Granted Patent
B2
US 12,320,802 · App. 17/440,632 · Granted Jun 3, 2025
Microbial nanowires modified to contain peptides and methods of making
Assignee:
UNIVERSITY OF MASSACHUSETTS
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Abstract
The present invention provides, in various embodiments, electrically conductive protein nanowires (e-PNs) having surface exposed peptides that confer additional sensing capabilities and/or enhance binding to other materials, as well as fusion proteins and methods for making such nanowires. The present invention also provides sensor devices comprising the nanowires.
Claims (30)
1. An electrically conductive nanowire comprising a fusion protein comprising a type IV pilin monomer and a tag, wherein the tag is at the C-terminus of the type IV pilin monomer, and wherein the nanowire further comprises an untagged type IV pilin monomer.
2. The electrically conductive nanowire of claim 1 , wherein the molar ratio of fusion protein to untagged type IV pilin monomer is in the range of about 10:1 to about 1:10.
3. The electrically conductive nanowire of claim 1 , comprising a plurality of surface exposed peptide tags.
4. The electrically conductive nanowire of claim 1 , wherein the nanowire has a diameter of from about 3 nm to about 4 nm.
5. The electrically conductive nanowire of claim 1 , wherein the nanowire has a length of from about 0.5 μm to about 20 μm.
6. The electrically conductive nanowire of claim 1 , wherein the nanowire has a conductivity of from about 40 μS/cm to about 300 S/cm at a temperature of about 25° C. and a pH of about 7.
7. A nanowire sensor device comprising:
the electrically conductive nanowire of claim 1 ;
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 electrode terminal and the second electrode terminal; and
an electrical current monitor in electrical communication with the electrical resistance, wherein the electrical current monitor is configured to measure an electrical current passing through the electrical resistance.
8. A polynucleotide encoding the electrically conductive nanowire of claim 1 .
9. An expression vector comprising the polynucleotide of claim 8 .
10. A host cell comprising the polynucleotide of claim 8 .
11. A method of producing electrically conductive protein nanowires, comprising the steps of:
a) introducing the polynucleotide of claim 8 into a host cell;
b) placing the host cell in a culture medium conditioned for producing type IV pili;
culturing the host cell for a time sufficient to produce a desired quantity of the c) type IV pili; and
d) isolating the type IV pili from the culture medium,
thereby producing the electrically conductive protein nanowires.
12. The electrically conductive nanowire of claim 1 , wherein the type IV pilin monomer in the fusion protein is a PilA monomer or a variant thereof.
13. The electrically conductive nanowire of claim 1 , wherein the type IV pilin monomer in the fusion protein comprises an amino acid sequence that has at least 90% sequence identity to the wildtype Geobacter sulfurreducens PilA monomer (SEQ ID NO: 2).
14. The electrically conductive nanowire of claim 1 , wherein the type IV pilin monomer in the fusion protein is wildtype Geobacter sulfurreducens PilA monomer (SEQ ID NO: 2).
15. The electrically conductive nanowire of claim 1 , wherein the type IV pilin monomer in the fusion protein is a Geobacter sulfurreducens PilA variant comprising a deletion or a substitution of an aromatic amino acid in the wildtype Geobacter sulfurreducens PilA (SEQ ID NO: 2).
16. The electrically conductive nanowire of claim 1 , wherein the tag comprises a binding motif.
17. The electrically conductive nanowire of claim 1 , wherein the tag comprises two to ten consecutive histidine amino acids.
18. The electrically conductive nanowire of claim 1 , wherein the tag comprises a human influenza hemagglutinin (HA) sequence (SEQ ID NO: 11).
19. The electrically conductive nanowire of claim 1 , wherein the untagged type IV pilin monomer comprises an amino acid sequence that has at least 90% sequence identity to wildtype Geobacter sulfurreducens PilA (SEQ ID NO: 2).
20. The electrically conductive nanowire of claim 1 , wherein the untagged type IV pilin monomer is a Geobacter sulfurreducens PilA variant comprising a deletion or a substitution of an aromatic amino acid in wildtype Geobacter sulfurreducens PilA (SEQ ID NO: 2).
Assignments (1)
ASSIGNMENT OF ASSIGNOR'S INTEREST
Recorded Jul 18, 2022
From: LOVLEY, DEREK R.; UEKI, TOSHIYUKI
To: UNIVERSITY OF MASSACHUSETTS
Reel/Frame 060532/0789 →
Continuity (2)
Provisional Application
62821208
· Mar 20, 2019
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