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Literature summary for 3.5.1.93 extracted from

  • Zhu, X.; Luo, H.; Chang, Y.; Su, H.; Li, Q.; Yu, H.; Shen, Z.
    Characteristic of immobilized cephalosporin C acylase and its application in one-step enzymatic conversion of cephalosporin C to 7-aminocephalosporanic acid (2011), World J. Microbiol. Biotechnol., 27, 823-829.
    View publication on PubMed

Application

Application Comment Organism
synthesis immobilzation on LX-1000EP, an epoxy support, with a recovery of 33.8% and an activity of 81 U/g wet resin, suggesting a potential in industrial 7-aminocephalosporanic acid production. Under optimal conditions, a very high 7-aminocephalosporanic acid yield of 96.6% is obtained within 60 min Pseudomonas sp. SE83

Cloned(Commentary)

Cloned (Comment) Organism
cloned in Escherichia coli BL21(DE3) using a pET28-acy plasmid haboring the CPC acylase gene Pseudomonas sp. SE83
expressed in Escherichia coli BL21(DE3) cells Pseudomonas sp.

General Stability

General Stability Organism
in the results of repeated batch production of 7-aminocephalosporanic acid, 50% activity of the initial cycle is maintained after recycled 24 times and the average conversion rate of cephalosporin reaches 98% Pseudomonas sp. SE83

Inhibitors

Inhibitors Comment Organism Structure
7-aminocephalosporanic acid product inhibition, competitive inhibitor Pseudomonas sp. SE83

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
additional information the apparent Km value of CPC acylase increases and Vmax is almost unaffected in the presence of 7-aminocephalosporanic acid Pseudomonas sp. SE83

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
28000
-
1 * 28000 + 1 * 58000, alpha- und beta-subunit, SDS-PAGE Pseudomonas sp. SE83
28000
-
1 * 58000 + 1 * 28000, SDS-PAGE Pseudomonas sp.
58000
-
1 * 28000 + 1 * 58000, alpha- und beta-subunit, SDS-PAGE Pseudomonas sp. SE83
58000
-
1 * 58000 + 1 * 28000, SDS-PAGE Pseudomonas sp.

Organism

Organism UniProt Comment Textmining
Pseudomonas sp.
-
-
-
Pseudomonas sp. SE83
-
-
-

Purification (Commentary)

Purification (Comment) Organism
Cu2+-immobilized iminodiacetic acid column chromatography Pseudomonas sp.
recombinant protein, to high purity by immobilized metal affinity chromatography, 56% recovery Pseudomonas sp. SE83

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
additional information
-
immobilized enzyme, on LX-1000EP support, 81 U/g wet resin Pseudomonas sp. SE83
additional information
-
purified enzyme 10 U/mg wet resin Pseudomonas sp. SE83

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
cephalosporin C + H2O
-
Pseudomonas sp. SE83 7-aminocephalosporanic acid + 2-amino-5-hydroxypentanoate
-
?
cephalosporin C + H2O
-
Pseudomonas sp. 7-aminocephalosporanic acid + ?
-
?

Subunits

Subunits Comment Organism
dimer 1 * 28000 + 1 * 58000, alpha- und beta-subunit, SDS-PAGE Pseudomonas sp. SE83
heterodimer 1 * 58000 + 1 * 28000, SDS-PAGE Pseudomonas sp.

Synonyms

Synonyms Comment Organism
cephalosporin C acylase
-
Pseudomonas sp.
cephalosporin C acylase
-
Pseudomonas sp. SE83
CPC acylase
-
Pseudomonas sp.
CPC acylase
-
Pseudomonas sp. SE83

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
25
-
the production of 7-aminocephalosporanic acid is carried out at 25°C and pH 8.5 to avoid product degradation Pseudomonas sp. SE83
37
-
assay at Pseudomonas sp. SE83
50
-
free enzyme Pseudomonas sp.
50
-
free enzyme Pseudomonas sp. SE83
55
-
immobilized enzyme Pseudomonas sp. SE83
55
-
LX-1000EP-immobilized enzyme Pseudomonas sp.

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
15 40 the LX-1000EP-immobilized enzyme is stable at temperatures up to 40°C (8% loss of activity after 2 h of incubation at 40°C) Pseudomonas sp.
25
-
above 25°C the stability of the free enzyme decays sharply Pseudomonas sp. SE83
25
-
there is no fatal decrease of the enzyme stability up to 25°C. Above 25°C, the stability of the free enzyme decays sharply Pseudomonas sp.
40
-
immobilized enzyme, 92% activity remains, the free enzyme is nearly inactive Pseudomonas sp. SE83

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8
-
assay at Pseudomonas sp. SE83
8.5
-
free enzyme Pseudomonas sp.
8.5 9.5 broad pH optimum, immobilized enzyme Pseudomonas sp. SE83
8.5 9.5 LX-1000EP-immobilized enzyme Pseudomonas sp.

pH Stability

pH Stability pH Stability Maximum Comment Organism
7 8 both forms of the enzyme, free and immobilized are partly inactivated between pH 7 and 8, while the immobilized CPC acylase shows good stability in solutions at pH 8-9.5 Pseudomonas sp. SE83
8 9.5 immobilized enzyme Pseudomonas sp. SE83
8 9.5 LX-1000EP-immobilized enzyme shows good stability in solutions at pH 8.0-9.5. after 2 h of incubation Pseudomonas sp.