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Literature summary extracted from

  • Otten, L.G.; Sio, C.F.; van der Sloot, A.M.; Cool, R.H.; Quax, W.J.
    Mutational analysis of a key residue in the substrate specificity of a cephalosporin acylase (2004), Chembiochem, 5, 820-825.
    View publication on PubMed

Protein Variants

EC Number Protein Variants Comment Organism
3.5.1.93 N266A turnover number for adipyl-7-aminodeacetoxycephalosporanic acid is 2.3fold lower than wild-type value, KM-value for adipyl-7-aminodeacetoxycephalosporanic acid is 1.8fold higher than wild-type value, turnover-number for glutaryl-7-aminocephalosporanic acid is 2.9fold lower than wild-type value, KM-value for glutaryl-7-aminocephalosporanic acid is 8.4fold higher than wild-type value Pseudomonas sp. SY-77-1
3.5.1.93 N266C turnover number for adipyl-7-aminodeacetoxycephalosporanic acid is 1.3fold lower than wild-type value, KM-value for adipyl-7-aminodeacetoxycephalosporanic acid is 1.5fold lower than wild-type value, turnover-number for glutaryl-7-aminocephalosporanic acid is 2fold lower than wild-type value, KM-value for glutaryl-7-aminocephalosporanic acid is 5.2fold higher than wild-type value Pseudomonas sp. SY-77-1
3.5.1.93 N266D turnover number for adipyl-7-aminodeacetoxycephalosporanic acid is 205fold lower than wild-type value, KM-value for adipyl-7-aminodeacetoxycephalosporanic acid is 1.8fold higher than wild-type value. Less efficient cleavage of the spacer from the alpha-subunit compared to wild-type enzyme Pseudomonas sp. SY-77-1
3.5.1.93 N266E turnover number for adipyl-7-aminodeacetoxycephalosporanic acid is 17.8fold lower than wild-type value, KM-value for adipyl-7-aminodeacetoxycephalosporanic acid is 11.5fold higher than wild-type value, turnover-number for glutaryl-7-aminocephalosporanic acid is 160fold lower than wild-type value, KM-value for glutaryl-7-aminocephalosporanic acid is 9.4fold higher than wild-type value. Less efficient cleavage of the spacer from the alpha-subunit compared to wild-type enzyme Pseudomonas sp. SY-77-1
3.5.1.93 N266F turnover number for adipyl-7-aminodeacetoxycephalosporanic acid is 1.1fold lower than wild-type value, KM-value for adipyl-7-aminodeacetoxycephalosporanic acid is 2fold lower than wild-type value, turnover-number for glutaryl-7-aminocephalosporanic acid is 4.4fold lower than wild-type value, KM-value for glutaryl-7-aminocephalosporanic acid is 3.2fold higher than wild-type value Pseudomonas sp. SY-77-1
3.5.1.93 N266G turnover number for adipyl-7-aminodeacetoxycephalosporanic acid is 1.8fold lower than wild-type value, KM-value for adipyl-7-aminodeacetoxycephalosporanic acid is 1.2fold higher than wild-type value, turnover-number for glutaryl-7-aminocephalosporanic acid is 2fold lower than wild-type value, KM-value for glutaryl-7-aminocephalosporanic acid is 17.4fold higher than wild-type value Pseudomonas sp. SY-77-1
3.5.1.93 N266H turnover number for adipyl-7-aminodeacetoxycephalosporanic acid is 1.15fold higher than wild-type value, KM-value for adipyl-7-aminodeacetoxycephalosporanic acid is 8.6fold lower than wild-type value, turnover-number for glutaryl-7-aminocephalosporanic acid is 1.3fold lower than wild-type value, KM-value for glutaryl-7-aminocephalosporanic acid is 1.4fold higher than wild-type value. 100% improved conversion of cephalosporin C compared to wild type enzyme Pseudomonas sp. SY-77-1
3.5.1.93 N266L turnover number for adipyl-7-aminodeacetoxycephalosporanic acid is 1.64fold lower than wild-type value, KM-value for adipyl-7-aminodeacetoxycephalosporanic acid is 1.2fold lower than wild-type value, turnover-number for glutaryl-7-aminocephalosporanic acid is 48.8fold lower than wild-type value, KM-value for glutaryl-7-aminocephalosporanic acid is 13.9fold higher than wild-type value Pseudomonas sp. SY-77-1
3.5.1.93 N266M turnover number for adipyl-7-aminodeacetoxycephalosporanic acid is 4.6fold higher than wild-type value, KM-value for adipyl-7-aminodeacetoxycephalosporanic acid is 3.6fold lower than wild-type value, turnover-number for glutaryl-7-aminocephalosporanic acid is 2.2fold lower than wild-type value, KM-value for glutaryl-7-aminocephalosporanic acid is 5.4fold higher than wild-type value Pseudomonas sp. SY-77-1
3.5.1.93 N266P turnover number for adipyl-7-aminodeacetoxycephalosporanic acid is 6.7fold lower than wild-type value, KM-value for adipyl-7-aminodeacetoxycephalosporanic acid is 3.7fold higher than wild-type value, turnover-number for glutaryl-7-aminocephalosporanic acid is 30.8fold lower than wild-type value, KM-value for glutaryl-7-aminocephalosporanic acid is 41.9fold higher than wild-type value Pseudomonas sp. SY-77-1
3.5.1.93 N266Q turnover number for adipyl-7-aminodeacetoxycephalosporanic acid is 1.12fold higher than wild-type value, KM-value for adipyl-7-aminodeacetoxycephalosporanic acid is 1.9fold lower than wild-type value, turnover-number for glutaryl-7-aminocephalosporanic acid is 1.7fold lower than wild-type value, KM-value for glutaryl-7-aminocephalosporanic acid is 3.9fold higher wild-type value. 100% improved conversion of cephalosporin C compared to wild type enzyme Pseudomonas sp. SY-77-1
3.5.1.93 N266S turnover number for adipyl-7-aminodeacetoxycephalosporanic acid is 1.7fold lower than wild-type value, KM-value for adipyl-7-aminodeacetoxycephalosporanic acid is 1.2fold lower than wild-type value, turnover-number for glutaryl-7-aminocephalosporanic acid is 2.5fold lower than wild-type value, KM-value for glutaryl-7-aminocephalosporanic acid is 2.4fold higher than wild-type value Pseudomonas sp. SY-77-1
3.5.1.93 N266T turnover number for adipyl-7-aminodeacetoxycephalosporanic acid is 2.7fold lower than wild-type value, KM-value for adipyl-7-aminodeacetoxycephalosporanic acid is 4.75fold higher than wild-type value, turnover-number for glutaryl-7-aminocephalosporanic acid is 8.2fold lower than wild-type value, KM-value for glutaryl-7-aminocephalosporanic acid is 48.4fold higher than wild-type value Pseudomonas sp. SY-77-1
3.5.1.93 N266W turnover number for adipyl-7-aminodeacetoxycephalosporanic acid is 1.2fold lower than wild-type value, KM-value for adipyl-7-aminodeacetoxycephalosporanic acid is 2fold lower than wild-type value, turnover-number for glutaryl-7-aminocephalosporanic acid is 3fold lower than wild-type value, KM-value for glutaryl-7-aminocephalosporanic acid is 5.8fold higher than wild-type value. 30% improved conversion of cephalosporin C compared to wild type enzyme Pseudomonas sp. SY-77-1
3.5.1.93 N266Y turnover number for adipyl-7-aminodeacetoxycephalosporanic acid is 1.2fold lower than wild-type value, KM-value for adipyl-7-aminodeacetoxycephalosporanic acid is 1.8fold lower than wild-type value, turnover-number for glutaryl-7-aminocephalosporanic acid is 6.6fold lower than wild-type value, KM-value for glutaryl-7-aminocephalosporanic acid is 3.5fold higher than wild-type value Pseudomonas sp. SY-77-1

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.5.1.93 0.031
-
 glutaryl-7-aminocephalosporanic acid wild-type enzyme Pseudomonas sp. SY-77-1
3.5.1.93 0.044
-
 glutaryl-7-aminocephalosporanic acid mutant enzyme N266H Pseudomonas sp. SY-77-1
3.5.1.93 0.075
-
 glutaryl-7-aminocephalosporanic acid mutant enzyme N266S Pseudomonas sp. SY-77-1
3.5.1.93 0.1
-
 glutaryl-7-aminocephalosporanic acid mutant enzyme N266F Pseudomonas sp. SY-77-1
3.5.1.93 0.11
-
 glutaryl-7-aminocephalosporanic acid mutant enzyme N266Y Pseudomonas sp. SY-77-1
3.5.1.93 0.12
-
 glutaryl-7-aminocephalosporanic acid mutant enzyme N266Q Pseudomonas sp. SY-77-1
3.5.1.93 0.14
-
 adipyl-7-aminodeacetoxycephalosporanic acid mutant enzyme N266H Pseudomonas sp. SY-77-1
3.5.1.93 0.16
-
 glutaryl-7-aminocephalosporanic acid mutant enzyme N266C Pseudomonas sp. SY-77-1
3.5.1.93 0.17
-
 glutaryl-7-aminocephalosporanic acid mutant enzyme N266M Pseudomonas sp. SY-77-1
3.5.1.93 0.18
-
 glutaryl-7-aminocephalosporanic acid mutant enzyme N266W Pseudomonas sp. SY-77-1
3.5.1.93 0.26
-
 glutaryl-7-aminocephalosporanic acid mutant enzyme N266A Pseudomonas sp. SY-77-1
3.5.1.93 0.29
-
 glutaryl-7-aminocephalosporanic acid mutant enzyme N266E Pseudomonas sp. SY-77-1
3.5.1.93 0.33
-
 adipyl-7-aminodeacetoxycephalosporanic acid mutant enzyme N266M Pseudomonas sp. SY-77-1
3.5.1.93 0.43
-
 glutaryl-7-aminocephalosporanic acid mutant enzyme N266L Pseudomonas sp. SY-77-1
3.5.1.93 0.54
-
 glutaryl-7-aminocephalosporanic acid mutant enzyme N266G Pseudomonas sp. SY-77-1
3.5.1.93 0.59
-
 adipyl-7-aminodeacetoxycephalosporanic acid mutant enzyme N266W Pseudomonas sp. SY-77-1
3.5.1.93 0.6
-
 adipyl-7-aminodeacetoxycephalosporanic acid mutant enzyme N266F Pseudomonas sp. SY-77-1
3.5.1.93 0.62
-
 adipyl-7-aminodeacetoxycephalosporanic acid mutant enzyme N266Q Pseudomonas sp. SY-77-1
3.5.1.93 0.65
-
 adipyl-7-aminodeacetoxycephalosporanic acid mutant enzyme N266Y Pseudomonas sp. SY-77-1
3.5.1.93 0.8
-
 adipyl-7-aminodeacetoxycephalosporanic acid mutant enzyme N266C Pseudomonas sp. SY-77-1
3.5.1.93 0.99
-
 adipyl-7-aminodeacetoxycephalosporanic acid mutant enzyme N266S Pseudomonas sp. SY-77-1
3.5.1.93 1
-
 adipyl-7-aminodeacetoxycephalosporanic acid mutant enzyme N266L Pseudomonas sp. SY-77-1
3.5.1.93 1.2
-
 adipyl-7-aminodeacetoxycephalosporanic acid wild-type enzyme Pseudomonas sp. SY-77-1
3.5.1.93 1.3
-
 glutaryl-7-aminocephalosporanic acid mutant enzyme N266P Pseudomonas sp. SY-77-1
3.5.1.93 1.4
-
 adipyl-7-aminodeacetoxycephalosporanic acid mutant enzyme N266G Pseudomonas sp. SY-77-1
3.5.1.93 1.5
-
 glutaryl-7-aminocephalosporanic acid mutant enzyme N266T Pseudomonas sp. SY-77-1
3.5.1.93 2.2
-
 adipyl-7-aminodeacetoxycephalosporanic acid mutant enzyme N266A Pseudomonas sp. SY-77-1
3.5.1.93 2.2
-
 adipyl-7-aminodeacetoxycephalosporanic acid mutant enzyme N266D Pseudomonas sp. SY-77-1
3.5.1.93 4.4
-
 adipyl-7-aminodeacetoxycephalosporanic acid mutant enzyme N266P Pseudomonas sp. SY-77-1
3.5.1.93 5.7
-
 adipyl-7-aminodeacetoxycephalosporanic acid mutant enzyme N266T Pseudomonas sp. SY-77-1
3.5.1.93 13.8
-
 adipyl-7-aminodeacetoxycephalosporanic acid mutant enzyme N266E Pseudomonas sp. SY-77-1

Organism

EC Number Organism UniProt Comment Textmining
3.5.1.93 Pseudomonas sp. SY-77-1
-
-
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.5.1.93 adipyl-7-aminodeacetoxycephalosporanic acid + H2O
-
 Pseudomonas sp. SY-77-1 adipate + 7-aminodeacetoxycephalosporanic acid
-
 ?
3.5.1.93 cephalosporin C + H2O low activity Pseudomonas sp. SY-77-1 cephalosporanic acid + 2-amino-5-hydroxypentanoate
-
 ?
3.5.1.93 glutaryl-7-aminocephalosporanic acid + H2O
-
 Pseudomonas sp. SY-77-1 7-aminocephalosporanate + glutarate
-
 ?

Synonyms

EC Number Synonyms Comment Organism
3.5.1.93 cephalosporin acylase
-
 Pseudomonas sp. SY-77-1

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.5.1.93 0.002
-
 adipyl-7-aminodeacetoxycephalosporanic acid mutant enzyme N266D Pseudomonas sp. SY-77-1
3.5.1.93 0.023
-
 adipyl-7-aminodeacetoxycephalosporanic acid mutant enzyme N266E Pseudomonas sp. SY-77-1
3.5.1.93 0.025
-
 glutaryl-7-aminocephalosporanic acid mutant enzyme N266E Pseudomonas sp. SY-77-1
3.5.1.93 0.061
-
 adipyl-7-aminodeacetoxycephalosporanic acid mutant enzyme N266P Pseudomonas sp. SY-77-1
3.5.1.93 0.082
-
 glutaryl-7-aminocephalosporanic acid mutant enzyme N266L Pseudomonas sp. SY-77-1
3.5.1.93 0.13
-
 glutaryl-7-aminocephalosporanic acid mutant enzyme N266P Pseudomonas sp. SY-77-1
3.5.1.93 0.15
-
 adipyl-7-aminodeacetoxycephalosporanic acid mutant enzyme N266T Pseudomonas sp. SY-77-1
3.5.1.93 0.18
-
 adipyl-7-aminodeacetoxycephalosporanic acid mutant enzyme N266A Pseudomonas sp. SY-77-1
3.5.1.93 0.23
-
 adipyl-7-aminodeacetoxycephalosporanic acid mutant enzyme N266G Pseudomonas sp. SY-77-1
3.5.1.93 0.24
-
 adipyl-7-aminodeacetoxycephalosporanic acid mutant enzyme N266S Pseudomonas sp. SY-77-1
3.5.1.93 0.25
-
 adipyl-7-aminodeacetoxycephalosporanic acid mutant enzyme N266L Pseudomonas sp. SY-77-1
3.5.1.93 0.31
-
 adipyl-7-aminodeacetoxycephalosporanic acid mutant enzyme N266C Pseudomonas sp. SY-77-1
3.5.1.93 0.33
-
 adipyl-7-aminodeacetoxycephalosporanic acid mutant enzyme N266W Pseudomonas sp. SY-77-1
3.5.1.93 0.33
-
 adipyl-7-aminodeacetoxycephalosporanic acid mutant enzyme N266Y Pseudomonas sp. SY-77-1
3.5.1.93 0.37
-
 adipyl-7-aminodeacetoxycephalosporanic acid mutant enzyme N266F Pseudomonas sp. SY-77-1
3.5.1.93 0.41
-
 adipyl-7-aminodeacetoxycephalosporanic acid wild-type enzyme Pseudomonas sp. SY-77-1
3.5.1.93 0.46
-
 adipyl-7-aminodeacetoxycephalosporanic acid mutant enzyme N266Q Pseudomonas sp. SY-77-1
3.5.1.93 0.47
-
 adipyl-7-aminodeacetoxycephalosporanic acid mutant enzyme N266H Pseudomonas sp. SY-77-1
3.5.1.93 0.49
-
 glutaryl-7-aminocephalosporanic acid mutant enzyme N266T Pseudomonas sp. SY-77-1
3.5.1.93 0.61
-
 glutaryl-7-aminocephalosporanic acid mutant enzyme N266Y Pseudomonas sp. SY-77-1
3.5.1.93 0.9
-
 glutaryl-7-aminocephalosporanic acid mutant enzyme N266F Pseudomonas sp. SY-77-1
3.5.1.93 1.3
-
 glutaryl-7-aminocephalosporanic acid mutant enzyme N266W Pseudomonas sp. SY-77-1
3.5.1.93 1.4
-
 glutaryl-7-aminocephalosporanic acid mutant enzyme N266A Pseudomonas sp. SY-77-1
3.5.1.93 1.6
-
 glutaryl-7-aminocephalosporanic acid mutant enzyme N266S Pseudomonas sp. SY-77-1
3.5.1.93 1.8
-
 glutaryl-7-aminocephalosporanic acid mutant enzyme N266M Pseudomonas sp. SY-77-1
3.5.1.93 1.9
-
 adipyl-7-aminodeacetoxycephalosporanic acid mutant enzyme N266M Pseudomonas sp. SY-77-1
3.5.1.93 2
-
 glutaryl-7-aminocephalosporanic acid mutant enzyme N266C Pseudomonas sp. SY-77-1
3.5.1.93 2
-
 glutaryl-7-aminocephalosporanic acid mutant enzyme N266G Pseudomonas sp. SY-77-1
3.5.1.93 2.4
-
 glutaryl-7-aminocephalosporanic acid mutant enzyme N266Q Pseudomonas sp. SY-77-1
3.5.1.93 3.1
-
 glutaryl-7-aminocephalosporanic acid mutant enzyme N266H Pseudomonas sp. SY-77-1
3.5.1.93 4
-
 glutaryl-7-aminocephalosporanic acid wild-type enzyme Pseudomonas sp. SY-77-1