Enhancing 3-hydroxypropionic acid production in Saccharomyces cerevisiae through enzyme localization within...

Area of Science:

• Biotechnology
• Metabolic Engineering
• Synthetic Biology

Background:

• Microbial 3-hydroxypropionic acid (3-HP) production presents a sustainable alternative to petroleum-based acrylic acid synthesis.
• Targeting intracellular compartments like mitochondria can optimize metabolic pathways.
• Mitochondria offer a unique environment with key intermediates such as acetyl-CoA for enhanced biosynthesis.

Purpose of the Study:

• To engineer a yeast strain for enhanced 3-HP production by targeting mitochondrial biosynthesis.
• To investigate the impact of mitochondrial localization of 3-HP producing enzymes.
• To further optimize 3-HP production through the overexpression of cMCR in the mitochondria.

Main Methods:

• Construction of yeast strains expressing 3-HP biosynthesis enzymes in mitochondria (YPH-mtA3HP) and cytosol (YPH-cyA3HP).
• Overexpression of cMCR, a rate-limiting enzyme in 3-HP synthesis, using the δ-integration strategy in mitochondria.
• Flask-scale cultivation and comparative analysis of 3-HP production yields between engineered strains.

Main Results:

• Mitochondrial expression of 3-HP producing enzymes (YPH-mtA3HP) resulted in higher 3-HP production compared to cytosolic expression (YPH-cyA3HP).
• Overexpression of cMCR in mitochondria using δ-integration significantly increased 3-HP production, with yields up to 257 mg/L.
• The YPH-mtA3HP22* strain achieved a 3.2-fold higher 3-HP titer than the cytosolic strain, demonstrating successful optimization.

Conclusions:

• Mitochondrial engineering in Saccharomyces cerevisiae is a viable strategy for enhancing microbial 3-HP production.
• Localization of metabolic pathways within mitochondria can significantly improve the yield of target compounds.
• This approach holds promise for the sustainable and efficient bio-based production of chemicals like 3-HP.