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Student Class

2029

Student Affiliation

Thayer School First Year Research in Engineering

Author ORCID Identifier

0009-0001-0275-1348

First Advisor

Daniel Olson

First Advisor Department

Department of Engineering Sciences—Thayer School of Engineering

Second Advisor

Marybeth Maloney

Second Advisor Department

Department of Engineering Sciences—Thayer School of Engineering

Abstract

Ethanol stress assays are commonly used to evaluate microbial tolerance, metabolic adaptation, and fermentation performance. However, manual liquid handling introduces variability across replicate wells and small-volume pipetting steps, limiting reproducibility and throughput. This study developed an automated OT-2 robotic workflow to generate replicated ethanol concentration gradients for high-throughput inhibition assays in engineered thermophilic biofuel strains. Kinetic plate-reader measurements were used to quantify ethanol-dependent growth responses under anaerobic fermentation conditions. The reasearch question is: How do engineered thermophilic biofuel strains differ in ethanol-dependent growth inhibition under anaerobic fermentation conditions, and can automated robotic assays improve the reproducibility of these measurements? Can high-throughput robotic ethanol inhibition assays reliably generate reproducible ethanol gradients and reveal strain-specific tolerance responses in engineered thermophilic biofuel organisms?

Publication Date

Spring 5-26-2026

Keywords

Microbial Metabolism, Alcohol Dehydrogenase E, Ethanol Stress, Enzyme Kinetics NADH Assays, High-Throughput Screening, Laboratory Automation, Robotic Liquid Handling, Bioprocess Engineering, Biofuel Production

Disciplines

Artificial Intelligence and Robotics | Biochemistry | Biological Engineering | Bioresource and Agricultural Engineering | Biotechnology | Molecular Biology | Molecular, Cellular, and Tissue Engineering | Numerical Analysis and Scientific Computing | Oil, Gas, and Energy | Systems and Integrative Engineering

High-Throughput Robotic Ethanol Inhibition Assays for Engineered Thermophilic Biofuel Strains

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