The improvement of amorpha‐4,11‐diene production by a yeast‐conform variant

Aims:  To investigate the effect of the yeast‐conform variant of the Artemisia annua gene encoding for amorpha‐4,11‐diene synthase (ADS) on the production of amorpha‐4,11‐diene in a transformed yeast. Methods and Results:  The ADS gene was mutated to the yeast‐conform variant ADSm . The ADSm synthesis was performed based on step‐by‐step extension of a short region of the gene through a series of polymerase chain reactions (PCR). The artificial ADSm gene contained codons preferred by the yeast translation machinery. The sequence was then integrated into a yeast expression vector pYeDP60. The fusion construct was active and the transformed yeast cells produced higher level of amorpha‐4,11‐diene compared with the plant gene‐transformed yeast cells. Conclusions:  Strains transformed with the yeast‐conform allele ( ADSm ) were more efficient in terms of production of amorpha‐4,11‐diene than those transformed with the plant gene. Significance and Impact of the Study:  We demonstrated that yeast‐conform allele of foreign genes by serial PCR reactions can be a solution to low efficiency of heterologous gene expression in Saccharomyces cerevisiae cells.