Part:BBa_K4701302

SpyCatcher002-FAST-PETase

FAST-PETase is an PET hydrolase that has been engineered to improve degradation efficiency [1]. It has also been observed, that creating a fusion protein with MHETase, another enzyme in involved in PET depolymerization, overall performance can be improved [2]. This part contains a SpyCatcher002 sequence codon optimized for E. coli, that has been fused to the N-terminus of FAST-PETase. The intended use is to form a fusion with part BBa_K4701303 which contains the MHETase domain. The predicted conformation created with ColabFold [4] is shown in Figure 1.

Figure 1: SpyCatcher/SpyTag fusion between this part and BBa_K4701303. (cyan: MHETase, magenta: linkers, yellow: SpyCatcher002, red: SpyTag, orange: FAST-PETase, gray: His-tags)

References

[1] Lu, H., et al. (2022). Machine learning-aided engineering of hydrolases for PET depolymerization. Nature, 604(7907), 662–667. https://doi.org/10.1038/s41586-022-04599-z

[2] Knott, BC., et al. (2020). Characterization and engineering of a two-enzyme system for plastics depolymerization. Proceedings of the National Academy of Sciences, 117(41), 25476–25485. https://doi.org/10.1073/pnas.2006753117

[3] Zakeri, B., et al. (2012) Peptide tag forming a rapid covalent bond to a protein, through engineering a bacterial adhesin. Proceedings of the National Academy of Sciencesm, 109(12). https://doi.org/10.1073/pnas.1115485109

[4] Mirdita, M., et al. (2022) ColabFold: making protein folding accessible to all. Nature Methods. 19(6), 679–682, https://doi.org/10.1038/s41592-022-01488-1

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]