High level expression and purification of antimicrobial human cathelicidin LL-37 in Escherichia coli

Applied Microbiology and Biotechnology
Ján KrahulecJana Machálková

Abstract

The human antimicrobial peptide LL-37 is a cationic peptide with antimicrobial activity against both Gram-positive and Gram-negative microorganisms. This work describes the development of an expression system based on Escherichia coli capable of high production of the recombinant LL-37. The fusion protein Trx-LL-37 was expressed under control of T7 promoter. The expression of T7 polymerase in the E. coli strain constructed in this work was controlled by regulation mechanisms of the arabinose promoter. The expression plasmid was stabilized by the presence of parB locus which ensured higher homology of the culture during cultivation without antibiotic selection pressure. This system was capable of producing up to 1 g of fusion protein per 1 l of culture. The subsequent semipreparative HPLC allowed us to isolate 40 mg of pure LL-37. LL-37 showed high antimicrobial activity against both Gram-negative and Gram-positive microorganisms. Its activity against Candida albicans was practically nonexistent. Minimal Inhibition Concentration (MIC) determined for E. coli was 1.65 microM; for Staphylococcus aureus 2.31 microM, and for Enterococcus faecalis 5.54 microM. The effects of cathelicidin on E. coli included the ability to permeabilize...Continue Reading

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Citations

Jun 1, 2014·Applied Microbiology and Biotechnology·Michaela OsadskáJán Turňa
Aug 17, 2011·Protein Expression and Purification·Yifeng Li
Dec 19, 2012·Cellular Immunology·Dieter VandammeLiliane Schoofs
Aug 9, 2016·Journal of Biotechnology·Diana SzéliováJán Turňa
Oct 6, 2015·BioFactors·Katarzyna BandurskaPiotr Krupa
May 18, 2020·Brazilian Journal of Microbiology : [publication of the Brazilian Society for Microbiology]·Na ZhanAnshan Shan
Jul 13, 2021·Frontiers in Microbiology·Viktor VargaStanislav Stuchlík
Sep 9, 2019·ACS Biomaterials Science & Engineering·Sergio AcostaJosé Carlos Rodríguez-Cabello

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