Engineering a versatile yeast platform for sesquiterpene production from glucose or methanol

Area of Science:

• Biotechnology
• Synthetic Biology
• Metabolic Engineering

Background:

• Natural sesquiterpenes have industrial applications but their production is challenging.
• Microbial synthesis using sustainable carbon sources like methanol is a promising alternative.
• Pichia pastoris is a well-established yeast for protein production and is being engineered for chemical synthesis.

Purpose of the Study:

• To engineer Pichia pastoris for sesquiterpene synthesis using methanol and glucose as carbon sources.
• To analyze the impact of mevalonate pathway modifications on sesquiterpene production.
• To establish a versatile platform for producing various sesquiterpenes.

Main Methods:

• Expression of α-bisabolene synthase gene under constitutive promoters in P. pastoris.
• Systematic analysis of mevalonate pathway components with methanol and glucose.
• Replacement of α-bisabolene synthase with β-farnesene synthase for alternative sesquiterpene production.

Main Results:

• Successful synthesis of α-bisabolene in P. pastoris utilizing dual carbon sources.
• Sesquiterpene synthase module enhanced methanol-based production.
• Metabolic modules MK and PMK improved glucose utilization, cell growth, and titer.
• Demonstrated synthesis of β-farnesene.

Conclusions:

• Established a P. pastoris platform strain for sesquiterpene synthesis from diverse carbon sources.
• Paved the way for P. pastoris as a high-efficiency microbial cell factory for chemicals.
• Laid the foundation for large-scale, efficient methanol-based synthesis of high-value chemicals.