Methodology for enabling high-throughput simultaneous saccharification and fermentation screening of yeast using...

Area of Science:

• Biotechnology
• Bioenergy
• Chemical Engineering

Background:

• High-throughput (HTP) screening is crucial for efficient biological data acquisition, particularly in second-generation biofuel production.
• Traditional methods are resource-intensive; HTP screening offers speed and cost-effectiveness.
• Developing scalable methodologies is key to advancing HTP biofuel research.

Purpose of the Study:

• To establish a 96-well plate scale methodology for hydrolyzing and fermenting solid lignocellulosic substrates.
• To facilitate high-throughput screening of ethanol production with high repeatability.
• To enable efficient screening of biomass substrates for biorefining applications.

Main Methods:

• Developed a 96-well plate scale process for lignocellulosic substrate hydrolysis and fermentation.
• Utilized consistent density biomass sheets and pipettable slurries for standardized transfers.
• Performed simultaneous saccharification and fermentation (SSF) at various scales (1 mL to 200 mL).

Main Results:

• Standardized and accurate biomass transfers to 96-well plates achieved (±3.1% and 1.7%).
• No significant difference in ethanol yields across different scales (1 mL, 10 mL, 200 mL).
• Successful trialing of substrate concentrations up to 10% (w/v) at 1 mL scale.
• Demonstrated methodology effectiveness on steam explosion pretreated oilseed rape and wheat straws.

Conclusions:

• The 96-well plate SSF assay can replace larger scale reactions for HTP experimentation.
• Method is compatible with colorimetric, HPLC, and NMR assays.
• Practical applications in biorefining for second-generation biofuels and biobased chemicals through HTP SSF screening.