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BRENDA support

Literature summary for 3.5.1.93 extracted from

  • Wei, Y.; Luo, H.; Chang, Y.; Yu, H.; Shen, Z.
    Reversible immobilization of cephalosporin C acylase on epoxy supports coated with polyethyleneimine (2015), Biocatal. Biotransform., 33, 250-259 .
No PubMed abstract available

Cloned(Commentary)

Cloned (Comment) Organism
recombinant expression of His6-tagged enzyme in Escherichia coli strain BL21(DE3) Pseudomonas sp.

Protein Variants

Protein Variants Comment Organism
additional information the purified recombinant His-tagged enzyme is immobilized on different supports, i.e. an epoxy-activated support LX1000-EPC4 (EP) or its derivatives, EP-polyethyleneimine (EP-PEI) and EP-ethylenediamine (EP-EDA) with cationic groups on the surface, validation and optimization. The CCA immobilized on EP-PEI has the best stability and its activity declines very slowly at 45°C. The enzyme immobilized on the cationic support EP-EDA has a half-life of 100 min at 45°C and pH 8.0, a value much lower than on EP-PEI. Stabilization factors of enzyme immobilized on EP-PEI and EP-EDA are 35.6fold and 5.5fold, respectively. The Km values of CCA immobilized on EP-PEI and EP-EDA are even lower than that of the free enzyme, indicating that a higher substrate affinity is obtained after the immobilization on the cationic supports. The reusability of the EP-PEI support significantly reduces the cost of the biocatalyst preparation Pseudomonas sp.

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
additional information Michaelis constant (Km) and maximum reaction velocity (Vmax) for the free and immobilized enzyme are calculated according to Lineweaver-Burk plots Pseudomonas sp.

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
cephalosporin C + H2O Pseudomonas sp.
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7-aminocephalosporanic acid + 2-amino-5-hydroxypentanoate
-
?

Organism

Organism UniProt Comment Textmining
Pseudomonas sp. A0A1D8GRD5
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
cephalosporin C + H2O
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Pseudomonas sp. 7-aminocephalosporanic acid + 2-amino-5-hydroxypentanoate
-
?
additional information substrate chephalosporin has a similar negative charge in pH 8.5 solution Pseudomonas sp. ?
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-

Synonyms

Synonyms Comment Organism
CCA
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Pseudomonas sp.
cephalosporin C acylase
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Pseudomonas sp.

Temperature Optimum [°C]

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

pH Optimum

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