Difference between revisions of "Part:BBa K3629000"
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===Usage and Biology=== | ===Usage and Biology=== | ||
| − | <i>Yarrowia lipolytica</i> 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 <i>Saccharomyces cerevisiae</i> 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 | + | <i>Yarrowia lipolytica</i> 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 <i>Saccharomyces cerevisiae</i> 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 signal peptide is one of the most commonly used and well studied signal peptide that can be attached to heterologous proteins for high secretion. Since cellulases must also be secreted at high levels, it was well suited for use in our parts.The XPR2 is shorter than the other commonly used Lip2 signal peptide [https://parts.igem.org/Part:BBa_K1592000 (BBa_K1592000)], which was also used in a few expression constructs from [https://2020.igem.org/Team:Calgary/Part_Collection our collection] (BBa_K33629012-14, BBa_K3629016-18, and BBa_K3629027). |
The XPR2 signal peptide is derived from the XPR2 extracellular protease in <i>Y. lipolytica.</i> 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). | The XPR2 signal peptide is derived from the XPR2 extracellular protease in <i>Y. lipolytica.</i> 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 | + | This signal peptide was used in the TrEGII expression construct [https://parts.igem.org/Part:BBa_K3629017 (BBa_K3629017)] as this secretion tag, in combination with the EXP promoter [https://parts.igem.org/Part:BBa_K3629002 (BBa_K3629002),] has been shown to secrete high levels of TrEGII in <i>Y. lipolytica</i> (up to 132mg/L) (2). |
===Sequence and Features=== | ===Sequence and Features=== | ||
<partinfo>BBa_K3629000 SequenceAndFeatures</partinfo> | <partinfo>BBa_K3629000 SequenceAndFeatures</partinfo> | ||
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| + | Coding sequence includes the start codon for easy addition to the N-terminus of coding sequences. | ||
===References=== | ===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 | 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 | ||
| + | |||
| + | 2. Wei H, Wang W, Alper HS, et al. Ameliorating the Metabolic Burden of the Co-expression of Secreted Fungal Cellulases in a High Lipid-Accumulating Yarrowia lipolytica Strain by Medium C/N Ratio and a Chemical Chaperone. Front Microbiol. 2019;9:3276. Published 2019 Jan 9. doi:10.3389/fmicb.2018.03276 | ||
<!-- Uncomment this to enable Functional Parameter display | <!-- Uncomment this to enable Functional Parameter display | ||
Latest revision as of 22:37, 27 October 2020
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 signal peptide is one of the most commonly used and well studied signal peptide that can be attached to heterologous proteins for high secretion. Since cellulases must also be secreted at high levels, it was well suited for use in our parts.The XPR2 is shorter than the other commonly used Lip2 signal peptide (BBa_K1592000), which was also used in a few expression constructs from 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 (BBa_K3629017) as this secretion tag, in combination with the EXP promoter (BBa_K3629002), has been shown to secrete high levels of TrEGII in Y. lipolytica (up to 132mg/L) (2).
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Coding sequence includes the start codon for easy addition to the N-terminus of coding sequences.
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
2. Wei H, Wang W, Alper HS, et al. Ameliorating the Metabolic Burden of the Co-expression of Secreted Fungal Cellulases in a High Lipid-Accumulating Yarrowia lipolytica Strain by Medium C/N Ratio and a Chemical Chaperone. Front Microbiol. 2019;9:3276. Published 2019 Jan 9. doi:10.3389/fmicb.2018.03276