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A molecular approach to optimize hIFN α2b expression and secretion in Yarrowia lipolytica.

Abstract : In this work, we investigated the effect of codon bias and consensus sequence (CACA) at the translation initiation site on the expression level of heterologous proteins in Yarrowia lipolytica; human interferon alpha 2b (hIFN-α2b) was studied as an example. A codon optimized hIFN-α2b gene was synthesized according to the frequency of codon usage in Y. lipolytica. Both wild-type (IFN-wt) and optimized hIFN-α2b (IFN-op) genes were expressed under the control of a strong inducible promoter acyl-co-enzyme A oxidase (POX2). Protein secretion was directed by the targeting sequence of the extracellular lipase (LIP2): pre-proLIP2. Codon optimization increased protein production by 11-fold, whereas the insertion of CACA sequence upstream of the initiation codon of IFN-op construct resulted in 16.5-fold increase of the expression level; this indicates that translational efficiency plays an important part in the increase of hIFN-α2b production level. The replacement of the pre-proLIP2 signal secretion with the LIP2 pre-region sequence followed by the X-Ala/X-Pro stretch but without the pro-region also increased the secretion of the target protein by twofold, suggesting therefore that the LIP2 pro-region is not necessary for extracellular secretion of small heterologous proteins in Yarrowia lipolytica.
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Submitted on : Thursday, July 7, 2011 - 11:54:57 AM
Last modification on : Friday, August 5, 2022 - 2:38:10 PM

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Najla Gasmi, Franck Fudalej, Héla Kallel, Jean-Marc Nicaud. A molecular approach to optimize hIFN α2b expression and secretion in Yarrowia lipolytica.. Applied Microbiology and Biotechnology, 2011, 89 (1), pp.109-19. ⟨10.1007/s00253-010-2803-0⟩. ⟨pasteur-00606851⟩



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