Production of human antimicrobial peptide LL-37 in Escherichia coli using a thioredoxin-SUMO dual fusion system.
Li. Yifeng Y
Key Findings
- LL-37 was expressed in E. coli using a thioredoxin‑SUMO dual‑tag fusion
- SUMO protease cleavage produced LL-37 with its native N‑terminus
- The purified peptide showed expected mass and antimicrobial activity, yielding ~2.4 mg/L
Practical Outcomes
- This protocol offers a relatively low‑cost, scalable route for labs or skilled DIY biohackers to produce active LL-37 without buying it commercially. It still requires molecular‑biology tools and purification steps, so it’s most useful for those with the necessary equipment and expertise.
Summary
Scientists figured out a way to make the human antimicrobial peptide LL-37 in ordinary bacteria by attaching it to a removable tag, then cutting it off to get the natural form, which still kills E. coli and gives about 2.4 mg of peptide per liter of culture.
Abstract
LL-37 is a human antimicrobial peptide that has been shown to possess multiple functions in host defense. In this report, the peptide was expressed as a fusion with a thioredoxin-SUMO dual-tag. Upon SUMO protease mediated cleavage at the SUMO/peptide junction, LL-37 with its native N-terminus was generated. The released peptide was separated from the dual-tag and cleavage enzyme by size-exclusion chromatography. Mass spectrometry analysis proves that the recombinant peptide has a molecular weight as theoretically expected for its native form. The produced peptide displayed antimicrobial activity against Escherichia coli K-12. On average, 2.4 mg peptide was obtained from one liter of bacterial culture. Thus, the described approach provides an effective alternative for producing active recombinant LL-37 with its natural amino acid sequence in E. coli.
Study Information
pubmed
2012
2012-11-07T00:00:00.000Z
10.1016/j.pep.2012.10.008
28
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