Patent Publication Number: US-2023160885-A1

Title: Microbial Nanowires Modified to Contain Peptides and Methods of Making

Description:
RELATED APPLICATION(S) 
     This application is the U.S. National Stage of International Application No. PCT/US2020/023824, filed Mar. 20, 2020, published in English, which claims the benefit of U.S. Provisional Application No. 62/821,208, filed on Mar. 20, 2019. The entire teachings of the above applications are incorporated herein by reference. 
    
    
     INCORPORATION BY REFERENCE OF MATERIAL IN ASCII TEXT FILE 
     This application incorporates by reference the Sequence Listing contained in the following ASCII text file being submitted concurrently herewith: 
     a) File name: 46821013002_Sequence_Listing_revised.txt; created Apr. 15, 2022, 51,005 Bytes in size. 
     BACKGROUND 
     Microbially produced, electrically conductive protein nanowires (e-PNs) possess properties and possibilities for functionalization not found in other electronic nanowire materials. Diverse microorganisms in both the Bacteria and Archaea produce e-PNs. Functional analysis of Geobacter e-PNs have demonstrated that, when attached to cells, they serve as conduits for long-range electron exchange with other cells or minerals. 
     The most intensively studied e-PNs are the electrically conductive pili of the microorganism  Geobacter sulfurreducens. G. sulfurreducens  assembles the thin (3 nm), long (10-30 μm) e-PNs from multiple copies of just one short (61 amino acids) monomer peptide. Even though they are comprised of protein, the e-PNs produced with  G. sulfurreducens  are remarkably robust. They retain function under conditions required for the fabrication of electronic materials, including stability in a range of organic solvents and temperatures greater than 100° C. The  G. sulfurreducens  e-PNs are produced from renewable feedstocks. No harsh chemicals are required for e-PN production and there are no toxic components in the final product. Unlike silicon nanowires, e-PNs do not dissolve in water or bodily fluids, a distinct advantage for wearable and environmental electronic sensor applications, as well as implantable electronics. e-PNs have evolved for making cell-to-cell electrical connections, suggesting they may be more biocompatible than other nanowire materials. The dramatic change in e-PN conductivity in response to pH suggests that they may be readily adapted for diverse sensor functions. 
     SUMMARY 
     The potential applications of e-PNs may be expanded if their outer surface is modified to confer additional sensing capabilities, enhance binding to other materials, and/or confer other functions. Accordingly, there is a need for modifying naturally occurring e-PNs to confer additional functionalities. 
     The invention described herein generally relates to electrically conductive protein nanowires having peptide ligands (e.g., surface-exposed peptides) that confer additional functionalities (e.g., a sensing functionality, a binding functionality) to the nanowires. 
     One aspect of the invention relates to a fusion protein, wherein the fusion protein comprises a type IV pilin monomer and a tag at the C-terminus of the type IV pilin monomer. In some embodiments, the tag is a peptide. In other embodiments, the tag is a single amino acid. 
     Another aspect of the invention relates to an electrically conductive nanowire, wherein the electrically conductive nanowire comprises a fusion protein comprising a type IV pilin monomer and a tag at the C-terminus of the type IV pilin monomer. 
     Another aspect of the invention relates to a nanowire sensor device comprising: an electrically conductive nanowire that comprises a fusion protein comprising a type IV pilin monomer and a tag at the C-terminus of the type IV pilin monomer; a first electrode having a first electrode terminal, wherein the first electrode is configured to support and is in physical contact with the electrically conductive nanowire; a second electrode having a second electrode terminal, wherein the second electrode is configured to support and is in physical contact with the electrically conductive nanowire; an electrical resistance connected between the first and the second electrode terminals; and an electrical current monitor in electrical communication with the electrical resistance. The electrical current monitor is configured to measure an electrical current passing through the electrical resistance in the device. 
     Another aspect of the invention relates to a polynucleotide that encodes a fusion protein comprising a type IV pilin monomer and a tag at the C-terminus of the type IV pilin monomer. 
     Another aspect of the invention relates to an expression vector comprising a polynucleotide that encodes a fusion protein comprising a type IV pilin monomer and a tag at the C-terminus of the type IV pilin monomer. 
     Another aspect of the invention relates to a host cell comprising a polynucleotide or an expression vector comprising a polynucleotide, wherein the polynucleotide encodes a fusion protein comprising a type IV pilin monomer and a tag at the C-terminus of the type IV pilin monomer. 
     Another aspect of the invention relates to a method of producing electrically conductive protein nanowires, comprising the steps of:
     a) introducing a polynucleotide or an expression vector comprising the polynucleotide into a host cell, wherein the polynucleotide encodes a fusion protein comprising a type IV pilin monomer and a tag at the C-terminus of the type IV pilin monomer;   b) placing the host cell in a culture medium conditioned for producing type IV pili;   c) culturing the host cell for a time sufficient to produce a desired quantity of the type IV pili; and   d) isolating the type IV pili from the culture medium.   

    
    
     
       BRIEF DESCRIPTION OF THE DRAWINGS 
         FIG.  1    depicts the DNA sequence of the pilA, pilA-6His and pilA HA coding sequences. The coding sequence for the tag is underlined in pilA-6His and pilA-HA sequences. 
         FIGS.  2 A- 2 C  depict construction of  G. sulfurreducens  strain PilA-WT/PilA-6His and expression of pilin monomers.  FIG.  2 A  depicts amino acid sequences of wild-type PilA and PilA-6His.  FIG.  2 B  shows the location of wild-type pilA and pilA-6His genes on the chromosome. Numbers are gene numbers from the genome sequence (NCBI, NC_017454.1). gen R  is the gentamicin resistance gene.  FIG.  2 C  is a Western blot of wild-type (lane 1) and PilA-WT/PilA-6His (lane 2) strains. Lane M is the molecular weight standard markers. The headings designate the antibodies employed. 
         FIGS.  3 A- 3 E  are transmission electron micrographs showing that 6His-tag is incorporated into the e-PNs of  G. sulfurreducens  strain PilA-WT/PilA-6His.  FIGS.  3 A- 3 B and  3 C- 3 E  are immunogold labeling and Ni 2+ -NTA-gold labeling of 6His-tag, respectively. 
         FIGS.  4 A- 4 D  are transmission electron micrographs showing a lack of filaments labeling of 6His-tag in the wild-type strain.  FIGS.  4 A- 4 B and  4 C- 4 D  are immunogold labeling and Ni 2+ -NTA-gold labeling of 6His-tag, respectively. 
         FIG.  5    depicts current production of wild-type  G. sulfurreducens  and  G. sulfurreducens  strains PilA-WT/PilA-6His and PilA-WT/PilA-6His/PilA-HA. 
         FIGS.  6 A- 6 G  show conductance of synthetic e-PNs decorated with peptide tags.  FIGS.  6 A- 6 F  are representative contact topological imaging of e-PNs from  G. sulfurreducens  strains PilA-WT/PilA-6His and PilA-WT/PilA-6His/PilA-HA.  FIG.  6 G  Point-mode current response (I-V) spectroscopy of e-PNs from PilA-WT/PilA-6His and PilA-WT/PilA-6His/PilA-HA, compared to previously published data on e-PNs from the wild-type strain of  G. sulfurreducens  and strain Aro-5, which expresses poorly conductive pili. 
         FIGS.  7 A- 7 C  show conductance for  G. sulfurreducens  strain PilA-WT/PilA-6His using point-mode current response (I-V) spectroscopy. Three individual wires ( FIGS.  7 A,  7 B and  7 C ) were measured at three independent points (1-3). Calculations were made using a linear fit model between −0.2 V and 0.2 V. S: Siemen. 
         FIGS.  8 A- 8 C  depict the construction of  G. sulfurreducens  strain PilA-WT/PilA-6His/PilA-HA and expression of pilin monomers.  FIG.  8 A  depicts the amino acid sequence of the PHA-HA.  FIG.  8 B  depicts the location of wild-type pilA, pilA-6His, and pilA-HA genes on the chromosome. lad is the Lac repressor gene. kan R  is the kanamycin resistance gene.  FIG.  8 C  Western blot analysis of cell lysates of wild-type (lane 1), PilA-WT/PilA-6His (lane 2), and PilA-WT/PilA-6His/PilA-HA (lane 3) strains. Lane M is the molecular weight standard markers. Headings designate the antibodies employed. 
         FIGS.  9 A- 9 F  are transmission electron microscopy images of immunogold labeling of  G. sulfurreducens  strain PilA-WT/PilA-6His/PilA-HA for 6His-tag ( FIGS.  9 A- 9 B ), HA-tag ( FIGS.  9 C- 9 D ), and both tags ( FIGS.  9 E- 9 F ). The diameters of the gold nanoparticles were 10 nm in  FIGS.  9 A- 9 D  and 40 nm in  FIGS.  9 E- 9 F . 
         FIGS.  10 A and  10 B  are transmission electron microscopy images demonstrating a lack of immunogold labeling of the HA-tag in the PilA-WT ( FIG.  10 A ) and PilA-6His ( FIG.  10 B ) strains. 
         FIGS.  11 A- 11 C  show conductance for  G. sulfurreducens  strain PilA-WT/PilA-6His/PilA-HA filaments using point-mode current response (I-V) spectroscopy. Three individual wires ( FIGS.  11 A,  11 B and  11 C ) were measured at three independent points (1-3). Calculations were made using a linear fit model between −0.2 V and 0.2 V. S: Siemen. 
         FIGS.  12 A- 12 D  depict the transcriptional control of PHA-HA expression and incorporation into e-PNs in strain PilA-WT/PilA-6His/PilA-HA evaluated with an anti-HA antibody.  FIG.  12 A  is a Western blot of cell lysates. Lane M is the molecular weight standard markers.  FIGS.  12 B- 12 D  are images showing immunogold labeling of HA-tag in e-PNs during growth in the absence or presence of the designated concentration of IPTG. 
     
    
    
     DETAILED DESCRIPTION 
     A description of example embodiments follows. 
     Several aspects of the invention are described below, with reference to examples for illustrative purposes only. It should be understood that numerous specific details, relationships, and methods are set forth to provide a full understanding of the invention. One having ordinary skill in the relevant art, however, will readily recognize that the invention can be practiced without one or more of the specific details or practiced with other methods, protocols, reagents, cell lines and animals. The present invention is not limited by the illustrated ordering of acts or events, as some acts may occur in different orders and/or concurrently with other acts or events. Furthermore, not all illustrated acts, steps or events are required to implement a methodology in accordance with the present invention. Many of the techniques and procedures described, or referenced herein, are well understood and commonly employed using conventional methodology by those skilled in the art. 
     Unless otherwise defined, all terms of art, notations and other scientific terms or terminology used herein are intended to have the meanings commonly understood by those of skill in the art to which this invention pertains. In some cases, terms with commonly understood meanings are defined herein for clarity and/or for ready reference, and the inclusion of such definitions herein should not necessarily be construed to represent a substantial difference over what is generally understood in the art. It will be further understood that terms, such as those defined in commonly-used dictionaries, should be interpreted as having a meaning that is consistent with their meaning in the context of the relevant art and/or as otherwise defined herein. 
     The terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting. 
     As used herein, the indefinite articles “a,” “an” and “the” should be understood to include plural reference unless the context clearly indicates otherwise. 
     The terms “protein,” “peptide” and “polypeptide” are used interchangeably herein to denote a polymer of at least two amino acids covalently linked by an amide bond, regardless of length or post-translational modification (e.g., glycosylation or phosphorylation). A protein, peptide or polypeptide can comprise any suitable L- and/or D-amino acid, for example, common a-amino acids (e.g., alanine, glycine, valine), non-a-amino acids (e.g., b-alanine, 4-aminobutyric acid, 6-aminocaproic acid, sarcosine, statine), and unusual amino acids (e.g., citrulline, homocitruline, homoserine, norleucine, norvaline, ornithine). The amino, carboxyl and/or other functional groups on a peptide can be free (e.g., unmodified) or protected with a suitable protecting group. Suitable protecting groups for amino and carboxyl groups, and methods for adding or removing protecting groups are known in the art and are disclosed in, for example, Green and Wuts, “ Protecting Groups in Organic Synthesis ,” John Wiley and Sons, 1991. The functional groups of a protein, peptide or polypeptide can also be derivatized (e.g., alkylated) or labeled (e.g., with a detectable label, such as a fluorogen or a hapten) using methods known in the art. A protein, peptide or polypeptide can comprise one or more modifications (e.g., amino acid linkers, acylation, acetylation, amidation, methylation, terminal modifiers (e.g., cyclizing modifications), N-methyl-a-amino group substitution), if desired. In addition, a protein, peptide or polypeptide can be an analog of a known and/or naturally-occurring peptide, for example, a peptide analog having conservative amino acid residue substitution(s). 
     As used herein, the term “sequence identity,” refers to the extent to which two nucleotide sequences, or two amino acid sequences, have the same residues at the same positions when the sequences are aligned to achieve a maximal level of identity, expressed as a percentage. For sequence alignment and comparison, typically one sequence is designated as a reference sequence, to which a test sequences are compared. The sequence identity between reference and test sequences is expressed as the percentage of positions across the entire length of the reference sequence where the reference and test sequences share the same nucleotide or amino acid upon alignment of the reference and test sequences to achieve a maximal level of identity. As an example, two sequences are considered to have 70% sequence identity when, upon alignment to achieve a maximal level of identity, the test sequence has the same nucleotide or amino acid residue at 70% of the same positions over the entire length of the reference sequence. 
     Alignment of sequences for comparison to achieve maximal levels of identity can be readily performed by a person of ordinary skill in the art using an appropriate alignment method or algorithm. In some instances, the alignment can include introduced gaps to provide for the maximal level of identity. Examples include the local homology algorithm of Smith &amp; Waterman,  Adv. Appl. Math.  2:482 (1981), the homology alignment algorithm of Needleman &amp; Wunsch,  J. Mol. Biol.  48:443 (1970), the search for similarity method of Pearson &amp; Lipman,  Proc. Nat&#39;l. Acad. Sci. USA  85:2444 (1988), computerized implementations of these algorithms (GAP, BESTFIT, FASTA, and TFASTA in the Wisconsin Genetics Software Package, Genetics Computer Group, 575 Science Dr., Madison, Wis.), and visual inspection (see generally Ausubel et al.,  Current Protocols in Molecular Biology ). 
     When using a sequence comparison algorithm, test and reference sequences are input into a computer, subsequent coordinates are designated, if necessary, and sequence algorithm program parameters are designated. The sequence comparison algorithm then calculates the percent sequence identity for the test sequence(s) relative to the reference sequence, based on the designated program parameters. A commonly used tool for determining percent sequence identity is Protein Basic Local Alignment Search Tool (BLASTP) available through National Center for Biotechnology Information, National Library of Medicine, of the United States National Institutes of Health. (Altschul et al., 1990). 
     Fusion Proteins Comprising a Type IV Pilin Monomer and a Tag(s) 
     In one aspect, the present invention provides a fusion protein comprising a type IV pilin monomer and a tag. In some embodiments, the tag is at the C-terminus of the type IV pilin monomer. 
     The term “fusion protein” refers to a synthetic, semi-synthetic or recombinant single protein molecule. A fusion protein can comprise all or a portion of two or more different proteins and/or peptides that are attached by covalent bonds (e.g., peptide bonds). 
     Fusion proteins of the invention can be produced recombinantly or synthetically, using routine methods and reagents that are well known in the art. For example, a fusion protein of the invention can be produced recombinantly in a suitable host cell (e.g., bacteria) according to methods known in the art. See, e.g.,  Current Protocols in Molecular Biology , Second Edition, Ausubel et al. eds., John Wiley &amp; Sons, 1992; and  Molecular Cloning: a Laboratory Manual,  2nd edition, Sambrook et al., 1989, Cold Spring Harbor Laboratory Press. For example, a nucleic acid molecule comprising a nucleotide sequence encoding a fusion protein described herein can be introduced and expressed in suitable host cell (e.g.,  E. coli ), and the expressed fusion protein can be isolated/purified from the host cell (e.g., in inclusion bodies) using routine methods and readily available reagents. For example, DNA fragments coding for different protein sequences (e.g., a light-responsive domain, a heterologous peptide component) can be ligated together in-frame in accordance with conventional techniques. In another embodiment, the fusion gene can be synthesized by conventional techniques including automated DNA synthesizers. Alternatively, PCR amplification of nucleic acid fragments can be carried out using anchor primers that give rise to complementary overhangs between two consecutive nucleic acid fragments that can subsequently be annealed and re-amplified to generate a chimeric nucleic acid sequence (see Ausubel et al., Current Protocols in Molecular Biology, 1992). 
     In some embodiments, the fusion protein is encoded by a nucleic acid sequence that comprises SEQ ID NO: 12. In some embodiments, the fusion protein comprises, consists of, or consists essentially of SEQ ID NO: 13. 
     In some embodiments, the fusion protein is encoded by a nucleic acid sequence that comprises SEQ ID NO: 14. In some embodiments, the fusion protein comprises, consists of, or consists essentially of SEQ ID NO: 15. 
     In some embodiments, the fusion protein comprises, consists of, or consists essentially of SEQ ID NO: 16. 
     In some embodiments, the fusion protein comprises, consists of, or consists essentially of SEQ ID NO: 17. 
     In various embodiments, the fusion protein comprises a type IV pilin monomer. A “type IV pilin monomer” refers to a constituent part of a type IV pilus structure that can made by several different species of bacteria. The term “type IV pilin monomer” encompasses naturally occurring (e.g., wildtype) and artificial (e.g, variant, truncated) type IV pilin monomers. One of ordinary skill in the art to which this invention pertains can readily identify type IV pilin monomers from any of several bacterial species that make type IV pili. 
     In some embodiments, the type IV pilin monomer is selected from the group consisting of PilA, PilE, GspG, EspG, OxpG, NE1308, SO0854, PulG, HofG, Yts1G, and combinations thereof. Additional non-limiting examples of type IV pilin monomers include type IV pilin monomers from  Geobacter metallireducens  (SEQ ID NO: 18),  Calditerrivibrio nitroreducens  (SEQ ID NO: 19),  Desulfurvibrio alkaliphilus  (SEQ ID NO: 20),  Felxistipes sinusarabici  (SEQ ID NO: 21),  Synthrophus aciditrophicus  (SEQ ID NO: 22),  Syntrophus gentianae  (SEQ ID NO: 23),  Smithella  sp. F21 (SEQ ID NO: 24),  Syntrophobacter fumaroxidans  (SEQ ID NO: 25),  Syntrophobacter  sp. DG 60 (SEQ ID NO: 26),  Syntrophobacter  sp. SbD1 (SEQ ID NO: 27),  Syntrophorhabdus aromaticivorans  (SEQ ID NO: 28),  Desulfatibacillum alkenivorans  (SEQ ID NO: 29),  Syntrophomonas zehnderi  (SEQ ID NO: 30),  Syntrophaceticus schinkii  (SEQ ID NO: 31),  Tepidanaerobacter acetatoxydans  (SEQ ID NO: 32),  Thermacetogenium phaeum  (SEQ ID NO: 33), or a combination thereof (Table 2). 
     In some embodiments, the type IV pilin monomer comprises, consists of, or consists essentially of a sequence selected from the group consisting of SEQ ID NOs: 18-33 (Table 2). In some embodiments, the type IV pilin monomer has an amino acid sequence that has at least 80% sequence identity to a sequence selected from the group consisting of SEQ ID NOs: 18-33 (Table 2). In some embodiments, the type IV pilin monomer has an amino acid sequence that has at least 85%, at least 90%, at least 95%, at least 98%, or about 99%, sequence identity to a sequence selected from the group consisting of SEQ ID NOs: 18-33 (Table 2). 
     In some embodiments, the type IV pilin monomer is a wildtype type IV pilin monomer produced in a gram-negative bacterium. In some embodiments, the type IV pilin monomer is a variant of the wildtype type IV pilin monomer produced in a gram-negative bacterium. In some embodiments, the variant comprises an amino acid sequence that has at least 80% sequence identity to the amino acid sequence of a wildtype type IV pilin monomer. In some embodiments, the type IV pilin monomer variant comprises an amino acid sequence that has at least 85%, at least 90%, at least 95%, at least 98%, or about 99%, sequence identity to the amino acid sequence of a wildtype type IV pilin monomer. 
     In some embodiments, the type IV pilin monomer is the wildtype  Geobacter sulfurreducens  PilA monomer (SEQ ID NO: 2) (Table 1). 
     In some embodiments, the type IV pilin monomer is a variant of the wildtype  Geobacter sulfurreducens  PilA monomer (SEQ ID NO: 2). 
     In some embodiments, the variant of the wildtype  Geobacter sulfurreducens  PilA monomer is a N-terminal truncation lacking from 1-5 (e.g., 1, 2, 3, 4, or 5) amino acids at the N-terminus of the wildtype  Geobacter sulfurreducens  PilA (SEQ ID NO: 2). In some embodiments, the variant is a C-terminal truncation lacking from 1-5 (e.g., 1, 2, 3, 4, or 5) amino acids at the C-terminus of the wildtype  Geobacter sulfurreducens  PilA (SEQ ID NO: 2). In some embodiments, the variant is a N-terminal addition having from 1-5 (e.g., 1, 2, 3, 4, or 5) additional amino acids at the N-terminus of the wildtype  Geobacter sulfurreducens  PilA (SEQ ID NO: 2). 
     In some embodiments, the variant of the wildtype  Geobacter sulfurreducens  PilA monomer comprises an addition of an aromatic amino acid to the wildtype  Geobacter sulfurreducens  PilA (SEQ ID NO: 2). In some embodiments, about 1-10 aromatic amino acids are added to SEQ ID NO: 2. The number of aromatic amino acids added in SEQ ID NO: 2 may be 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 amino acids; at least 1, 2, 3, 4, 5, 6, 7, 8 or 9 amino acids; or about 1-8, about 2-8, about 2-6, about 3-6 or about 4-6 amino acids. 
     In some embodiments, the variant of the wildtype  Geobacter sulfurreducens  PilA monomer comprises a deletion of one or more aromatic amino acids in the wildtype  Geobacter sulfurreducens  PilA (SEQ ID NO: 2). In some embodiments, about 1-10 aromatic amino acids are deleted from SEQ ID NO: 2. The number of aromatic amino acids deleted in SEQ ID NO: 2 may be 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 amino acids; at least 1, 2, 3, 4, 5, 6, 7, 8 or 9 amino acids; or about 1-8, about 2-8, about 2-6, about 3-6 or about 4-6 amino acids. 
     In some embodiments, the variant of the wildtype  Geobacter sulfurreducens  PilA monomer comprises a substitution of an aromatic amino acid. In some embodiments, about 1-10 aromatic amino acids are substituted in SEQ ID NO: 2. The number of aromatic amino acids substituted in SEQ ID NO: 2 may be 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 amino acids; at least 1, 2, 3, 4, 5, 6, 7, 8 or 9 amino acids; or about 1-8, about 2-8, about 2-6, about 3-6 or about 4-6 amino acids. In some embodiments, the aromatic amino acid is substituted with a non-aromatic amino acid (e.g., alanine (A)). In some embodiments, the aromatic amino acid is substituted with a different aromatic amino acid (e.g., phenylalanine (F)-to-tryptophan (W) or tyrosine (Y)-to-W). 
     In some embodiments, the deleted or substituted aromatic amino acid is F24, F51, Y27, Y32, Y57, or a combination thereof in SEQ ID NO: 2. In some embodiments, the substitution is F24A, F51A, Y27A, Y32A, Y57A, or a combination thereof in SEQ ID NO: 2. In some embodiments, the substitution is F24W, F51W, Y27W, Y32W, Y57W, or a combination thereof in SEQ ID NO: 2. 
     In some embodiments, the variant of the wildtype  Geobacter sulfurreducens  PilA monomer comprises a substitution of a non-aromatic amino acid with an aromatic amino acid. In some embodiments, about 1-10 non-aromatic amino acids are substituted in SEQ ID NO: 2. The number of non-aromatic acids substituted in SEQ ID NO: 2 may be 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 amino acids; at least 1, 2, 3, 4, 5, 6, 7, 8 or 9 amino acids; or about 1-8, about 2-8, about 2-6, about 3-6 or about 4-6 amino acids. 
     In some embodiments, the type IV pilin monomer comprises an amino acid sequence that has at least 80% sequence identity to the amino acid sequence of wildtype  Geobacter sulfurreducens  PilA (SEQ ID NO: 2). In some embodiments, the type IV pilin monomer comprises an amino acid sequence that has at least 85%, at least 90%, at least 95%, at least 98%, or about 99%, sequence identity to the amino acid sequence of wildtype  Geobacter sulfurreducens  PilA (SEQ ID NO: 2). 
     In various embodiments, the fusion protein comprises a tag at the C-terminus of a type IV pilin monomer. As used herein, the term “tag” refers to one or more amino acids that are covalently attached to a type IV pilin monomer (e.g., at the C-terminus of a type IV pilin monomer). In some embodiments, the tag is covalently attached to a type IV pilin monomer by a peptide bond. 
     In some embodiments, the tag is a single amino acid. In some embodiments, the single amino acid is cysteine. 
     In some embodiments, the peptide tag has a length of about (e.g., consists of) 2-200 amino acids, e.g., about 2-180, about 3-180, about 3-160, about 4-160, about 4-140, about 5-140, about 5-120, about 6-120, about 6-100, about 7-100, about 7-80, about 8-80, about 8-60, about 9-60, about 9-50, about 10-50, about 10-40, about 12-40, about 12-35, about 15-35, about 15-30, or about 20-30 amino acids. In some embodiments, the peptide tag consists of about 2-100 amino acids, e.g., about 2-90, about 3-90, about 3-80, about 4-80, about 4-70, about 5-70, about 5-60, about 6-60, about 6-50, about 7-50, about 7-40, about 8-40, about 8-30, about 9-30, about 9-20, or about 10-20 amino acids. In some embodiments, the peptide tag consists of about 2-50 amino acids. In some embodiments, the peptide tag consists of about 5-15 amino acids. 
     In some embodiments, the tag is a peptide. In some embodiments, the peptide tag comprises, consists of, or consists essentially of a polyhistidine sequence, for example, 2-10 consecutive histidine amino acids, e.g., a 2×His tag, 3×His tag, 4×His tag (SEQ ID NO: 3), 5×His tag (SEQ ID NO: 4), 6×His (SEQ ID NO: 5), 7×His tag (SEQ ID NO: 6), 8×His tag (SEQ ID NO: 7), 9×His tag (SEQ ID NO: 8), or 10×His tag (SEQ ID NO: 9). In some embodiments, the peptide tag comprises, consists of, or consists essentially of a 6×His tag (SEQ ID NO: 5). 
     In some embodiments, the peptide tag comprises, consists of, or consists essentially of HHHHHHC (SEQ ID NO: 10). 
     In some embodiments, the peptide tag comprises, consists of, or consists essentially of a human influenza hemagglutinin (HA) sequence (SEQ ID NO: 11). 
     In some embodiments, the peptide tag comprises or consists of a binding motif. Non-limiting examples of the binding motif include nucleic acid (e.g., DNA or RNA)-binding sequences, protein-binding sequences (e.g., an epitope tag or calmodulin binding protein (CBP)), and chemical-binding sequences, etc. Non-limiting examples of epitope tags include HA, FLAG, AU1, AUS, Myc, Glu-Glu, OLLAS, T7, V5, VSV-G, E-Tag, S-Tag, Avi, HSV, KT3, and TK15, etc. Non-limiting examples of chemical-binding sequences include 6His, beta-galactosidase, Strep-tag, Strep-tag II, maltose binding protein, glutathione S transferase (GST), etc. Additional non-limiting examples of tags can be found in Table 3. 
     Electrically Conductive Nanowires 
     The fusion proteins described herein can be joined covalently (e.g., by peptide bonds) to one another to make a polymer. The resultant polymer is referred to herein as a nanowire. In various embodiments, the nanowire is an electrically conductive nanowire. 
     Accordingly, in another aspect, the present invention provides an electrically conductive nanowire, wherein the electrically conductive nanowire comprises one or more fusion proteins described herein. 
     In some embodiments, the electrically conductive nanowire comprises one fusion protein described herein. In some embodiments, the electrically conductive nanowire comprises at least 2 fusion proteins described herein, e.g., 2, 3, 4, 5, 6, 7, 8, 9 or 10; at least 3, 4, 5, 6, 7, 8 or 9; or about 2-10, about 2-8, about 2-6, about 3-6 or about 4-6 fusion proteins described herein. 
     In some embodiments, the electrically conductive nanowire further comprises one or more untagged type IV pilin monomers. The untagged type IV pilin monomer can be any one of the type IV pilin monomers described herein (e.g., a wildtype type IV pilin monomer, a N- or C-terminal truncation, a N-terminal addition, or a variant of the wildtype type IV pilin monomer). In some embodiments, the electrically conductive nanowire comprises one untagged type IV pilin monomer. In some embodiments, the electrically conductive nanowire comprises at least 2 untagged type IV pilin monomers, e.g., 2, 3, 4, 5, 6, 7, 8, 9 or 10; at least 3, 4, 5, 6, 7, 8 or 9; or about 2-10, about 2-8, about 2-6, about 3-6 or about 4-6 untagged type IV pilin monomers. 
     In some embodiments, the molar ratio of fusion protein to untagged type IV pilin monomer in the nanowire is in the range of about 2:1-1:20, e.g., about 2:1-1:18, about 3:2-1:18, about 3:2-1:15, about 1:1-1:15, about 1:1-1:10, about 1:1-1:9, about 1:1-1:8, about 1:1-1:7, about 1:1-1:6, about 1:1-1:5, about 1:1-1:4, about 1:1-1:3, or about 1:1-1:2. In some embodiments, the molar ratio of fusion protein to untagged type IV pilin monomer in the nanowire is at least 1:20, e.g., at least 1:18, at least 1:16, at least 1:15, at least 1:14, at least 1:12, at least 1:10, at least 1:9, at least 1:8, at least 1:7, at least 1:6, at least 1:5, at least 1:4, at least 1:3, at least 1:2, at least 1:1, at least 3:2, or at least 2:1. In some embodiments, the molar ratio of fusion protein to untagged type IV pilin monomer in the nanowire is in the range of about 20:1-1:20, e.g., about 20:1-1:18, about 18:1-1:18, about 18:1-1:16, about 16:1-1:16, about 16:1-1:14, about 14:1-1:14, about 12:1-1:14, about 12:1-1:12, about 10:1-1:12, about 8:1-1:10, about 8:1-1:8, about 6:1-1:8, about 6:1-1:6, about 4:1-1:6, about 4:1-1:4, about 2:1-1:4, about 2:1-1:2, or about 1:1-1:2. In some embodiments, the molar ratio of fusion protein to untagged type IV pilin monomer in the nanowire is in the range of about 10:1 to about 1:10. In some embodiments, the molar ratio of fusion protein to untagged type IV pilin monomer in the nanowire is in the range of about 1:1 to about 1:10. 
     In some embodiments, the electrically conductive nanowire contains no untagged type IV pilin monomers (e.g., the wire consists of fusion proteins described herein). 
     In some embodiments, a plurality of the tags (e.g., peptide tags) are surface exposed. In some embodiments, at least 2% of the tags (e.g., peptide tags), e.g., at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95% or 100% of tags (e.g., peptide tags) are surface exposed. 
     In some embodiments, the electrically conductive nanowire has a diameter of about 3-10 nm, e.g., about 3 nm, about 4 nm, about 5 nm, about 6 nm, about 7 nm, about 8 nm, about 9 nm, about 10 nm, about 3-9 nm, about 3-8 nm, about 3-7 nm, about 3-6 nm, or about 3-5 nm. In some embodiments, the electrically conductive nanowire has a diameter of from about 3-4 nm. 
     In some embodiments, the electrically conductive nanowire has a length of about 0.1-100 μm, e.g., about 0.2-100 μm, about 0.2-80 μm, about 0.3-80 μm, about 0.3-60 μm, about 0.4-60 μm, about 0.4-40 μm, about 0.5-40 μm, about 0.6-20 μm, about 0.6-18 μm, about 0.7-18 μm, about 0.7-16 μm, about 0.8-16 μm, about 0.8-14 μm, about 0.9-14 μm, about 0.9-12 μm, about 1-12 μm, about 1-10 μm, about 1.5-9 μm, about 1.5-8 μm, about 2-8 μm, about 2-6 μm, about 3-6 μm, or about 3-4 μm. In some embodiments, the electrically conductive nanowire has a length of about 0.5 μm to about 20 μm. 
     In some embodiments, the electrically conductive nanowire has a conductivity of about 10 μS/cm-500 S/cm (e.g., at about 25° C. and pH 7), e.g., about 20 μS/cm-500 S/cm, about 20 μS/cm-400 S/cm, about 30 μS/cm-400 S/cm, about 30 μS/cm-300 S/cm, about 40 μS/cm-200 S/cm, about 50 μS/cm-200 S/cm, about 50 μS/cm-100 S/cm, about 60 μS/cm-100 S/cm, about 60 μS/cm-50 S/cm, about 70 μS/cm-50 S/cm, about 80 μS/cm-20 S/cm, about 80 μS/cm-10 S/cm, about 90 μS/cm-10 S/cm, about 9 μS/cm-5 S/cm, or about 100 μS/cm-50 S/cm. In some embodiments, the electrically conductive nanowire has a conductivity of about 40 μS/cm to about 300 S/cm (e.g., at about 25° C. and pH 7). 
     Polynucleotides, Expression Vectors, Host Cells 
     In another aspect, the present invention provides a polynucleotide, wherein the polynucleotide encodes a fusion protein comprising a type IV pilin monomer and a tag, and wherein the tag is at the C-terminus of the type IV pilin monomer. The fusion protein, the type IV pilin monomer and the tag can be any of the fusion proteins, type IV pilin monomers and tags described herein. In some embodiments, the polynucleotide comprises a nucleotide sequence that is at least 70% identical to SEQ ID NO: 1, SEQ ID NO: 12 or SEQ ID NO: 14 (Table 1), e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% identical to SEQ ID NO: 1, SEQ ID NO: 12 or SEQ ID NO: 14. In some embodiments, the polynucleotide comprises SEQ ID NO: 1, SEQ ID NO: 12 or SEQ ID NO: 14. 
     In another aspect, the present invention provides an expression vector comprising a polynucleotide. The polynucleotide is any one of the polynucleotides described herein. In some embodiments, the expression vector further comprises an expression control polynucleotide sequence operably linked to the polynucleotide, a polynucleotide sequence encoding a selectable marker, or both. In some embodiments, the expression control polynucleotide sequence comprises a promoter sequence, an enhancer sequence, or both. In some embodiments, the expression control polynucleotide sequence comprises an inducible promoter sequence. In some embodiments, transcription of the fusion protein can be regulated by an inducer. In some embodiments, the inducer is Isopropyl β-D-1-thiogalactopyranoside (IPTG). 
     The term “promoter” refers to a region of DNA to which RNA polymerase binds and initiates the transcription of a gene. 
     The term “operably linked” means that the nucleic acid is positioned in the recombinant polynucleotide, e.g., vector, in such a way that enables expression of the nucleic acid under control of the element (e.g., promoter) to which it is linked. 
     The term “selectable marker element” is an element that confers a trait suitable for artificial selection. Selectable marker elements can be negative or positive selection markers. 
     In another aspect, the present invention provides a host cell, wherein the host cell comprises a polynucleotide or an expression vector. The polynucleotide and the expression vector can be any of the polynucleotides and expression vectors described herein. 
     In some embodiments, the host cell is a bacterium. In some embodiments, the bacterium normally expresses type IV pili. In some embodiments, the bacterium is a Geobacter cell or a genetically modified Geobacter cell. In some embodiments, the bacterium is a  Geobacter sulfurreducens  cell or a genetically modified  Geobacter sulfurreducens  cell. In some embodiments, the bacterium is a  Geobacter sulfurreducens  KN400 cell or a genetically modified  Geobacter sulfurreducens  KN400 cell. 
     In some embodiments, the bacterium is an  Escherichia coli  cell or a genetically modified  Escherichia coli . In some embodiments, the genetically modified  Escherichia coli  cell expresses a gene selected from the group consisting of: hofB, hofC, hofM, hofN, hofO, hovP, hofQ, ppdA, ppdB, ygdB, ppdC, gspO and combinations thereof. 
     In some embodiments, the bacterium is selected from the group consisting of:  Geobacter sulfurreducens, Escherichia coli, Geobacter metallireducens, Calditerrivibrio nitroreducens, Desulfurvibrio alkahphilus, Felxistipes sinusarabici, Synthrophus aciditrophicus, Syntrophus gentianae, Smithella  sp. F21 , Syntrophobacter fumaroxidans, Syntrophobacter  sp. DG 60 , Syntrophobacter  sp. SbD1 , Syntrophorhabdus aromaticivorans, Desulfatibacillum alkenivorans, Syntrophomonas zehnderi, Syntrophaceticus schinkii, Tepidanaerobacter acetatoxydans, Thermacetogenium phaeum , or a genetically modified variant thereof. 
     In some embodiments, the polynucleotide is integrated into the chromosome of the bacterium (e.g.,  Geobacter sulfurreducens  or  Escherichia coli ). In some embodiments, the polynucleotide is located on an extrachromosomal plasmid in the bacterium (e.g.,  Geobacter sulfurreducens  or  Escherichia coli ). 
     In some embodiments, the host cell further comprises a polynucleotide encoding any one of the untagged type IV pilin monomer described herein. In some embodiments, the polynucleotide encoding the untagged type IV pilin monomer is integrated into the chromosome of the host cell. In some embodiments, the polynucleotide encoding the untagged type IV pilin monomer is located on an extrachromosomal plasmid in the host cell. 
     Methods of Producing Electrically Conductive Protein Nanowires 
     In another aspect, the present invention provides a method of producing the modified, electrically conductive protein nanowires described herein, comprising the steps of:
     a) introducing a polynucleotide or an expression vector into a host cell;   b) placing the host cell in a culture medium conditioned for producing type IV pili;   c) culturing the host cell for a time sufficient to produce a desired quantity of the type IV pili; and   d) isolating the type IV pili from the culture medium,
 
thereby producing the electrically conductive protein nanowires.
   

     The electrically conductive protein nanowires, the polynucleotide, the expression vector, and the host cells employed in the method are as described herein. 
     In some embodiments, the culture medium comprises an Fe(III) compound. In some embodiments, the Fe(III) compound is Fe(III) oxide. In some embodiments, the culture medium comprises about 20 mM acetate and about 40 mM fumarate. In some embodiments, the host cell is cultured under an anaerobic condition at about 30° C. In some embodiments, the method further comprises culturing the host cell in the presence of an inducing molecule. In some embodiments, the inducing molecule is Isopropyl β-D-1-thiogalactopyranoside (IPTG). 
     In another aspect, the present invention provides  Geobacter sulfurreducens  PilA nanowires functionalized to include surface exposed peptide ligands are provided. In one embodiment, the nanowires are electrically conductive. 
     In another aspect, the present invention provides a method of making synthetic electrically conductive protein nanowires having surface exposed peptide ligands, the method including modifying the  Geobacter sulfurreducens  gene for the monomer that assembles into e-PNs to include peptide ligands at the carboxyl terminus of the monomer. 
     Nanowire Sensor Devices 
     In another aspect, the present invention provides a nanowire sensor device comprising:
         an electrically conductive nanowire described herein, comprising one or more of the fusion proteins described herein;   a first electrode having a first electrode terminal;   a second electrode having a second electrode terminal;   an electrical resistance connected between the first and the second electrode terminals; and   an electrical current monitor in electrical communication with the electrical resistance,
 
wherein:
   the first and second electrodes are each configured to support and are each in physical contact with the electrically conductive nanowire; and   the electrical current monitor is configured to measure an electrical current passing through the electrical resistance.       

     The electrically conductive nanowire in the sensor device can be any one of the electrically conductive nanowires described herein, comprising one or more of the fusion proteins described herein. 
     In some embodiments, the first electrode is an anode electrode configured to be imbedded in a material; and the second electrode is a cathode electrode in electrical contact with the material. In some embodiments, the first electrode is a cathode electrode configured to be imbedded in a material and the second electrode is an anode electrode in electrical contact with the materials. 
     In some embodiments, the material is a liquid (e.g., solution) or a solid. 
     In some embodiments, the nanowire sensor device measures the pH of the material. In some embodiments, the peptide tag can detect and/or quantify a chemical or biological analyte. Non-limiting examples of the analytes include aluminium, Ca 2+ , Cobalt, Cu 2+ , Fe—Pt Alloy, germania, Ni 2+ , palladium, Pb 2+ , platinum, stainless steel, Zn 2+ , H 2 O 2 , acetone, ATP, Anthrax protective antigen, atrazine, BRCA1, cardiac troponin I, caspase-3, carbonic anhydrase, CEA, DA, Dioxins, Dioxins and PCBs, diuron, EDCs Enterotoxin B, HA, hCG, HPV, Hydroxyapatite, IgY, methotrexate, MDB, MMP-2, MMP-7, myofibers, prolactin, porphyrin, PSA, streptavidin, substance P, TNF-α, TNT, VOCs, K562 cells,  Ps. aeruginosa  whole cells, and SW620 metastatic cells, etc. Additional non-limiting examples of the analytes can be found in Table 3. 
     EXAMPLES 
     Fabrication of electrically conductive protein nanowires (e-PNs) with  G. sulfurreducens  offers tight, reproducible, and consistent control of nanowire structure and electronic properties, combined with the potential for broad possibilities in the design of e-PNs through genetic modification of the monomer peptide. For example, the conductivity of e-PNs produced with  G. sulfurreducens  have been tuned over six orders of magnitude (ca. 1 mS/cm tol kS/cm) by genetically manipulating the abundance of aromatic amino acids in the monomer peptide. 
     As described herein, a strategy for functionalizing e-PNs with surface-exposed peptide ligands has been developed. The  G. sulfurreducens  gene for the monomer that assembles into e-PNs was modified to add known peptide ligands at the carboxyl terminus of the monomer. Strains of  G. sulfurreducens  were constructed that fabricated synthetic e-PNs with a six-histidine ‘His-tag’ or both the His-tag and a nine-peptide ‘HA-tag’ exposed on the outer surface. The abundance in HA-tag in e-PNs was controlled by placing expression of the gene for the synthetic monomer with the HA-tag under inducible transcriptional regulation. 
       G. sulfurreducens  express e-PNs containing monomers in which peptide ligands were added to the carboxyl terminus. The peptide ligands introduced are accessible on the outer surface of the e-PNs. The potential impact of the added peptide ligands on e-PN conductivity was studied. Addition of the peptide ligands did not diminish e-PN conductivity. 
     Thus, genetic modification of e-PN structure makes it feasible to decorate the outer surface of e-PNs with short peptides. The results also indicate that  G. sulfurreducens  e-PNs can be decorated with a diversity of outer surface peptide ligands to introduce new binding properties. The present disclosure suggests broad possibilities for tailoring e-PN properties for diverse applications. 
     Example 1. Methods and Materials 
     Strains and Growth Conditions 
       G. sulfurreducens  strains were grown under anaerobic conditions at 30° C. in a defined medium with acetate as the electron donor and fumarate as the electron acceptor as previously described.  Escherichia coli  was cultivated with Luria-Bertani medium with or without antibiotics. 
     Construction of  G. sulfurreducens  PilA-WT/PilA-6his Strain 
       G. sulfurreducens  PilA-WT/PilA-6His strain was constructed with  G. sulfurreducens  KN400. A gene for PilA-6His, the gene KN400_3442, which is located downstream of the PilA gene (KN400_1523) on the chromosome, and the putative transcription terminator were integrated at a non-coding region between KN400_0788 and KN400_0787 in the chromosome. The primer pair upKpnI (CTAGGTACCGTGGTGGACCCCCTTACCGGT) (SEQ ID NO: 134) /upSpeI (CGAACTAGTTGTGACCGCTGCCGGCTCCG) (SEQ ID NO: 135) was used to amplify by PCR ca. 550 bp of KN400_0788 upstream of the integration site with the genomic DNA as template. This PCR product was digested with KpnI/SpeI and ligated with the vector pCR2.1GmrloxP 40, resulting in pCR2.1UP-GmrloxP. The 6His tag was fused to the C-terminal end of PilA by PCR with the primer pair PilAdnNotI (CACGCGGCCGCAAGAGGAGCCAGTGACGAAAATC) (SEQ ID NO: 136) /PilACHis (GAGTTAGTGGTGGTGGTGGTGGTGACTTTCGGGCGGATAGGTTTGATC) (SEQ ID NO: 137). For the construction of PilA-6His-KN400_3442, two PCR products were generated before being combined by recombinant PCR. PilA-6His was amplified with the primer pair PilAdnNotI/PilAHisrecup (CTCCAGTATGTATTTAATCAATTAGTGGTGGTGGTGGTGGTG) (SEQ ID NO: 138) while KN400_3442 was amplified with the primer pair PilAHisrecdn (CACCACCACCACCACCACTAATTGATTAAATACATACTGGAG) (SEQ ID NO: 139) /GSU1497XhoIAvrII (CTGCTCGAGGATACCTAGGCTATTCCGACAACTACGAGAC) (SEQ ID NO: 140). The primer pair PilAdnNotI/GSU1497XhoIAvrII was then used to amplify PilA-6His-KN400_3442 by recombinant PCR. PilA-6His-KN400_3442 was cloned at NotI/XhoI sites in pCR2.1UP-GmrloxP downstream of GmrloxP, resulting in pCR2.1UP-GmrloxP-PilAHis-3442. The primer pair dnAvrII (CATCCTAGGAGGGCAGACATTGCGGAACGT) (SEQ ID NO: 141) /dnXhoI (CATCTCGAGCGGGTTCCGCTGCCGTCGTAC) (SEQ ID NO: 142) was used to amplify by PCR ca. 530 bp of KN400_0787 downstream of the integration site. This PCR product was cloned at AvrII/XhoI sites in pCR2.1UP-GmrloxP-PilAHis-3442, resulting in pCR2.1UP-GmrloxP-PilAHis-3442-DN. The final plasmid was linearized with XhoI for transformation as previously described. Transformants were selected with the medium containing gentamicin (20 μg/ml) and were verified by PCR. 
     Construction of  G. sulfurreducens  PilA-WT/PilA-6his/PilA-HA Strain 
       G. sulfurreducens  PilA-WT/PHA-6His/PHA-HA strain was constructed by introducing a gene encoding PilA monomer with the HA tag (PHA-HA) together with the gene KN400_3442 in the chromosome of the  G. sulfurreducens  PilA-WT/PHA-6His strain. The PHA-HA gene was amplified by PCR with a primer pair TCTGGATCCAGGAGGAGACACTTATGCTTCAGAAAC/GTATTTAATCAATTACGCGTAGTCCGGCACGTCGTACGGGTAACTTTCGGGCGGATAG (SEQ ID NO: 143). The gene KN400_3442 was amplified by PCR with a primer pair CTATCCGCCCGAAAGTTACCCGTACGACGTGCCGGACTACGCGTAATTGATTAAATAC (SEQ ID NO: 144) /TCTGAATTCCGATATGACTACTGCGAC (SEQ ID NO: 145). The PHA-HA gene and the KN400_3442 gene were connected by PCR with a primer pair TCTGGATCCAGGAGGAGACACTTATGCTTCAGAAAC/TCTGAATTCCGATATGACTACTGCGAC (SEQ ID NO: 146). The PCR product of PHA-HA/KN400_3442 was digested with BamHI/EcoRI and cloned in the plasmid pKIkan, which is a derivative of pKIapr and has a kanamycin-resistance gene instead of the apramycin-resistance gene. The sequences of KN400_1082 and 1083 used for homologous recombination for introduction of PHA-HA/KN400_3442 are same as those of GSU1106 and 1107 for homologous recombination sequences in pKIkan, respectively. The plasmid thus constructed was linearized with XhoI for electroporation. 
     Western Blot Analysis 
     The wild-type, PilA-WT/PHA-6His, and PilA-WT/PHA-6His/PHA-HA strains were grown with acetate and fumarate at 25° C. IPTG was added at 1 mM for the PHA-WT/PHA-6His/PHA-HA strain with the exception of the study of the impact on IPTG concentrations on incorporation of PHA-HA in filaments. Cell extracts were prepared with B-PER Complete Bacterial Protein Extraction Reagent (Thermo Fisher Scientific) and the amount of protein was measured with the Bradford Protein Assay (Bio-Rad) as instructed by the manufacturer. Cell extracts were separated on 16.5% Tris-Tricine gel (Bio-Rad). An anti-PilA antibody was obtained against peptide, ESAFADDQTYPPES (SEQ ID NO: 147), corresponding to the C-terminal end of PilA (New England Peptide). An anti-6His antibody (6×-His Tag Polyclonal Antibody) and an anti-HA antibody (HA Tag Polyclonal Antibody) were purchased from Invitrogen. Western blot analysis was conducted as described previously. (Leang, C.; Qian, X.; Mester, T.; Lovley, D. R., Alignment of the c-type cytochrome OmcS along pili of  Geobacter sulfurreducens . Appl Environ Microbiol 2010, 76, 4080-4084.) 
     Immunogold Labeling 
     The strains were grown with acetate and fumarate at 25° C. The PilA-WT/PilA-6His/PilA-HA strain was grown with 1 mM IPTG unless otherwise specified. Immunogold labeling was conducted as previously described. (Leang, C.; Qian, X.; Mester, T.; Lovley, D. R., Alignment of the c-type cytochrome OmcS along pili of  Geobacter sulfurreducens . Appl Environ Microbiol 2010, 76, 4080-4084.) For immunogold labeling of just one type of ligand, the 6×-His Tag Polyclonal Antibody or HA Tag Polyclonal Antibody was the primary antibody and the anti-rabbit IgG-gold (10 nm) antibody (Sigma-Aldrich) was the secondary. Dual immunogold labeling was conducted with 6×-His Tag Monoclonal Antibody (Invitrogen) and the HA Tag Polyclonal Antibody as primary antibodies and an anti-mouse IgG-gold (40 nm) antibody (40 nm Goat Anti-Mouse IgG gold conjugate, Expedeon) and the anti-rabbit IgG-gold (10 nm) antibody as secondary antibodies. Samples were examined with transmission electron microscopy. 
     Ni 2+ -Binding Assay 
     The wild-type and PilA-WT/PilA-6His strains were grown with acetate and fumarate at 25° C. Ni 2+ -binding assay was conducted with Ni-NTA-Nanogold (5 nm) (Nanoprobes). Seven μl of the culture was placed on a copper grid and incubated for 5 min. The grid was floated upside down in phosphate-buffered saline (PBS) for 5 min, in PBS containing 3% bovine serum albumin (BSA) and 40 mM imidazole for 15 min, and in PBS containing 0.3% BSA, 40 mM imidazole, and the Ni-NTA-Nanogold for 30 min at room temperature. The grid was washed with PBS containing 40 mM imidazole three times and with water once. Samples were stained with 2% uranyl acetate and examined by transmission electron microscopy. 
     Current Production 
     The capacity to produce current was determined in the two-chambered H-cell system with a continuous flow of medium with acetate (10 mM) as the electron donor and graphite stick anode (65 cm 2 ) poised at 300 mV versus Ag/AgCl as the electron acceptor. 
     Conductance of Individual e-PNs 
     An aliquot (100 μl) of cell culture was drop-cast onto highly oriented pyrolytic graphite (HOPG). Conducting probe atomic force microscopy was preformed using an Oxford Instruments/Asylum Research Cypher ES atomic force microscope with a Pt/Ir-coated Arrow-ContPT tip (NanoWorld AG, Neuchatel, Switzerland). Topographical imaging was performed in contact mode with a force of 0.1 nN. Point-mode current response (I-V) spectroscopy was achieved by applying a 1 nN force to the top of the wire and conducting quadruplicate voltage sweeps of −0.6-0.6 V at 0.99 Hz. The voltage sweep was averaged for each of the I-V curves and conductance was calculated from the linear portion of the I-V curve (−0.2-0.2 V). Average conductance and standard deviation were calculated using 3 independent points on 3 independent e-PNs of each strain. Average height and standard deviation were calculated from 6 independent points on 3 independent e-PNs. 
     Example 2. Wires Decorated with Histidine Ligand 
     To evaluate displaying peptide ligands on the outer surface of e-PNs, the wild-type  G. sulfurreducens  gene for the pilin monomer (PilA) was modified to encode six histidines (i.e., a ‘His-tag’) at the carboxyl end.  FIG.  1    shows the DNA sequence of wild-type pilA, pilA-6His, and pilA-HA genes. Sequences for the His-tag and HA-tag are indicated in solid and dashed underline, respectively. 
       FIG.  2 A  shows the amino acid sequence of the wild-type PilA and of the PHA-6His. As shown in  FIG.  2 B , the synthetic gene was inserted into the chromosome of  G. sulfurreducens  strain KN400, along with the gene for the protein Spc (gene KN400_3442) that may be required for pilin monomer stability. The resultant strain, which contained genes for the wild-type PilA as well as the histidine-modified PilA pilin monomer (PilA-6His), was designated strain PilA-WT/PilA-6His. Western blot analysis with anti-6His antibody of cell lysates of strain PilA-WT/PilA-6His separated with SDS-PAGE revealed a single protein band at the molecular weight expected for the PilA-6His monomer ( FIG.  2 C ). There was no corresponding band in lysates of wild-type cells. Western blot analysis with antibody that detected wild-type PilA detected a single band in wild-type cell lysates and two bands in lysates of strain PilA-WT/PilA-6His ( FIG.  2 C ). The additional band in the strain PHA-WT/PilA-6His lysate was positioned at the higher molecular weight position detected with the anti-6His antibody. 
     Referring to  FIGS.  3 A- 3 B , transmission electron microscopy of cells labeled with the anti-6His antibody and a secondary antibody conjugated with gold revealed abundant His-tag loci along the wires that were accessible to the antibody. As shown in  FIGS.  4 A- 4 B , there was no gold labeling of wild-type cells. 
     When strain PilA-WT/PHA-6His cells were treated with a Ni 2+ -NTA-gold reagent designed to label His-tags, the gold nanoparticles were specifically localized along the wires ( FIGS.  3 C- 3 E ). As shown in  FIGS.  4 C- 4 D , wild-type cells were not labeled. These results further demonstrate that the His-tag ligand was accessible on the outer surface of the wires. 
       G. sulfurreducens  can only produce high current densities on graphite electrodes if its pili are electrically conductive.  G. sulfurreducens  strain PilA-WT/PHA-6His produced maximum currents comparable to the wild-type strain with just a slightly longer lag in the initiation of current production ( FIG.  5   ). This result indicates that introducing the His-tag did not substantially decrease pili conductivity. 
     Conductivity of individual wires was more directly evaluated with conducting tip atomic force microscopy. The wires were readily identified in topographical imaging in contact mode and had a diameter of 3.1+0.3 nm (mean+standard deviation; n=18, 6 points on 3 wires). The conductive tip was lightly applied to the top of the wire (1 nN), and point-mode current response (I-V) spectroscopy revealed a conductance of 7.2+1.5 nS (mean+standard deviation; n=9) ( FIGS.  6  and  7   ). This is comparable to previously observed conductance of 4.5+0.3 nS for e-PNs comprised solely of the wild-type monomer and much higher than the previously reported conductance of the e-PNs from strain  G. sulfurreducens  strain Aro-5, which lacks key aromatic amino acids required for high conductivity. 
     Example 3. Wires Decorated with Two Different Peptide Ligands 
     To determine whether two peptide ligands with different functions could be displayed on one e-PN, a gene ( FIG.  1   ) encoding the nine-peptide ‘HA-tag’ (YPYDVPDYA) (SEQ ID NO: 15) at the carboxyl end of the wild-type PilA pilin monomer ( FIG.  8 A ) was incorporated into the chromosome along with the PilA-6His and wild-type (WT) genes ( FIG.  8 B ). The gene for the PilA with the HA-tag (PHA-HA) was located downstream of the IPTG-inducible lac promoter/operator to provide the option of controlling the stoichiometry of incorporation of the PHA-HA monomer in the e-PNs ( FIG.  8 B ). This strain was designated  G. sulfurreducens  strain PilA-WT/PHA-6His/PHA-HA. Western blot analysis demonstrated that, in the presence of 1 mM IPTG, monomers of WT-PilA, PHA-6His, and PHA-HA were expressed in this strain ( FIG.  8 C ). 
     Immunogold labeling for just the His-tag ( FIGS.  9 A- 9 B ) or the HA-tag ( FIGS.  9 C- 9 D ) demonstrated that both tags were abundant in the e-PNs from strain PilA-WT/PilA-6His/PilA-HA grown with 1 mM IPTG. Dual labeling with secondary antibodies with different size gold particles demonstrated that both tags were present in the same e-PNs ( FIGS.  9 E- 9 F ). e-PNs of strain PilA-WT/PilA-6His cells were not immunogold labeled with the anti-HA antibody ( FIG.  10   ). The current production of strain PilA-WT/PilA-6His/PilA-HA was similar to that of strain PilA-WT/PilA-6His, indicating the addition of the HA-tag did not significantly diminish pili conductivity ( FIG.  5   ). Analysis of individual e-PNs of strain PilA-WT/PilA-6His/PilA-HA ( FIGS.  5  and  11 A- 11 C ) yielded higher currents at equivalent applied voltages than observed with the e-PNs with just the His-tag, with an estimated conductance of 27.2+1.0 nS (n=9). A potential explanation for this is that the HA-tag contains multiple aromatic amino acids, which may promote electron transport. 
     Some PHA-HA was expressed in strain PilA-WT/PilA-6His/PilA-HA even in the absence of the IPTG inducer ( FIG.  12 A ). However, the concentration of PHA-HA monomer in the cells was greater with added IPTG ( FIG.  12 A ). Increased pools of PHA-HA were associated with e-PNs that labeled more heavily with immunogold labeling for the HA-tag ( FIGS.  12 B- 12 D ). These results demonstrate that it is possible to control the abundance of a specific peptide ligand displayed on e-PNs with transcriptional control of the expression of the monomer with that ligand. 
     The results demonstrate that e-PNs produced with  G. sulfurreducens  can be decorated with one or more peptide ligands while maintaining or increasing their conductivity. The stoichiometry of ligand density can be controlled with transcriptional regulation. These capabilities greatly expand the potential applications of e-PNs in electronic devices and for the fabrication of electrically conductive composite materials. 
     For example, sensors developed from other nanowire materials can provide highly sensitive and specific, real-time electrical response for detection of diverse chemicals and biologics. Analytes of interest are detected as a change in nanowire conductivity that results from changes in pH associated with the activity of enzymes incorporated into the sensors, or binding of analytes to nanowires functionalized with antibodies, peptides, or other ligands. The conductivity of  G. sulfurreducens  e-PNs have already been shown to be highly responsive to pH. Short peptides for binding enzymes and antibodies displayed on the outer-surface of e-PNs could be an effective method for functionalizing e-PN-based sensors. Furthermore, peptides can be designed to function as ligands for a wide diversity of chemical and biological analytes or to enhance attachment to cells. Thus, the simplicity of modifying the peptides displayed on e-PNs and controlling the abundance of peptide display provide unprecedented flexibility in nanowire sensor design not readily achieved with other nanowire materials. In a similar manner modifying the surface chemistry of e-PNs with short peptides or unnatural amino acids may enable chemical linkages with polymers or enhance binding to materials to aid in e-PN alignment in electronic devices. 
     Peptides of up to 9 amino acids can be added to the 61-amino acid monomer backbone of  G. sulfurreducens  e-PNs. However, much larger peptide can be used to decorate e-PNs because the monomers of other conductive pili have an N-terminal end homologous to the  G. sulfurreducens  monomer, but are comprised of over 100 amino acids. These broad possibilities for modifying e-PNs with peptides coupled with the advantages of e-PNs as a “green” sustainable material can lead to the development of e-PN-based electronic devices and materials. 
     While example embodiments have been particularly shown and described, it will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the scope of the embodiments encompassed by the appended claims. 
     
       
         
           
               
             
               
                 TABLE 1 
               
               
                   
               
               
                   Geobacter sulfurreducens  PilA, 
               
               
                 Tag, and Fusion Protein Sequences 
               
               
                   
               
             
            
               
                 Wildtype  Geobacter sulfurreducens   
               
               
                 PilA DNA and Protein Sequences 
               
            
           
           
               
               
            
               
                 SEQ ID 
                 ATGCTTCAGAAACTCAGAAACAGGAAAGGTTTCACCCTTATCGAG 
               
               
                 NO: 1 
                 CTGCTGATCGTCGTTGCGATCATCGGTATTCTCGCTGCAATTGCG 
               
               
                   
                 ATTCCGCAGTTCTCGGCGTATCGTGTCAAGGCGTACAACAGCGCG 
               
               
                   
                 GCGTCAAGCGACTTGAGAAACCTGAAGACTGCTCTTGAGTCCGCA 
               
               
                   
                 TTTGCTGATGATCAAACCTATCCGCCCGAAAGTTAA 
               
               
                   
               
               
                 SEQ ID 
                 FTLIELLIVVAIIGILAAIAIPQFSAYRVKAYNSAASSDLRNLKT 
               
               
                 NO: 2 
                 ALESAFADDQTYPPES 
               
               
                   
               
            
           
           
               
            
               
                 Example Histidine (His) Tags 
               
            
           
           
               
               
            
               
                 SEQ ID 
                 HHHH 
               
               
                 NO: 3 
                   
               
               
                   
               
               
                 SEQ ID 
                 HHHHH 
               
               
                 NO: 4 
                   
               
               
                   
               
               
                 SEQ ID 
                 HHHHHH 
               
               
                 NO: 5 
                   
               
               
                   
               
               
                 SEQ ID 
                 HHHHHHH 
               
               
                 NO: 6 
                   
               
               
                   
               
               
                 SEQ ID 
                 HHHHHHHH 
               
               
                 NO: 7 
                   
               
               
                   
               
               
                 SEQ ID 
                 HHHHHHHHH 
               
               
                 NO: 8 
                   
               
               
                   
               
               
                 SEQ ID 
                 HHHHHHHHHH 
               
               
                 NO: 9 
                   
               
               
                   
               
               
                 SEQ ID 
                 HHHHHHC 
               
               
                 NO: 10 
                   
               
               
                   
               
            
           
           
               
            
               
                 Human Influenza Hemagglutinin (HA) Tag 
               
            
           
           
               
               
            
               
                 SEQ ID 
                 YPYDVPDYA 
               
               
                 NO: 11 
                   
               
               
                   
               
            
           
           
               
            
               
                   Geobacter sulfurreducens  PilA-6His Fusion Protein 
               
            
           
           
               
               
            
               
                 SEQ ID 
                 ATGCTTCAGAAACTCAGAAACAGGAAAGGTTTCACCCTTATCGAG 
               
               
                 NO: 12 
                 CTGCTGATCGTCGTTGCGATCATCGGTATTCTCGCTGCAATTGCG 
               
               
                   
                 ATTCCGCAGTTCTCGGCGTATCGTGTCAAGGCGTACAACAGCGCG 
               
               
                   
                 GCGTCAAGCGACTTGAGAAACCTGAAGACTGCTCTTGAGTCCGCA 
               
               
                   
                 TTTGCTGATGATCAAACCTATCCGCCCGAAAGTCACCACCACCAC 
               
               
                   
                 CACCACTAA 
               
               
                   
               
               
                 SEQ ID 
                 FTLIELLIVVAIIGILAAIAIPQFSAYRVKAYNSAASSDLRNLKT 
               
               
                 NO: 13 
                 ALESAFADDQTYPPESHHHHHH 
               
               
                   
               
            
           
           
               
            
               
                   Geobacter sulfurreducens  PilA-HA Fusion Protein 
               
            
           
           
               
               
            
               
                 SEQ ID 
                 ATGCTTCAGAAACTCAGAAACAGGAAAGGTTTCACCCTTATCGAG 
               
               
                 NO: 14 
                 CTGCTGATCGTCGTTGCGATCATCGGTATTCTCGCTGCAATTGCG 
               
               
                   
                 ATTCCGCAGTTCTCGGCGTATCGTGTCAAGGCGTACAACAGCGCG 
               
               
                   
                 GCGTCAAGCGACTTGAGAAACCTGAAGACTGCTCTTGAGTCCGCA 
               
               
                   
                 TTTGCTGATGATCAAACCTATCCGCCCGAAAGTTACCCGTACGAC 
               
               
                   
                 GTGCCGGACTACGCGTAA 
               
               
                   
               
               
                 SEQ ID 
                 FTLIELLIVVAIIGILAAIAIPQFSAYRVKAYNSAASSDLRNLKT 
               
               
                 NO: 15 
                 ALESAFADDQTYPPESYPYDVPDYA 
               
               
                   
               
            
           
           
               
            
               
                   Geobacter sulfurreducens  PilA-Cys Fusion Protein 
               
            
           
           
               
               
            
               
                 SEQ ID 
                 FTLIELLIVVAIIGILAAIAIPQFSAYRVKAYNSAASSDLRNLKT 
               
               
                 NO: 16 
                 ALESAFADDQTYPPEC 
               
               
                   
               
            
           
           
               
            
               
                 
                   Geobacter sulfurreducens 
                 
               
               
                 PilA-6His-Cys Fusion Protein 
               
            
           
           
               
               
            
               
                 SEQ ID 
                 FTLIELLIVVAIIGILAAIAIPQFSAYRVKAYNSAASSDLRNLKT 
               
               
                 NO: 17 
                 ALESAFADDQTYPPESHHHHHHC 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 2 
               
               
                   
               
               
                 Non-limiting Examples of 
               
               
                 Type IV Pilin Monomer Sequences 
               
               
                   
               
             
            
               
                 
                   Geobacter metallireducens 
                 
               
            
           
           
               
               
               
            
               
                   
                 SEQ ID 
                 FTLIELLIVVAIIGILAAIAIPQFAAYRQKAFNSAAESD 
               
               
                   
                 NO: 18 
                 LKNTKTNLESYYSEHQFYPN 
               
               
                   
               
            
           
           
               
            
               
                 
                   Calditerrivibrio nitroreducens 
                 
               
            
           
           
               
               
               
            
               
                   
                 SEQ ID 
                 FTLIELLVVVAIIAILAAIAIPQFAKYRENAAKASAVAD 
               
               
                   
                 NO: 19 
                 AKNIATAIESYYADTQSFPSSISDGSIVPLGTQTFSLSK 
               
               
                   
                   
                 NNSFKGYYYNNPSYTFVVSNTAFNRSVTFNSATGGVDVN 
               
               
                   
                   
                 VW 
               
               
                   
               
            
           
           
               
            
               
                 Desulfurvibrio alkaliphilus 
               
            
           
           
               
               
               
            
               
                   
                 SEQ ID 
                 FTLVELMIVVAIIGILAAVAIPQFAQYRIRGFNSSALSD 
               
               
                   
                 NO: 20 
                 VRNLTTAQEAFFADWLRYAVTHEAADVTEVKATGDLLEG 
               
               
                   
                   
                 PSTGAMVLAQWARQAHQQLPLAIGNGVVMQADVIPATAV 
               
               
                   
                   
                 SYVAISKHLQGNTMYGATNTSTAIHRDQETLVPGQGGDV 
               
               
                   
                   
                 LPITGYMPEPHETDDPFIDHEEFEAQ 
               
               
                   
               
            
           
           
               
            
               
                 
                   Felxistipes sinusarabici 
                 
               
            
           
           
               
               
               
            
               
                   
                 SEQ ID 
                 FTLIELLVVVAIIGILAAIAIPQFAKYRINAFNSAAQSD 
               
               
                   
                 NO: 21 
                 LANVKSALESYYAENFTYPSP 
               
               
                   
               
            
           
           
               
            
               
                 
                   Synthrophus aciditrophicus 
                 
               
            
           
           
               
               
               
            
               
                   
                 SEQ ID 
                 FTLIELMIVIAIIGILAAIAIPQFQQYRTRGYNTAAKAD 
               
               
                   
                 NO: 22 
                 AKNAYTAAQAYFSDHPSVTISSITDLANYGFKASADVTT 
               
               
                   
                   
                 TAAGDMDGLAITAKHSASDTTYQVDSQGTITP 
               
               
                   
               
            
           
           
               
            
               
                 
                   Syntrophus gentianae 
                 
               
            
           
           
               
               
               
            
               
                   
                 SEQ ID 
                 FTLIELMIVIAIIGILAAIAIPQFTQYRKRAYDASSKAD 
               
               
                   
                 NO: 23 
                 LKSAYTAAQAWFNDNPSGTLPAATITSATTTGELPGNGF 
               
               
                   
                   
                 KASTGVTATVTSGTMGTFSIATTHSQGTKTYNITAGGTL 
               
               
                   
                   
                 TES 
               
               
                   
               
            
           
           
               
            
               
                   Smithella  sp. F21 
               
            
           
           
               
               
               
            
               
                   
                 SEQ ID 
                 FTLIELMIVVAIIGILAAIAIPQFANYRTKGYNTKAKAE 
               
               
                   
                 NO: 24 
                 LKSAYTACQAYFSDNPGATACANATLGGFNNSSDVNIAV 
               
               
                   
                   
                 GLSTPTGWTATASHIGGNQTFTVDNGGRITP 
               
               
                   
               
            
           
           
               
            
               
                 
                   Syntrophobacter fumaroxidans 
                 
               
            
           
           
               
               
               
            
               
                   
                 SEQ ID 
                 FTLVELMIVVAIIGILAAVAVPYYQKYIQKSRMVSKVFP 
               
               
                   
                 NO: 25 
                 GMHAIETNMGTYFSFKNTLLDVGSTATFGQFVQDADTKC 
               
               
                   
                   
                 FSPSWAGEYLLITIKDPTLCQELKALTGMTLSATPRMDT 
               
               
                   
                   
                 SRTKIRGWALAGPLAVQLGLEGEQ 
               
               
                   
               
            
           
           
               
            
               
                   Syntrophobacter  sp. DG_60 
               
            
           
           
               
               
               
            
               
                   
                 SEQ ID 
                 FTLIELMIVVAIIAILAAIAIPQYKKFQLKAKTSEAKAN 
               
               
                   
                 NO: 26 
                 LGSIRSCEEAYSAETDNYVFCGWTPSNAPTAAGQAWVTT 
               
               
                   
                   
                 SGHEGFVSIGFAPAGTSRYCYCVGGSVNTAGTDAATNAF 
               
               
                   
                   
                 NEGNVDIYMTAKGDLDGDGSNQWFYATDEDLKVMADYSQ 
               
               
                   
                   
                 DDF 
               
               
                   
               
            
           
           
               
            
               
                   Syntrophobacter  sp. SbD1 
               
            
           
           
               
               
               
            
               
                   
                 SEQ ID 
                 FTLVELMIVVAIIGILAAVAVPYYQKYIQKARLTSKVIP 
               
               
                   
                 NO: 27 
                 GIHSIQTDLATYFSFQQMFPGAGATVNAMFTDANTHCFT 
               
               
                   
                   
                 PTVTSAAGATSNFKITFAIVGAGCTELSSLYNQTITASP 
               
               
                   
                   
                 ILGNNAQVITGWTFGGTLAANMGLAGAQ 
               
               
                   
               
            
           
           
               
            
               
                 
                   Syntrophorhabdus aromaticivorans 
                 
               
            
           
           
               
               
               
            
               
                   
                 SEQ ID 
                 FTLIELLIVIAIIGVLAAIAIPAYTGYTKKAKVGEIIHA 
               
               
                   
                 NO: 28 
                 LGAIKSAVSVYYSEAGATTDATTADLIRTTYGVDVPTGR 
               
               
                   
                   
                 ASFQYTATSREIQATSKITGVTGTMTLTGSTDFKTWTWD 
               
               
                   
                   
                 GTMDKAYIPKN 
               
               
                   
               
            
           
           
               
            
               
                   Desulfatibacillum alkenivorans  PilA 
               
            
           
           
               
               
               
            
               
                   
                 SEQ ID 
                 FTLIELMIVIAIIGILAAIAIPNFVSYRKKAYNRTAQAD 
               
               
                   
                 NO: 29 
                 LSSAYSTVMAYYADEKHKEIDNFTLDNLKDAGFKQTVGV 
               
               
                   
                   
                 AVTVTSVNFQDFELTARHSQGDIVYTIDAAGARSHN 
               
               
                   
               
            
           
           
               
            
               
                   Syntrophomonas zehnderi  PilA 
               
            
           
           
               
               
               
            
               
                   
                 SEQ ID 
                 FTLIEILVALFLAILVASSLVTVFQMSHHIFYRDFSRSE 
               
               
                   
                 NO: 30 
                 LQYMARKAMEDIIDYVVQAQPDSLAVNGAEGSQLEFILS 
               
               
                   
                   
                 SGAKIQYSQGANYWLYRKGPDSGPPQPIVEQIAKVKFCL 
               
               
                   
                   
                 SGPHILTVDVVAGNEKNSFTLTQMIVPRAEIDENDWLNS 
               
               
                   
                   
                 L 
               
               
                   
               
            
           
           
               
            
               
                   Syntrophaceticus schinkii  PilA 
               
            
           
           
               
               
               
            
               
                   
                 SEQ ID 
                 FTLVELMVVLLIIGILVAIAIPIYNKTQENAQKRACQSN 
               
               
                   
                 NO: 31 
                 LRTLDSAAAQYGAATGNYPTDPLNNANFVGENGYVKTKP 
               
               
                   
                   
                 TCPAGGVYNYDATNGKFSCNVPDHVYP 
               
               
                   
               
            
           
           
               
            
               
                   Tepidanaerobacter acetatoxydans  PilA 
               
            
           
           
               
               
               
            
               
                   
                 SEQ ID 
                 FTLIELILALGLLSLIMTTSFTIYSAGQKTYEYENSKIF 
               
               
                   
                 NO: 32 
                 VQQNARQAFLWLSTSIKQARSVEVMSENSIKTVAGDGET 
               
               
                   
                   
                 IIYYFKNGVLYREKNNGINPIAELSQLKFKQPKDKQYIE 
               
               
                   
                   
                 IFLAAQGKEGDDIIIKTKATPFGLWVN 
               
               
                   
               
            
           
           
               
            
               
                   Thermacetogenium phaeum  PilA 
               
            
           
           
               
               
               
            
               
                   
                 SEQ ID 
                 FTMIEMMVVLIIIAVLIAGGIRFYLGYVERAKVTKAKSE 
               
               
                   
                 NO: 33 
                 ITTMQAALDSYYAEKGEYPDDENDRELVKAGLATDRISI 
               
               
                   
                   
                 STEGNDSIQYIYEGGGNSYKIITTATFRAGKLVGEGQDG 
               
               
                   
                   
                 KSTEPDFGSGE 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 3 
               
             
            
               
                   
               
               
                 Non-limiting Examples of Tags and their Corresponding Targets 
               
            
           
           
               
               
               
            
               
                 Target/ 
                   
                 SEQ 
               
               
                 Analyte 
                 Peptide 
                 ID NO: 
               
               
                   
               
            
           
           
               
            
               
                 Table 1 in Karimzadeh et al. (2018) 
               
            
           
           
               
               
               
            
               
                 MMP-2 
                 Biotin-Gly-Pro-Leu-Gly-Val-Arg-Gly-Lys-Gly-Gly-Cys 
                 34 
               
               
                   
               
               
                 PSA 
                 CEHSSKLQLAK-NH2 
                 35 
               
               
                   
               
               
                 HPV 
                 Ac-(Glu)3-CATACACCTCCAGC-Lys(AQ)NH2 
                 36 
               
               
                   
               
               
                 PSA 
                 Diphenylalanine 
                   
               
               
                   
               
               
                 MMP-7 
                 Fc-Lys-Thr-Phe-Lys-Gly-Gly-Gly-Gly-Gly-Gly-Cys 
                 37 
               
               
                   
               
               
                 MMP-7 
                 NH2-KKKRPLALWRSCCC-SH 
                 38 
               
               
                   
               
               
                 Caspase-3 
                 Asp-Glu-Val-Asp 
                 39 
               
               
                   
               
               
                 ATP 
                 EKEKEKE-PPPPC 
                 40 
               
               
                   
               
               
                 hCG 
                 PPLRINRHILTR 
                 41 
               
               
                   
               
               
                 HA 
                 YPYDVPDYA 
                 42 
               
               
                   
               
               
                 Caspase-3 
                 Phe-Arg-Arg 
                   
               
               
                   
               
               
                 H 2 O 2   
                 Diphenylalanine 
                   
               
               
                   
               
               
                 PSA 
                 CHSSLKQK 
                 44 
               
               
                   
               
               
                 K562 cells 
                 Diphenylalanine 
                   
               
               
                   
               
               
                 MMP-7 
                 biotin-RPLALWRSCCC-SH 
                 46 
               
               
                   
               
               
                 Hg 2+   
                 peptide nucleic acid 
                   
               
               
                   
               
               
                 Substance P 
                 Phe-Phe 
                   
               
               
                   
               
               
                 TNF-α 
                 Phe-Phe 
                   
               
               
                   
               
               
                 IgY 
                 H2N-QSPSYPDREYSDEDRQIKQMLHQECPRL-CONH2 
                 47 
               
               
                   
               
               
                 Pb 2+   
                 Cys-Thr-Asn-Thr-Leu-Ser-Asn-Asn-Cys (-S-S-) 
                 48 
               
               
                   
               
               
                 Prolactin 
                 peptide hormone prolactin 
                   
               
               
                   
               
               
                 BRCA1 
                 EKEKEKE 
                 49 
               
               
                   
               
               
                 MMP-2 
                 biotin-GPLGVRGKGGC 
                 50 
               
               
                   
               
               
                 MDB 
                 peptide nucleic acid 
                   
               
               
                   
               
               
                 CEA 
                 peptide-conjugated hemin/G-quadruplex 
                   
               
               
                   
               
               
                 DA 
                 Phe-Phe 
                   
               
               
                   
               
               
                 MMP-7 
                 NH2-KKKRPLALWRSCCC-SH 
                 51 
               
               
                   
               
            
           
           
               
            
               
                 Table 1 in Pavan &amp; Berti (2012) 
               
            
           
           
               
               
               
            
               
                 Cu 2+   
                 3-mer GGH 
                   
               
               
                   
               
               
                 Cu 2+   
                 3-mer GGH 
                   
               
               
                   
               
               
                 Cu 2+   
                 3-mer GGH 
                   
               
               
                   
               
               
                 Cu 2+   
                 8-mer NCGAITIG 
                 52 
               
               
                   
               
               
                 Zn 2+   
                 Zinc-finger peptide 
                   
               
               
                   
               
               
                 Ni 2+   
                 Tetanus toxin (P12) 
                   
               
               
                   
               
               
                 Ni 2+   
                 HCV sequence (NS4/7) 
                   
               
               
                   
               
               
                 Ni 2+   
                 poly-H sequence 
                   
               
               
                   
               
               
                 VOCs 
                 Short-chain polypeptide: PAC1 
                   
               
               
                   
               
               
                 VOCs 
                 Short-chain polypeptide: PAC2 
                   
               
               
                   
               
               
                 VOCs 
                 Short-chain polypeptide: PAM1 
                   
               
               
                   
               
               
                 VOCs 
                 Short-chain polypeptide: PAM2 
                   
               
               
                   
               
               
                 Dioxins and 
                 [N]NFQG-aa-[C] 
                 53 
               
               
                 PCBs 
                   
                   
               
               
                   
               
               
                 Dioxins 
                 FLDQPhenylglycine (and other designed pentamers) 
                 54 
               
               
                   
               
               
                 Atrazine, 
                 EYYY (and other 4-mers from combinatorial libraries) 
                 55 
               
               
                 diuron 
                   
                   
               
               
                   
               
               
                 Carbonic 
                 42-mer helix-loop-helix peptide 
                   
               
               
                 anhydrase 
                   
                   
               
               
                   
               
            
           
           
               
            
               
                 Table 2 in Pavan &amp; Berti (2012) 
               
            
           
           
               
               
               
            
               
                 Murine 
                 7-mer ASSLNIA 
                 56 
               
               
                 myofibers 
                   
                   
               
               
                   
               
               
                 
                   Ps. Aeruginosa 
                 
                 9-mer QRKLAAKLT resembling the sequence of natural CPP 
                 57 
               
               
                 whole cells 
                   
                   
               
               
                   
               
               
                 SW620 
                 12-mer 
                   
               
               
                 metastatic 
                   
                   
               
               
                 cells 
                   
                   
               
               
                   
               
               
                 
                   Anthrax 
                 
                 16-mer HKHAHNYRLPASGGGKK 
                 58 
               
               
                 protective 
                   
                   
               
               
                 antigen 
                   
                   
               
               
                   
               
               
                 Enterotoxin B 
                 12-mer 
                   
               
               
                   
               
               
                 Cardiac 
                 12-mer FYSHSFHENWPS 
                 59 
               
               
                 troponin I 
                   
                   
               
               
                   
               
               
                 Streptavidin 
                 12-mer from filamentous R5C2 phages 
                   
               
               
                   
               
               
                 Methotrexate 
                 11-mer SIFPLCNSGAL selected directly on a QCM device 
                 60 
               
               
                   
               
               
                 TNT 
                 12-mers sharing a HR consensus dyad 
                   
               
               
                   
               
               
                 Porphyrin 
                 5-mer HASYS peptide immobilized on Latex beads to allow 
                 61 
               
               
                   
                 2:1 peptide-to-target complex formation 
                   
               
               
                   
               
               
                 EDCs 
                 Peptide α/β I 
                   
               
               
                   
               
            
           
           
               
            
               
                 Table 3 in Pavan &amp; Berti (2012) 
               
            
           
           
               
               
               
            
               
                 Taq DNA 
                 58-mer affibodies from phage display 
                   
               
               
                 polymerase and 
                   
                   
               
               
                 IgA 
                   
                   
               
               
                   
               
               
                 IgA 
                 (58-mer) n  multimeric affibodies from phage display, by 
                   
               
               
                   
                 oligomerization of the peptides over the microarray slides 
                   
               
               
                   
               
               
                 IgE 
                 (58-mer) n  multimeric affibodies from phage display, by 
                   
               
               
                   
                 oligomerization of the peptides over the microarray slides 
                   
               
               
                   
               
               
                 IgG 
                 (58-mer) n  multimeric affibodies from phage display, by 
                   
               
               
                   
                 oligomerization of the peptides over the microarray slides 
                   
               
               
                   
               
               
                 TNF-α 
                 (58-mer) n  multimeric affibodies from phage display, by 
                   
               
               
                   
                 oligomerization of the peptides over the microarray slides 
                   
               
               
                   
               
               
                 insulin 
                 (58-mer) n  multimeric affibodies from phage display, by 
                   
               
               
                   
                 oligomerization of the peptides over the microarray slides 
                   
               
               
                   
               
               
                 Taq DNA 
                 (58-mer) n  multimeric affibodies from phage display, by 
                   
               
               
                 polymerase 
                 oligomerization of the peptides over the microarray slides 
                   
               
               
                   
               
               
                 HER-2 
                 58-mer affibodies from phage display 
                   
               
               
                 expressing 
                   
                   
               
               
                 tumor cells 
                   
                   
               
               
                   
               
               
                 HIV-1 
                 25-mer coiled coil from gp41 HIV protein. The peptide acts as 
                   
               
               
                 pseudopeptide 
                 an ion channel 
                   
               
               
                 adhesion 
                   
                   
               
               
                 inhibitors 
                   
                   
               
               
                   
               
               
                 Antibodies 
                 15-mer gramicidin A. 6.3 helical ion channel displaying an 
                   
               
               
                   
                 antigen 
                   
               
               
                   
               
               
                 C-reactive 
                 35-mer helix-loop-helix chemically modified coiled coils 
                   
               
               
                 protein 
                   
                   
               
               
                   
               
               
                 Caffeine 
                 35-mer K coiled-coil randomized peptide from phage display 
                   
               
               
                   
               
               
                 
                   Botulinum 
                 
                 22-mer synaptotagmin-derived peptide 
                   
               
               
                 neurotoxin 
                   
                   
               
               
                   
               
               
                 Phosphoryl 
                 33-mer WW phosphoprotein binding domain 
                   
               
               
                 peptides 
                   
                   
               
               
                   
               
               
                 Nitric oxide 
                 12-mers derived from the integin-binding fibronectin domain 
                   
               
               
                 released by 
                 and laminins 
                   
               
               
                 captured cells 
                   
                   
               
               
                   
               
               
                 Alcohols 
                 LUSH protein sequence (81-93) 
                   
               
               
                 associated with 
                   
                   
               
               
                 
                   Salmonella 
                 
                   
                   
               
               
                   typhimurium  in 
                   
                   
               
               
                 beef 
                   
                   
               
               
                   
               
               
                 VOCs xylene 
                 Human olfactory receptor protein (P30953) fragment: 
                   
               
               
                   
                 horp61(61-72) 
                   
               
               
                   
               
               
                 ammonia 
                 Human olfactory receptor protein (P30953) fragment: 
                   
               
               
                   
                 horp103(103-8) 
                   
               
               
                   
               
               
                 trimethylamine 
                 Human olfactory receptor protein (P30953) fragment: 
                   
               
               
                   
                 horp109(109-14) 
                   
               
               
                   
               
               
                 acetic acid 
                 Human olfactory receptor protein (P30953) fragment: 
                   
               
               
                   
                 horp193(193-99) 
                   
               
               
                   
               
               
                 VOCs 1-hexanol 
                 Mouse olfactory receptor (OR744) sequence 210-216 
                   
               
               
                   
               
               
                 1-pentanol 
                 Mouse olfactory receptor (OR744) sequence 210-216 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                   
               
             
            
               
                 Table 4 in Pavan &amp; Berti (2012) 
               
            
           
           
               
               
               
            
               
                 mAb against the 
                 CDYKDDDDK (FLAG) peptide 
                  62 
               
               
                 FLAG peptide 
                   
                   
               
               
                   
               
               
                 mAb (12 F10) 
                 Polypeptide from HCVARFP 
                  63 
               
               
                 against HCV 
                   
                   
               
               
                 protein ARFP 
                   
                   
               
               
                 (A97) 
                   
                   
               
               
                   
               
               
                 HIV-1 PR 
                 Protease substrate DABCYL-SQNYPIVQ-EDANS 
                  64 
               
               
                   
               
               
                 Trypsin 
                 Trypsin substrate DABCYL-GPAXAALAIG-EDANS 
                  65 
               
               
                   
               
               
                 
                   Botulinum 
                 
                 SNAP-25 (187-203 and derivates) 
                   
               
               
                 neurotoxins A 
                   
                   
               
               
                   
               
               
                 
                   Botulinum 
                 
                 VAMP-2 (35-70 and derivates) 
                   
               
               
                 neurotoxins B 
                   
                   
               
               
                   
               
               
                 
                   Botulinum 
                 
                 VAMP (60-94 and derivates) 
                   
               
               
                 neurotoxins F 
                   
                   
               
               
                   
               
               
                 Caspase 3 
                 Peptide substrate SGDEVDSG 
                  66 
               
               
                   
               
               
                 
                   Botulinum 
                 
                 SNAP-25 (187-202) modified 
                   
               
               
                 neurotoxin LcA 
                   
                   
               
               
                   
               
               
                 Subtilisin 
                 Decapeptide substrate 
                   
               
               
                   
               
               
                 MMP-7 
                 Helix peptide with sequence RPLALWRSC 
                  67 
               
               
                   
               
               
                 Proteases 
                 Fc-peptide monolayers 
                   
               
               
                   
               
               
                 Plasmin 
                 Fc-peptide conjugate (KTFK) 
                  68 
               
               
                   
               
               
                 Papain 
                 Fc-peptide conjugates 
                   
               
               
                   
               
               
                 Caspase 3 
                 Fc-peptide conjugate (GDGDEVDGC) 
                  69 
               
               
                 (detection of 
                   
                   
               
               
                 apoptotic cells) 
                   
                   
               
               
                   
               
               
                 HIV-1 PR 
                 Fc-pepstatin 
                   
               
               
                   
               
               
                 HIV-1 PR 
                 Fc-pepstatin conjugates 
                   
               
               
                   
               
               
                 Protein kinases 
                 Peptide substrates for tyrosine kinases Abl and Src 
                   
               
               
                   
               
               
                 Protein kinases 
                 ARCs. ARCs are potent inhibitors of protein kinases 
                   
               
               
                   
               
               
                 MMP-1 
                 Peptide substrate of catalytic domain (AMFLEA) 
                  70 
               
               
                   
               
               
                 β-galactosidase 
                 20-mer YHNN and QYHH selected from a microarray based 
                  71, 
               
               
                   
                 on β-galactosidase binding 
                  72 
               
               
                   
               
            
           
           
               
            
               
                 Table 1 in Puiu &amp; Bala (2018) 
               
            
           
           
               
               
               
            
               
                 Matrix 
                 GPLGMWSRC 
                  73 
               
               
                 metalloproteinase- 
                   
                   
               
               
                 9 (MMP-9) 
                   
                   
               
               
                   
               
               
                 Trypsin 
                 GRPS-PEG disulfide 
                  74 
               
               
                   
               
               
                 α-thrombin 
                 RFSRPQL-PEG disulfide 
                  75 
               
               
                   
               
               
                 Plasmin 
                 KTKTC 
                  76 
               
               
                   
               
               
                 Epidermal growth 
                 YHWYGYTPQNVI 
                  77 
               
               
                 factor receptor 
                   
                   
               
               
                   
               
               
                 Prostate specific 
                 HSSKLQL 
                  78 
               
               
                 antigen 
                   
                   
               
               
                   
               
               
                 Amyloid 1-42 
                 RGTWEGKWK 
                  79 
               
               
                   
               
               
                 HIV anti-p24 
                 EAAEWDRVHP 
                  80 
               
               
                 antibody 
                   
                   
               
               
                   
               
               
                 Legumain 
                 H2N-(CH2)4-CO-AAAN-NH-CH2- 
                  81 
               
               
                   
               
               
                 Cathepsin B 
                 H2N-(CH2)4-CO-LRFG-NH-CH2- 
                  82 
               
               
                   
               
               
                 SW620 colorectal 
                 DDAGNRQP 
                  83 
               
               
                 carcinoma cells 
                   
                   
               
               
                   
               
            
           
           
               
            
               
                 Table 1 in Seker &amp; Demir (2011) 
               
            
           
           
               
               
               
            
               
                 Gold 
                 #VSGSSPDS 
                  84 
               
               
                   
               
               
                 Gold 
                 #LKAHLPPSRLPS 
                  85 
               
               
                   
               
               
                 Gold 
                 *TGTSVLIATPYV 
                  86 
               
               
                   
               
               
                 Silver 
                 *AYSSGAPPMPPF 
                  87 
               
               
                   
               
               
                 Silver 
                 *IRPAIHIIPISH 
                  88 
               
               
                   
               
               
                 Silver 
                 *WSWRSPTPHVVT 
                  89 
               
               
                   
               
               
                 Silica 
                 #MSPHPHPRHHHT 
                  90 
               
               
                   
               
               
                 Silica 
                 #RGRRRRLSCRLL 
                  91 
               
               
                   
               
               
                 Silica 
                 RLNPPSQMDPPF 
                  92 
               
               
                   
               
               
                 Silica 
                 QTWPPPLWFSTS 
                  93 
               
               
                   
               
               
                 Silica 
                 HPPMNASHPHMH 
                  94 
               
               
                   
               
               
                 Silica 
                 HTKHSHTSPPPL 
                  95 
               
               
                   
               
               
                 Silica 
                 CHKKPSKSC 
                  96 
               
               
                   
               
               
                 Titania/Titanium 
                 *RKLPDAPGMHTW 
                  97 
               
               
                   
               
               
                 Titania/Titanium 
                 *YPSAPPQWLTNT 
                  98 
               
               
                   
               
               
                 Titania/Titanium 
                 *STPLVTGTNNLM 
                  99 
               
               
                   
               
               
                 Titania/Titanium 
                 *QSGSHVTGDLRL 
                 100 
               
               
                   
               
               
                 Titania/Titanium 
                 *ATTLHPPRTSLP 
                 101 
               
               
                   
               
               
                 Titania/Titanium 
                 #SCSDCLKSVDFIPSSLASS 
                 102 
               
               
                   
               
               
                 Titania/Titanium 
                 #LNAAVPFTMAGS 
                 103 
               
               
                   
               
               
                 Titania/Titanium 
                 #ATWVSPY 
                 104 
               
               
                   
               
               
                 Titania/Titanium 
                 *RKKRTKNPTHKLGGGW 
                 105 
               
               
                   
               
               
                 Titania/Titanium 
                 *KSLSRHDHIHHHGGGW 
                 106 
               
               
                   
               
               
                 Titania/Titanium 
                 *TQHLSHPRYATKGGGW 
                 107 
               
               
                   
               
               
                 Zinc Oxide 
                 *EAHVMHKVAPRP 
                 108 
               
               
                   
               
               
                 Zinc Oxide 
                 *GLHVMHLVAPPR 
                 109 
               
               
                   
               
               
                 Zinc Oxide 
                 *VRTRDDARTHRK 
                 110 
               
               
                   
               
               
                 Iridium Oxide 
                 #AGETQQAM 
                 111 
               
               
                   
               
               
                 Iron Oxide 
                 #LSTVQTISPSNH 
                 112 
               
               
                   
               
               
                 Germania 
                 *TGHQSPGAYAAH 
                 113 
               
               
                   
               
               
                 Germania 
                 *SLKMPHWPHLLP 
                 114 
               
               
                   
               
               
                 Platinum 
                 *CPTSTGQAC 
                 115 
               
               
                   
               
               
                 Platinum 
                 *CTLHVSSYC 
                 116 
               
               
                   
               
               
                 Palladium 
                 *QQSWPIS 
                 117 
               
               
                   
               
               
                 Palladium 
                 *NFMSLPRLGHMH 
                 118 
               
               
                   
               
               
                 Palladium 
                 #SVTQNKY 
                 119 
               
               
                   
               
               
                 Palladium 
                 #SPHPGPY 
                 120 
               
               
                   
               
               
                 Palladium 
                 #HAPTPML 
                 121 
               
               
                   
               
               
                 Aluminium 
                 #VPSSGPQDTRTT 
                 122 
               
               
                   
               
               
                 Aluminium 
                 #YSPDPRPWSSRY 
                 123 
               
               
                   
               
               
                 Stainless Steel 
                 *MTWDPSLASPRS 
                 124 
               
               
                   
               
               
                 Stainless Steel 
                 *ATIHDAFYSAPE 
                 125 
               
               
                   
               
               
                 Stainless Steel 
                 *NLNPNTASAMHV 
                 126 
               
               
                   
               
               
                 Fe-Pt Alloy 
                 #HNKHLPSTQPLA 
                 127 
               
               
                   
               
               
                 Fe-Pt Alloy 
                 SVSVGMKPSPRP 
                 128 
               
               
                   
               
               
                 Fe-Pt Alloy 
                 VISNHRESSRPL 
                 129 
               
               
                   
               
               
                 Cobalt 
                 #HSVRWLLPGAHP 
                 130 
               
               
                   
               
               
                 Cobalt 
                 KLHSSPHTLPVQ, 
                 131 
               
               
                   
               
               
                 Hydroxyapatite 
                 #SVSVGMKPSPRP 
                 132 
               
               
                   
               
               
                 Hydroxyapatite 
                 *CMLPHHGAC 
                 133 
               
               
                   
               
            
           
         
       
     
     REFERENCES 
     
         
         1. Lovley, D. R. (2017) Electrically conductive pili: biological function and potential applications in electronics. Curr. Opin. Electrochem. 4, 190-198. 
         2. Lovley, D. R. (2017) e-Biologics: Fabrication of sustainable electronics with ‘green’ biological materials. mBio 8, e00695. 
         3. Guterman, T., and Gazit, E. (2018) Toward peptide-based bioelectronics: reductionist design of conductive pili mimetics. Bioelectron. Med. 1, 131-137. 
         4. Creasey, R. C. G., Mostert, A. B., Nguyen, T. A. H., Virdis, B., Freguia, S., and Laycock, B. (2018) Microbial nanowires—electron transport and the role of synthetic analogues. Acta Biomater. 69, 1-30. 
         5. Ing, N. L., El-Naggar, M. Y., and Hochbaum, A. I. (2018) Going the distance: long-range conductivity in protein and peptide bioelectronic materials. J. Phys. Chem. B 122, 10403-10423. 
         6. Zhou, W., Dai, X., Fu, T.-M., Xie, C., Liu, J., and Lieber, C. M. (2014) Long term stability of nanowire nanoelectronics in physiological environments. Nano Lett. 14, 1614-1619. 
         7. Malvankar, N. S., Vargas, M., Nevin, K. P., Franks, A. E., Leang, C., Kim, B.-C., Inoue, K., Mester, T., Covalla, S. F., Johnson, J. P., Rotello, V. M., Tuominen, M. T., and Lovley, D. R. (2011) Tunable metallic-like conductivity in nanostructured biofilms comprised of microbial nanowires. Nat. Nanotechnol. 6, 573-579. 
         8. Adhikari, R. Y., Malvankar, N. S., Tuominen, M. T., and Lovley, D. R. (2016) Conductivity of individual Geobacter pili. RSC Adv. 6, 8354-8357. 
         9. Lovley, D. R. (2017) Syntrophy goes electric: direct interspecies electron transfer. Annu. Rev. Microbiol. 71, 643-664. 
         10. Reguera, G., McCarthy, K. D., Mehta, T., Nicoll, J. S., Tuominen, M. T., and Lovley, D. R. (2005) Extracellular electron transfer via microbial nanowires. Nature 435, 1098-1101. 
         11. Tan, Y., Adhikari, R. Y., Malvankar, N. S., Ward, J. E., Woodard, T. L., Nevin, K. P., and Lovley, D. R. (2017) Expressing the  Geobacter metallireducens  PilA in  Geobacter sulfurreducens  yields pili with exceptional conductivity. mBio 8, e02203. 
         12. Holmes, D. E., Dang, Y., Walker, D. J. F., and Lovley, D. R. The electrically conductive pili of Geobacter species are a recently evolved feature for extracellular electron transfer. Microb. Genomics 2016, 2. 
         13. Sure, S., Ackland, M. L., Torriero, A. J., Adholeya, A., and Kochar, M. (2016) Microbial nanowires: an electrifying tale. Microbiology 162, 2017-2028. 
         14. Walker, D. J. F., Adhikari, R. Y., Holmes, D. E., Ward, J. E., Woodard, T. L., Nevin, K. P., and Lovley, D. R. (2018) Electrically conductive pili from genes of phylogenetically diverse microorganisms. ISME J. 12, 48-58. 
         15. Walker, D. J. F., Martz, E., Holmes, D. E., Zhou, Z., Nonnenmann, S. S., and Lovley, D. R. (2019) The archaellum of  Methanospirillum hungatei  is electrically conductive. mBio 10, e00579. 
         16. Cologgi, D. L., Lampa-Pastirk, S., Speers, A. M., Kelly, S. D., and Reguera, G. (2011) Extracellular reduction of uranium via Geobacter conductive pili as a protective cellular mechanism. Proc. Natl. Acad. Sci. U.S.A 108, 15248-15252. 
         17. Veazey, J. P., Reguera, G., and Tessmer, S. H. (2011) Electronic properties of conductive pili of the metal-reducing bacterium  Geobacter sulfurreducens  probed by scanning tunneling microscopy. Phys. Rev. 84, 060901. 
         18. Vargas, M., Malvankar, N. S., Tremblay, P.-L., Leang, C., Smith, J. A., Patel, P., Snoeyenbos-West, O., Nevin, K. P., and Lovley, D. R. (2013) Aromatic amino acids required for pili conductivity and longrange extracellular electron transport in  Geobacter sulfurreducens . mBio 4, e00105. 
         19. Lampa-Pastirk, S., Veazey, J. P., Walsh, K. A., Feliciano, G. T., Steidl, R. J., Tessmer, S., and Reguera, G. (2016) Thermally activated charge transport in microbial protein nanowires. Sci. Rep. 6, 23517. 
         20. Malvankar, N. S., Yalcin, S. E., Tuominen, M. T., and Lovley, D. R. (2014) Visualization of charge propagation along individual pili proteins using ambient electrostatic force microscopy. Nat. Nanotechnol. 9, 1012-1017. 
         21. Malvankar, N. S., Vargas, M., Nevin, K. P., Tremblay, P.-L., Evans-Lutterodt, K., Nykypanchuk, D., Martz, E., Tuominen, M. T., and Lovley, D. R. (2015) Structural basis for metallic-like conductivity in microbial nanowires. mBio 6, e00084. 
         22. Tan, Y., Adhikari, R. Y., Malvankar, N. S., Pi, S., Ward, J. E., Woodard, T. L., Nevin, K. P., Xia, Q., Tuominen, M. T., and Lovley, D. R. (2016) Synthetic biological protein nanowires with high conductivity. Small 12, 4481-4485. 
         23. Ing, N. L., Nusca, T. D., and Hochbaum, A. I. (2017)  Geobacter sulfurreducens  pili support ohmic electronic conduction in aqueous solution. Phys. Chem. Chem. Phys. 19, 21791-21799. 
         24. Wang, F., Gu, Y., O&#39;Brien, J. P., Yi, S. M., Yalcin, S. E., Srikanth, V., Shen, C., Vu, D., Ing, N. L., Hochbaum, A. I., Egelman, E. H., and Malvankar, N. S. (2019) Structure of microbial nanowires reveals stacked hemes that transport electrons over micrometers. Cell 177, 361-369. 
         25. Filman, D. J., Marino, S. F., Ward, J. E., Yang, L., Mester, Z., Bullitt, E., Lovley, D. R., and Strauss, M. (2019) Cryo-EM reveals the structural basis of long-range electron transport in a cytochromebased bacterial nanowire. Commun. Biol. 2, 219. 
         26. Lovley, D. R., and Walker, D. J. F. (2019) Geobacter protein nanowires. Peed. Prepr. 7, e27773v1. 
         27. Yi, H., Nevin, K. P., Kim, B.-C., Franks, A. E., Klimes, A., Tender, L. M., and Lovley, D. R. (2009) Selection of a variant of  Geobacter sulfurreducens  with enhanced capacity for current production in microbial fuel cells. Biosens. Bioelectron. 24, 3498-3503. 
         28. Malvankar, N. S., Tuominen, M. T., and Lovley, D. R. (2012) Lack of involvement of c-type cytochromes in long-range electron transport in microbial biofilms and nanowires. Energy Environ. Sci. 5, 8651-8659. 
         29. Tan, Y., Adhikari, R. Y., Malvankar, N. S., Ward, J. E., Nevin, K. P., Woodard, T. L., Smith, J. A., Snoeyenbos-West, O. L., Franks, A. E., Tuominen, M. T., and Lovley, D. R. (2016) The low conductivity of  Geobacter uraniireducens  pili suggests a diversity of extracellular electron transfer mechanisms in the genus  Geobacter . Front. Microbiol. 7, 980. 
         30. Nguyen, P. Q., Botyanszki, Z., Tay, P. K. R., and Joshi, N. S. (2014) Programmable biofilm-based materials from engineered curli nanofibres. Nat. Commun. 5, 4945. 
         31. Chen, A. Y., Deng, Z., Billings, A. N., Seker, U. O. S., Lu, M. Y., Citorik, R. J., Zakeri, B., and Lu, T. K. (2014) Synthesis and patterning of tunable multiscale materials with engineered cells. Nat. Mater. 13, 515-523. 
         32. Wang, Y., Pu, J., An, B., Lu, T. K., and Zhong, C. (2018) Emerging paradigms for synthetic design of functional amyloids. J. Mol. Biol. 430, 3720-3734. 
         33. Nguyen, P. Q., Dorval Courchesne, N.-M., Duraj-Thatte, A., Praveschotinunt, P., and Joshi, N. S. (2018) Engineered living materials: prospects and challenges for using biological systems to direct the assembly of smart Materials. Adv. Mater. 30, 1704847. 
         34. Hospenthal, M. K., Costa, T. R. D., and Waksman, G. (2017) A comprehensive guide to pilus biogenesis in Gram-negative bacteria. Nat. Rev. Microbiol. 15, 365-379. 
         35. Liu, X., Zhou, S., and Lovley, D. R. (2019) A pilin chaperone required for the expression of electrically conductive  Geobacter sulfurreducens  pili. Environ. Microbiol. 21, 2511-2522. 
         36. Malvankar, N. S., Mester, T., Tuominen, M. T., and Lovley, D. R. (2012) Supercapcitors based on c-type cytochromes using conductive nanostructured networks of living bacteria. ChemPhysChem 13, 463-468. 
         37. Liu, X., Tremblay, P.-L., Malvankar, N. S., Nevin, K. P., Lovley, D. R., and Vargas, M. (2014) A  Geobacter sulfurreducens  strain expressing  Pseudomonas aeruginosa  type IV pili localizes OmcS on pili but is deficient in Fe(III) oxide reduction and current production. Appl. Environ. Microbiol. 80, 1219-1224. 
         38. Smith, J. A., Tremblay, P.-L., Shrestha, P. M., Snoeyenbos-West, O. L., Franks, A. E., Nevin, K. P., and Lovley, D. R. (2014) Going wireless: Fe(III) oxide reduction without pili by  Geobacter sulfurreducens  strain JS-1. Appl. Environ. Microbiol. 80, 4331-4340. 
         39. Liu, X., Wang, S., Xu, A., Zhang, L., Liu, H., and Ma, L. Z. (2019) Biological synthesis of high-conductive pili in aerobic bacterium  Pseudomonas aeruginosa . Appl. Microbiol. Biotechnol. 103, 1535-1544. 
         40. Field, J., Nikawa, J., Broek, D., MacDonald, B., Rodgers, L., Wilson, I. A., Lerner, R. A., and Wigler, M. (1988) Purification of a RAS-responsive adenylyl cyclase complex from  Saccharomyces cerevisiae  by use of an epitope addition method. Mol. Cell. Biol. 8, 2159-65. 
         41. Ueki, T., Nevin, K. P., Woodard, T. L., and Lovley, D. R. (2016) Genetic switches and related tools for controlling gene expression and electrical outputs of  Geobacter sulfurreducens . J. Ind. Microbiol. Biotechnol. 43, 1561-1575. 
         42. Ahmad, R., Mahmoudi, T., Ahn, M. S., and Hahn, Y. B. (2018) Recent advances in nanowires-based field-effect transistors for biological sensor applications. Biosens. Bioelectron. 100, 312-325. 
         43. Zhang, A., and Lieber, C. M. (2016) Nano-Bioelectronics. Chem. Rev. 116, 215-57. 
         44. Zhu, C., Yang, G., Li, H., Du, D., and Lin, Y. (2015) Electrochemical sensors and biosensors based on nanomaterials and nanostructures. Anal. Chem. 87, 230-49. 
         45. Lu, Y., Huang, F., Wang, J., and Xia, J. (2014) Affinity-guided covalent conjugation reactions based on PDZ-peptide and SH3-peptide interactions. Bioconjugate Chem. 25, 989-99. 
         46. Yan, X., Zhou, H., Zhang, J., Shi, C., Xie, X., Wu, Y., Tian, C., Shen, Y., and Long, J. (2009) Molecular mechanism of inward rectifier potassium channel 2.3 regulation by tax-interacting protein-1. J. Mol. Biol. 392, 967-76. 
         47. Huang, J., Nagy, S. S., Koide, A., Rock, R. S., and Koide, S. (2009) A peptide tag system for facile purification and singlemolecule immobilization. Biochemistry 48, 11834-6. 
         48. Gaidamakova, E. K., Backer, M. V., and Backer, J. M. (2001) Molecular vehicle for target-mediated delivery of therapeutics and diagnostics. J. Controlled Release 74, 341-7. 
         49. Zakeri, B., Fierer, J. O., Celik, E., Chittock, E. C., Schwarz-Linek, U., Moy, V. T., and Howarth, M. (2012) Peptide tag forming a rapid covalent bond to a protein, through engineering a bacterial adhesin. Proc. Natl. Acad. Sci. U.S.A 109 (12), E690-7. 
         50. Skerra, A., and Schmidt, T. G. (2000) Use of the Strep-Tag and streptavidin for detection and purification of recombinant proteins. Methods Enzymol. 326, 271-304. 
         51. Pavan, S., and Berti, F. (2012) Short peptides as biosensor transducers. Anal. Bioanal. Chem. 402, 3055. 
         52. Liu, Q., Wang, J., and Boyd, B. J. (2015) Peptide-based biosensors. Talanta 136, 114-127. 
         53. Chen, H., Huang, J., Palaniappan, A., Wang, Y., Liedberg, B., Platt, M., and Tok, A. L. Y. (2016) A review on electronic bio-sensing approaches based on non-antibody recognition elements. Analyst 141, 2335. 
         54. Walker, D. J. F., Nevin, K. P., Nonnenmann, S. S., Holmes, D. E., Woodard, T. L., Ward, J. E., Rotaru, A.-E., McInerney, M. J., and Lovley, D. R. Syntrophus conductive pili demonstrate that common hydrogen-donating syntrophs can have a direct electron transfer option. bioRxiv 2018. 
         55. Coppi, M. V., Leang, C., Sandler, S. J., and Lovley, D. R. (2001) Development of a genetic system for  Geobacter sulfurreducens . Appl. Environ. Microbiol. 67, 3180-3187. 
         56. Sambrook, J., and Russell, D. W. Molecular Cloning: A Laboratory Manual, 3rd ed.; Cold Spring Harbor: Cold Spring Harbor, N.Y., 2001. 
         57. Aklujkar, M., and Lovley, D. R. (2010) Interference with histidyl-tRNA synthetase by a CRISPR spacer sequence as a factor in the evolution of  Pelobacter carbinolicus . BMC Evol. Biol. 10, 230. 
         58. Ueki, T., DiDonato, L. N., and Lovley, D. R. (2017) Toward establishing minimum requirements for extracellular electron transfer in  Geobacter sulfurreducens . FEMS Microbiol. Lett. 364, fnx093. 
         59. Leang, C., Qian, X., Mester, T., and Lovley, D. R. (2010) Alignment of the c-type cytochrome OmcS along pili of  Geobacter sulfurreducens . Appl. Environ. Microbiol. 76, 4080-4084. 
         60. Nevin, K. P., Kim, B.-C., Glaven, R. H., Johnson, J. P., Woodard, T. L., Methe, B. A., DiDonato, R. J., Jr, Covalla, S. F., Franks, A. E., Liu, A., and Lovley, D. R. (2009) Anode biofilm transcriptomics reveals outer surface components essential for high current power production in  Geobacter sulfurreducens  fuel cells. PLoS One 4, e5628. 
         61. Chen H, Huang J, Palaniappan A, Wang Y, Liedberg B, Platt M et al (2016). A review on electronic bio-sensing approaches based on non-antibody recognition elements. Analyst 141: 2335. 
         62. Gaidamakova E K, Backer M V, Backer J M (2001). Molecular vehicle for target-mediated delivery of therapeutics and diagnostics. J Control Release 74: 341-347. 
         63. Huang J, Nagy S S, Koide A, Rock R S, Koide S (2009). A peptide tag system for facile purification and single-molecule immobilization. Biochemistry 48: 11834-11836. 
         64. Karimzadeh A, Hasanzadeh M, Shadjou N, de la Guardia M (2018). Peptide based biosensors. Trends Anal Chem 107: 1-20. 
         65. Lee S H, Song K B (2009). Isolation of a Calcium-binding Peptide from Enzymatic Hydrolysates of Porcine Blood Plasma Protein. J Korean Soc Appl Biol Chem 52: 290-294. 
         66. Liu Q, Wang J, Boyd BJ (2015). Peptide-based biosensors. Talanta 136: 114-127. 
         67. Lu Y, Huang F, Wang J, Xia J (2014). Affinity-guided covalent conjugation reactions based on PDZ-peptide and SH3-peptide interactions. Bioconjug Chem 25: 989-999. 
         68. Mascini M, Pizzoni D, Perez G, Chiarappa E, Di Natale C, Pittia P et al (2017). Tailoring gas sensor arrays via the design of short peptides sequences as binding elements. Biosens Bioelectron 93: 161-169. 
         69. Pavan S, Berti F (2012). Short peptides as biosensor transducers. Anal Bioanal Chem 402. 
         70. Pizzoni D, Mascini M, Lanzone V, Del Carlo M, Di Natale C, Compagnone D (2014). Selection of peptide ligands for piezoelectric peptide based gas sensors arrays using a virtual screening approach. Biosens Bioelectron 52: 247-254. 
         71. Puiu M, Bala C (2018). Peptide-based biosensors: From self-assembled interfaces to molecular probes in electrochemical assays. Bioelectrochemistry 120: 66-75. 
         72. Seker U O S, Demir H V (2011). Material Binding Peptides for Nanotechnology. Molecules 16: 1426-1451. 
         73. Skerra A, Schmidt T G (2000). Use of the Strep-Tag and streptavidin for detection and purification of recombinant proteins. Methods in enzymology 326: 271-304. 
         74. Tang N, Skibsted L H (2016). Calcium Binding to Amino Acids and Small Glycine Peptides in Aqueous Solution: Toward Peptide Design for Better Calcium Bioavailability. J Agric Food Chem 64: 4376-4389. 
         75. Yan X, Zhou H, Zhang J, Shi C, Xie X, Wu Y et al (2009). Molecular mechanism of inward rectifier potassium channel 2.3 regulation by tax-interacting protein-1. Journal of molecular biology 392: 967-976. 
         76. Zakeri B, Fierer J O, Celik E, Chittock E C, Schwarz-Linek U, Moy V T et al (2012). Peptide tag forming a rapid covalent bond to a protein, through engineering a bacterial adhesin. Proc Natl Acad Sci USA 109: E690-697. 
       
    
     The teachings of all patents, published applications and references cited herein are incorporated by reference in their entirety. 
     While example embodiments have been particularly shown and described, it will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the scope of the embodiments encompassed by the appended claims.