Off-Colony Screening of Biosynthetic Libraries by Rapid Laser-Enabled Mass Spectrometry

• Published in: ACS synthetic biology 2019-11, Vol.8 (11), p.2566-2575
• Main Authors: Gowers, Glen-Oliver F, Cameron, Simon J. S, Perdones-Montero, Alvaro, Bell, David, Chee, Soo Mei, Kern, Marcelo, Tew, David, Ellis, Tom, Takáts, Zoltan
• Format: Article
• Language: English
• ISSN: 2161-5063; 2161-5063
• DOI: 10.1021/acssynbio.9b00243

By leveraging advances in DNA synthesis and molecular cloning techniques, synthetic biology increasingly makes use of large construct libraries to explore large design spaces. For biosynthetic pathway engineering, the ability to screen these libraries for a variety of metabolites of interest is essential. If the metabolite of interest or the metabolic phenotype is not easily measurable, screening soon becomes a major bottleneck involving time-consuming culturing, sample preparation, and extraction. To address this, we demonstrate the use of automated laser-assisted rapid evaporative ionization mass spectrometry (LA-REIMS)a form of ambient laser desorption ionization mass spectrometryto perform rapid mass spectrometry analysis direct from agar plate yeast colonies without sample preparation or extraction. We use LA-REIMS to assess production levels of violacein and betulinic acid directly from yeast colonies at a rate of 6 colonies per minute. We then demonstrate the throughput enabled by LA-REIMS by screening over 450 yeast colonies within