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

doi:10.1007/s12010-013-0311-2

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

Department of Chemical Engineering & Materials Science

Research output: Contribution to journal › Article › peer-review

24 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 language	English
Pages (from-to)	1767-1779
Number of pages	13
Journal	Applied Biochemistry and Biotechnology
Volume	170
Issue number	7
DOIs	https://doi.org/10.1007/s12010-013-0311-2
State	Published - Aug 2013

Bibliographical note

Funding Information:
Acknowledgments 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.

Keywords

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

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10.1007/s12010-013-0311-2

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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.",

keywords = "BODIPY, Escherichia coli, FACS, Nile red, Poly(3-hydroxybutyrate)",

author = "Lee, {Jae Hyung} and Lee, {Seung Hwan} and Yim, {Sung Sun} and Kang, {Kyoung Hee} and Lee, {Sang Yup} and Park, {Si Jae} and Jeong, {Ki Jun}",

note = "Funding Information: Acknowledgments 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.",

year = "2013",

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doi = "10.1007/s12010-013-0311-2",

language = "English",

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AU - Lee, Jae Hyung

AU - Lee, Seung Hwan

AU - Yim, Sung Sun

AU - Kang, Kyoung Hee

AU - Lee, Sang Yup

AU - Park, Si Jae

AU - Jeong, Ki Jun

N1 - Funding Information: Acknowledgments 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.

PY - 2013/8

Y1 - 2013/8

N2 - 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.

AB - 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.

KW - BODIPY

KW - Escherichia coli

KW - FACS

KW - Nile red

KW - Poly(3-hydroxybutyrate)

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JO - Applied Biochemistry and Biotechnology

JF - Applied Biochemistry and Biotechnology

IS - 7

ER -

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

Abstract

Bibliographical note

Keywords

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