Engineering caveolin-mediated endocytosis in Saccharomyces cerevisiae

Area of Science:

• Biotechnology
• Cell Biology
• Metabolic Engineering

Background:

• Oils are hydrophobic and cannot be directly transported into cells for industrial applications.
• Low-cost oils are potential feedstocks for producing acetyl-CoA derivatives in industrial settings.
• Efficient uptake of hydrophobic substrates is crucial for optimizing microbial production processes.

Purpose of the Study:

• To investigate the potential of caveolin in Saccharomyces cerevisiae for absorbing exogenous oils.
• To establish a caveolin-mediated system for transporting hydrophobic substrates into yeast cells.
• To enhance the production of naringenin by facilitating oil uptake and hydrolysis in engineered yeast.

Main Methods:

• Expressed caveolin fused with green fluorescent protein in Saccharomyces cerevisiae to observe vesicle formation.
• Utilized 5,6-carboxyfluorescein to confirm the ability of caveolae to transport substances into cells.
• Co-expressed caveolin and lipase from Bacillus pumilus in a naringenin-producing yeast strain.

Main Results:

• Caveolin expression mediated the formation of microvesicles in yeast, demonstrating its role in transport.
• Caveolae were shown to absorb exogenous soybean oil, confirmed by Nile Red staining and triacylglycerol detection.
• Co-expression of caveolin and lipase increased naringenin production from 222 mg/g DCW to 269 mg/g DCW.

Conclusions:

• Caveolin facilitates a transporter-independent system for the uptake of hydrophobic oils in Saccharomyces cerevisiae.
• This caveolin-mediated system offers a promising strategy for improving the transport of hydrophobic substrates in industrial biotechnology.
• The enhanced naringenin production highlights the potential of this approach for optimizing the biosynthesis of valuable compounds.