Part:BBa_K3570001

Sweet rose taste induction system in S. cerevisiae

Introduction

This biobrick is purposed to make S. cerevisiae to have a sweet rose taste. This is obtained by producing a brazzein, a sweet taste protein. The rose flavor is obtained by expressing the geraniol synthase (GES), which produces geraniol, a rose odorant monoterpenoid. This biobrick can be directly integrated into the yeast genome thanks to YPRcdelta15 homology arms. The experimentator will be able to counter-select the cells that have integrated the construction since they are ment to acquire a prototroph character for leucine.

Design

Fig. 1: Brazzein-CrGES-pUC19. The integrative locus used is yprcΔ15. The selective locus used is LEU2. The gene coding for geraniol synthase (CrGES) comes from C. roseus. The gene coding for brazzein comes from an artificial sequence (LG263246.1). CYC1 and PGK terminators are used. The bidirectional promoter Gal 1/10 is used.

Geraniol (3,7-dimethylocta-trans-2,6-dien-1-ol), an acyclic monoterpene alcohol, is widely used in the flavor and fragrance industries[1]. It is produced by GES, which is a monoterpene synthase, that catalyzes a reaction from geranyl pyrophosphate (GPP) to geraniol. Most of the GES come from Plantae and contain N-terminal transit peptide that targets the translation product towards the plastids[2].

Experiments

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Team iGEM Toulouse 2020 did not have sufficient time to complete the cloning and hence, to test this part functionality.

References

• [1]- Chen, W., & Viljoen, A. M. (2010). Geraniol — A review of a commercially important fragrance material. South African Journal of Botany, 76(4), 643–651. https://doi.org/10.1016/j.sajb.2010.05.008
• [2]- Turner, G., Gershenzon, J., Nielson, E. E., Froehlich, J. E., & Croteau, R. (1999). Limonene Synthase, the Enzyme Responsible for Monoterpene Biosynthesis in Peppermint, Is Localized to Leucoplasts of Oil Gland Secretory Cells. Plant Physiology, 120(3), 879–886. https://doi.org/10.1104/pp.120.3.879

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal EcoRI site found at 4894
  Illegal XbaI site found at 3746
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 4894
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 4894
  Illegal BamHI site found at 3752
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal EcoRI site found at 4894
  Illegal XbaI site found at 3746
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal EcoRI site found at 4894
  Illegal XbaI site found at 3746
  Illegal AgeI site found at 1551
  Illegal AgeI site found at 4507
  
• 1000
  COMPATIBLE WITH RFC[1000]