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Literature summary for 3.5.1.93 extracted from

  • Huang, X.; Zeng, R.; Ding, X.; Mao, X.; Ding, Y.; Rao, Z.; Xie, Y.; Jiang, W.; Zhao, G.
    Affinity alkylation of the Trp-B4 residue of the beta -subunit of the glutaryl 7-aminocephalosporanic acid acylase of Pseudomonas sp. 130 (2002), J. Biol. Chem., 277, 10256-10264.
    View publication on PubMed
Search for more literature for 3.5.1.93

Protein Variants

Protein Variants Comment Organism
W4A beta-subunit mutant, KM-value for glutaryl-7-aminocephalosporanic acid is 1.6fold higher than wild-type value, turnover number for glutaryl-7-aminocephalosporanic acid is 2.2fold lower than wild-type value, t1/2 at 37°C is 9.8fold lower than wild-type value Pseudomonas sp. 130
W4F beta-subunit mutant, KM-value for glutaryl-7-aminocephalosporanic acid is 1.8fold higher than wild-type value, turnover number for glutaryl-7-aminocephalosporanic acid is 1.6fold lower than wild-type value, t1/2 at 37°C is 37.5fold lower than wild-type value Pseudomonas sp. 130
W4H beta-subunit mutant, KM-value for glutaryl-7-aminocephalosporanic acid is 1.9fold higher than wild-type value, turnover number for glutaryl-7-aminocephalosporanic acid is 1.4fold lower than wild-type value, t1/2 at 37°C is 1.9fold lower than wild-type value Pseudomonas sp. 130
W4L beta-subunit mutant, KM-value for glutaryl-7-aminocephalosporanic acid is 1.8fold higher than wild-type value, turnover number for glutaryl-7-aminocephalosporanic acid is 2.1fold lower than wild-type value, t1/2 at 37°C is 6.1fold lower than wild-type value Pseudomonas sp. 130
W4T beta-subunit mutant, KM-value for glutaryl-7-aminocephalosporanic acid is 1.7fold higher than wild-type value, turnover number for glutaryl-7-aminocephalosporanic acid is 2.4fold lower than wild-type value, t1/2 at 37°C is 50fold lower than wild-type value Pseudomonas sp. 130
W4Y beta-subunit mutant, KM-value for glutaryl-7-aminocephalosporanic acid is 1.6fold higher than wild-type value, turnover number for glutaryl-7-aminocephalosporanic acid is 1.3fold lower than wild-type value, t1/2 at 37°C is 12fold lower than wild-type value Pseudomonas sp. 130

Inhibitors

Inhibitors Comment Organism Structure
7beta-3-bromopropionyl aminocephalosporanic acid Trp4 of the beta-subunit is alkylated Pseudomonas sp. 130
7beta-bromoacetyl aminocephalosporanic acid Trp4 of the beta-subunit is alkylated Pseudomonas sp. 130

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.21
-
 glutaryl-7-aminocephalosporanic acid pH 8.0, 37°C, wild-type enzyme Pseudomonas sp. 130
0.33
-
 glutaryl-7-aminocephalosporanic acid pH 8.0, 37°C, beta-subunit mutant W4A Pseudomonas sp. 130
0.33
-
 glutaryl-7-aminocephalosporanic acid pH 8.0, 37°C, beta-subunit mutant W4Y Pseudomonas sp. 130
0.36
-
 glutaryl-7-aminocephalosporanic acid pH 8.0, 37°C, beta-subunit mutant W4T Pseudomonas sp. 130
0.37
-
 glutaryl-7-aminocephalosporanic acid pH 8.0, 37°C, beta-subunit mutant W4F Pseudomonas sp. 130
0.37
-
 glutaryl-7-aminocephalosporanic acid pH 8.0, 37°C, beta-subunit mutant W4L Pseudomonas sp. 130
0.4
-
 glutaryl-7-aminocephalosporanic acid pH 8.0, 37°C, beta-subunit mutant W4H Pseudomonas sp. 130

Organism

Organism UniProt Comment Textmining
Pseudomonas sp. 130
-
-
-

Purification (Commentary)

Purification (Comment) Organism
-
 Pseudomonas sp. 130

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
glutaryl-7-aminocephalosporanic acid + H2O
-
 Pseudomonas sp. 130 7-aminocephalosporanate + glutarate
-
 ?

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
37
-
 pH 8.0, t1/2: 600 min for wild-type enzyme, 51 min for beta-subunit mutant enzyme W4A, 98 min for beta-subunit mutant enzyme W4L, 12 min for beta-subunit mutant enzyme W4T, 16 min for beta-subunit mutant enzyme W4F, 307 min for beta-subunit mutant enzyme W4H, 50 min for beta-subunit mutant enzyme W4Y Pseudomonas sp. 130

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.3
-
 glutaryl-7-aminocephalosporanic acid pH 8.0, 37°C, beta-subunit mutant W4T Pseudomonas sp. 130
3.6
-
 glutaryl-7-aminocephalosporanic acid pH 8.0, 37°C, beta-subunit mutant W4A Pseudomonas sp. 130
3.8
-
 glutaryl-7-aminocephalosporanic acid pH 8.0, 37°C, beta-subunit mutant W4L Pseudomonas sp. 130
5
-
 glutaryl-7-aminocephalosporanic acid pH 8.0, 37°C, beta-subunit mutant W4F Pseudomonas sp. 130
5.5
-
 glutaryl-7-aminocephalosporanic acid pH 8.0, 37°C, beta-subunit mutant W4H Pseudomonas sp. 130
5.9
-
 glutaryl-7-aminocephalosporanic acid pH 8.0, 37°C, beta-subunit mutant W4Y Pseudomonas sp. 130
7.8
-
 glutaryl-7-aminocephalosporanic acid pH 8.0, 37°C, wild-type enzyme Pseudomonas sp. 130

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
0.87
-
 7beta-bromoacetyl aminocephalosporanic acid pH 8.0, 37°C, beta-subunit mutant W4Y Pseudomonas sp. 130
5.4
-
 7beta-bromoacetyl aminocephalosporanic acid pH 8.0, 37°C, beta-subunit mutant W4H Pseudomonas sp. 130
5.4
-
 7beta-bromoacetyl aminocephalosporanic acid pH 8.0, 37°C, beta-subunit mutant W4T Pseudomonas sp. 130
5.5
-
 7beta-bromoacetyl aminocephalosporanic acid pH 8.0, 37°C, wild-type enzyme Pseudomonas sp. 130
6.3
-
 7beta-bromoacetyl aminocephalosporanic acid pH 8.0, 37°C, beta-subunit mutant W4F Pseudomonas sp. 130
6.4
-
 7beta-bromoacetyl aminocephalosporanic acid pH 8.0, 37°C, beta-subunit mutant W4A Pseudomonas sp. 130
6.8
-
 7beta-bromoacetyl aminocephalosporanic acid pH 8.0, 37°C, beta-subunit mutant W4L Pseudomonas sp. 130
14
-
 7beta-3-bromopropionyl aminocephalosporanic acid pH 8.0, 37°C, wild-type enzyme Pseudomonas sp. 130
20
-
 7beta-3-bromopropionyl aminocephalosporanic acid pH 8.0, 37°C, beta-subunit mutant W4Y Pseudomonas sp. 130