Efficient diterpene production in yeast by engineering Erg20p into a geranylgeranyl diphosphate synthase

Area of Science:

• Biotechnology and synthetic biology
• Microbial engineering
• Metabolic engineering

Background:

• Terpenes are versatile compounds with applications in pharmaceuticals, fragrances, and biofuels.
• Sustainable and cost-effective microbial terpene production is a growing area of interest.
• Current methods in Saccharomyces cerevisiae show lower efficiency for 20-carbon diterpenes compared to 15-carbon sesquiterpenes.

Purpose of the Study:

• To identify and address limitations in yeast-based diterpene production.
• To engineer yeast to enhance the synthesis of geranylgeranyl diphosphate (GGPP).
• To improve the yields of various isoprenoids derived from GGPP.

Main Methods:

• Identified modular structure of GGPP synthesis in yeast as a key limitation.
• Engineered the yeast farnesyl diphosphate synthase (Erg20p) to produce GGPP.
• Employed a combination of protein and genetic engineering techniques.
• Developed novel yeast strains for efficient isoprenoid production.

Main Results:

• Achieved significant improvements in the production of sclareol and other isoprenoids like cis-abienol, abietadiene, and β-carotene.
• Demonstrated enhanced yields of diterpenes through engineered Erg20p.
• Developed yeast strains suitable as dedicated platforms for diterpene production.

Conclusions:

• Engineering Erg20p effectively overcomes limitations in GGPP synthesis for improved diterpene yields.
• The developed yeast strains provide a robust chassis for GGPP-derived isoprenoid production.
• This approach is broadly applicable to various GGPP-derived isoprenoids and compatible with existing terpene production platforms.