Inoculate 100 ml of starter medium (Table 1) with single colony. Shake at 37 degree.
Inoculate 10-20 litre fermenter containing prewarmed fermentation medium (Table 2) with the 100 ml starter inoculum in mid-log phase.
Monitor cell growth in order to identify times for induction and harvest.
Table 1
Starter medium
Ingredients | Concentration |
1. Minimal medium
2. All amino acids except methionine 3. Adenine, guanine, thymine and uracil 4. Selenomethionine 5. LB |
Minimal medium A [27], with carbon source at 5g/l
40 mg/l 0.5 g/l 10-50 mg/l for 100 ml starter 5% (v/v) LB for 20 l fermenter |
Table 2
Fermentation medium
Ingredients | Concentration |
1. Minimal medium
2. All amino acids except methionine 3. Adenine, guanine, thymine and uracil 4. Selenomethionine 5. Thiamine 6. Biotin (optional) |
Minimal medium A [27], with carbon source at 5g/l
40 mg/l 0.5 g/l 10-50 mg/l 2 mg/l 2 mg/l |