Cell-directed synthesis of multifunctional nanopatterns and nanomaterials

• US 9,580,480 B2
• Filed: 05/31/2012
• Issued: 02/28/2017
• Est. Priority Date: 05/31/2011
• Status: Active Grant

• Current Assignee: Massachusetts Institute of Technology
• Original Assignee: Massachusetts Institute of Technology
• Inventors: Lu, Timothy Kuan-Ta, Chen, Allen Yuyin
• Primary Examiner(s): DESAI, ANAND U
• Application Number:
  US14/119,319
• Publication Number:
  US 20150118712A1
• Time in Patent Office: 1,734 Days
• Field of Search: None
• US Class Current: 1/1
• CPC Class Codes: B82Y 40/00 Manufacture or treatment of nanostructures
• CPC Class Codes: B82Y 5/00 Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery
• CPC Class Codes: C07K 14/245 Escherichia (G)
• CPC Class Codes: C07K 14/4711 Alzheimer's disease; Amyloid plaque core protein
• CPC Class Codes: C07K 2319/00 Fusion polypeptide
• CPC Class Codes: C07K 2319/20 containing a tag with affinity for a non-protein ligand
• CPC Class Codes: C07K 2319/21 containing a His-tag
• CPC Class Codes: C07K 2319/24 containing a MBP (maltose binding protein)-tag
• CPC Class Codes: C07K 2319/43 containing a FLAG-tag
• CPC Class Codes: C07K 2319/60 containing spectroscopic/fluorescent detection, e.g. green fluorescent protein [GFP]
• CPC Class Codes: C07K 2319/735 containing a domain for self-assembly, e.g. a viral coat protein (includes phage display)

Abstract

Aspects of the invention relate to the engineering of biological nanostructures and materials.

Claims

1. A bacterial cell that is genetically engineered to express at least one modified amyloid fiber that comprises at least one inorganic-binding peptide.

2. The bacterial cell of claim 1, wherein the at least one modified amyloid fiber is bound through the inorganic-binding peptide to at least one metal or metal nanoparticle.

3. The bacterial cell of claim 2, wherein the at least one metal or metal nanoparticle comprises a gold nanoparticle, a silver nanoparticle, zinc sulfate, cadmium sulfate, iron oxide, cobalt oxide, cobalt platinum, iron platinum or a quantum dot.

4. The bacterial cell of claim 1, wherein expression of the at least one modified amyloid fiber is under the control of an inducible promoter.

5. The bacterial cell of claim 1, wherein the bacterial cell is an Escherichia coli (E. coli) cell.

6. The bacterial cell of claim 1, wherein the at least one modified amyloid fiber comprises a curli fiber.

7. The bacterial cell of claim 6, wherein the bacterial cell expresses a csgBAC operon, a csgDEFG operon, or both a csgBAC operon and a csgDEFG operon, and wherein one or more components of the csgBAC operon, the csgDEFG operon, or both the csgBAC operon and the csgDEFG operon is/are genetically engineered.

8. The bacterial cell of claim 7, wherein one or more components of the csgBAC operon, the csgDEFG operon, or both the csgBAC operon and the csgDEFG operon is/are expressed on a plasmid, wherein csgA, csgB, or both csgA and csgB is/are expressed on a plasmid under the control of an inducible promoter, wherein one or more heterologous polypeptide domains are attached to CsgA, and/or wherein csgA, csgB, or both csgA and csgB is/are genetically engineered to express charged residues.

9. The bacterial cell claim 1, wherein the at least one modified amyloid fiber is genetically engineered to promote release of the at least one modified amyloid fiber from the bacterial cell.

10. The bacterial cell of claim 9, wherein exogenous D-amino acids enable detachment of amyloid fibers from the bacterial cell surface and/or wherein the bacterial cell overexpresses a gene encoding for an enzyme that converts L-amino acids to D-amino acids.

11. The bacterial cell of claim 1, wherein the at least one inorganic-binding peptide comprises a metal-binding peptide or a semiconductor-binding peptide.

12. The bacterial cell of claim 11, wherein the at least one inorganic-binding peptide comprises a gold-binding peptide (Au-BP), a ZnS-binding peptide (ZnS-BP), or a CdS-binding peptide (CdS-BP).

13. The bacterial cell of claim 12, wherein the Au-BP comprises the sequence LKAHLPPSRLPS (SEQ ID NO:

1), wherein the ZnS-BP comprises the sequence CNNPMHQNC (SEQ ID NO;

2), or wherein the CdS-BP comprises the sequence SLTPLTTSHLRS (SEQ ID NO;

3).

14. The bacterial cell of claim 6, wherein the curli fiber binds to and nucleates inorganic nanomaterials, comprises tetra-glutamate peptides, and/or displays a conductive cytochrome on the surface of the curli fiber.

15. The bacterial cell of claim 4, wherein the inducible promoter is a visible-light-inducible promoter or comprises a UV-inducible toggle switch.

16. A combination of two or more of the bacterial cells of claim 1, wherein the bacterial cells are spatially patterned.

17. A bacterial cell that is genetically engineered to express at least one modified amyloid fiber that comprises at least one heterologous polypeptide domain, wherein the bacterial cell expresses a csgBAC operon, a csgDEFG operon, or both a csgBAC operon and a csgDEFG operon, and wherein at least one component of the csgBAC operon, the csgDEFG operon or both the csgBAC operon and the csgDEFG is genetically engineered.

18. The bacterial cell of claim 17, wherein the csgBAC operon, the csgDEFG operon, or both the csgBAC operon and the csgDEFG operon is/are expressed on a plasmid.

19. The bacterial cell of claim 17, the csgBAC operon, the csgDEFG operon, or both the csgBAC operon and the csgDEFG operon is/are under the control of an inducible promoter.

20. The bacterial cell of claim 17, wherein at least one of the heterologous polypeptide domains is attached to CsgA.

21. The bacterial cell of claim 17, wherein csgA, csgB, or both csgA and csgB is/are genetically engineered to express charged residues.