Difference between revisions of "Part:BBa K3570000"

Revision as of 13:21, 13 October 2020

GGPP production enhancement in S. cerevisiae

The ultimate goal of this biobrick is to enhance the mevalonate pathway in S. Cerevisiae to increase the pool of geranylgeranyl pyrophosphate (GGPP). The surplus of GGPP can be used to make S. Cerevisiae produce provitamin A (𝛽-carotene), geraniol or limonene using BBa_K3570001, BBa_K3570002 or BBa_K3570003 biobricks respectively. BBa_K3570002 and then, produce β-carotene. This construction shall be used in tight synergy with to achieve this goal. Precisely, β-carotene derives from the mevalonate pathway as do limonene and geraniol (figure 1).

Fig. 1: Metabolic pathway of 𝛽-carotene, limonene and geraniol from Acetyl-CoA in engineered yeast.

We, therefore, decided to enhance the mevalonate pathway and the production of GGPP. According to Rabeharindranto et al. 2019, the enhancement of the mevalonate pathway can be achieved by overexpressing the gene HMG1 and CrtE. Our design differs from this publication in the choice of the promoter. The promoter TDH1 has been chosen instead of Gal1/10 because we planned to use this promoter for the regulation of other genes. We thus created the plasmids containing a truncated version of the HMG1 gene from S. Cerevisiae and the CrtE gene from X. Dendrorhous as on the figure 2.

Fig. 2: HMG1-CrtE-pUC19. The integrative locus used is DPP. The selective locus used is HIS. The genes coding for the truncated version of HMG1 comes from S. cerevisiae. The genes coding for CrtE comes from X. dendrorhous. We use the bidirectional TDH3-TEF1 promoter and the terminators tCYC1 and tPGK.

Literature: Hery Rabeharindranto, Sara Castaño-Cerezo, Thomas Lautier, Luis F. Garcia-Alles, Christian Treitz, Andreas Tholey, Gilles Truan, 2019. Enzyme-fusion strategies for redirecting and improving carotenoid synthesis in S. cerevisiae. Metab Eng Commun. 2019 Jun; 8: e00086.

Sequence and Features