Composite

Part:BBa_K5089007:Design

Designed by: Camille Prefontaine   Group: iGEM24_UManitoba   (2024-09-29)


MGS0156-CsgA-LCI


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NotI site found at 457
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 1059
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 159
    Illegal NgoMIV site found at 330
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 843


Design Notes

This expression system is codon optimized for expression in E.coli.

Source

MGS0156 is a serine-dependent ⍺/β hydrolase originally identified from an environmental metagenomic analysis study to look for enzymes to degrade PLA[1]. LCI (Liquid Chromatography peak I) is a binding peptide for polystyrene, polypropylene and polylactic acid [2]. CsgA is a protein component of E.coli biofilms, which can be used as an anchor to surface display proteins [3].

References

1. Hajighasemi, M., et al., 2016. Biochemical and Structural Insights into Enzymatic. Depolymerization of Polylactic Acid and Other Polyesters by Microbial Carboxylesterases. Biomacromolecules, 17(6), 2027–2039. https://doi.org/10.1021/acs.biomac.6b00223

2. Lu, Y., Hintzen, K.-W., Kurkina, T., Ji, Y., and Schwaneberg, U. 2023. Directed Evolution of Material Binding Peptide for Polylactic Acid-specific Degradation in Mixed Plastic Wastes. ACS Catalysis, 13(19), 12746–12754. https://doi.org/10.1021/acscatal.3c02142

3. Fei Li, Luona Ye, Longyu Zhang, Xiaoyan Li, Xiaoxiao Liu, Jiarui Zhu, Huanhuan Li, Huimin Pang, Yunjun Yan, Li Xu, Min Yang, Jinyong Yan, Design of a genetically programmed barnacle-curli inspired living-cell bioadhesive, Materials Today Bio, Volume 14, 2022.