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Literature summary for 3.5.3.6 extracted from

  • Zhu, L.; Tee, K.L.; Roccatano, D.; Sonmez, B.; Ni, Y.; Sun, Z.H.; Schwaneberg, U.
    Directed evolution of an antitumor drug (arginine deiminase PpADI) for increased activity at physiological pH (2010), ChemBioChem, 11, 691-697.
    View publication on PubMed
Search for more literature for 3.5.3.6

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia BL21-Gold (DE3) cells Pseudomonas plecoglossicida

Protein Variants

Protein Variants Comment Organism
H404R the mutant shows higher activity (1.8fold) and improved activity ratio at pH 7.4 to 6.4 (4.7fold) compared to the wild type ADI Pseudomonas plecoglossicida
K5T/D44E/H404R at pH 7.4, the mutant has a 4times higher kcat value than the wild type ADI and retains about 50% of its activity relative to its pH optimum (pH 7.0), compared to about 10% in the case of the wild type ADI Pseudomonas plecoglossicida

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.7
-
 L-arginine wild type enzyme, in 0.5 M phosphate buffer, pH 7.4, at 37°C Pseudomonas plecoglossicida
1.2
-
 L-arginine mutant enzyme H404R, in 0.5 M phosphate buffer, pH 7.4, at 37°C Pseudomonas plecoglossicida
2.5
-
 L-arginine mutant enzyme K5T/D44E/H404R, in 0.5 M phosphate buffer, pH 7.4, at 37°C Pseudomonas plecoglossicida

Organism

Organism UniProt Comment Textmining
Pseudomonas plecoglossicida
-
-
-

Purification (Commentary)

Purification (Comment) Organism
Super-Q column chromatography and Toyoperl HW-55S gel filtration Pseudomonas plecoglossicida

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
L-arginine + H2O
-
 Pseudomonas plecoglossicida L-citrulline + NH3
-
 ?

Synonyms

Synonyms Comment Organism
ADI
-
 Pseudomonas plecoglossicida

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
5.6
-
 L-arginine wild type enzyme, in 0.5 M phosphate buffer, pH 7.4, at 37°C Pseudomonas plecoglossicida
8.1
-
 L-arginine mutant enzyme H404R, in 0.5 M phosphate buffer, pH 7.4, at 37°C Pseudomonas plecoglossicida
22.5
-
 L-arginine mutant enzyme K5T/D44E/H404R, in 0.5 M phosphate buffer, pH 7.4, at 37°C Pseudomonas plecoglossicida

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
6.5
-
-
 Pseudomonas plecoglossicida

pH Range

pH Minimum pH Maximum Comment Organism
6.5 7.5 a pH shift from 6.5 to 7.5 results in an approximately 80% drop in activity Pseudomonas plecoglossicida

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
6.73
-
 L-arginine mutant enzyme H404R, in 0.5 M phosphate buffer, pH 7.4, at 37°C Pseudomonas plecoglossicida
7.48
-
 L-arginine wild type enzyme, in 0.5 M phosphate buffer, pH 7.4, at 37°C Pseudomonas plecoglossicida
9.05
-
 L-arginine mutant enzyme K5T/D44E/H404R, in 0.5 M phosphate buffer, pH 7.4, at 37°C Pseudomonas plecoglossicida