Synthesis of monoterpene geraniol by peroxisome from Saccharomyces cerevisiae

Abstract

Geraniol, an acyclic monoterpenoid compound, is widely used in the production of cosmetics and spices due to its rose-like odor. In addition, its medicinal value has also attracted wide attention. Geraniol, as a precursor for the synthesis of some clinical anticancer drugs, plays a key role in the biosynthesis of monoterpene indole alkaloids such as camptothecin and vincristine. It is also its wide application field and development prospect. By using the microbial cell factory method, this study selected a Saccharomyces cerevisiae strain 051H with good upstream and downstream metabolic pathways as a chassis cell to overcome the disadvantages of traditional methods such as low yield and easy to be affected by the environment, and to increase the production of geranyl alcohol. Meet the current high market demand. In this study, by constructing a recombinant plasmid expression frame, the gene encoding peroxisome whole geraniol metabolic pathway was introduced into the genome of S. cerevisiae by homologous recombination technology, so that it could be stably expressed. The recombinant strain P0515 was constructed to avoid the metabolic burden caused by plasmid insertion and improve the production of geraniol without affecting the normal growth of the strain. The research results of this paper are as follows: (1) Firstly, the efficiency of localization of peroxisome signal peptide was verified. tVoGES was localized into the peroxisome of Xylobacter spp. to construct strain 051P. The yield of geranyl alcohol was 15.46 mg/L, which proved that there was more abundant GPP precursor in peroxisome and this organelles had significant advantages. Subsequently, in order to realize the synthesis of geraniol in the peroxisomal compartment of S. cerevisiae, other enzymes in the geraniol synthesis pathway 5 were integrated into S. cerevisiae in turn to complete the reconstruction of the whole geraniol synthesis pathway in peroxisomes. Strain P0515 was obtained, and the yield was increased to 43.96 mg/L, and the growth effect was less than that of the control strain. It is proved that peroxisomes are not essential for cell growth and therefore more suitable for the construction of heterologous compound synthesis pathways