Part:BBa_K3629000

Yarrowia lipolytica XRP2 signal peptide (secretion tag)

Signal peptide sequence from the Yarrowia lipolytica alkaline extracellular protease XRP2 gene.

Usage and Biology

Yarrowia lipolytica is an emerging chassis in the molecular biology community. Its unique metabolic properties and efficient protein production and secretion mechanisms make it a desirable chassis for heterologous protein expression/secretion (1). In fact, it has been shown to have better secretory mechanisms than Saccharomyces cerevisiae as it uses co-translational translocation of polypeptides to the ER lumen and expresses secretory genes at a high level (1). To date the XPR2 and Lip2 signal peptides <a href="BBa_K1592000">(BBa_K1592000)</a> are two of the most commonly used and well studied signal peptides that can be attached to heterologous proteins for high secretion. However, XPR2 is shorter than the Lip2 signal peptide, which was used extensively in the other expression constructs of our collection (BBa_K33629012-14, BBa_K3629016-18, and BBa_K3629027)

The XPR2 signal peptide is derived from the XPR2 extracellular protease in Y. lipolytica. The signal peptide is located at the N-terminus of the protein and directs the section of the protein to the extracellular space. The general structure of this signal peptide (and Lip2) follows the “Sec-type” signal peptide structure with a positive amino acid in the N domain (K for XPR2 and Lip2), followed by hydrophobic residues that form an alpha-helix necessary for translocation, and finally a C-domain which is “helix-breaking” with a polar residue for the signal peptidase to recognize (consensus = A-X-A where X is any amino acid) (1).

This signal peptide was used in the TrEGII expression construct <a href="BBa_K3629017">(BBa_K3629017)</a> as this secretion tag, in combination with the EXP promoter <a href="BBa_K3629002">(BBa_K3629002)</a>, has been shown to secrete high levels of TrEGII in Y. lipolytica (up to 132mg/L) (2).

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]

References

1. Celińska, E., Borkowska, M., Białas, W., Korpys, P., & Nicaud, J. M. (2018). Robust signal peptides for protein secretion in Yarrowia lipolytica: identification and characterization of novel secretory tags. Applied microbiology and biotechnology, 102(12), 5221–5233. https://doi.org/10.1007/s00253-018-8966-9