Table 4 The advantages and disadvantages of different synthesis and expression strategies for AMPs17.
Strategies for improving antimicrobial peptide production | |||
|---|---|---|---|
Production of AMPs | Method | Advantages | Disadvantages |
Non-ribosomal peptide synthesis | solid-phase-peptide synthesis | • No proteases during production • Comparable or improved activity • High purity • The synthesis of libraries of new AMPs in one experimental method | • High costs • Select peptides difficult to synthesize |
Synthesis by using D-peptides | • The enantiomeric form of amino acids • Increase stabilityand protection against degradation | • Is still costly when compared to production in bacterial systems • Probable of secondary structure disruption and potentially cause a loss of activity of the respective protein | |
Ribosomal protein synthesis—expression and purification of AMPs using heterologous systems | Bacterial expression systems | • High purity • High expression levels • Low costs • Well established method | • Prone to protease degradation • Endotoxin contaminants • No post-translational modifications |
Fungal expression systems | • Efficient secretion if utilized • Post-translational modifications • High expression levels • Low cost | • Fermentation production • Hyperglycosylation may occur | |
Plant based expression systems | • Large scale production • Low cost • Post-translational Modifications • Option for cell suspension | • Genetic modification is difficult • Long growth time • Low yields and low stability | |
Insect based expression systems | • Genomic or plasmid expression • Post-translational modifications | • Low yields • High cost • Difficult to upscale • Potential issues with Lytic cycle | |
Fusion-protein based approaches for recombinant production of AMPs | Fusion-protein based secretion | • Improved purification • Improved peptide stability • Decreased toxicity | • Degradation during purification • Potential folding issues • Potential toxicity |
Fusion-protein based production using inclusion bodies | • Easy purification • Decreased cytotoxicity | • Lower yields • Protein misfolding • No post- translational modifications | |
Fusion-protein based enhanced solubility | • Higher intracellular concentrations • Less protein misfolding • Improved production method | • Low expression yields • High toxicity in the cell | |
Fusion-protein based masking of AMP toxicity | •Higher yields • Low production costs • Prevents inclusion body formation | • Some strategies cause protein to enter inclusion bodies | |
Hybridization expression of AMPs | • Potential novel AMPs • Coproduction of two AMPs • Decreased cytotoxicity • Improved yields, selectivity, activity, and stability | • Inactive hybrids • Need for characterization of hybrid AMPs | |
Cleaving the fusion-protein | • Ability to use autocleavage • Removal of fusion proteins after use • High yields • Used with other methods | • Extra purification steps • Dependent on protein solubility • Dependent on cleavage site Accessibility | |
Multimeric expression | • Improved yields • Improved stability | • Expression based on copy number cannot be predicted • Expression system can affect expression | |
