Part:BBa_K4380003

mutated LCI peptide

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]

Introduction

Vilnius-Lithuania Igem 2022 project NanoFind was working to create an easily accessible nanoplastic detection tool, using peptides, whose interaction with nanoplastic particles would lead to an easily interpretable response. The system itself focused on smaller protein molecules, peptides, which are modified to acquire the ability to connect to the surface of synthetic polymers – plastics. The detection system works when peptides and nanoplastic particles combine and form a sandwich complex - one nanoplastic particle is surrounded by two peptides, attached to their respective protein. The sandwich complex consisted of two main parts – one is a peptide bound to a fluorescent protein, and another peptide is immobilized on a cellulose membrane by a cellulose binding domain. This peptide played a huge role in the team's project because this mutated peptide has been proven to bind to polypropylene plastic with high affinity.

Design

This sequence is a mutated variant of LCI (liquid chromatography peak I), a 47-residue cationic antimicrobial peptide (AMP) found in Bacillus subtilis, and is one of the main effective components that have strong antimicrobial activity against Xanthomonas campestris pv Oryzea and Pseudomonas solanacearum PE1, etc. [1]
The peptide was created by the Ultrahigh‐throughput screening system for directed polymer binding peptide evolution by Apitius et al., 2018 [2]. The polypropylene (PP)‐binding peptide liquid chromatography peak I (LCI) was simultaneously saturated and screened for improved PP‐binding in the presence of the anionic surfactant sodium dodecyl benzenesulfonate (LAS; 0.25 mM). The cell surface system enabled efficient screening of the generated LCI diversity (in total ~10 million clones were screened) and therefore only variants with the strongest binding affinity were chosen as characterization of identified LCI binders revealed an up to 12‐fold improvement when comparing the peptide with its unmutated self in PP‐binding strength in the presence of the surfactant.[2]

References

[1] Gong, W., Wang, J., Chen, Z., Xia, B., & Lu, G. (2011). Solution structure of LCI, a novel antimicrobial peptide from Bacillus subtilis. Biochemistry, 50(18), 3621–3627. https://doi.org/10.1021/bi200123w
[2] Apitius, L., Rübsam, K., Jakesch, C., Jakob, F., & Schwaneberg, U. (2019). Ultra‐high‐throughput screening system for directed polymer binding peptide evolution. Biotechnology and Bioengineering. doi:10.1002/bit.26990