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Table 1 Summary of the expression and purification schemes in the three different systems

From: Rapid and efficient production of cecropin A antibacterial peptide in Escherichia coli by fusion with a self-aggregating protein

Expression systems AT-HIS expression system CF expression system SA-ELK16 expression system
Required steps Step 1: Expression of fusion protein CeA-Mxe-PT-His6 Step 1: Expression of antibacterial peptide His6-CeA Step 1: Expression of fusion protein CeA-Mxe-ELK16
Step 2: Cell disruption and supernatant collection Step 2: Ni2+ affinity chromatography Step 2: Cell disruption and acquisition of protein aggregates
Step 3: Ni2+ affinity chromatography Step 3: Intein-mediated cleavage
Step 4: Intein-mediated cleavage Step 4: acetic acid treatment and collection of CeA peptide.
Step 5: The second Ni2+ affinity chromatography
Time consuming 5 days 2 days 2 days
Final yield of peptide production (μg/mg wet cell pellet) a 0.41 0.93 b 6.20
Purity of bio-produced Cecropin A peptide (%) 92.1 90.4 99.8
  1. aYield of cecropin A peptide 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
  2. bYield of CeA peptide in CF system, 0.93 μg/ml of reaction mixture