Quantified High-Throughput Screening of Escherichia coli Producing Poly(3-hydroxybutyrate) Based on FACS.

Lee, Jae; Lee, Seung; Yim, Sung; Kang, Kyoung-Hee; Lee, Sang; Park, Si; Jeong, Ki

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.

ESCHERICHIA coli; FLUORESCENT dyes; FLOW cytometry; HYDROXY acids; BIODEGRADABLE plastics; STAINS & staining (Microscopy)

Applied Biochemistry & Biotechnology, 2013, Vol 170, Issue 7, p1767

0273-2289

Academic Journal

10.1007/s12010-013-0311-2