Part:BBa_K4380004

mutated TA2 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
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 49
  

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 peptide bound to a fluorescent protein, other peptide immobilized on cellulose membrane by cellulose binding domain. This peptide played a huge role in the teams project due to the fact that this mutated variant has been prooven to bind to polystyrene plastic with high affinity.

Design

This sequence is a mutated variant of TA2 (Tachystatin 2). Originally, the unmutated TA2 variant comes from a series of antimicrobial peptides, named tachystatins A, B, and C, that were identified from hemocytes of the horseshoe crab Tachypleus tridentatus. Tachystatins exhibited a broad spectrum of antimicrobial activity against Gram-negative and Gram-positive bacteria and fungi.
The unmutated peptide has been found to bind polystyrene and polypropylene plastics (see Part:BBa_K3013001). This peptide is a mutated version of TA2 designed by Rübsam et al., 2018 [1]. Peptide evolution was performed employing a high mutagenesis frequency (59,4 mutations/kb) and a 96-well-based assay was developed using fluorescence quantification to find the best TA2 variants for polystyrene plastic binding.

References

[1] Rübsam, K., Weber, L., Jakob, F., & Schwaneberg, U. (2017). Directed evolution of polypropylene and polystyrene binding peptides. Biotechnology and Bioengineering, 115(2), 321–330. doi:10.1002/bit.26481