Expression of electrically conductive protein nanowires in Escherichia coli
AssigneeUniversity of Massachusetts
InventorsDerek R. Lovley, Toshiyuki Ueki, David Walker, Trevor Woodard, Kelly Nevin Lovley
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Abstract
The present invention provides, in various embodiments, genetically modified aerobic bacteria, polynucleotides and methods for expressing and/or harvesting electrically conductive protein nanowires (e-PNs). The present invention also provides e-PNs produced using the genetically modified aerobic bacteria, polynucleotides and methods.
Claims16
A genetically modified aerobic bacterium, comprising a polynucleotide encoding an electrically conductive fusion protein that comprises a non-native pilin monomer, wherein the non-native pilin monomer comprises an amino acid sequence that has at least 90% sequence identity to the amino acid sequence of wildtype Geobacter sulfurreducens PilA monomer (SEQ ID NO:16), wherein the bacterium is an E. coli delta type I fimbriae major subunit (ΔfimA), delta flagellin (ΔfliC) cell.
A polynucleotide comprising an artificial operon that comprises a cluster of two or more polynucleotide sequences, each polynucleotide sequence encoding a pilus assembly protein.
A method of producing electrically conductive protein nanowires, comprising the steps of:
- a) placing a genetically modified aerobic bacterium comprising a polynucleotide encoding an electrically conductive fusion protein that comprises a non-native pilin monomer in a culture medium conditioned for producing pili comprising the non-native pilin monomers, wherein the non-native pilin monomer comprises an amino acid sequence that has at least 90% sequence identity to the amino acid sequence of wildtype Geobacter sulfurreducens PilA monomer (SEQ ID NO:16), and wherein the bacterium is an E. coli delta type I fimbriae major subunit (ΔfimA), delta flagellin (ΔfliC) cell;
- b) culturing the bacterium for a time sufficient to produce a desired quantity of the pili;
- and
- c) isolating the pili from the culture medium, thereby producing the electrically conductive protein nanowires.
Dependencies are inferred from the claim text.