Service interruption on Monday 11 July from 12:30 to 13:00: all the sites of the CCSD (HAL, EpiSciences, SciencesConf, AureHAL) will be inaccessible (network hardware connection).
Skip to Main content Skip to Navigation
Journal articles

Molecular optimization of rabies virus glycoprotein expression in Pichia pastoris

Abstract : In this work, different approaches were investigated to enhance the expression rabies virus glycoprotein (RABV-G) in the yeast Pichia pastoris; this membrane protein is responsible for the synthesis of rabies neutralizing antibodies. First, the impact of synonymous codon usage bias was examined and an optimized RABV-G gene was synthesized. Nevertheless, data showed that the secretion of the optimized RABV-G gene was not tremendously increased as compared with the non-optimized one. In addition, similar levels of RABV-G were obtained when α-factor mating factor from Saccharomyces cerevisiae or the acid phosphatase PHO1 was used as a secretion signal. Therefore, sequence optimization and secretion signal were not the major bottlenecks for high-level expression of RABV-G in P. pastoris. Unfolded protein response (UPR) was induced in clones containing high copy number of RABV-G expression cassette indicating that folding was the limiting step for RABV-G secretion. To circumvent this limitation, co-overexpression of five factors involved in oxidative protein folding was investigated. Among these factors only PDI1, ERO1 and GPX1 proved their benefit to enhance the expression. The highest expression level of RABV-G reached 1230 ng ml(-1). Competitive neutralizing assay confirmed that the recombinant protein was produced in the correct conformational form in this host.
Document type :
Journal articles
Complete list of metadata

Cited literature [64 references]  Display  Hide  Download
Contributor : AbdelHakim Ben Hassine Connect in order to contact the contributor
Submitted on : Friday, February 8, 2019 - 1:38:27 PM
Last modification on : Thursday, March 31, 2022 - 1:00:20 PM
Long-term archiving on: : Thursday, May 9, 2019 - 2:48:32 PM


Molecular optimization of rabi...
Publication funded by an institution




Safa Ben Azoun, Aicha Eya Belhaj, Rebecca Göngrich, Brigitte Gasser, Héla Kallel. Molecular optimization of rabies virus glycoprotein expression in Pichia pastoris. Microbial Biotechnology, Wiley, 2016, 9 (3), pp.355-368. ⟨10.1111/1751-7915.12350⟩. ⟨pasteur-02012033⟩



Record views


Files downloads