Biosynthesis of catharanthine in engineered Pichia pastoris

A roadmap to establish a comprehensive platform for sustainable manufacturing of natural products in yeast

“…Furthermore, it shows a higher expression level of heterologous protein than S. cerevisiae. For example, recombinant proteins expressed under the control of the yeast alcohol oxidase 1 (AOX1) promoter can contribute more than 30% of the total proteins in P. pastoris 23 . A recent study has shown that a cytochrome P450 enzyme, which catalyzes the conversion of scoulerine (a product downstream of reticuline) to cheilanthifoline, exhibits a higher conversion rate in P. pastoris cells (70%) than S. cerevisiae cells (20%) 24 .…”

Advances in Metabolic Engineering of Plant Monoterpene Indole Alkaloids

“…The methods and tools, such as gene expression cassettes for the simultaneous integration of multiple genes in engineering P. pastoris, are more readily available [16,70,72]. Remarkably, the de novo biosynthesis of catharanthine in P. pastoris was recently described, allowing for improved production of catharanthine from simple carbon sources at a maximum titer of 2.57 mg/L in comparison to previously reported yield from S. cerevisiae [73,74]. This study highlights the significance of optimization strategies developed for reconstitution, such as the complex long MIA pathway in P. pastoris leading to catharanthine, by addressing various approaches.…”

“…This study highlights the significance of optimization strategies developed for reconstitution, such as the complex long MIA pathway in P. pastoris leading to catharanthine, by addressing various approaches. This includes the selection of suitable plant enzymes with enhanced functionality, facilitating the trafficking of substrates by expressing fusion protein, and the amplification of genes involved in rate-limiting steps [73,74]. The rapid development of tools in synthetic biology, including the application of CRISPR/Cas9, boosts the prospect of overcoming challenges in metabolic regulation to improve precursor supplies and ensures balanced microbial cell growth in the future industrial production of high-value natural products.…”

Yeast Metabolic Engineering for Biosynthesis of Caffeic Acid-Derived Phenethyl Ester and Phenethyl Amide

“…However, as the structural complexity of the synthesized natural products increases, the synthetic pathways become much longer, consisting of more heterologous reactions. For example, the opioid and vinblastine synthesis pathways were constructed in yeast by modifying dozens of endogenous and exogenous genes. − In the construction of such complex pathways, microbial chassis are often required to simultaneously provide multiple intermediates, such as phenolic compounds, as key precursors for subsequent reactions. As a secondary metabolism, these precursors are also difficult to synthesize and accumulate in high amounts.…”