to K at position 165 Escherichia coli
E221A alanine racemase Alr from Escherichia coli with single point mutation from E to A at position 221 Escherichia coli
E221K alanine racemase Alr from Escherichia coli with single point mutation from E to K at position 221 Escherichia coli
E221P alanine racemase Alr from Escherichia coli with single point mutation from E to P at position 221 Escherichia coli
P219A alanine racemase Alr from Escherichia coli with single point mutation from P to A at position 219 Escherichia coli

Inhibitors

Inhibitors Comment Organism Structure
D-cycloserine
-
 Escherichia coli

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.304
-
 D-alanine +/-0.034, Alr P219A, 30°C, pH 8.0, spectrophotometrically measured at 340 nm Escherichia coli
0.311
-
 D-alanine +/-0.008, Alr wildtype, 30°C, pH 8.0, spectrophotometrically measured at 340 nm Escherichia coli
0.402
-
 D-alanine +/-0.055, Alr E221K, 30°C, pH 8.0, spectrophotometrically measured at 340 nm Escherichia coli
0.439
-
 D-alanine +/-0.080, Alr E221A, 30°C, pH 8.0, spectrophotometrically measured at 340 nm Escherichia coli
0.513
-
 D-alanine +/-0.084, Alr E221P, 30°C, pH 8.0, spectrophotometrically measured at 340 nm Escherichia coli
0.528
-
 D-alanine +/-0.079, Alr E165K, 30°C, pH 8.0, spectrophotometrically measured at 340 nm Escherichia coli
0.592
-
 D-alanine +/-0.085, Alr E165A, 30°C, pH 8.0, spectrophotometrically measured at 340 nm Escherichia coli
0.604
-
 D-alanine +/-0.070, Alr D164K, 30°C, pH 8.0, spectrophotometrically measured at 340 nm Escherichia coli
0.615
-
 D-alanine +/-0.032, Alr D164A, 30°C, pH 8.0, spectrophotometrically measured at 340 nm Escherichia coli
1.008
-
 D-alanine +/-0.069, Alr wildtype, 30°C, pH 8.0, spectrophotometrically measured at 340 nm Escherichia coli
1.049
-
 L-alanine +/-0.131, Alr P219A, 30°C, pH 8.0, spectrophotometrically measured at 340 nm Escherichia coli
1.401
-
 L-alanine +/-0.209, Alr E221P, 30°C, pH 8.0, spectrophotometrically measured at 340 nm Escherichia coli
1.516
-
 L-alanine +/-0.083, Alr E221A, 30°C, pH 8.0, spectrophotometrically measured at 340 nm Escherichia coli
1.562
-
 L-alanine +/-0.256, Alr E165A, 30°C, pH 8.0, spectrophotometrically measured at 340 nm Escherichia coli
1.993
-
 L-alanine +/-0.269, Alr E221K, 30°C, pH 8.0, spectrophotometrically measured at 340 nm Escherichia coli
2.057
-
 L-alanine +/-0.038, Alr E165K, 30°C, pH 8.0, spectrophotometrically measured at 340 nm Escherichia coli
3.03
-
 L-alanine +/-0.114, Alr D164A, 30°C, pH 8.0, spectrophotometrically measured at 340 nm Escherichia coli
3.603
-
 L-alanine +/-0.180, Alr D164K, 30°C, pH 8.0, spectrophotometrically measured at 340 nm Escherichia coli

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
L-alanine Escherichia coli
-
 R-alanine
-
 r

Organism

Organism UniProt Comment Textmining
Escherichia coli P0A6B4
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
L-alanine
-
 Escherichia coli R-alanine
-
 r

Subunits

Subunits Comment Organism
dimer 4 EcAlr monomers form 2 identical dimers, crystallography Escherichia coli

Synonyms

Synonyms Comment Organism
EcAlr
-
 Escherichia coli

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.333
-
 D-alanine +/-0.033, Alr D164K, 30°C, pH 8.0, spectrophotometrically measured at 340 nm Escherichia coli
1.317
-
 D-alanine +/-0.100, Alr E165K, 30°C, pH 8.0, spectrophotometrically measured at 340 nm Escherichia coli
2.8
-
 L-alanine +/-0.250, Alr D164K, 30°C, pH 8.0, spectrophotometrically measured at 340 nm Escherichia coli
4
-
 D-alanine +/-0.417, Alr E165A, 30°C, pH 8.0, spectrophotometrically measured at 340 nm Escherichia coli
4.466
-
 D-alanine +/-0.200, Alr D164A, 30°C, pH 8.0, spectrophotometrically measured at 340 nm Escherichia coli
5.267
-
 D-alanine +/-0.333, Alr P219A, 30°C, pH 8.0, spectrophotometrically measured at 340 nm Escherichia coli
5.783
-
 D-alanine +/-0.483, Alr wildtype, 30°C, pH 8.0, spectrophotometrically measured at 340 nm Escherichia coli
6.35
-
 D-alanine +/-0.333, Alr E221K, 30°C, pH 8.0, spectrophotometrically measured at 340 nm Escherichia coli
6.817
-
 D-alanine +/-0.650, Alr E221A, 30°C, pH 8.0, spectrophotometrically measured at 340 nm Escherichia coli
7.617
-
 D-alanine +/-0.750, Alr E221P, 30°C, pH 8.0, spectrophotometrically measured at 340 nm Escherichia coli
16.72
-
 L-alanine +/-0.850, Alr E165K, 30°C, pH 8.0, spectrophotometrically measured at 340 nm Escherichia coli
22.47
-
 L-alanine +/-2.667, Alr E165A, 30°C, pH 8.0, spectrophotometrically measured at 340 nm Escherichia coli
41.82
-
 L-alanine +/-2.600, Alr D164A, 30°C, pH 8.0, spectrophotometrically measured at 340 nm Escherichia coli
51.77
-
 L-alanine +/-6.067, Alr P219A, 30°C, pH 8.0, spectrophotometrically measured at 340 nm Escherichia coli
53.98
-
 L-alanine +/-3.217, Alr wildtype, 30°C, pH 8.0, spectrophotometrically measured at 340 nm Escherichia coli
67.07
-
 L-alanine +/-7.633, Alr E221P, 30°C, pH 8.0, spectrophotometrically measured at 340 nm Escherichia coli
68.23
-
 L-alanine +/-7.550, Alr E221K, 30°C, pH 8.0, spectrophotometrically measured at 340 nm Escherichia coli
70.92
-
 L-alanine +/-2.333, Alr E221A, 30°C, pH 8.0, spectrophotometrically measured at 340 nm Escherichia coli

Cofactor

Cofactor Comment Organism Structure
pyridoxal 5'-phosphate
-
 Escherichia coli