Metabolite trafficking enables membrane-impermeable-terpene secretion by yeast

Area of Science:

• Synthetic Biology
• Metabolic Engineering
• Biotechnology

Background:

• Metabolites accumulate intracellularly due to membrane impermeability, limiting their extraction.
• Sustainable secretion of intracellular metabolites is crucial for efficient, continuous production of high-value chemicals.
• Current methods for microbial metabolite production often face challenges with product recovery and yield.

Purpose of the Study:

• To engineer a synthetic pathway for intracellular metabolite trafficking and secretion in yeast.
• To demonstrate the feasibility of secreting lipophilic metabolites, specifically terpenes, using this novel system.
• To establish an efficient and selective microbial secretion system for valuable chemical compounds.

Main Methods:

• Development of a synthetic metabolite trafficking system by coupling metabolite-binding proteins with signal peptides.
• Systematic fusion of a terpene-binding protein with an export signal peptide in yeast.
• Quantification of secreted terpene compounds (squalene and β-carotene) using analytical techniques.

Main Results:

• Demonstration of efficient and selective terpene secretion by engineered yeast cells.
• Achieved secretion yields of approximately 225 mg/L for squalene and 1.6 mg/L for β-carotene.
• Established a versatile platform for tailoring metabolite trafficking pathways by fusing different carrier proteins with signal peptides.

Conclusions:

• The developed synthetic pathway represents a highly efficient cognate system for metabolite secretion by microorganisms.
• This engineered system holds significant potential for the sustainable mass-production of high-value chemicals.
• The modular nature of the system allows for broad applicability across different microbes and metabolites.