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 (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 | 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 | Comment | Organism | Structure |
---|---|---|---|
7-aminocephalosporanic acid | product inhibition, competitive inhibitor | Pseudomonas sp. SE83 |
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 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 | UniProt | Comment | Textmining |
---|---|---|---|
Pseudomonas sp. | - |
- |
- |
Pseudomonas sp. SE83 | - |
- |
- |
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 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 | 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 | 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 | 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 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 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 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 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. |