Quantified high-throughput screening of Escherichia coli producing poly(3-hydroxybutyrate) based on FACS

Jae Hyung Lee, Seung Hwan Lee, Sung Sun Yim, Kyoung Hee Kang, Sang Yup Lee, Si Jae Park, Ki Jun Jeong

Research output: Contribution to journalArticlepeer-review

21 Scopus citations

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.

Original languageEnglish
Pages (from-to)1767-1779
Number of pages13
JournalApplied Biochemistry and Biotechnology
Volume170
Issue number7
DOIs
StatePublished - Aug 2013

Keywords

  • BODIPY
  • Escherichia coli
  • FACS
  • Nile red
  • Poly(3-hydroxybutyrate)

Fingerprint

Dive into the research topics of 'Quantified high-throughput screening of Escherichia coli producing poly(3-hydroxybutyrate) based on FACS'. Together they form a unique fingerprint.

Cite this