Application | Comment | Organism |
---|---|---|
biotechnology | since commercially available D-Trp is chemically synthesized and expensive a mutant BAR protein might represent an effective method to synthesize D-Trp | Pseudomonas putida |
Cloned (Comment) | Organism |
---|---|
recombinant BAR is cloned in Escherichia coli BL-21 as His-tagged BAR | Pseudomonas putida |
Protein Variants | Comment | Organism |
---|---|---|
I384M | I384M mutant BAR shows the highest activity for Trp. Using I384M mutant BAR the proportion of D-Trp reaches 43% while wildtype BAR racemizes only 6% of initial L-Trp | Pseudomonas putida |
I384M | specific activity: 0.124 (substrate Trp), 0.0744 (substrate Phe), 0.223 (substrate Lys), 0.026 (substrate Ala) | Pseudomonas putida |
I83L/D361V/Y396C | specific activity: 0.0303 (substrate Trp), 0.0257 (substrate Phe), 0.473 (substrate Lys), 0.0121 (substrate Ala) | Pseudomonas putida |
L126H/Y396C | specific activity: 0.0524 (substrate Trp), 0.0436 (substrate Phe), 0.477 (substrate Lys), 0.0144 (substrate Ala) | Pseudomonas putida |
additional information | random mutagenesis on bar gene is performed to obtain mutant BAR derivates with high activity for Trp. Five positive mutant are isolated after the two-step-screening of the randomly mutated BAR | Pseudomonas putida |
additional information | substitutions at Y396 and I384 increases the Trp specific racemization activity and the racemazation activity for overall amino acids, respectively | Pseudomonas putida |
Y293A/Y301S/Y396C | specific activity: 0.0267 (substrate Trp), 0.0249 (substrate Phe), 0.458 (substrate Lys), 0.0141 (substrate Ala) | Pseudomonas putida |
Y396C | specific activity: 0.0487 (substrate Trp), 0.00706 (substrate Phe), 0.03215 (substrate Lys), 0.825 (substrate Ala) | Pseudomonas putida |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
45000 | - |
SDS-PAGE | Pseudomonas putida |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Pseudomonas putida | - |
IFO 12996 | - |
Purification (Comment) | Organism |
---|---|
His6-tagged BAR is applied onto a His-Trap HP 1-ml column | Pseudomonas putida |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
additional information | - |
wildtype BAR, specific activity for L-Lys 100%, L-Arg 65%, L-Ala 33%, L-Ser 20%, L-Met 14%, L-Cys 14%, L-Leu 3.3%, L-His 1.6%, L-Phe 0.29%, L-Pro 0.1.3%, L-Thr 0.069%, L-Asn 0.054%, L-Asp 0.01%, L-Trp 0.006%, L-Val 0.003% | Pseudomonas putida |
0.00588 | - |
substrate Trp, wildtype BAR | Pseudomonas putida |
0.00706 | - |
substrate Phe, mutant Y396C | Pseudomonas putida |
0.00782 | - |
substrate Ala, wildtype BAR | Pseudomonas putida |
0.00838 | - |
substrate Phe, wildtype BAR | Pseudomonas putida |
0.0121 | - |
substrate Ala, mutant I83L, D361V, Y396C | Pseudomonas putida |
0.0141 | - |
substrate Ala, mutant Y293A, Y301S, Y396C | Pseudomonas putida |
0.0144 | - |
substrate Ala, mutant L126H, Y396C | Pseudomonas putida |
0.0249 | - |
substrate Phe, mutant Y293A, Y301S, Y396C | Pseudomonas putida |
0.0257 | - |
substrate Phe, mutant I83L, D361V, Y396C | Pseudomonas putida |
0.026 | - |
substrate Ala, mutant I384M | Pseudomonas putida |
0.0267 | - |
substrate Trp, mutant Y293A, Y301S, Y396C | Pseudomonas putida |
0.0303 | - |
substrate Trp, mutant I83L, D361V, Y396C | Pseudomonas putida |
0.03215 | - |
substrate Lys, mutant Y396C | Pseudomonas putida |
0.0436 | - |
substrate Phe, mutant L126H, Y396C | Pseudomonas putida |
0.0487 | - |
substrate Trp, mutant Y396C | Pseudomonas putida |
0.0524 | - |
substrate Trp, mutant L126H, Y396C | Pseudomonas putida |
0.0744 | - |
substrate Phe, mutant I384M | Pseudomonas putida |
0.124 | - |
substrate Trp, mutant I384M | Pseudomonas putida |
0.132 | - |
substrate Lys, wildtype BAR | Pseudomonas putida |
0.223 | - |
substrate Lys, mutant I384M | Pseudomonas putida |
0.458 | - |
substrate Lys, mutant Y293A, Y301S, Y396C | Pseudomonas putida |
0.473 | - |
substrate Lys, mutant I83L, D361V, Y396C | Pseudomonas putida |
0.477 | - |
substrate Lys, mutant L126H, Y396C | Pseudomonas putida |
0.825 | - |
substrate Ala, mutant Y396C | Pseudomonas putida |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
L-Trp | - |
Pseudomonas putida | D-Trp | - |
r |
Synonyms | Comment | Organism |
---|---|---|
broad specificity amino acid racemase | - |
Pseudomonas putida |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Pseudomonas putida |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8.5 | - |
assay at | Pseudomonas putida |