Cell Extracts from Bacteria and Yeast Retain Metabolic Activity after Extended Storage and Repeated Thawing

Blake J. Rasor, Ashty S. Karim, Hal S. Alper, Michael C. Jewett*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

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.

Original languageEnglish (US)
Pages (from-to)904-908
Number of pages5
JournalACS synthetic biology
Volume12
Issue number3
DOIs
StatePublished - Mar 17 2023

Keywords

  • biocatalysis
  • cell-free
  • extract
  • in vitro
  • metabolism
  • stability

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology (miscellaneous)
  • Biomedical Engineering

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