Part:BBa_K3989002

eTEV, efficiency-enhanced TEV protease variant

eTEV protease is a variant from the original TEV protease. Seven amino acids(S3I, P8Q, S31T, E79G, T173A, V219R, A231V) are mutated in this variant and the catalytic efficiency is higher than the original one.

Structure and binding site

This variant is generated via a directed evaluation system called YESS 2.0 (Yeast Endoplasmic Reticulum (ER) Sequestration Screening). The structure is shown below.(Fig.A) Particularly, the sub-structure of at the binding site is shown as well.(Fig. B) The blue amino acid residues represent the changing sites of the variant and the purple residue represent the binding and cutting site of the ligand.

Fig. A Structural model of eTEV

Fig. B Sub-structure at the binding site of eTEV.

Performance

Besides this variant, the author also reported some other variants(for instance, BBa_K3989001) with different mutations. Different mutations in the TEV protease ligand-binding site show different catalytic efficincy. The results are shown in Fig. C.

Fig. C Catalytic efficiencies of different TEV protease variants.

Sequence and Features

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]

Reference

1) Denard, C. A., Paresi, C., Yaghi, R., McGinnis, N., Bennett, Z., Yi, L., ... & Iverson, B. L. (2021). YESS 2.0, a Tunable Platform for Enzyme Evolution, Yields Highly Active TEV Protease Variants. ACS Synthetic Biology, 10(1), 63-71.