Table 2.
Expression systems | MW (kDa) | Fusions yielda (μg/mg wet cell pellet) | Peptide yieldb (μg/mg wet cell pellet) | Cleavage effciencyc (%) | Percent recoveryd (%) | Peptide puritye (%) |
---|---|---|---|---|---|---|
CF system | 4.6 | ─ | 0.93f | ─ | ─ | 90.4 |
AT-HIS system | 4.4 | 4.85 | 0.41 | 100 | 52.14 | 92.1 |
SA-ELK16 system | 4.4 | 51.16 | 6.20 | 79.50 | 78.38 | 99.9 |
aYield of fusion proteins in the form of aggregates or soluble fractions
bYield of cecropin A peptides after intein-mediated cleavage in LB culture: 2.87 ± 0.62 mg/ml wet cell weight in the SA-ELK16 system and 1.48 ± 0.48 mg/ml wet cell weight in the AT-HIS system, estimated with the densitometric analysis software Gel-Pro Analyzer or BCA Protein Assay Kit
cCleavage efficiency was calculated by dividing the amount of cleaved protein aggregate by the amount of total aggregate before cleavage
dPercentage recovery of cecropin A peptide was calculated by dividing the mass of cecropin A peptide after cleavage by the mass that could be theoretically obtained from the fusion protein
ePurity of CeA peptide was calculated with the BandScan software
fYield of CeA peptide in the CF system was 0.93 μg/ml of reaction mixture