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Literature summary for 3.5.1.93 extracted from

  • Zhang, J.; Yu, H.; Wang, Y.; Luo, H.; Shen, Z.
    Determination of the second autoproteolytic cleavage site of cephalosporin C acylase and the effect of deleting its flanking residues in the alpha-C-terminal region (2014), J. Biotechnol., 184, 138-145.
    View publication on PubMed
Search for more literature for 3.5.1.93

Cloned(Commentary)

Cloned (Comment) Organism
recombinant expression in in Escherichia coli JM109(DE3)/pET28, the enzyme performs self-activation Pseudomonas sp.

Protein Variants

Protein Variants Comment Organism
additional information construction of 14 mutants with deletion in the alpha-C-terminal region, the majority of the fragment-deleted mutant proteins completely lose their activity due to failure of the first autocleavage, while mutant proteins D2 (227-AM-228 deletion) and D4 (212-ADLA-215 deletion) formally activate into mature enzyme with high activity. The Kcat/Kmvalues of mutant proteins D2 and D4 are 46% and 102% higher than that of wild-type control, respectively Pseudomonas sp.

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
51
-
 cephalosporin C deletion mutant D4(212-ADLA-215), pH 8.5, 37°C Pseudomonas sp.
51.6
-
 cephalosporin C wild-type enzyme, pH 8.5, 37°C Pseudomonas sp.
74.8
-
 cephalosporin C deletion mutant D2(227-AM-228), pH 8.5, 37°C Pseudomonas sp.

Organism

Organism UniProt Comment Textmining
Pseudomonas sp.
-
-
-

Posttranslational Modification

Posttranslational Modification Comment Organism
proteolytic modification a first autoproteolytic cleavage site of the pro-enzyme is G239-S240, a second cleavage site is A232-S233, determined by tandem MS/MS analysis of the purified alpha-subunit C-terminus Pseudomonas sp.

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
cephalosporin C + H2O
-
 Pseudomonas sp. (7R)-7-aminocephalosporanate + 2-aminoadipate
-
 ?

Subunits

Subunits Comment Organism
heterodimer alphabeta Pseudomonas sp.

Synonyms

Synonyms Comment Organism
cephalosporin C acylase
-
 Pseudomonas sp.
CPCAcy
-
 Pseudomonas sp.

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
 assay at Pseudomonas sp.

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.022
-
 cephalosporin C wild-type enzyme and deletion mutant D4(212-ADLA-215), pH 8.5, 37°C Pseudomonas sp.
0.023
-
 cephalosporin C deletion mutant D2(227-AM-228), pH 8.5, 37°C Pseudomonas sp.

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8.5
-
 assay at Pseudomonas sp.

General Information

General Information Comment Organism
evolution the enzyme is a member of the N-terminal nucleophile (Ntn) hydrolase superfamily, in which the precursor gene is translated into a single polypeptide chain and then folded into a self-activating pre-protein activated by intramolecular double cleavages Pseudomonas sp.

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.000315
-
 cephalosporin C deletion mutant D2(227-AM-228), pH 8.5, 37°C Pseudomonas sp.
0.000419
-
 cephalosporin C wild-type enzyme, pH 8.5, 37°C Pseudomonas sp.
0.000424
-
 cephalosporin C deletion mutant D4(212-ADLA-215), pH 8.5, 37°C Pseudomonas sp.