Off-colony screening of biosynthetic libraries by rapid laser-enabled mass spectrometry
| Autor: | David G. Tew, Marcelo Kern, Glen-Oliver F. Gowers, Simon J S Cameron, Tom Ellis, Derek Bell, Alvaro Perdones-Montero, Zoltan Takats, Soo Mei Chee |
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| Přispěvatelé: | Biotechnology and Biological Sciences Research Council (BBSRC) |
| Jazyk: | angličtina |
| Rok vydání: | 2019 |
| Předmět: |
0106 biological sciences
High-throughput screening Metabolite Biomedical Engineering Computational biology ambient mass spectrometry Saccharomyces cerevisiae Mass spectrometry 01 natural sciences Biochemistry Genetics and Molecular Biology (miscellaneous) Agar plate Metabolic engineering 03 medical and health sciences Synthetic biology chemistry.chemical_compound Transformation Genetic 010608 biotechnology Sample preparation Betulinic Acid 030304 developmental biology 0303 health sciences REIMS General Medicine Yeast Triterpenes Culture Media High-Throughput Screening Assays Agar chemistry Metabolic Engineering Spectrometry Mass Matrix-Assisted Laser Desorption-Ionization Synthetic Biology Pentacyclic Triterpenes high throughput screening Chromatography Liquid Plasmids |
| Zdroj: | Gowers, G-O F, Cameron, S J S, Perdones-Montero, A, Bell, D, Chee, S M, Kern, M, Tew, D, Ellis, T & Takats, Z 2019, ' Off-colony screening of biosynthetic libraries by rapid laser-enabled mass spectrometry ', ACS Synthetic Biology . https://doi.org/10.1021/acssynbio.9b00243 |
| Popis: | 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 |
| Databáze: | OpenAIRE |
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