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  • Microdroplet enabled cultivation of single yeast cells correlates with bulk growth and reveals subpopulation phenomena.

Microdroplet enabled cultivation of single yeast cells correlates with bulk growth and reveals subpopulation phenomena.

Biotechnology and bioengineering (2020-10-07)
Hangrui Liu, Xin Xu, Kai Peng, Yuxin Zhang, Lianmei Jiang, Thomas C Williams, Ian T Paulsen, James A Piper, Ming Li
摘要

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 experiences the same trend 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.

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Sigma-Aldrich
OptiPrep密度梯度培养基, used for cell and subcellular organelle isolation
Sigma-Aldrich
不含氨基酸的酵母氮源基, Yeast classification medium used for selecting yeasts based on amino acid and carbohydrate requirements
Sigma-Aldrich
酵母营养缺陷型培养基补充剂, without uracil
Sigma-Aldrich
三氯(十八烷基)硅烷, ≥90%