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Literature summary extracted from

  • Luo, H.; Zhu, L.; Chang, Y.; Liu, X.; Liu, Z.; Sun, H.; Li, X.; Yu, H.; Shen, Z.
    Microenvironmental pH changes in immobilized cephalosporin C acylase during a proton-producing reaction and regulation by a two-stage catalytic process (2017), Biores. Technol., 223, 157-165 .
    View publication on PubMed

Application

EC Number Application Comment Organism
3.5.1.93 industry cephalosporin C acylase (CCA), a proton-forming enzyme, is an important industrial enzyme that can directly catalyze the substrate cephalosporin C (CPC) to 7-aminocephalosporanic acid (7-ACA), which is an intermediate in many types of synthetic cephalosporins. Immobilized CCA on porous carriers is applied in industry for the production of 7-ACA because of its high efficiency and environmentally friendly nature compared with the chemical process and two-step enzymatic process involving two enzymes. Mutational improvement of catalytic efficiency and operational stability Pseudomonas sp.
3.5.1.93 synthesis cephalosporin C acylase (CCA), a proton-forming enzyme, is an important industrial enzyme that can directly catalyze the substrate cephalosporin C (CPC) to 7-aminocephalosporanic acid (7-ACA), which is an intermediate in many types of synthetic cephalosporins. Immobilized CCA on porous carriers is applied in industry for the production of 7-ACA because of its high efficiency and environmentally friendly nature compared with the chemical process and two-step enzymatic process involving two enzymes Pseudomonas sp.

Protein Variants

EC Number Protein Variants Comment Organism
3.5.1.93 additional information proton-producing enzyme cephalosporin C acylase (CCA) is covalently bound on an epoxy-activated porous support. The microenvironmental pH change in immobilized CCA during the reaction is detected using pH-sensitive fluorescein labeling (pH-sensitive fluorescein molecule co-immobilized on the carrier). The high catalytic velocity of the initial stage of conversion results in a sharp intraparticle pH gradient, which is likely the key factor relating to low operational stability. Mass transfer in the immobilized CCA is relatively severe during CPC hydrolysis. Reducing the intraparticle pH gradient, i.e. diminishing the diffusion limitation, is important for the catalysis of CCA. Accordingly, another strategy for a two-stage catalytic process is developed to reduce the reaction rate of stage I at a low temperature to preserve enzymatic activity and to shorten the duration of catalysis at a high reaction temperature in stage II. The reaction using the two-stage catalytic process (10-37°C shift at 30 min) shows significantly improved stability compared with that of the single-temperature reaction at 37°C (29 batches versus five batches, respectively) and a shorter catalytic period than the reaction at 10°C (40 min versus 70 min, respectively) Pseudomonas sp.

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.5.1.93 19.2
-
 cephalosporin C recombinant free enzyme, pH 8.5, 37°C Pseudomonas sp.
3.5.1.93 95.1
-
 cephalosporin C recombinant immobilized enzyme, pH 8.5, 37°C Pseudomonas sp.

Natural Substrates/ Products (Substrates)

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

Organism

EC Number Organism UniProt Comment Textmining
3.5.1.93 Pseudomonas sp. A0A1D8GRD5
-
-

Substrates and Products (Substrate)

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

Synonyms

EC Number Synonyms Comment Organism
3.5.1.93 CCA
-
 Pseudomonas sp.
3.5.1.93 cephalosporin C acylase
-
 Pseudomonas sp.

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.5.1.93 37
-
 immobilized enzyme Pseudomonas sp.

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
3.5.1.93 10 37 purified recombinant enzyme, the immobilized enzyme CCA is more labile at 37°C than at 20°C or 10°C Pseudomonas sp.

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.5.1.93 8
-
 immobilized enzyme Pseudomonas sp.

pH Stability

EC Number pH Stability pH Stability Maximum Comment Organism
3.5.1.93 6 8.5 the purified wild-type enzyme is not stable, stability is increased by immobilization Pseudomonas sp.