Table 1.
Purification step | Total protein/La | Purification-fold | Purityb | Yield/L |
---|---|---|---|---|
Bacterial lysate | 2.7 g | 1 | 0 | 2.7 g |
Amylose | 2.7 g | 19.3 | 45.2 % | 0.14 g |
Thermolysin digestion | 0.12 g | 1 | 1.4 % | 0.12 g |
IMAC | 0.12 g | 171.4 | 96.5 % | 0.70 mg |
Ion-exchange | 0.70 mg | 1.43 | >98.0 % | 0.49 mg |
Gel filtration | 0.49 mg | 1.40 | >98.0 % | 0.34 mg |
Protein concentration and total amount of protein was determined by measuring its absorbance at 280 nm.
Purity of MBP-proMeCPAHis6PR (up until thermolysin digestion) and MeCPA- His6PR (after thermolysin digestion) was estimated by SDS-PAGE using ImageQuant software.