Display of Functionally Active PHB Depolymerase on Escherichia Coli Cell Surface

Tomohiro Hiraishi; Koichi Yamashita; Masafumi Sakono; Jun Nakanishi; Liu Tzea Tan; Kumar Sudesh; Hideki Abe; Mizuo Maeda

doi:10.1002/mabi.201100273

Display of Functionally Active PHB Depolymerase on Escherichia Coli Cell Surface

Tomohiro Hiraishi^*, Koichi Yamashita, Masafumi Sakono, Jun Nakanishi, Liu Tzea Tan, Kumar Sudesh, Hideki Abe, Mizuo Maeda

^*Corresponding author for this work

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

5 Citations (Scopus)

Abstract

The display of PHB depolymerase (PhaZ _RpiT1) from R. pickettii T1 on the surface of E. coli JM109 cells is realized using OprI of P. aeruginosa as the anchoring motif. The fusion protein is stably expressed and its surface localization is verified by immunofluorescence microscopy. The displayed PhaZ _RpiT1 retains its cleaving ability for soluble substrates as well as its ability to adsorb to the PHB surface, and also remains catalycically active in the degradation of insoluble polyester materials, in spite of the possible suppression of the enzyme movement on the polymer surface. The results demonstrate that PhaZ _RpiT1-displaying E. coli shows potential for use as a whole-cell biocatalyst for the production of (R)-3-hydroxybutyrate monomers from insoluble PHB materials.

Original language	English
Pages (from-to)	218-224
Number of pages	7
Journal	Macromolecular Bioscience
Volume	12
Issue number	2
DOIs	https://doi.org/10.1002/mabi.201100273
Publication status	Published - 2012 Feb
Externally published	Yes

Keywords

Bioengineering
Cell surface display
Degradation
PHB depolymerase
Poly[(R)-3-hydroxybutyrate]

ASJC Scopus subject areas

Biotechnology
Bioengineering
Biomaterials
Polymers and Plastics
Materials Chemistry

Access to Document

10.1002/mabi.201100273

Cite this

@article{e0cb50ce4b234d2f975f401726bb8d6f,

title = "Display of Functionally Active PHB Depolymerase on Escherichia Coli Cell Surface",

abstract = "The display of PHB depolymerase (PhaZ RpiT1) from R. pickettii T1 on the surface of E. coli JM109 cells is realized using OprI of P. aeruginosa as the anchoring motif. The fusion protein is stably expressed and its surface localization is verified by immunofluorescence microscopy. The displayed PhaZ RpiT1 retains its cleaving ability for soluble substrates as well as its ability to adsorb to the PHB surface, and also remains catalycically active in the degradation of insoluble polyester materials, in spite of the possible suppression of the enzyme movement on the polymer surface. The results demonstrate that PhaZ RpiT1-displaying E. coli shows potential for use as a whole-cell biocatalyst for the production of (R)-3-hydroxybutyrate monomers from insoluble PHB materials.",

keywords = "Bioengineering, Cell surface display, Degradation, PHB depolymerase, Poly[(R)-3-hydroxybutyrate]",

author = "Tomohiro Hiraishi and Koichi Yamashita and Masafumi Sakono and Jun Nakanishi and Tan, {Liu Tzea} and Kumar Sudesh and Hideki Abe and Mizuo Maeda",

year = "2012",

month = feb,

doi = "10.1002/mabi.201100273",

language = "English",

volume = "12",

pages = "218--224",

journal = "Macromolecular Bioscience",

issn = "1616-5187",

publisher = "Wiley-VCH Verlag",

number = "2",

}

TY - JOUR

T1 - Display of Functionally Active PHB Depolymerase on Escherichia Coli Cell Surface

AU - Hiraishi, Tomohiro

AU - Yamashita, Koichi

AU - Sakono, Masafumi

AU - Nakanishi, Jun

AU - Tan, Liu Tzea

AU - Sudesh, Kumar

AU - Abe, Hideki

AU - Maeda, Mizuo

PY - 2012/2

Y1 - 2012/2

N2 - The display of PHB depolymerase (PhaZ RpiT1) from R. pickettii T1 on the surface of E. coli JM109 cells is realized using OprI of P. aeruginosa as the anchoring motif. The fusion protein is stably expressed and its surface localization is verified by immunofluorescence microscopy. The displayed PhaZ RpiT1 retains its cleaving ability for soluble substrates as well as its ability to adsorb to the PHB surface, and also remains catalycically active in the degradation of insoluble polyester materials, in spite of the possible suppression of the enzyme movement on the polymer surface. The results demonstrate that PhaZ RpiT1-displaying E. coli shows potential for use as a whole-cell biocatalyst for the production of (R)-3-hydroxybutyrate monomers from insoluble PHB materials.

AB - The display of PHB depolymerase (PhaZ RpiT1) from R. pickettii T1 on the surface of E. coli JM109 cells is realized using OprI of P. aeruginosa as the anchoring motif. The fusion protein is stably expressed and its surface localization is verified by immunofluorescence microscopy. The displayed PhaZ RpiT1 retains its cleaving ability for soluble substrates as well as its ability to adsorb to the PHB surface, and also remains catalycically active in the degradation of insoluble polyester materials, in spite of the possible suppression of the enzyme movement on the polymer surface. The results demonstrate that PhaZ RpiT1-displaying E. coli shows potential for use as a whole-cell biocatalyst for the production of (R)-3-hydroxybutyrate monomers from insoluble PHB materials.

KW - Bioengineering

KW - Cell surface display

KW - Degradation

KW - PHB depolymerase

KW - Poly[(R)-3-hydroxybutyrate]

UR - http://www.scopus.com/inward/record.url?scp=84856580974&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84856580974&partnerID=8YFLogxK

U2 - 10.1002/mabi.201100273

DO - 10.1002/mabi.201100273

M3 - Article

C2 - 22095689

AN - SCOPUS:84856580974

SN - 1616-5187

VL - 12

SP - 218

EP - 224

JO - Macromolecular Bioscience

JF - Macromolecular Bioscience

IS - 2

ER -

Display of Functionally Active PHB Depolymerase on Escherichia Coli Cell Surface

Abstract

Keywords

ASJC Scopus subject areas

Access to Document

Other files and links

Fingerprint

Cite this