Cell Extracts from Bacteria and Yeast Retain Metabolic Activity after Extended Storage and Repeated ThawingClick to copy article linkArticle link copied!
- Blake J. RasorBlake J. RasorDepartment of Chemical and Biological Engineering, Chemistry of Life Processes Institute and Center for Synthetic Biology, Northwestern University, Evanston, Illinois 60208, United StatesMore by Blake J. Rasor
- Ashty S. KarimAshty S. KarimDepartment of Chemical and Biological Engineering, Chemistry of Life Processes Institute and Center for Synthetic Biology, Northwestern University, Evanston, Illinois 60208, United StatesMore by Ashty S. Karim
- Hal S. AlperHal S. AlperInstitute for Cellular and Molecular Biology and McKetta Department of Chemical Engineering, The University of Texas at Austin, Austin, Texas 78712, United StatesMore by Hal S. Alper
- Michael C. Jewett*Michael C. Jewett*E-mail: [email protected]Department of Chemical and Biological Engineering, Chemistry of Life Processes Institute and Center for Synthetic Biology, Northwestern University, Evanston, Illinois 60208, United StatesRobert H. Lurie Comprehensive Cancer Center and Simpson Querrey Institute, Northwestern University, Chicago, Illinois 60611, United StatesMore by Michael C. Jewett
Abstract

Cell-free synthetic biology enables rapid prototyping of biological parts and synthesis of proteins or metabolites in the absence of cell growth constraints. Cell-free systems are frequently made from crude cell extracts, where composition and activity can vary significantly based on source strain, preparation and processing, reagents, and other considerations. This variability can cause extracts to be treated as black boxes for which empirical observations guide practical laboratory practices, including a hesitance to use dated or previously thawed extracts. To better understand the robustness of cell extracts over time, we assessed the activity of cell-free metabolism during storage. As a model, we studied conversion of glucose to 2,3-butanediol. We found that cell extracts from Escherichia coli and Saccharomyces cerevisiae subjected to an 18-month storage period and repeated freeze–thaw cycles retain consistent metabolic activity. This work gives users of cell-free systems a better understanding of the impacts of storage on extract behavior.
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