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Gilles Truan - Jeudi 30 novembre 2017 11h

Salle de l’Erist - Bât. 400 - Séminaire Micalis

 Gilles Truan -  Jeudi 30 novembre 2017 11h
Enzyme-fusion strategies for redirecting carotenoid synthesis in S. cerevisiae

Enzyme-fusion strategies for redirectingcarotenoid synthesis in S. cerevisiae

Gilles Truan

LISBP - INSA de Toulouse

Spatial clustering of metabolic enzymes has proven an efficient approach for the optimization of metabolic pathways. Among the multiple methods used to promote colocalisation, enzyme fusion is probably the simplest. Inspired by natural enzymes fusions, we have explored different spatial reorganizations of the carotenoids enzymatic domains from Xanthophyllomyces dendrorhous using an enzymatic domain fusion strategy followed by the heterologous expression in the yeast Saccharomyces cerevisiae. Various synthetic genes were constructed encoding bidomain enzymes composed of CrtI and a truncated CrtB domain from the natural CrtYB fusion. Domains were connected with the two possible orientations using a series of different natural and synthetic linkers. Genes encoding tridomain enzymes harboring the full β-carotene producing pathway were also designed. All these constructs were integrated in the genome of S. cerevisiae and tested for both the level of enzyme expression as well as for the production of carotenoid molecules. Our results demonstrate that enzyme orientation and linker selection impact considerably enzyme expression and/or stability, domain functionality and product yield. Furthermore, we demonstrate that specific enzyme fusions can be used to produce carotenoid molecules that are not present when using heterologous expression of the natural X. dendrorhous enzymes in S. cerevisiae. Our data strengthen the idea that fusion of enzymatic domains is an appropriate technique to achieve spatial confinement and modify the production of molecules not easily attainable with optimized natural systems.

Jeudi 30 novembre 2017

11h

Salle de l’Erist - Bât. 400

INRA, Jouy en Josas

Invité par Anne-Gaëlle Planson