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High-throughput optofluidic screening for improved microbial cell factories via real-time micron-scale productivity monitoring.
Rienzo, Matthew; Lin, Ke-Chih; Mobilia, Kellen C; Sackmann, Eric K; Kurz, Volker; Navidi, Adam H; King, Jarett; Onorato, Robert M; Chao, Lawrence K; Wu, Tony; Jiang, Hanxiao; Valley, Justin K; Lionberger, Troy A; Leavell, Michael D.
Afiliación
  • Rienzo M; Research and Development, Amyris, Inc., 5885 Hollis St., Suite 100, Emeryville, CA 94608, USA. leavell@amyris.com.
  • Lin KC; Technology and Business Development, Berkeley Lights, Inc., 5858 Horton St., Unit 320, Emeryville, CA 94608, USA. troy.lionberger@berkeleylights.com.
  • Mobilia KC; Technology and Business Development, Berkeley Lights, Inc., 5858 Horton St., Unit 320, Emeryville, CA 94608, USA. troy.lionberger@berkeleylights.com.
  • Sackmann EK; Technology and Business Development, Berkeley Lights, Inc., 5858 Horton St., Unit 320, Emeryville, CA 94608, USA. troy.lionberger@berkeleylights.com.
  • Kurz V; Technology and Business Development, Berkeley Lights, Inc., 5858 Horton St., Unit 320, Emeryville, CA 94608, USA. troy.lionberger@berkeleylights.com.
  • Navidi AH; Research and Development, Amyris, Inc., 5885 Hollis St., Suite 100, Emeryville, CA 94608, USA. leavell@amyris.com.
  • King J; Research and Development, Amyris, Inc., 5885 Hollis St., Suite 100, Emeryville, CA 94608, USA. leavell@amyris.com.
  • Onorato RM; Technology and Business Development, Berkeley Lights, Inc., 5858 Horton St., Unit 320, Emeryville, CA 94608, USA. troy.lionberger@berkeleylights.com.
  • Chao LK; Research and Development, Amyris, Inc., 5885 Hollis St., Suite 100, Emeryville, CA 94608, USA. leavell@amyris.com.
  • Wu T; Research and Development, Amyris, Inc., 5885 Hollis St., Suite 100, Emeryville, CA 94608, USA. leavell@amyris.com.
  • Jiang H; Research and Development, Amyris, Inc., 5885 Hollis St., Suite 100, Emeryville, CA 94608, USA. leavell@amyris.com.
  • Valley JK; Research and Development, Amyris, Inc., 5885 Hollis St., Suite 100, Emeryville, CA 94608, USA. leavell@amyris.com.
  • Lionberger TA; Technology and Business Development, Berkeley Lights, Inc., 5858 Horton St., Unit 320, Emeryville, CA 94608, USA. troy.lionberger@berkeleylights.com.
  • Leavell MD; Research and Development, Amyris, Inc., 5885 Hollis St., Suite 100, Emeryville, CA 94608, USA. leavell@amyris.com.
Lab Chip ; 21(15): 2901-2912, 2021 08 07.
Article en En | MEDLINE | ID: mdl-34160512
The industrial synthetic biology sector has made huge investments to achieve relevant miniaturized screening systems for scalable fermentation. Here we present the first example of a high-throughput (>103 genotypes per week) perfusion-based screening system to improve small-molecule secretion from microbial strains. Using the Berkeley Lights Beacon® system, the productivity of each strain could be directly monitored in real time during continuous culture, yielding phenotypes that correlated strongly (r2 > 0.8, p < 0.0005) with behavior in industrially relevant bioreactor processes. This method allows a much closer approximation of a typical fed-batch fermentation than conventional batch-like droplet or microplate culture models, in addition to rich time-dependent data on growth and productivity. We demonstrate these advantages by application to the improvement of high-productivity strains using whole-genome random mutagenesis, yielding mutants with substantially improved (by up to 85%) peak specific productivities in bioreactors. Each screen of ∼5 × 103 mutants could be completed in under 8 days (including 5 days involving user intervention), saving ∼50-75% of the time required for conventional microplate-based screening methods.
Asunto(s)

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Reactores Biológicos / Ensayos Analíticos de Alto Rendimiento Tipo de estudio: Diagnostic_studies / Prognostic_studies / Screening_studies Idioma: En Revista: Lab Chip Asunto de la revista: BIOTECNOLOGIA / QUIMICA Año: 2021 Tipo del documento: Article País de afiliación: Estados Unidos Pais de publicación: Reino Unido

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Reactores Biológicos / Ensayos Analíticos de Alto Rendimiento Tipo de estudio: Diagnostic_studies / Prognostic_studies / Screening_studies Idioma: En Revista: Lab Chip Asunto de la revista: BIOTECNOLOGIA / QUIMICA Año: 2021 Tipo del documento: Article País de afiliación: Estados Unidos Pais de publicación: Reino Unido