All reactions were transferred to an Eppendorf Thermomixer ® and spun at 300 RPM for 10 minutes and were then transferred to ice. The reactions were loaded into pre-washed (100 mM NaOH then RXN buffer) 0.5 mL Amicon™ Ultra 10 kDa MWCO centrifugal filter units, concentrated to ~50 µL, then diluted to 500 µL, and concentrated again. This washing step was performed two more times. All centrifugation steps occurred at 4°C to prevent thioester cleavage. TycF thioesterase (final concentration of 5 µM) was added to each centricon for product cleavage in a final volume of 150 µL. The product cleavage reaction was run for 90 minutes at 25°C. The centricon volume was diluted to 400 µL with RXN buffer and the flowthrough was collected into deactivated, silanized Waters vials. Three total rounds of dilution and collection were performed to collect all cleaved product. The vials containing ~1.2 mL of cleaved product in RXN buffer were lyophilized overnight.