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Literature summary extracted from

  • Gu, J.; Liu, M.; Guo, F.; Xie, W.; Lu, W.; Ye, L.; Chen, Z.; Yuan, S.; Yu, H.
    Virtual screening of mandelate racemase mutants with enhanced activity based on binding energy in the transition state (2014), Enzyme Microb. Technol., 55, 121-127.
    View publication on PubMed

Protein Variants

EC Number Protein Variants Comment Organism
5.1.2.2 additional information development and evaluation of a virtual mutant screening method for non-natural substrates based on the binding energy in the transition state, the method is beneficial in enzyme rational redesign and helps to better understand the catalytic properties of the enzyme, and it is effective in predicting the trend of mutational effects on catalysis, molecular dynamic simulation, overview Pseudomonas putida
5.1.2.2 V26I site-directed mutagenesis, the mutant shows 2fold higher catalytic efficiency towards R-mandelamide than the wild-type enzyme Pseudomonas putida
5.1.2.2 V26I/Y54V site-directed mutagenesis, the mutant shows 5.2fold higher catalytic efficiency towards (3R)-3-chloromandelic acid than the wild-type enzyme Pseudomonas putida
5.1.2.2 V29L site-directed mutagenesis, the mutant shows 2fold higher catalytic efficiency towards R-2-naphthylglycolate than the wild-type enzyme Pseudomonas putida

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
5.1.2.2 1.13
-
 (R)-mandelamide pH 7.5, 25°C, wild-type enzyme Pseudomonas putida
5.1.2.2 1.2
-
 (2R)-2-naphthylglycolate pH 7.5, 25°C, wild-type enzyme Pseudomonas putida
5.1.2.2 1.24
-
 (R)-mandelamide pH 7.5, 25°C, mutant V26L Pseudomonas putida
5.1.2.2 1.27
-
 (R)-3-chloromandelate pH 7.5, 25°C, mutant V29L Pseudomonas putida
5.1.2.2 1.6
-
 (2R)-2-naphthylglycolate pH 7.5, 25°C, mutant V29L Pseudomonas putida
5.1.2.2 1.6 2 (R)-3-chloromandelate pH 7.5, 25°C, mutant Y54L Pseudomonas putida
5.1.2.2 1.7 2 (R)-3-chloromandelate pH 7.5, 25°C, mutant L319F Pseudomonas putida
5.1.2.2 1.87
-
 (R)-3-chloromandelate pH 7.5, 25°C, mutant V26L Pseudomonas putida
5.1.2.2 1.89
-
 (R)-3-chloromandelate pH 7.5, 25°C, wild-type enzyme Pseudomonas putida
5.1.2.2 1.9
-
 (R)-3-chloromandelate pH 7.5, 25°C, mutant V26S Pseudomonas putida
5.1.2.2 1.95
-
 (R)-3-chloromandelate pH 7.5, 25°C, mutant Y54F Pseudomonas putida
5.1.2.2 1.99
-
 (R)-3-chloromandelate pH 7.5, 25°C, mutant V26I/Y54I Pseudomonas putida
5.1.2.2 2.12
-
 (R)-3-chloromandelate pH 7.5, 25°C, mutant V22I Pseudomonas putida
5.1.2.2 2.17
-
 (R)-3-chloromandelate pH 7.5, 25°C, mutant Y54V Pseudomonas putida
5.1.2.2 2.22
-
 (R)-3-chloromandelate pH 7.5, 25°C, mutant Y54A Pseudomonas putida
5.1.2.2 2.4
-
 (R)-3-chloromandelate pH 7.5, 25°C, mutant V22L Pseudomonas putida
5.1.2.2 2.46
-
 (R)-3-chloromandelate pH 7.5, 25°C, mutant V26I Pseudomonas putida
5.1.2.2 3.02
-
 (R)-3-chloromandelate pH 7.5, 25°C, mutant Y54I Pseudomonas putida
5.1.2.2 3.5
-
 (2R)-2-naphthylglycolate pH 7.5, 25°C, wild-type enzyme, in presence of 20 mM MgCl2 Pseudomonas putida
5.1.2.2 3.92
-
 (R)-3-chloromandelate pH 7.5, 25°C, mutant V26I/Y54V Pseudomonas putida
5.1.2.2 4.2
-
 (2R)-2-naphthylglycolate pH 7.5, 25°C, mutant V22I Pseudomonas putida

Metals/Ions

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

Organism

EC Number Organism UniProt Comment Textmining
5.1.2.2 Pseudomonas putida P11444
-
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
5.1.2.2 (2R)-2-naphthylglycolate a non-natural substrate Pseudomonas putida (2S)-2-naphthylglycolate
-
 ?
5.1.2.2 (R)-3-chloromandelate a non-natural substrate Pseudomonas putida (S)-3-chloromandelate
-
 ?
5.1.2.2 (R)-mandelamide a non-natural substrate Pseudomonas putida (S)-mandelamide
-
 ?

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 11.03
-
 (R)-mandelamide pH 7.5, 25°C, wild-type enzyme Pseudomonas putida
5.1.2.2 21.17
-
 (R)-mandelamide pH 7.5, 25°C, mutant V26L Pseudomonas putida
5.1.2.2 46
-
 (2R)-2-naphthylglycolate pH 7.5, 25°C, wild-type enzyme Pseudomonas putida
5.1.2.2 70
-
 (2R)-2-naphthylglycolate pH 7.5, 25°C, wild-type enzyme, in presence of 20 mM MgCl2 Pseudomonas putida
5.1.2.2 137
-
 (2R)-2-naphthylglycolate pH 7.5, 25°C, mutant V29L Pseudomonas putida
5.1.2.2 147
-
 (2R)-2-naphthylglycolate pH 7.5, 25°C, mutant V22I Pseudomonas putida
5.1.2.2 388
-
 (R)-3-chloromandelate pH 7.5, 25°C, mutant V29L Pseudomonas putida
5.1.2.2 469
-
 (R)-3-chloromandelate pH 7.5, 25°C, wild-type enzyme Pseudomonas putida
5.1.2.2 507
-
 (R)-3-chloromandelate pH 7.5, 25°C, mutant Y54F Pseudomonas putida
5.1.2.2 510
-
 (R)-3-chloromandelate pH 7.5, 25°C, mutant V26I/Y54V Pseudomonas putida
5.1.2.2 559
-
 (R)-3-chloromandelate pH 7.5, 25°C, mutant V26S Pseudomonas putida
5.1.2.2 565
-
 (R)-3-chloromandelate pH 7.5, 25°C, mutant Y54L Pseudomonas putida
5.1.2.2 637
-
 (R)-3-chloromandelate pH 7.5, 25°C, mutant V22I Pseudomonas putida
5.1.2.2 913
-
 (R)-3-chloromandelate pH 7.5, 25°C, mutant L319F Pseudomonas putida
5.1.2.2 953
-
 (R)-3-chloromandelate pH 7.5, 25°C, mutant Y54A Pseudomonas putida
5.1.2.2 1030
-
 (R)-3-chloromandelate pH 7.5, 25°C, mutant V26I/Y54I Pseudomonas putida
5.1.2.2 1160
-
 (R)-3-chloromandelate pH 7.5, 25°C, mutant Y54V Pseudomonas putida
5.1.2.2 1220
-
 (R)-3-chloromandelate pH 7.5, 25°C, mutant V22L Pseudomonas putida
5.1.2.2 1520
-
 (R)-3-chloromandelate pH 7.5, 25°C, mutant Y54I Pseudomonas putida
5.1.2.2 1710
-
 (R)-3-chloromandelate pH 7.5, 25°C, mutant V26L Pseudomonas putida
5.1.2.2 2020
-
 (R)-3-chloromandelate pH 7.5, 25°C, mutant V26I 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 additional information development and evaluation of a virtual mutant screening method based on the binding energy in the transition state, the method is beneficial in enzyme rational redesign and helps to better understand the catalytic properties of the enzyme, and it is effective in predicting the trend of mutational effects on catalysis, molecular dynamic simulation, overview 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 9.79
-
 (R)-mandelamide pH 7.5, 25°C, wild-type enzyme Pseudomonas putida
5.1.2.2 17.1
-
 (R)-mandelamide pH 7.5, 25°C, mutant V26L Pseudomonas putida
5.1.2.2 20.1
-
 (2R)-2-naphthylglycolate pH 7.5, 25°C, wild-type enzyme, in presence of 20 mM MgCl2 Pseudomonas putida
5.1.2.2 34
-
 (2R)-2-naphthylglycolate pH 7.5, 25°C, mutant V22I Pseudomonas putida
5.1.2.2 38
-
 (2R)-2-naphthylglycolate pH 7.5, 25°C, wild-type enzyme Pseudomonas putida
5.1.2.2 85
-
 (2R)-2-naphthylglycolate pH 7.5, 25°C, mutant V29L Pseudomonas putida
5.1.2.2 249
-
 (R)-3-chloromandelate pH 7.5, 25°C, wild-type enzyme Pseudomonas putida
5.1.2.2 259
-
 (R)-3-chloromandelate pH 7.5, 25°C, mutant Y54F Pseudomonas putida
5.1.2.2 294
-
 (R)-3-chloromandelate pH 7.5, 25°C, mutant V26S Pseudomonas putida
5.1.2.2 300
-
 (R)-3-chloromandelate pH 7.5, 25°C, mutant V22I Pseudomonas putida
5.1.2.2 305
-
 (R)-3-chloromandelate pH 7.5, 25°C, mutant V29L Pseudomonas putida
5.1.2.2 349
-
 (R)-3-chloromandelate pH 7.5, 25°C, mutant Y54L Pseudomonas putida
5.1.2.2 429
-
 (R)-3-chloromandelate pH 7.5, 25°C, mutant Y54A Pseudomonas putida
5.1.2.2 502
-
 (R)-3-chloromandelate pH 7.5, 25°C, mutant Y54I Pseudomonas putida
5.1.2.2 509
-
 (R)-3-chloromandelate pH 7.5, 25°C, mutant V22L Pseudomonas putida
5.1.2.2 515
-
 (R)-3-chloromandelate pH 7.5, 25°C, mutant V26I/Y54I Pseudomonas putida
5.1.2.2 531
-
 (R)-3-chloromandelate pH 7.5, 25°C, mutant L319F Pseudomonas putida
5.1.2.2 533
-
 (R)-3-chloromandelate pH 7.5, 25°C, mutant Y54V Pseudomonas putida
5.1.2.2 821
-
 (R)-3-chloromandelate pH 7.5, 25°C, mutant V26I Pseudomonas putida
5.1.2.2 916
-
 (R)-3-chloromandelate pH 7.5, 25°C, mutant V26L Pseudomonas putida
5.1.2.2 1300
-
 (R)-3-chloromandelate pH 7.5, 25°C, mutant V26I/Y54V Pseudomonas putida