Engineering energy-efficient Saccharomyces cerevisiae for methanol and CO₂ assimilation

Area of Science:

• Biotechnology
• Synthetic Biology
• Microbial Engineering

Background:

• Methanol is a key C1 feedstock for microbial bioconversion.
• Engineered Saccharomyces cerevisiae often experiences energy limitations during methanol assimilation.

Purpose of the Study:

• To develop an energy-efficient methylotrophic Saccharomyces cerevisiae strain for improved methanol utilization.
• To enhance ATP and NADH generation during methanol metabolism.
• To enable co-assimilation of methanol-derived intermediates and CO₂.

Main Methods:

• Engineering of heterologous methanol-formaldehyde-formate (MFF) oxidation pathways.
• Adaptive laboratory evolution of Saccharomyces cerevisiae.
• Characterization of key energy modules (Fdh1sc, Adh2m, Aoxm, Rgi2m).
• Investigation of formaldehyde-induced DNA damage.

Main Results:

• Developed SC-AOX25, an energy-efficient methylotrophic S. cerevisiae strain.
• SC-AOX25 efficiently generates ATP and NADH during methanol metabolism.
• Co-assimilation of methanol intermediates and CO₂ via native and non-native pathways (Calvin cycle).
• Identified strategies for methanol detoxification and phenotype enhancement based on DNA damage analysis.

Conclusions:

• SC-AOX25 is a robust and energy-efficient methylotrophic platform for C1 engineering.
• The engineered strain facilitates CO₂ assimilation during methanol fermentation.
• This work advances the potential of microbial bioconversion using methanol as a feedstock.