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Literature summary extracted from

  • Pollegioni, L.; Lorenzi, S.; Rosini, E.; Marcone, G.L.; Molla, G.; Verga, R.; Cabri, W.; Pilone, M.S.
    Evolution of an acylase active on cephalosporin C (2005), Protein Sci., 14, 3064-3076.
    View publication on PubMedView publication on EuropePMC

Application

EC Number Application Comment Organism
3.5.1.93 synthesis bioconversion of cephalosporin C into 7-aminocephalosporanate Pseudomonas sp.

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.5.1.93 expression of mutant enzymes in Escherichia coli Pseudomonas sp.

Protein Variants

EC Number Protein Variants Comment Organism
3.5.1.93 A215E Vmax/Km for cephalosporin C is 1.2fold lower than wild-type value, Vmax/Km for glutaryl-7-amino cephalosporanic acid is 7.6fold lower than wild-type value Pseudomonas sp.
3.5.1.93 A215F Vmax/Km for cephalosporin C is 2fold lower than wild-type value, Vmax/Km for glutaryl-7-amino cephalosporanic acid is 3.9fold lower than wild-type value Pseudomonas sp.
3.5.1.93 A215L Vmax/Km for cephalosporin C is 1.3fold higher than wild-type value, Vmax/Km for glutaryl-7-amino cephalosporanic acid is 4.2fold lower than wild-type value Pseudomonas sp.
3.5.1.93 A215V Vmax/Km for cephalosporin C is 1.5fold higher than wild-type value, Vmax/Km for glutaryl-7-amino cephalosporanic acid is 1.5fold lower than wild-type value Pseudomonas sp.
3.5.1.93 A215Y Vmax/Km for cephalosporin C is 4.3fold higher than wild-type value, Vmax/Km for glutaryl-7-amino cephalosporanic acid is 1.6fold lower than wild-type value Pseudomonas sp.
3.5.1.93 A215Y/H296S Vmax/Km for cephalosporin C is 1.2fold lower than wild-type value, Vmax/Km for glutaryl-7-amino cephalosporanic acid is 25.1fold lower than wild-type value Pseudomonas sp.
3.5.1.93 A215Y/H296S/H309S Vmax/Km for cephalosporin C is 3.5fold lower than wild-type value, Vmax/Km for glutaryl-7-amino cephalosporanic acid is 13.7fold lower than wild-type value Pseudomonas sp.
3.5.1.93 A215Y/H309S Vmax/Km for cephalosporin C is 3.8fold higher than wild-type value, Vmax/Km for glutaryl-7-amino cephalosporanic acid is 2.9fold higher than wild-type value Pseudomonas sp.
3.5.1.93 D416Y Vmax/Km for cephalosporin C is 1.5fold higher than wild-type value, Vmax/Km for glutaryl-7-amino cephalosporanic acid is 5.2fold lower than wild-type value Pseudomonas sp.
3.5.1.93 D416Y/H417Y Vmax/Km for cephalosporin C is 5.3fold higher than wild-type value, Vmax/Km for glutaryl-7-amino cephalosporanic acid is 151fold lower than wild-type value Pseudomonas sp.
3.5.1.93 E89A/A215Y Vmax/Km for cephalosporin C is 6fold lower than wild-type value, Vmax/Km for glutaryl-7-amino cephalosporanic acid is 25.2fold lower than wild-type value Pseudomonas sp.
3.5.1.93 F270M Vmax/Km for cephalosporin C is 1.8fold higher than wild-type value, Vmax/Km for glutaryl-7-amino cephalosporanic acid is 1.7fold lower than wild-type value Pseudomonas sp.
3.5.1.93 H296F Vmax/Km for cephalosporin C is 6fold lower than wild-type value, Vmax/Km for glutaryl-7-amino cephalosporanic acid is 50.3fold lower than wild-type value Pseudomonas sp.
3.5.1.93 H296N Vmax/Km for cephalosporin C is 1.2fold lower than wild-type value, Vmax/Km for glutaryl-7-amino cephalosporanic acid is fold lower than wild-type value Pseudomonas sp.
3.5.1.93 H296S Vmax/Km for cephalosporin C is 1.7fold higher than wild-type value, Vmax/Km for glutaryl-7-amino cephalosporanic acid is 10.1fold lower than wild-type value Pseudomonas sp.
3.5.1.93 H296S/H309S Vmax/Km for cephalosporin C is 4fold higher than wild-type value, Vmax/Km for glutaryl-7-amino cephalosporanic acid is 21.6fold lower than wild-type value Pseudomonas sp.
3.5.1.93 H296S/H417M Vmax/Km for cephalosporin C is 2fold lower than wild-type value, Vmax/Km for glutaryl-7-amino cephalosporanic acid is 151fold lower than wild-type value Pseudomonas sp.
3.5.1.93 H296T Vmax/Km for cephalosporin C is 1.5fold lower than wild-type value, Vmax/Km for glutaryl-7-amino cephalosporanic acid is 5.2fold lower than wild-type value Pseudomonas sp.
3.5.1.93 H309S Vmax/Km for cephalosporin C is 1.5fold higher than wild-type value, Vmax/Km for glutaryl-7-amino cephalosporanic acid is 8.9fold lower than wild-type value Pseudomonas sp.
3.5.1.93 H309Y Vmax/Km for cephalosporin C is 1.5fold lower than wild-type value, Vmax/Km for glutaryl-7-amino cephalosporanic acid is 16.8fold lower than wild-type value Pseudomonas sp.
3.5.1.93 H417Y Vmax/Km for cephalosporin C is more than 6fold lower than wild-type value, Vmax/Km for glutaryl-7-amino cephalosporanic acid is 4.4fold lower than wild-type value Pseudomonas sp.
3.5.1.93 I44V/E49stop/D416Y/H417Y Vmax/Km for cephalosporin C is more than 6fold lower than wild-type value, Vmax/Km for glutaryl-7-amino cephalosporanic acid is more than 1510fold lower than wild-type value Pseudomonas sp.
3.5.1.93 S22P/T394P/D416Y/H417Y Vmax/Km for cephalosporin C is more than 6fold lower than wild-type value, Vmax/Km for glutaryl-7-amino cephalosporanic acid is more than 1510fold lower than wild-type value Pseudomonas sp.
3.5.1.93 Y215Y/F270S Vmax/Km for cephalosporin C is 2.5fold higher than wild-type value, Vmax/Km for glutaryl-7-amino cephalosporanic acid is 1.2fold lower than wild-type value Pseudomonas sp.
3.5.1.93 Y271F Vmax/Km for cephalosporin C is 1.5fold lower than wild-type value, Vmax/Km for glutaryl-7-amino cephalosporanic acid is 5.4fold lower than wild-type value Pseudomonas sp.
3.5.1.93 Y27V Vmax/Km for cephalosporin C is 1.5fold lower than wild-type value, Vmax/Km for glutaryl-7-amino cephalosporanic acid is 2.1fold lower than wild-type value Pseudomonas sp.

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.5.1.93 0.7
-
 glutaryl-7-amino cephalosporanic acid pH 8.0, 37°C, mutant enzyme H296S Pseudomonas sp.
3.5.1.93 0.9
-
 glutaryl-7-amino cephalosporanic acid pH 8.0, 37°C, mutant enzyme A215F Pseudomonas sp.
3.5.1.93 0.9
-
 glutaryl-7-amino cephalosporanic acid pH 8.0, 37°C, mutant enzyme A215Y/H309S Pseudomonas sp.
3.5.1.93 1
-
 glutaryl-7-amino cephalosporanic acid pH 8.0, 37°C, mutant enzyme A215E Pseudomonas sp.
3.5.1.93 1
-
 glutaryl-7-amino cephalosporanic acid pH 8.0, 37°C, mutant enzyme H309Y Pseudomonas sp.
3.5.1.93 1.1
-
 glutaryl-7-amino cephalosporanic acid pH 8.0, 37°C, mutant enzyme E89A/A215Y Pseudomonas sp.
3.5.1.93 1.2
-
 glutaryl-7-amino cephalosporanic acid pH 8.0, 37°C, mutant enzyme A215V Pseudomonas sp.
3.5.1.93 1.2
-
 glutaryl-7-amino cephalosporanic acid pH 8.0, 37°C, mutant enzyme Y271F Pseudomonas sp.
3.5.1.93 1.6
-
 glutaryl-7-amino cephalosporanic acid pH 8.0, 37°C, wild-type enzyme Pseudomonas sp.
3.5.1.93 1.7
-
 glutaryl-7-amino cephalosporanic acid pH 8.0, 37°C, mutant enzyme A215L Pseudomonas sp.
3.5.1.93 1.7
-
 glutaryl-7-amino cephalosporanic acid pH 8.0, 37°C, mutant enzyme A215Y Pseudomonas sp.
3.5.1.93 1.7
-
 glutaryl-7-amino cephalosporanic acid pH 8.0, 37°C, mutant enzyme D416Y Pseudomonas sp.
3.5.1.93 1.7
-
 cephalosporin C pH 8.0, 37°C, mutant enzyme H296S/H417M Pseudomonas sp.
3.5.1.93 1.9
-
 glutaryl-7-amino cephalosporanic acid pH 8.0, 37°C, mutant enzyme H296F Pseudomonas sp.
3.5.1.93 1.9
-
 glutaryl-7-amino cephalosporanic acid pH 8.0, 37°C, mutant enzyme Y27V Pseudomonas sp.
3.5.1.93 2
-
 glutaryl-7-amino cephalosporanic acid pH 8.0, 37°C, mutant enzyme H296T Pseudomonas sp.
3.5.1.93 2
-
 glutaryl-7-amino cephalosporanic acid pH 8.0, 37°C, mutant enzyme Y215Y/F270S Pseudomonas sp.
3.5.1.93 2.5
-
 glutaryl-7-amino cephalosporanic acid pH 8.0, 37°C, mutant enzyme A215Y/H296S/H309S Pseudomonas sp.
3.5.1.93 2.7
-
 glutaryl-7-amino cephalosporanic acid pH 8.0, 37°C, mutant enzyme F270M Pseudomonas sp.
3.5.1.93 2.7
-
 cephalosporin C pH 8.0, 37°C, mutant enzyme Y271V Pseudomonas sp.
3.5.1.93 2.8
-
 glutaryl-7-amino cephalosporanic acid pH 8.0, 37°C, mutant enzyme H296N Pseudomonas sp.
3.5.1.93 3.2
-
 cephalosporin C pH 8.0, 37°C, mutant enzyme A215E Pseudomonas sp.
3.5.1.93 4
-
 glutaryl-7-amino cephalosporanic acid pH 8.0, 37°C, mutant enzyme A215Y/H296S Pseudomonas sp.
3.5.1.93 4.1
-
 cephalosporin C pH 8.0, 37°C, mutant enzyme H296F Pseudomonas sp.
3.5.1.93 4.3
-
 glutaryl-7-amino cephalosporanic acid pH 8.0, 37°C, mutant enzyme H417Y Pseudomonas sp.
3.5.1.93 4.6
-
 glutaryl-7-amino cephalosporanic acid pH 8.0, 37°C, mutant enzyme H309S Pseudomonas sp.
3.5.1.93 4.8
-
 cephalosporin C pH 8.0, 37°C, mutant enzyme H296T Pseudomonas sp.
3.5.1.93 5.2
-
 cephalosporin C pH 8.0, 37°C, mutant enzyme H296N Pseudomonas sp.
3.5.1.93 5.5
-
 glutaryl-7-amino cephalosporanic acid pH 8.0, 37°C, mutant enzyme H296S/H417M Pseudomonas sp.
3.5.1.93 6.3
-
 cephalosporin C pH 8.0, 37°C, mutant enzyme H309Y Pseudomonas sp.
3.5.1.93 6.5
-
 cephalosporin C pH 8.0, 37°C, mutant enzyme F270M Pseudomonas sp.
3.5.1.93 6.7
-
 cephalosporin C pH 8.0, 37°C, mutant enzyme H296S Pseudomonas sp.
3.5.1.93 6.9
-
 cephalosporin C pH 8.0, 37°C, mutant enzyme A215Y Pseudomonas sp.
3.5.1.93 6.9
-
 glutaryl-7-amino cephalosporanic acid pH 8.0, 37°C, mutant enzyme H296S/H309S Pseudomonas sp.
3.5.1.93 7.2
-
 cephalosporin C pH 8.0, 37°C, mutant enzyme D416Y Pseudomonas sp.
3.5.1.93 7.7
-
 cephalosporin C pH 8.0, 37°C, mutant enzyme A215F Pseudomonas sp.
3.5.1.93 8
-
 cephalosporin C pH 8.0, 37°C, mutant enzyme Y271F Pseudomonas sp.
3.5.1.93 8.3
-
 cephalosporin C pH 8.0, 37°C, mutant enzyme A215V Pseudomonas sp.
3.5.1.93 9
-
 glutaryl-7-amino cephalosporanic acid pH 8.0, 37°C, mutant enzyme D416Y/H417Y Pseudomonas sp.
3.5.1.93 9.5
-
 cephalosporin C pH 8.0, 37°C, mutant enzyme A215L Pseudomonas sp.
3.5.1.93 10
-
 cephalosporin C pH 8.0, 37°C, mutant enzyme A215Y/H296S Pseudomonas sp.
3.5.1.93 10.1
-
 cephalosporin C pH 8.0, 37°C, mutant enzyme Y215Y/F270S Pseudomonas sp.
3.5.1.93 11
-
 cephalosporin C pH 8.0, 37°C, wild-type enzyme Pseudomonas sp.
3.5.1.93 12.2
-
 cephalosporin C pH 8.0, 37°C, mutant enzyme H296S/H309S Pseudomonas sp.
3.5.1.93 12.3
-
 cephalosporin C pH 8.0, 37°C, mutant enzyme H417Y Pseudomonas sp.
3.5.1.93 13
-
 cephalosporin C pH 8.0, 37°C, mutant enzyme D416Y/H417Y Pseudomonas sp.
3.5.1.93 17.2
-
 cephalosporin C pH 8.0, 37°C, mutant enzyme A215Y/H309S Pseudomonas sp.
3.5.1.93 17.8
-
 cephalosporin C pH 8.0, 37°C, mutant enzyme A215Y/H296S/H309S Pseudomonas sp.
3.5.1.93 18.9
-
 cephalosporin C pH 8.0, 37°C, mutant enzyme E89A/A215Y Pseudomonas sp.
3.5.1.93 24.4
-
 cephalosporin C pH 8.0, 37°C, mutant enzyme H309S Pseudomonas sp.
3.5.1.93 24.5
-
 cephalosporin C pH 8.0, 37°C, mutant enzyme S22P/T394P/D416Y/H417Y Pseudomonas sp.
3.5.1.93 25
-
 glutaryl-7-amino cephalosporanic acid pH 8.0, 37°C, mutant enzyme I44V/E49stop/D416Y/H417Y Pseudomonas sp.
3.5.1.93 32
-
 cephalosporin C pH 8.0, 37°C, mutant enzyme I44V/E49stop/D416Y/H417Y Pseudomonas sp.
3.5.1.93 37.2
-
 glutaryl-7-amino cephalosporanic acid pH 8.0, 37°C, mutant enzyme S22P/T394P/D416Y/H417Y Pseudomonas sp.

Organism

EC Number Organism UniProt Comment Textmining
3.5.1.93 Pseudomonas sp.
-
 N176
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.5.1.93 cephalosporin C + H2O
-
 Pseudomonas sp. cephalosporanic acid + 2-amino-5-hydroxypentanoate
-
 ?
3.5.1.93 glutaryl-7-amino cephalosporanic acid + H2O
-
 Pseudomonas sp. 7-aminocephalosporanate + glutarate
-
 ?