Abstract
Here, we report on a highly sensitive method for the detection of P(3HB) accumulation in Escherichia coli cells based on the automated flow cytometry system using fluorescent dyes. E. coli containing P(3HB) were stained with either BODIPY or Nile red fluorescent dye, and their staining properties were analyzed under a variety of conditions. Compared with Nile red, BODIPY was much more sensitive in staining P(3HB) and overall demonstrated a more rapid staining of cells, a greater resistance to photobleaching, and greater cell viability. In addition, we also successfully monitored heterogeneity in P(3HB) accumulation within a cell population using BODIPY staining and flow cytometry. We believe this optimized staining method using BODIPY in combination with screening by high-speed flow cytometer will be helpful in the engineering of host cells toward an enhanced production of bioplastics.
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This work was supported by the Intelligent Synthetic Biology Center of Global Frontier Project funded by the Ministry of Science, ICT, and Future Planning (MSIP). Further support from the 2012 Research Fund of Myongji University is appreciated.
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Jae Hyung Lee and Seung Hwan Lee equally contributed to this work.
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Lee, J.H., Lee, S.H., Yim, S.S. et al. Quantified High-Throughput Screening of Escherichia coli Producing Poly(3-hydroxybutyrate) Based on FACS. Appl Biochem Biotechnol 170, 1767–1779 (2013). https://doi.org/10.1007/s12010-013-0311-2
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DOI: https://doi.org/10.1007/s12010-013-0311-2