It produces 6-APA (6-aminopenicillanic acid) from Penicillin G.
Standard
Activity
≥200 U/g wet weight
Size of immobilization support
150~300 μm (> 95%)
Water content
< 60%
Enzyme assay
One unit (U) represents the amount of enzyme required for adding 1μmol NaOH per min to adjust pH with degradation of Pen G to 6-APA in 50mM K-phosphate, pH 8.0, at 28℃.
Application
6-APA production from Penicillin G.
Characteristics
Very high specific activity
High substrate selectivity
High stability (reusable more than 800 cycles)
High production rate of 6-APA (more than 98.5%)
Reaction condition
pH
8.0 ± 0.2
Temperature
28~29℃
Solvent
Phosphate buffer or water
Enzyme amount
7~15 U/ml
Reaction time
2~3 hours
Scheme for enzyme reaction
Production instruction
Category
Contents
Materials
- Catabolic enzyme: Immobilized Penicillin G acylase (200U/g wet, 60% water content) - Pen-G (penicillin G potassium) - Water - 3M ammonium hydroxide - HPLC - Chromatography column (Hypersil ODS, 5μm, 125mm*4.6mm)
1. Remove the water after washing immobilized enzyme with water more than five times. 2. Dissolve 215mM Pen G in 130ml water. 3. Leave substrate in reactor at 28℃ for 10 minutes. 4. Pour the catabolic enzyme(Immobilized Penicillin G acylase 7~15 U/ml) in conversion reactor 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 in 100ml flask. 7. Measure the yield of 6-APA with HPLC by calculating peak area.
One unit (U) represents the amount of enzyme required for adding 1µ mol NaOH per min to adjust pH with degradation of Pen G to 6-APA in 50mM K-phosphate, pH 8.0, at 28℃.
ZJ26F0434623
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