Coordinated induction of multi-gene pathways in Saccharomyces cerevisiae

Abstract

Bacterial operons are nature’s tool for regulating and coordinating multi-gene expression in prokaryotes. They are also a gene architecture commonly used in the biosynthesis of many pharmaceutically important compounds and industrially useful chemicals. Despite being an important eukaryotic production host, Saccharomyces cerevisiae has never had such gene architecture. Here, we report the development of a system to assemble and regulate a multi-gene pathway in S. cerevisiae. Full pathways can be constructed using pre-made parts from a plasmid toolbox. Subsequently, through the use of a yeast strain containing a stably integrated gene switch, the assembled pathway can be regulated using a readily available and inexpensive compound—estradiol—with extremely high sensitivity (10 nM). To demonstrate the use of the system, we assembled the five-gene zeaxanthin biosynthetic pathway in a single step and showed the ligand-dependent coordinated expression of all five genes as well as the tightly regulated production of zeaxanthin. Compared with a previously reported constitutive zeaxanthin pathway, our inducible pathway was shown to have 50-fold higher production level.

Development and characterization of AND‐gate dynamic controllers with a modular synthetic GAL1 core promoter in Saccharomyces cerevisiae

“…To overcome the glucose repression barrier, we constructed synthetic hybrid promoters with UEEs from the copper inducible CUP1 promoter and phosphate starvation inducible PHO5 promoter. Recently, the use of hormone based gene expression system was characterized (Liang et al, ; McIsaac et al, ). This will enable the use of inducers with very low costs and even single gene‐specificity induction.…”

“…Error bars represent standard deviation from three replicates. system was characterized (Liang et al, 2013;McIsaac et al, 2013). This will enable the use of inducers with very low costs and even single gene-specificity induction.…”

Engineering Microbial Cells for the Biosynthesis of Natural Compounds of Pharmaceutical Significance

“…New methods for the rapid cloning of single genes together with the availability of synthetic operons, such as bacterial operons (commonly used in the biosynthesis of many medically and pharmaceutically important compounds), have accelerated the construction of synthetic multigene pathways [88, 89]. A very recent application was the coordinated induction of a five-gene pathway leading to zeaxanthin production in tailored S. cerevisiae [90]. …”

Rational design and evolutional fine tuning of Saccharomyces cerevisiae for biomass breakdown

“…Recent advances in molecular biology have made it easier to express multiple genes or delete marker-reusable genes in S. cerevisiae, making it possible to test a wider range of genetic modifications [50][51][52]. As engineering transcription factors and activators can trigger global changes in transcription, this is an attractive strategy for enhancing stress tolerance in yeast [53].…”