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Literature summary for 5.1.2.2 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
Search for more literature for 5.1.2.2

Protein Variants

Protein Variants Comment Organism
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
V26I site-directed mutagenesis, the mutant shows 2fold higher catalytic efficiency towards R-mandelamide than the wild-type enzyme Pseudomonas putida
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
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]

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

Metals/Ions

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

Organism

Organism UniProt Comment Textmining
Pseudomonas putida P11444
-
-

Substrates and Products (Substrate)

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

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
11.03
-
 (R)-mandelamide pH 7.5, 25°C, wild-type enzyme Pseudomonas putida
21.17
-
 (R)-mandelamide pH 7.5, 25°C, mutant V26L Pseudomonas putida
46
-
 (2R)-2-naphthylglycolate pH 7.5, 25°C, wild-type enzyme Pseudomonas putida
70
-
 (2R)-2-naphthylglycolate pH 7.5, 25°C, wild-type enzyme, in presence of 20 mM MgCl2 Pseudomonas putida
137
-
 (2R)-2-naphthylglycolate pH 7.5, 25°C, mutant V29L Pseudomonas putida
147
-
 (2R)-2-naphthylglycolate pH 7.5, 25°C, mutant V22I Pseudomonas putida
388
-
 (R)-3-chloromandelate pH 7.5, 25°C, mutant V29L Pseudomonas putida
469
-
 (R)-3-chloromandelate pH 7.5, 25°C, wild-type enzyme Pseudomonas putida
507
-
 (R)-3-chloromandelate pH 7.5, 25°C, mutant Y54F Pseudomonas putida
510
-
 (R)-3-chloromandelate pH 7.5, 25°C, mutant V26I/Y54V Pseudomonas putida
559
-
 (R)-3-chloromandelate pH 7.5, 25°C, mutant V26S Pseudomonas putida
565
-
 (R)-3-chloromandelate pH 7.5, 25°C, mutant Y54L Pseudomonas putida
637
-
 (R)-3-chloromandelate pH 7.5, 25°C, mutant V22I Pseudomonas putida
913
-
 (R)-3-chloromandelate pH 7.5, 25°C, mutant L319F Pseudomonas putida
953
-
 (R)-3-chloromandelate pH 7.5, 25°C, mutant Y54A Pseudomonas putida
1030
-
 (R)-3-chloromandelate pH 7.5, 25°C, mutant V26I/Y54I Pseudomonas putida
1160
-
 (R)-3-chloromandelate pH 7.5, 25°C, mutant Y54V Pseudomonas putida
1220
-
 (R)-3-chloromandelate pH 7.5, 25°C, mutant V22L Pseudomonas putida
1520
-
 (R)-3-chloromandelate pH 7.5, 25°C, mutant Y54I Pseudomonas putida
1710
-
 (R)-3-chloromandelate pH 7.5, 25°C, mutant V26L Pseudomonas putida
2020
-
 (R)-3-chloromandelate pH 7.5, 25°C, mutant V26I Pseudomonas putida

pH Optimum

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

General Information

General Information Comment Organism
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]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
9.79
-
 (R)-mandelamide pH 7.5, 25°C, wild-type enzyme Pseudomonas putida
17.1
-
 (R)-mandelamide pH 7.5, 25°C, mutant V26L Pseudomonas putida
20.1
-
 (2R)-2-naphthylglycolate pH 7.5, 25°C, wild-type enzyme, in presence of 20 mM MgCl2 Pseudomonas putida
34
-
 (2R)-2-naphthylglycolate pH 7.5, 25°C, mutant V22I Pseudomonas putida
38
-
 (2R)-2-naphthylglycolate pH 7.5, 25°C, wild-type enzyme Pseudomonas putida
85
-
 (2R)-2-naphthylglycolate pH 7.5, 25°C, mutant V29L Pseudomonas putida
249
-
 (R)-3-chloromandelate pH 7.5, 25°C, wild-type enzyme Pseudomonas putida
259
-
 (R)-3-chloromandelate pH 7.5, 25°C, mutant Y54F Pseudomonas putida
294
-
 (R)-3-chloromandelate pH 7.5, 25°C, mutant V26S Pseudomonas putida
300
-
 (R)-3-chloromandelate pH 7.5, 25°C, mutant V22I Pseudomonas putida
305
-
 (R)-3-chloromandelate pH 7.5, 25°C, mutant V29L Pseudomonas putida
349
-
 (R)-3-chloromandelate pH 7.5, 25°C, mutant Y54L Pseudomonas putida
429
-
 (R)-3-chloromandelate pH 7.5, 25°C, mutant Y54A Pseudomonas putida
502
-
 (R)-3-chloromandelate pH 7.5, 25°C, mutant Y54I Pseudomonas putida
509
-
 (R)-3-chloromandelate pH 7.5, 25°C, mutant V22L Pseudomonas putida
515
-
 (R)-3-chloromandelate pH 7.5, 25°C, mutant V26I/Y54I Pseudomonas putida
531
-
 (R)-3-chloromandelate pH 7.5, 25°C, mutant L319F Pseudomonas putida
533
-
 (R)-3-chloromandelate pH 7.5, 25°C, mutant Y54V Pseudomonas putida
821
-
 (R)-3-chloromandelate pH 7.5, 25°C, mutant V26I Pseudomonas putida
916
-
 (R)-3-chloromandelate pH 7.5, 25°C, mutant V26L Pseudomonas putida
1300
-
 (R)-3-chloromandelate pH 7.5, 25°C, mutant V26I/Y54V Pseudomonas putida