Difference between revisions of "Part:BBa K3930018"
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<h2>Introduction</h2> | <h2>Introduction</h2> | ||
| − | <p>This sequence codes for the CrtY enzyme, that transforms Lycopene into β-carotene. The <i>CrtY</i> sequence comes from <i>Pantoea ananatis</i> and was codon optimized for its expression into <i>S.cerevisiae</i>. Its sequence is described into the publication of López et al. (2020). | + | <p>This sequence codes for the CrtY enzyme, that transforms Lycopene into β-carotene. The <i>CrtY</i> sequence comes from <i>Pantoea ananatis</i> and was codon optimized for its expression into <i>S. cerevisiae</i>. Its sequence is described into the publication of López et al. (2020). |
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<h2>Characterisation</h2> | <h2>Characterisation</h2> | ||
<h3>Production of β-carotene</h3> | <h3>Production of β-carotene</h3> | ||
<p>To demonstrate the CrtY activity, carotenoids contained in the cells were extracted using the method described by López et al. (2020). Yeast cells were lysed in acetone using glass beads and the supernatant obtained after this lysis was analyzed by RP-HPLC using a C18 column. In the LycoYeast-pFRAMBOISE-notfused strains, lycopene is converted into a new product with a higher retention time upon induction (Figure 1). Considering the yellow color of pFRAMBOISE-notfused strains and resulting the β-ionone production, this new peak most likely corresponds to β-carotene, i.e., the expected precursor. | <p>To demonstrate the CrtY activity, carotenoids contained in the cells were extracted using the method described by López et al. (2020). Yeast cells were lysed in acetone using glass beads and the supernatant obtained after this lysis was analyzed by RP-HPLC using a C18 column. In the LycoYeast-pFRAMBOISE-notfused strains, lycopene is converted into a new product with a higher retention time upon induction (Figure 1). Considering the yellow color of pFRAMBOISE-notfused strains and resulting the β-ionone production, this new peak most likely corresponds to β-carotene, i.e., the expected precursor. | ||
| − | The negative control with no inducer also presents the same activity, meaning the induction system did not work here.Nonetheless, CrtY is functional to produce β-carotene</p> | + | The negative control with no inducer also presents the same activity, meaning the induction system did not work here.Nonetheless, CrtY is functional to produce β-carotene.</p> |
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Revision as of 22:23, 16 October 2021
Gene coding for the lycopene cyclase CrtY
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]
Introduction
This sequence codes for the CrtY enzyme, that transforms Lycopene into β-carotene. The CrtY sequence comes from Pantoea ananatis and was codon optimized for its expression into S. cerevisiae. Its sequence is described into the publication of López et al. (2020).
Characterisation
Production of β-carotene
To demonstrate the CrtY activity, carotenoids contained in the cells were extracted using the method described by López et al. (2020). Yeast cells were lysed in acetone using glass beads and the supernatant obtained after this lysis was analyzed by RP-HPLC using a C18 column. In the LycoYeast-pFRAMBOISE-notfused strains, lycopene is converted into a new product with a higher retention time upon induction (Figure 1). Considering the yellow color of pFRAMBOISE-notfused strains and resulting the β-ionone production, this new peak most likely corresponds to β-carotene, i.e., the expected precursor. The negative control with no inducer also presents the same activity, meaning the induction system did not work here.Nonetheless, CrtY is functional to produce β-carotene.
We concluded the CrtY (BBa_K3930018) part works under those lab conditions
References
- López J, Bustos D, Camilo C, Arenas N, Saa PA, Agosin E. 2020. Engineering Saccharomyces cerevisiae for the Overproduction of β-Ionone and Its Precursor β-Carotene. Front Bioeng Biotechnol. 8:578793. doi:10.3389/fbioe.2020.578793.