Recombinant DNA procedures for producing small antimicrobial cationic peptides in bacteria

Construction of recombinant E. coli Nissle 1917 (EcN) strains for the expression and secretion of defensins

“…These include the potential ability of the cationic peptides to kill the producing strain and the susceptibility of the cationic peptides to proteolytic degradation. Piers and coworkers found that they could overcome such barriers by using a fusion protein expression system (Piers et al 1993). …”

“…Small cationic antimicrobial peptides have already been successfully synthesized in E. coli by recombinant DNA methods (Piers et al 1993). However, there are several pitfalls using E. coli as the host cell for cationic antimicrobial peptide expression, such as the host-killing activity of the product and its susceptibility to proteolytic degradation.…”

“…; Piers et al 1993). It has been reported that a fusion expression system with thioredoxin (TrxA) resulted in an increased yield of soluble products (LaVallie et al 2000).…”

Molecular insight into mechanism of antimicrobial action of the β‐hairpin peptide arenicin: Specific oligomerization in detergent micelles

“…It cannot be said in advance which carrier protein will be suited well for this task in the case of each specific AMP. Incomplete list of the carrier proteins tested for this purpose includes: L-ribulokinase fragment, 15 glutathione-S-transferase, 16 Pseudomonas aeruginosa outer membrane protein, 16 Staphylococcus aureus protein A, and the dimer of its IgG-binding domain, 17 subtilisin inhibitor, 18 b-galactosidase, 18 maltose-binding protein, 18 prochymosin, 19 E.coli replication protein RepA and its fragment, 20 cellulose-binding domains, 20 human defensin HNP-1 preprodomain, 20 modified magainin intervening sequence (MIS), 21 GABA-transaminase, 22 truncated E.coli amidophosphoribosyltransferase, 23 Pseudomonas testosteroni ketosteroid isomerase, 24 fluorescent proteins GFP and obelin, 25 31 and ubiquitin. 32 An intein-based expression and purification system was also described.…”

High‐level expression of cecropin CMIV in E. coli from a fusion construct containing the human tumor necrosis factor

“…These include 0) the potential ability of the cationic peptide to kill the host bacteria and Q the susceptibility of the cationic peptides to endogenous proteolytic digestion and degradation. Use of fusion protein expression systems may overcome such barriers, although it was found in some cases that some fusion proteins were still unstable (15).…”