title: Surface display of a redox enzyme and its site-specific wiring to gold electrodes.
abstract: The generation of a current through interaction between bacteria and electrodes has been explored by various methods. We demonstrate the attachment of living bacteria through a surface displayed redox enzyme, alcohol dehydrogenase II. The unnatural amino acid para-azido-L-phenylalanine was incorporated into a specific site of the displayed enzyme, facilitating electron transfer between the enzyme and an electrode. In order to attach the bacteria carrying the surface displayed enzyme to a surface, a linker containing an alkyne and a thiol moiety on opposite ends was synthesized and attached to the dehydrogenase site specifically through a copper(I)-catalyzed azide-alkyne cycloaddition reaction. Using this approach we were able to covalently link bacteria to gold-coated surfaces and to gold nanoparticles, while maintaining viability and catalytic activity. We show the performance of a biofuel cell using these modified bacteria at the anode, which resulted in site-specific dependent fuel cell performance for at least a week. This is the first example of site-specific attachment of a true living biohybrid to inorganic material.
authors.affiliation: The Avram and Stella Goldstein-Goren Department of Biotechnology Engineering, Ben-Gurion University of the Negev, Beer-Sheva 84105, Israel.
authors.forename: Liron
authors.initials: L
authors.lastname: Amir
authors.forename: Stewart A
authors.initials: SA
authors.lastname: Carnally
authors.forename: Josep
authors.initials: J
authors.lastname: Rayo
authors.forename: Shaked
authors.initials: S
authors.lastname: Rosenne
authors.forename: Sarit
authors.initials: S
authors.lastname: Melamed Yerushalmi
authors.forename: Orr
authors.initials: O
authors.lastname: Schlesinger
authors.forename: Michael M
authors.initials: MM
authors.lastname: Meijler
authors.forename: Lital
authors.initials: L
authors.lastname: Alfonta
mesh_terms: D000426:Alcohol Dehydrogenase / Q000737:chemistry* / Q000378:metabolism; D000480:Alkynes / Q000737:chemistry; D001386:Azides / Q000737:chemistry / Q000378:metabolism; D002384:Catalysis; D003300:Copper / Q000737:chemistry; D003500:Cyclization; D004566:Electrodes; D004926:Escherichia coli / Q000737:chemistry / Q000378:metabolism; D006046:Gold / Q000737:chemistry* / Q000378:metabolism; D010084:Oxidation-Reduction; D010649:Phenylalanine / Q000031:analogs & derivatives / Q000737:chemistry / Q000378:metabolism; D013499:Surface Properties; D017355:Zymomonas / Q000201:enzymology / Q000378:metabolism
chemical_list: D000480:Alkynes; D001386:Azides; C027162:4-azidophenylalanine; D010649:Phenylalanine; D006046:Gold; D003300:Copper; D000426:Alcohol Dehydrogenase