It produces cefradine from 7-ADCA (7-amino-3-deacetoxy-cephalosporanic acid) and DHME (D-dihydrophenylglycine methyl ester).
Standard
Activity: >50 U/g wet weight
Size of immobilization support: 250~500μm (>95%)
Water content <65%
Enzyme assay: One unit (U) represents the amount of enzyme required for producing1μmol cefradine per min in 0.1M K-phosphatebuffer containing 20mM 7-ADCA and (7-ami-no-3-deacetoxy-cephalosporanic acid) and 20mM DHME(D-dihy-drophenylglycine methyl ester), , pH 7.5 at 28°C.
Application
Cefradine production from 7-ACDA and DHME
Characteristics
·High synthesis activity of cefradiner
·Low degradation activity of ccefradine and DHME
·Fast enzyme reaction(reaction completed in 90 mins)
·High yield synthesis of cefradine (>95%)
Reaction condition
| pH | 8.0 ~ 8.3 |
| Temperature | 10℃ |
| Solvent | Water |
| 7-APRA：HPGM | 1:1.05-1.10 |
| Enzyme amount | 7U/ml |
Scheme for enzyme reaction
Optional reaction conditions (Temperature, pH)
Production instruction
| Category | Contents |
|---|
| Materials | - Synthesizing enzyme: Immobilized Cefradine Acylase (80U/g wet, 65% water content)
- 7-ADCA, DHME·HCl
- Water
- Ammonium hydroxide (28%) |
| Condition | - pH: 8.0-8.2
- Temperature: 10℃
- 7-ADCA：DHME: 350mM:385mM(1:1.10)
- Enzyme: 7U/ml
- Reaction solution: Water |
| Step | 1. Remove the water after washing immobilized enzyme with water more than five times.
2. Dissolve 7-ADCA(350mM) with ammonium hydroxide and water , and then add DHME·HCl (385mM). Adjust pH to 8.0 and volume to 70ml.
3. Pour 6.13g wet enzyme in conversion reactor.
4. Proceed synthetic reaction at 10℃ and 550rpm after pouring substrates.
5. Extract every 60 minutes after 1 hour. Stop the reaction if conversion rate of 7-ADCA is more than 98.5%.
6. Take the reaction solution in reactor. |
Fig 3. Cefradine production from Immobilized Cefradine Acylase
ZJ26F0434574