title: Real-Time Screening of Biocatalysts in Live Bacterial Colonies.
abstract: 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.
authors.affiliation: Manchester Synthetic Biology Research Centre for Fine and Speciality Chemicals (SYNBIOCHEM), Manchester Institute of Biotechnology, School of Chemistry, The University of Manchester , 131 Princess Street, M1 7DN, Manchester, United Kingdom.
authors.forename: Cunyu
authors.identifier: 0000-0002-3603-2421
authors.initials: C
authors.lastname: Yan
authors.affiliation: Manchester Synthetic Biology Research Centre for Fine and Speciality Chemicals (SYNBIOCHEM), Manchester Institute of Biotechnology, School of Chemistry, The University of Manchester , 131 Princess Street, M1 7DN, Manchester, United Kingdom.
authors.forename: Fabio
authors.identifier: 0000-0001-5861-9269
authors.initials: F
authors.lastname: Parmeggiani
authors.affiliation: Waters Corp., Stamford Avenue, Altrincham Road, SK9 4AX, Wilmslow, United Kingdom.
authors.forename: Emrys A
authors.initials: EA
authors.lastname: Jones
authors.affiliation: Waters Corp., Stamford Avenue, Altrincham Road, SK9 4AX, Wilmslow, United Kingdom.
authors.forename: Emmanuelle
authors.initials: E
authors.lastname: Claude
authors.affiliation: Manchester Synthetic Biology Research Centre for Fine and Speciality Chemicals (SYNBIOCHEM), Manchester Institute of Biotechnology, School of Chemistry, The University of Manchester , 131 Princess Street, M1 7DN, Manchester, United Kingdom.
authors.forename: Shaneela A
authors.initials: SA
authors.lastname: Hussain
authors.affiliation: Manchester Synthetic Biology Research Centre for Fine and Speciality Chemicals (SYNBIOCHEM), Manchester Institute of Biotechnology, School of Chemistry, The University of Manchester , 131 Princess Street, M1 7DN, Manchester, United Kingdom.
authors.forename: Nicholas J
authors.identifier: 0000-0002-8708-0781
authors.initials: NJ
authors.lastname: Turner
authors.affiliation: Manchester Synthetic Biology Research Centre for Fine and Speciality Chemicals (SYNBIOCHEM), Manchester Institute of Biotechnology, School of Chemistry, The University of Manchester , 131 Princess Street, M1 7DN, Manchester, United Kingdom.
authors.forename: Sabine L
authors.identifier: 0000-0003-3974-646X
authors.initials: SL
authors.lastname: Flitsch
authors.affiliation: Manchester Synthetic Biology Research Centre for Fine and Speciality Chemicals (SYNBIOCHEM), Manchester Institute of Biotechnology, School of Chemistry, The University of Manchester , 131 Princess Street, M1 7DN, Manchester, United Kingdom.
authors.forename: Perdita E
authors.identifier: 0000-0002-7720-586X
authors.initials: PE
authors.lastname: Barran
mesh_terms: D000642:Ammonia-Lyases / Q000737:chemistry / Q000378:metabolism*; D046870:Anabaena variabilis / Q000201:enzymology*; D055162:Biocatalysis; D004926:Escherichia coli / Q000166:cytology / Q000378:metabolism*; D006899:Mixed Function Oxygenases / Q000737:chemistry / Q000378:metabolism*; D015394:Molecular Structure; D021241:Spectrometry, Mass, Electrospray Ionization; D013997:Time Factors
chemical_list: D006899:Mixed Function Oxygenases; D000642:Ammonia-Lyases