title: Quantified high-throughput screening of Escherichia coli producing poly(3-hydroxybutyrate) based on FACS.
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.
authors.affiliation: Department of Chemical and Biomolecular Engineering (BK21 Program), KAIST, 291 Daehak-ro, Yuseong-gu, Daejeon 305-701, Republic of Korea.
authors.forename: Jae Hyung
authors.initials: JH
authors.lastname: Lee
authors.forename: Seung Hwan
authors.initials: SH
authors.lastname: Lee
authors.forename: Sung Sun
authors.initials: SS
authors.lastname: Yim
authors.forename: Kyoung-Hee
authors.initials: KH
authors.lastname: Kang
authors.forename: Sang Yup
authors.initials: SY
authors.lastname: Lee
authors.forename: Si Jae
authors.initials: SJ
authors.lastname: Park
authors.forename: Ki Jun
authors.initials: KJ
authors.lastname: Jeong
mesh_terms: D001896:Boron Compounds; D002469:Cell Separation / Q000379:methods*; D004926:Escherichia coli / Q000166:cytology / Q000302:isolation & purification* / Q000378:metabolism*; D005434:Flow Cytometry / Q000379:methods*; D006885:Hydroxybutyrates / Q000302:isolation & purification / Q000378:metabolism*; D008856:Microscopy, Fluorescence / Q000379:methods*; D010078:Oxazines; D011091:Polyesters / Q000302:isolation & purification / Q000378:metabolism*; D000094122:Polyhydroxybutyrates
chemical_list: D001896:Boron Compounds; D006885:Hydroxybutyrates; D010078:Oxazines; D011091:Polyesters; C095489:4,4-difluoro-4-bora-3a,4a-diaza-s-indacene; C003182:poly-beta-hydroxybutyrate; D000094122:Polyhydroxybutyrates; C044808:nile red