Journal: Nanomaterials
Article Title: The Electrosome: A Surface-Displayed Enzymatic Cascade in a Biofuel Cell’s Anode and a High-Density Surface-Displayed Biocathodic Enzyme
doi: 10.3390/nano7070153
Figure Lengend Snippet: Schematic representation of ( A ) ethanol oxidation cascade displayed on the surface of S. cerevisiae using a chimeric scaffoldin, including dockerin-containing ADH and FormDH; ( B ) oxygen-reduction complexes displayed on S. cerevisiae using different scaffoldins, including one to four dockerin-containing copper oxidase copies; ( C ) all the enzymes and scaffoldins used in this study. Top row: dockerin-containing oxidoreductases. zADH-Ac: ADH from Zymomonas mobilis with an Acetivibrio cellulolyticus dockerin. pFormDH-Ct: Pseudomonas putida formaldehyde dehydrogenase with a Clostridium thermocellum dockerin. CueO-Ct: E. coli copper oxidase with a C. thermocellum dockerin. Bottom row from left to right: scaffoldins used in this study. 2Ct: di-valent scaffoldin comprising 2 C. thermocellum cohesins for CueO-Ct binding. 3Ct: tri-valent scaffoldin comprising 3 C. thermocellum cohesins for CueO-Ct binding. 4Ct: tetra-valent scaffoldin comprising 4 C. thermocellum cohesins for CueO-Ct binding. L20(-cellulose-binding module (CBM)): tri-valent scaffoldin, Scaf20L, comprising cohesins from A. cellulolyticus , Bacteroides cellulosolvens , and C. thermocellum but lacking the CBM for zADH-Ac and pFormDH-Ct; this construct was also used as a mono-valent CueO-Ct binding site. All the different scaffoldin constructs contain the Aga2p (depicted as a green rectangle) at the N-terminus and the c-myc (depicted as a blue square) at the C-terminus. A complete description of the system is given in the experimental section.
Article Snippet: The genes encoding dockerins of Acetivibrio cellulolyticus (dockerin from ScaB scaffoldin strain ATCC 33288) and Clostridium thermocellum (dockerin from Cel48S strain ATCC 27405) were cloned and ligated to the C-terminus of Zymomonas mobilis alcohol dehydrogenase and to Pseudomonas putida formaldehyde dehydrogenase (zADH-Ac and pFormDH-Ct, respectively) by standard methods [ ].
Techniques: Binding Assay, Construct
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