De novo expression and antibacterial potential of four lactoferricin peptides in cell-free protein synthesis system
| Autor: | Maie Ahmed Elkhawaga, Hoda Reda Kholef, Nawal Abd El-Baky, Elrashdy Mustafa Redwan, Mona M Sharaf, Eman S. Abdelkhalek |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
0106 biological sciences
Peptide 3D three dimensional structures medicine.disease_cause 01 natural sciences Applied Microbiology and Biotechnology chemistry.chemical_compound Lf lactoferrin Lactoferricin chemistry.chemical_classification 0303 health sciences biology Lactoferrin Chemistry MRSA methicillin-resistant Staphylococcus aureus MICs minimum inhibitory concentrations cLFcin camel lactoferricin ELISA enzyme-linked immunosorbent assay Biochemistry Staphylococcus aureus Antibacterial activity Biotechnology Research Article MIC minimum inhibitory concentration lcsh:Biotechnology Antimicrobial peptides p-NPP p-Nitrophenyl phosphate 03 medical and health sciences Minimum inhibitory concentration 010608 biotechnology lcsh:TP248.13-248.65 LC50 concentration lethal to 50 % of the cells medicine Escherichia coli CAMH cation-adjusted Mueller-Hinton broth Bioactive peptides 030304 developmental biology CFPS cell-free protein synthesis PBMCs peripheral blood mononuclear cells hLf human lactoferrin In vitro protein synthesis ConLFcin consensus lactoferricin bLFcin bovine lactoferricin hLFcin human lactoferricin cLf camel lactoferrin LFcin lactoferricin SDS-PAGE sodium dodecyl sulfate polyacrylamide gel electrophoresis biology.protein HSV herpes simplex virus Antimicrobial SD Shine-Dalgarno sequence |
| Zdroj: | Biotechnology Reports, Vol 29, Iss, Pp e00583-(2021) Biotechnology Reports |
| Popis: | Highlights • Cell-free (in vitro) method is ideal for short bioactive peptides rapid expression. • Camel lactoferricin is the most active among natural lactoferricin peptides. • Consensus lactoferricin is a candidate for further development into therapeutic use. For the first time, we produced four lactoferricin (LFcin) peptides by a cell-free (in vitro) method. These short antimicrobial peptides were expressed in an E. coli cell-free protein synthesis (CFPS) system and the bioactivity of the produced peptides was demonstrated. Additionally, we designed a novel synthetic consensus peptide (ConLFcin). The genes of bovine Lfcin (bLFcin), human Lfcin (hLFcin), camel Lfcin (cLFcin), and ConLFcin were cloned into pET101/D-TOPO vector then peptides were synthesized in vitro by E. coli CFPS system. The antibacterial activity of these synthesized peptides was evaluated against Escherichia coli, Salmonella typhi, Pseudomonas aeruginosa, Staphylococcus aureus, and methicillin-resistant Staphylococcus aureus (MRSA). The four cell-free synthesized peptides showed significant antibacterial potency at minimum inhibitory concentration (MIC) values between 1.25 and 10 μg/mL. cLFcin and ConLFcin showed higher antibacterial effects than bLFcin and hLFcin. Thus, cell-free expression system is an ideal system for rapid expression of functionally active short bioactive peptides. |
| Databáze: | OpenAIRE |
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