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Literature summary for 3.4.17.11 extracted from

  • Al-Qahtani, A.D.; Bashraheel, S.S.; Rashidi, F.B.; OConnor, C.D.; Romero, A.R.; Domling, A.; Goda, S.K.
    Production of "biobetter" variants of glucarpidase with enhanced enzyme activity (2019), Biomed. Pharmacother., 112, 108725 .
    View publication on PubMed
Search for more literature for 3.4.17.11

Application

Application Comment Organism
diagnostics the enzyme is used in targeted cancer strategies such as antibody-directed enzyme prodrug therapy Pseudomonas sp. RS-16
medicine the enzyme is used in drug detoxification when cancer patients have excessive levels of the anti-cancer agent methotrexate Pseudomonas sp. RS-16

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli BL21(DE3) Pseudomonas sp. RS-16

Protein Variants

Protein Variants Comment Organism
G123S the variant has increased catalytic efficiency relative to wild type enzyme. The mutant enzyme has higher alpha-helical content relative to the wild type enzyme Pseudomonas sp. RS-16
I100T the variant has increased catalytic efficiency relative to wild type enzyme. The mutant enzyme has higher alpha-helical content relative to the wild type enzyme Pseudomonas sp. RS-16
T239A the variant has increased catalytic efficiency relative to wild type enzyme. The mutant enzyme has higher alpha-helical content relative to the wild type enzyme Pseudomonas sp. RS-16

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
62.68
-
 methotrexate pH 7.3, 37°C, mutant enzyme I100T Pseudomonas sp. RS-16
71.38
-
 methotrexate pH 7.3, 37°C, mutant enzyme G123S Pseudomonas sp. RS-16
82.41
-
 methotrexate pH 7.3, 37°C, mutant enzyme T329A Pseudomonas sp. RS-16
171.7
-
 methotrexate pH 7.3, 37°C, wild-type enzyme Pseudomonas sp. RS-16

Organism

Organism UniProt Comment Textmining
Pseudomonas sp. RS-16 P06621
-
-

Purification (Commentary)

Purification (Comment) Organism
-
 Pseudomonas sp. RS-16

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
methotrexate + H2O
-
 Pseudomonas sp. RS-16 4-[(2,4-diamino-6-(pteridinyl)methyl)-methylamino]-benzoic acid + L-glutamate
-
 ?

Synonyms

Synonyms Comment Organism
carboxypeptidase G2
-
 Pseudomonas sp. RS-16
CPG2
-
 Pseudomonas sp. RS-16
glucarpidase
-
 Pseudomonas sp. RS-16

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
24.83
-
 methotrexate pH 7.3, 37°C, wild-type enzyme Pseudomonas sp. RS-16
26.11
-
 methotrexate pH 7.3, 37°C, mutant enzyme I100T Pseudomonas sp. RS-16
26.93
-
 methotrexate pH 7.3, 37°C, mutant enzyme T329A Pseudomonas sp. RS-16
27.21
-
 methotrexate pH 7.3, 37°C, mutant enzyme G123S Pseudomonas sp. RS-16