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Display of Functionally Active PHB Depolymerase on Escherichia Coli Cell Surface
Author(s) -
Hiraishi Tomohiro,
Yamashita Koichi,
Sakono Masafumi,
Nakanishi Jun,
Tan LiuTzea,
Sudesh Kumar,
Abe Hideki,
Maeda Mizuo
Publication year - 2012
Publication title -
macromolecular bioscience
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.924
H-Index - 105
eISSN - 1616-5195
pISSN - 1616-5187
DOI - 10.1002/mabi.201100273
Subject(s) - escherichia coli , chemistry , polyester , biocatalysis , enzyme , adsorption , monomer , polymer , biophysics , biochemistry , biology , catalysis , gene , organic chemistry , ionic liquid
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.