It produces D-CPC (deacetyl-cephalosporic C) from CPC (Cephalosporin C) or D-7-ACA (deacetyl-7-aminocephalo-sporanic acid) from 7-ACA (7-aminocephalosporanic acid).
Standard
Activity: >300 U/g wet weight
Size of immobilization support: ≥70% passes 70-90 mesh sieve
Water content <60%
Enzyme assay: One unit (U) represents the amount of enzyme required for producing 1μmol D-CPC and acetic acid per min in 50mM K-phosphate buffer containing 5% CPC, pH 8.0, at 25°C.
Application
·D-CPC production from CPC
·D-7-ACA production from 7-ACA
Characteristics
·Very high specific activity
·High substrate selectivity
·High stability (reusable more than 800 cycles)
·High production rate of D-CPC (more than 98%)
·High production rate of D-7-ACA (more than 98%)
Reaction condition
| pH | 7.9~8.0 |
| Temperature | 20℃ |
| Solvent | Water |
| Enzyme amount | 7-10 U/ml |
| Reaction time | 1~1.5 hours |
Scheme for enzyme reaction
Optional reaction conditions (Temperature, pH)
Production instruction
| Category | Contents |
|---|
| Materials | ·Catabolic enzyme : Immobilized Cephalosporin C or 7-ACA esterase (300 U/g wet, 62% water content)
·CPC-Na or 7-ACA
·Water
·3M ammonium hydroxide
·HPLC
·Chromatography column (Nucleosil 120 3C18, 3μm, 125mm×4mm)
| | Condition | ·pH: 7.9-8.0
·Temperature: 20°C
·CPC-Na(or 7-ACA): 150mM
·Enzyme: 7~10 U/ml (130ml reaction)
·Reaction solution: Water
| | Step | 1. Remove the water after washing immobilized enzyme with watermore than five times.
2. Dissolve 150mM CPC-Na(or 7-ACA) in 130ml water.
3. Leave substrate in reactor at 20°C for 10 minutes.
4.Pour the catabolic enzyme (Immobilized Cephalosporin C or 7-ACA esterase 7~10 U/ml) in conversionreactor and measure time simultaneously.
5. Adjust pH to 8.0 with 3M ammonium hydroxide.
6. Extract 0.1ml reaction solution(supernatant) and dilute with water in100ml flask.
7. Measure the yield of D-CPC(or D-7-ACA) with HPLC by calculatingpeak area. |
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ZJ26F0434569