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.
authors.affiliation: Bioengineering Laboratory, RIKEN Advanced Science Institute, Wako-shi, Saitama, Japan. thiraish@riken.jp
authors.forename: Tomohiro
authors.initials: T
authors.lastname: Hiraishi
authors.forename: Koichi
authors.initials: K
authors.lastname: Yamashita
authors.forename: Masafumi
authors.initials: M
authors.lastname: Sakono
authors.forename: Jun
authors.initials: J
authors.lastname: Nakanishi
authors.forename: Liu-Tzea
authors.initials: LT
authors.lastname: Tan
authors.forename: Kumar
authors.initials: K
authors.lastname: Sudesh
authors.forename: Hideki
authors.initials: H
authors.lastname: Abe
authors.forename: Mizuo
authors.initials: M
authors.lastname: Maeda
mesh_terms: D020155:3-Hydroxybutyric Acid / Q000096:biosynthesis*; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D055162:Biocatalysis; D002265:Carboxylic Ester Hydrolases / Q000235:genetics / Q000378:metabolism*; D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics; D015870:Gene Expression; D006885:Hydroxybutyrates / Q000378:metabolism*; D007700:Kinetics; D008074:Lipoproteins / Q000235:genetics / Q000378:metabolism*; D008856:Microscopy, Fluorescence; D010957:Plasmids; D011091:Polyesters / Q000378:metabolism*; D011550:Pseudomonas aeruginosa / Q000737:chemistry / Q000235:genetics; D049920:Ralstonia pickettii / Q000737:chemistry / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D013379:Substrate Specificity; D013499:Surface Properties; D000094122:Polyhydroxybutyrates
chemical_list: D020155:3-Hydroxybutyric Acid; D001426:Bacterial Proteins; D002265:Carboxylic Ester Hydrolases; D006885:Hydroxybutyrates; D008074:Lipoproteins; D011091:Polyesters; D011993:Recombinant Fusion Proteins; C060507:outer membrane lipoprotein I, Bacteria; C003182:poly-beta-hydroxybutyrate; D000094122:Polyhydroxybutyrates; C022417:poly-beta-hydroxybutyrate depolymerase