Table 1.
c | Total Protiena(mg) |
Target proteinb(mg) | Approximate purityc(%) | Yieldd(%) |
---|---|---|---|---|
Cell lysis | 200.33 | 133.82 (CBD-intein-DPIP) | 66.8 | 100 |
Refolding of fusion protein | 156.26 | 116.73 (CBD-intein-DPIP) | 74.7 | 87.2 |
Chitin column and intein-mediated cleavage | 0.621 | 0.548 (DPIP) | 88.3 | 2.4 |
One liter of culture contained about 3.2 g wet weight cells
a: Total protein was quantified by Bradford assay
b: The amount of fusion protein was calculated based on the following formula: target protein = total protein amount x approximate purity
c: The purities of fusion protein were determined by densitometry analysis of their corresponding SDS gel bands
d: The relative low recovery of DPIP was partially due to the low molecular weight of DPIP (6 kDa) compared to the fusion precursor CBD-intein-DPIP (33 kDa)