Difference between revisions of "Part:BBa K3930018"
Line 16: Line 16:
 
         <div class="thumb tnone">
 
         <div class="thumb tnone">
 
             <div class="thumbinner" style="width:50%;">
 
             <div class="thumbinner" style="width:50%;">
−
                 <a href="https://2021.igem.org/wiki/images/2/22/T--Toulouse_INSA-UPS--2021_fig38_results_Activity_assay_of_DBR1.png" class="image">
+
                 <a href="https://2021.igem.org/wiki/images/a/ac/T--Toulouse_INSA-UPS--2021_results_Carotenoid_analysis.png" class="image">
−
                     <img alt="" src="https://2021.igem.org/wiki/images/2/22/T--Toulouse_INSA-UPS--2021_fig38_results_Activity_assay_of_DBR1.png" width="100%" height=auto class="thumbimage" /></a>                  <div class="thumbcaption">
+
                     <img alt="" src="https://2021.igem.org/wiki/images/a/ac/T--Toulouse_INSA-UPS--2021_results_Carotenoid_analysis.png" width="100%" height=auto class="thumbimage" /></a>                  <div class="thumbcaption">
 
                     <div class="magnify">
 
                     <div class="magnify">
−
                         <a href="https://2021.igem.org/wiki/images/2/22/T--Toulouse_INSA-UPS--2021_fig38_results_Activity_assay_of_DBR1.png" class="internal" title="Enlarge"></a>
+
                         <a href="https://2021.igem.org/wiki/images/a/ac/T--Toulouse_INSA-UPS--2021_results_Carotenoid_analysis.png" class="internal" title="Enlarge"></a>
 
                     </div>
 
                     </div>
 
                     <b>Figure 5: </b> <b>GC-MS analysis of the dodecane layer of the LycoYeast-pFRAMBOISE-notfused</b>  
 
                     <b>Figure 5: </b> <b>GC-MS analysis of the dodecane layer of the LycoYeast-pFRAMBOISE-notfused</b>  

Revision as of 08:26, 15 October 2021


Gene coding for the lycopene cyclase CrtY 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]

Introduction

This sequence codes for the CrtY enzyme, that transforms Lycopene into β-carotene. The CrtY sequence comes from Pantoea ananatis and is codon optimized for an expression into S.cerevisiae. Its sequence is described into the publication of (López et al. 2020).

Characterisation

Production of β-ionone

The β-ionone is very volatile. 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 5 shows the GC-MS spectrum for the LycoYeast-FRAMBOISE-notfused strain. A peak can be observed at the same retention time as the β-ionone standard for the induced LycoYeast-FRAMBOISE-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, the main molecule of the violet odour, was successfully achieved with this construction.


Figure 5: GC-MS analysis of the dodecane layer of the LycoYeast-pFRAMBOISE-notfused

β-ionone is produced in vivo by our strain LycoYeast-pFRAMBOISE-notfused. On the right are presented the mass spectra that correspond between the standard and the observed peak.


The CrtY (BBa_K3930018) part works under those lab conditions

References

  1. 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.
    2.