1 research outputs found
Lighting up Pyruvate Metabolism in Saccharomyces cerevisiae by a Genetically Encoded Fluorescent Biosensor
Monitoring
intracellular pyruvate is useful for the exploration
of fundamental metabolism and for guiding the construction of yeast
cell factories for chemical production. Here, we employed a genetically
encoded fluorescent Pyronic biosensor to light up the pyruvate metabolic
state in the cytoplasm, nucleus, and mitochondria of Saccharomyces cerevisiae BY4741. A strong correlation
was observed between the pyruvate fluctuation in mitochondria and
cytoplasm when exposed to different metabolites. Further metabolic
analysis of pyruvate uptake and glycolytic dynamics showed that glucose
and fructose dose-dependently activated cytoplasmic pyruvate levels
more effectively than direct exposure to pyruvate. Meanwhile, the
Pyronic biosensor could visually distinguish phenotypes of the wild-type S. cerevisiae BY4741 and the pyruvate-hyperproducing S. cerevisiae TAM at a single-cell resolution, having
the potential for high-throughput screening. Overall, Pyronic biosensors
targeting different suborganelles contribute to mapping and studying
the central carbon metabolism in-depth and guide the design and construction
of yeast cell factories