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Real-Time Screening of Biocatalysts in Live Bacterial Colonies.

Cunyu YanFabio ParmeggianiEmrys A JonesEmmanuelle ClaudeShaneela A HussainNicholas J TurnerSabine L FlitschPerdita E Barran
Published in: Journal of the American Chemical Society (2017)
Screening of bacterial colonies to identify new biocatalytic activities is a widely adopted tool in biotechnology, but is constrained by the requirements for colorimetric or tag-based detection methods. Herein we report a label-free screening platform for biotransformations in live colonies using desorption electrospray ionization coupled with ion mobility mass spectrometry imaging (DiBT-IMMS). The screening method is demonstrated for both ammonia lyases and P450 monooxygenases expressed within live bacterial colonies and is shown to enable multiplexing of enzyme variants and substrate libraries simultaneously.
Keyphrases
  • label free
  • mass spectrometry
  • high resolution
  • quantum dots
  • living cells
  • single cell
  • anaerobic digestion
  • fluorescence imaging
  • single molecule
  • simultaneous determination