Streamlined Production of Cell-Free Protein Synthesis Reactions Using Auto-Induction Media

AIChE  Poster Competition Abstract
Christina Muhlestein

Streamlined Production of Cell-free Protein Synthesis Reactions Using Auto-induction Media.

As protein engineering becomes increasingly popular, the benefits of Cell-free Protein Synthesis (CFPS) research are beginning to be realized.  Among the procedures required for CFPS is the production of the necessary enzymes, cofactors, chaperons from cells.  As more labs and industries would like to apply CFPS for their bioconversion application, the use of CFPS is often hindered by labor-intensive, complicated procedures and specialized equipment.  To address this constraint and reduce the time and machinery required by current labs preparing CFPS reagents, here we streamline and simplify the production of CFPS reagents.  Even shake-flask scale culture requires 25 hours, with eight of those hours requiring active participation for proper exogenous protein induction.  Here we show that using a CFPS-compatible auto-induction media (AIM) during cell growth, the requirement for monitoring (~5 hours) is eliminated.  In addition, the resulting cell extract has proven not only safe for use in a cell-free system, but when tested against the traditional extract protocol, showed a 24% increase in protein yields.