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Enabling high-throughput enzyme discovery and engineering with a low-cost, robot-assisted pipeline
Brenna Norton-Baker1,2,3, Mackenzie C R Denton1,2, Natasha P Murphy1,2
1Renewable Resources and Enabling Sciences Center, National Renewable Energy Laboratory, Golden, CO, USA.
Scientific Reports
|June 24, 2024
Summary
This study introduces a cost-effective, high-throughput protein purification pipeline using E. coli expression and a liquid-handling robot. The method efficiently purifies 96 proteins in parallel, yielding high-quality enzymes for characterization and enabling plastic-degrading enzyme comparisons.
Area of Science:
- Biochemistry
- Molecular Biology
- Bioengineering
Background:
- Expanding genomic databases necessitate efficient protein characterization.
- Advancements in artificial intelligence drive demand for high-throughput biological data.
- Existing protein purification methods can be costly and time-consuming.
Purpose of the Study:
- To develop a generalizable, low-cost pipeline for high-throughput protein purification.
- To enable parallel purification of numerous proteins with minimal waste.
- To create a standardized method for characterizing plastic-degrading enzymes.
Main Methods:
- Small-scale protein expression in Escherichia coli (E. coli).
- Utilization of an affordable liquid-handling robot for automated purification.
- Parallel purification of 96 protein samples.
Main Results:
- Successful purification of poly(ethylene terephthalate) hydrolases.
- Reproducible results with high enzyme purity and yields (up to 400 µg).
- Generation of a benchmark dataset for enzyme comparison.
Conclusions:
- The developed pipeline is cost-effective, scalable, and easy to implement.
- This platform is broadly applicable to various protein characterization needs in biological sciences.
- Enables efficient generation of standardized data for enzyme analysis.
