US 11,913,052 B2
Cell-free protein synthesis driven metabolic engineering
Michael Christopher Jewett, Evanston, IL (US); Ashty Stephen Karim, Chicago, IL (US); Jian Li, Evanston, IL (US); and Quentin Dudley, Evanston, IL (US)
Assigned to Northwestern University, Evanston, IL (US)
Filed by Northwestern University, Evanston, IL (US)
Filed on Mar. 2, 2020, as Appl. No. 16/807,147.
Application 16/807,147 is a continuation in part of application No. 15/175,497, filed on Jun. 7, 2016, granted, now 10,577,632.
Claims priority of provisional application 62/173,818, filed on Jun. 10, 2015.
Prior Publication US 2020/0291445 A1, Sep. 17, 2020
Int. Cl. C12N 15/67 (2006.01); C12P 21/02 (2006.01)
CPC C12P 21/02 (2013.01) [C12N 15/67 (2013.01)] 15 Claims
 
1. A cell-free method for synthesizing a non-ribosomal peptide target protein without the use of a transcription or translation template encoding the target protein, the method comprising:
a) contacting a translation template encoding a non-ribosomal peptide synthase (“NRPS”) that is specific for the synthesis of the target protein with a cellular extract from a host strain, the extract comprising natural enzyme metabolism from the host strain;
wherein the translation template is translated once it is contacted with the cellular extract, thereby providing a NRPS-containing extract;
b) contacting the NRPS-containing extract to a feedstock, the feedstock comprising components for the formation of the target protein.