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Microdroplet enabled cultivation of single yeast cells correlates with bulk growth and reveals subpopulation phenomena
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  • Hangrui Liu,
  • Xin Xu,
  • Kai Peng,
  • Yuxin Zhang,
  • Lianmei Jiang,
  • Thomas Williams,
  • Ian Paulsen,
  • James Piper,
  • Ming Li
Hangrui Liu
Macquarie University

Corresponding Author:hangrui.liu@hdr.mq.edu.au

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Xin Xu
Macquarie University
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Kai Peng
Macquarie University
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Yuxin Zhang
Macquarie University
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Lianmei Jiang
Macquarie University
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Thomas Williams
Macquarie University
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Ian Paulsen
Macquarie University
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James Piper
Macquarie University
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Ming Li
Macquarie University
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Abstract

Yeast has been engineered for cost-effective organic acid production through metabolic engineering and synthetic biology techniques. However, cell growth assays in these processes were performed in bulk at the population level, thus obscuring the dynamics of rare single cells exhibiting beneficial traits. Here, we introduce the use of monodisperse picolitre droplets as bioreactors to cultivate yeast at the single-cell level. We investigated the effect of acid stress on growth and the effect of potassium ions on propionic acid tolerance for single yeast cells of different species, genotypes and phenotypes. The results showed that the average growth of single yeast cells in microdroplets was identical to those of yeast populations grown in bulk, and microdroplet compartments do not significantly affect cell viability. This approach offers the prospect of detecting cell-to-cell variations in growth and physiology and is expected to be applied for the engineering of yeast to produce value-added bioproducts.
29 Jun 2020Submitted to Biotechnology and Bioengineering
29 Jun 2020Submission Checks Completed
29 Jun 2020Assigned to Editor
08 Jul 2020Reviewer(s) Assigned
07 Aug 2020Editorial Decision: Revise Minor
07 Aug 2020Review(s) Completed, Editorial Evaluation Pending
18 Sep 20201st Revision Received
19 Sep 2020Submission Checks Completed
19 Sep 2020Assigned to Editor
19 Sep 2020Reviewer(s) Assigned
04 Oct 2020Review(s) Completed, Editorial Evaluation Pending
04 Oct 2020Editorial Decision: Accept
21 Oct 2020Published in Biotechnology and Bioengineering. 10.1002/bit.27591