Improving 2-phenylethanol production via Ehrlich pathway using genetic engineered Saccharomyces cerevisiae strains

Area of Science:

• Biotechnology
• Synthetic Biology
• Microbial Engineering

Background:

• 2-phenylethanol (2-PE) is a valuable aromatic compound with a rose-like scent, widely utilized in the food and cosmetic industries.
• There is a growing demand for naturally sourced 2-PE, driving research into sustainable production methods.
• The Ehrlich pathway in yeast presents a potential metabolic route for 2-PE biosynthesis.

Purpose of the Study:

• To enhance the production of natural 2-phenylethanol (2-PE) using genetically modified Saccharomyces cerevisiae.
• To investigate the role of the Ehrlich pathway enzymes, transaminase (encoded by ARO8) and decarboxylase (encoded by ARO10), in 2-PE biosynthesis.
• To optimize 2-PE yield through the enhanced expression of key genes in yeast.

Main Methods:

• Genetic modification of Saccharomyces cerevisiae S288c to express the ARO8 and ARO10 genes.
• Construction of recombinant strains with enhanced activities of transaminase and decarboxylase.
• Development of a co-expression vector for simultaneous ARO8 and ARO10 expression.
• Fed-batch fermentation to evaluate 2-PE production in engineered yeast strains.

Main Results:

• Individual enhanced expression of ARO8 and ARO10 increased 2-PE yield by 9.3% and 16.3%, respectively, compared to the wild strain.
• Co-expression of ARO8 and ARO10 in the recombinant strain SPO810 resulted in a significant increase in 2-PE production.
• The SPO810 strain achieved a 2-PE yield of 2.61 g L(-1) after 60 hours of fed-batch fermentation, a 36.8% improvement over the wild strain.

Conclusions:

• Enhanced expression of ARO8 and ARO10 genes in the Ehrlich pathway significantly improves 2-phenylethanol production in Saccharomyces cerevisiae.
• Metabolic engineering of yeast offers a viable strategy for increasing the yield of natural 2-PE.
• These findings provide a foundation for developing efficient microbial platforms for the sustainable production of natural aromatic compounds.