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Literature summary for 3.4.21.4 extracted from

  • Page, M.J.; Wong, S.L.; Hewitt, J.; Strynadka, N.C.; MacGillivray, R.T.
    Engineering the primary substrate specificity of Streptomyces griseus trypsin (2003), Biochemistry, 42, 9060-9066.
    View publication on PubMed
Search for more literature for 3.4.21.4

Cloned(Commentary)

Cloned (Comment) Organism
expression in Bacillus subtilis in a soluble active form Streptomyces griseus

Crystallization (Commentary)

Crystallization (Comment) Organism
hanging drop vapour diffusion method, recombinant wild-type enzyme and mutant enzyme T190P at 1.5 and 1.9 A resolution Streptomyces griseus

Protein Variants

Protein Variants Comment Organism
T190A the ratio of turnover number to Km-value with tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin is 47% of that of the wild-type enzyme, the ratio of turnover number to Km-value with tosyl-Gly-Pro-Lys-7-amido-4-methylcoumarin is 55% of that of the wild-type enzyme Streptomyces griseus
T190P the ratio of turnover number to Km-value with tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin is 0.5% of that of the wild-type enzyme, the ratio of turnover number to Km-value with tosyl-Gly-Pro-Lys-7-amido-4-methylcoumarin is 0.7% of that of the wild-type enzyme Streptomyces griseus
T190S the ratio of turnover number to Km-value with tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin is 18% of that of the wild-type enzyme, the ratio of turnover number to Km-value with tosyl-Gly-Pro-Lys-7-amido-4-methylcoumarin is 9% of that of the wild-type enzyme Streptomyces griseus
T190V the ratio of turnover number to Km-value with tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin is 2.5% of that of the wild-type enzyme, the ratio of turnover number to Km-value with tosyl-Gly-Pro-Lys-7-amido-4-methylcoumarin is 0.6% of that of the wild-type enzyme Streptomyces griseus

Inhibitors

Inhibitors Comment Organism Structure
benzamidine
-
 Streptomyces griseus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.002
-
 tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin pH 7.6, recombinant enzyme Streptomyces griseus
0.0023
-
 tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin pH 7.6, wild-type enzyme Streptomyces griseus
0.0032
-
 tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin pH 7.6, recombinant enzyme Streptomyces griseus
0.0036
-
 tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin pH 7.6, wild-type enzyme Streptomyces griseus
0.0044
-
 tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin pH 7.6, mutant enzyme T190A Streptomyces griseus
0.0061
-
 tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin pH 7.6, mutant enzyme T190S Streptomyces griseus
0.0101
-
 tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin pH 7.6, mutant enzyme T190S Streptomyces griseus
0.0129
-
 tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin pH 7.6, mutant enzyme T190A Streptomyces griseus
0.067
-
 tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin pH 7.6, mutant enzyme T190P Streptomyces griseus
0.166
-
 tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin pH 7.6, mutant enzyme T190P Streptomyces griseus
0.224
-
 tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin pH 7.6, mutant enzyme T190V Streptomyces griseus
0.231
-
 tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin pH 7.6, mutant enzyme T190V Streptomyces griseus

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
23080
-
 mutant enzyme T190A, electrospray mass spectrometry analysis Streptomyces griseus
23090
-
 mutant enzyme T190S, electrospray mass spectrometry analysis Streptomyces griseus
23100
-
 mutant enzyme T190P, electrospray mass spectrometry analysis Streptomyces griseus
23100
-
 mutant enzyme T190V, electrospray mass spectrometry analysis Streptomyces griseus
23110
-
 wild-type enzyme and recombinant enzyme, electrospray mass spectrometry analysis Streptomyces griseus

Organism

Organism UniProt Comment Textmining
Streptomyces griseus P00775
-
-

Purification (Commentary)

Purification (Comment) Organism
-
 Streptomyces griseus

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
237300
-
-
 Streptomyces griseus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin + H2O
-
 Streptomyces griseus ?
-
 ?
tosyl-Gly-Pro-Lys-7-amido-4-methylcoumarin + H2O
-
 Streptomyces griseus ?
-
 ?

Synonyms

Synonyms Comment Organism
SGT
-
 Streptomyces griseus

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.04 1.97 tosyl-Gly-Pro-Lys-7-amido-4-methylcoumarin pH 7.6, mutant enzyme T190P Streptomyces griseus
0.5
-
 tosyl-Gly-Pro-Lys-7-amido-4-methylcoumarin pH 7.6, mutant enzyme T190V Streptomyces griseus
0.52 0.7 tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin pH 7.6, wild-type enzyme Streptomyces griseus
2.7
-
 tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin pH 7.6, recombinant enzyme Streptomyces griseus
3.2
-
 tosyl-Gly-Pro-Lys-7-amido-4-methylcoumarin pH 7.6, mutant enzyme T190A Streptomyces griseus
8
-
 tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin pH 7.6, mutant enzyme T190P Streptomyces griseus
8.78
-
 tosyl-Gly-Pro-Lys-7-amido-4-methylcoumarin pH 7.6, mutant enzyme T190P Streptomyces griseus
38.33
-
 tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin pH 7.6, mutant enzyme T190V Streptomyces griseus
82.5
-
 tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin pH 7.6, mutant enzyme T190A Streptomyces griseus

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
0.0027
-
 benzamidine pH 7.6, recombinant enzyme Streptomyces griseus
0.0048
-
 benzamidine pH 7.6, mutant enzyme T190S Streptomyces griseus
0.0099
-
 benzamidine pH 7.6, mutant enzyme T190A Streptomyces griseus
0.0164
-
 benzamidine pH 7.6, mutant enzyme T190P Streptomyces griseus
0.197
-
 benzamidine pH 7.6, mutant enzyme T190V Streptomyces griseus