Difference between revisions of "Part:BBa K3930015"
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<h3>Production of β-ionone</h3> | <h3>Production of β-ionone</h3> | ||
| − | <p>All the experiments that characterized this part are related to the final construct | + | <p>All the experiments that characterized this part are related to the final construct pFRAMBOISE-notfused <a href="https://parts.igem.org/Part:BBa_K3930002" class="pr-0" target="_blank">(BBa_K3930002)</a> which was cloned into the S. cerevisiae LycoYeast strain. For more information on the experimental background, please refer to this part</p> |
<p>In the part (BBa_K3930002), the TEF1 promoter control the expression of CrtY, an enzyme that produce β-ionone from β-carotene. The production of β-ionone can be so considered as a contol of functionality of the promoter TEF1. pFLEUR was transformed into the <i>S.cerevisiae</i> LycoYeast strain. The β-ionone is very volatile and a common strategy to avoid losing these molecules during the culture is to grow the engineered microorganisms in a culture medium supplemented with an organic phase to trap the molecules of interest.The most common organic solvent used is dodecane for ionones (Chen et al. 2019; López et al. 2020). Figure 1 shows the GC-MS spectrum for the LycoYeast-pFRAMBOISE-notfused strain. A peak can be observed at the same retention time as the β-ionone standard for the induced LycoYeast-pFRAMBOISE-notfused strain. The mass spectra associated with this peak matched with the one obtained with the analytical standard. The β-ionone attribution was further confirmed by the NIST mass spectral library (National Institute of Standards and Technology). The production of β-ionone, was successfully achieved with this construction. This mean that the TEF1 promoter is functional to express gene in S. cerevisiae</p> | <p>In the part (BBa_K3930002), the TEF1 promoter control the expression of CrtY, an enzyme that produce β-ionone from β-carotene. The production of β-ionone can be so considered as a contol of functionality of the promoter TEF1. pFLEUR was transformed into the <i>S.cerevisiae</i> LycoYeast strain. The β-ionone is very volatile and a common strategy to avoid losing these molecules during the culture is to grow the engineered microorganisms in a culture medium supplemented with an organic phase to trap the molecules of interest.The most common organic solvent used is dodecane for ionones (Chen et al. 2019; López et al. 2020). Figure 1 shows the GC-MS spectrum for the LycoYeast-pFRAMBOISE-notfused strain. A peak can be observed at the same retention time as the β-ionone standard for the induced LycoYeast-pFRAMBOISE-notfused strain. The mass spectra associated with this peak matched with the one obtained with the analytical standard. The β-ionone attribution was further confirmed by the NIST mass spectral library (National Institute of Standards and Technology). The production of β-ionone, was successfully achieved with this construction. This mean that the TEF1 promoter is functional to express gene in S. cerevisiae</p> | ||
Revision as of 15:21, 16 October 2021
Constitutive promoter TEF1
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 168
Introduction
The promoter TEF1 is a constitutive promoter that comes from the plasmid p405TEF1 from Nicolas Buchler & Fred Cross (unpublished). The TEF1 promoter coming from the yeast Y. lipolitica is already described and used in part (BBa_K2117000), but ours comes from the S. cerevisiae genome.
Results
Production of β-ionone
All the experiments that characterized this part are related to the final construct pFRAMBOISE-notfused (BBa_K3930002) which was cloned into the S. cerevisiae LycoYeast strain. For more information on the experimental background, please refer to this part
In the part (BBa_K3930002), the TEF1 promoter control the expression of CrtY, an enzyme that produce β-ionone from β-carotene. The production of β-ionone can be so considered as a contol of functionality of the promoter TEF1. pFLEUR was transformed into the S.cerevisiae LycoYeast strain. The β-ionone is very volatile and a common strategy to avoid losing these molecules during the culture is to grow the engineered microorganisms in a culture medium supplemented with an organic phase to trap the molecules of interest.The most common organic solvent used is dodecane for ionones (Chen et al. 2019; López et al. 2020). Figure 1 shows the GC-MS spectrum for the LycoYeast-pFRAMBOISE-notfused strain. A peak can be observed at the same retention time as the β-ionone standard for the induced LycoYeast-pFRAMBOISE-notfused strain. The mass spectra associated with this peak matched with the one obtained with the analytical standard. The β-ionone attribution was further confirmed by the NIST mass spectral library (National Institute of Standards and Technology). The production of β-ionone, was successfully achieved with this construction. This mean that the TEF1 promoter is functional to express gene in S. cerevisiae
This promoter TEF1 works under those lab conditions.