Decontamination of Environmental Water Sources Using Living Engineered Biofilm Materials

Abstract

A living engineered biofilm material comprises microbial cells embedded in a protective extracellular matrix comprising a fusion protein of an amyloid domain and a contaminant binding domain operative to bind a contaminant of a water source, and thereby facilitate decontamination of the water source.

Description

Claims

1. A living engineered biofilm material configured for decontamination of an environmental water source, wherein the biofilm material comprises microbial cells embedded in a protective extracellular matrix comprising a fusion protein expressed by the microbial cells, the fusion protein comprising an amyloid domain and a contaminant binding domain operative to bind a contaminant of the water source, and thereby facilitate decontamination of the water source.

2. The material of claim 1, wherein the contaminant is selected from a microbial (e.g. viral, bacterial, fungal or protozoan) pathogen, organic dye, antibiotic, artificial sweetener, pharmaceutical, perfluorinated compound, and flame retardant.

3. The material of claim 1, wherein the contaminant is a water-transmitted microbial pathogen that is a viral pathogen selected from Adenovirus, Astrovirus, Hepatitis A and E viruses, Rotavirns, Norovirus, Coxsackie viruses, Polioviruses, Polyomaviruses, Cytomegalovirus, Coronaviruses and Influenza viruses, or a bacterial pathogen selected from Aeromonas, Pseudomonas Salmonella, Aeromonas, Shigella, Vibrio spp., Enterobacter and Klebsiella, or a protozoan pathogen selected from Giardia lamblia and Schistosome.

4. The material of claim 1, wherein the contaminant is a harmful chemical in water selected from arsenic, heavy metals, halogenated aromatics, nitrosoamines, nitrates and phosphates.

5. The material of claim 1, wherein the contaminant binding domain and contaminant are selected from: Contaminant binding domainTarget contaminantCHKKPSKSC (SEQ ID NO:1)TiO2 bindingCHRRPSRSC (SEQ ID NO:2)TiO2 bindingAHKKPSKSA (SEQ ID NO:3)TiO2 bindingMHGKTQATSGTIQS (SEQ ID NO:4)Gold bindingWALRRSIRRQSY (SEQ ID NO:5)Gold bindingWAGAKRLVLRRE (SEQ ID NO:6)Gold bindingLKAHLPPSRLPS (SEQ ID NO:7)Gold bindingVSGSSPDS (SEQ ID NO:8)Gold bindingRRTVKHHVN (SEQ ID NO:9)Iron oxideACTARSPWICG (SEQ ID NO:10)Lanthanide oxide and upconversion nanocrystalsMSPHPHPRHHHT (SEQ ID NO:11)silicaSSKKSGSYSGSKGSKRRIL (SEQ ID NO:12)silicaHPPMNASHPHMH (SEQ ID NO:13)silicaRKLPDA (SEQ ID NO:14)silicaCTYSRKHKC (SEQ ID NO:15)Cadmium sulphideLRRSSEAHNSIV (SEQ ID NO:16)Zinc sulphideTSNAVHPTLRHL (SEQ ID NO:17)Palladium bindingPTSTGQA (SEQ ID NO:18)Platinum bindingTLTTLTN (SEQ ID NO:19)Platinum bindingSSFPQPN (SEQ ID NO:20)Platinum bindingCSQSVTSTKSC (SEQ ID NO:21)Platinum bindingAYSSGAPPMPPF (SEQ ID NO:22)silverNPSSLFRYLPSD (SEQ ID NO:23)silverRPRENRGRERGL (SEQ ID NO:24)TitaniumRKLPDA (SEQ ID NO:25)TitaniumPPPWLPYMPPWS (SEQ ID NO:26)QuartzVKTQATSREEPPRLPSKHRPG (SEQ ID NO:27)ZeolitesEAHVMHKVAPRP (SEQ ID NO:28)Zinc oxideEPLQLKM (SEQ ID NO:29)GrapheneHSSYWYAFNNKT (SEQ ID NO:30)Single-walled carbon nanotubesDYFSSPYYEQLF (SEQ ID NO:31)Single-walled carbon nanotubesDSPHTELP (SEQ ID NO:32)Single-walled carbon nanotubes.

6. The material of claim 1, wherein the contaminant is a viral pathogen and the contaminant binding domain and contaminant are selected from: Contaminant binding domain peptideContaminantLRNIRLRNIRLRNIRLRNIR (SEQ ID NO:33)hepatitis B virusIINNPITCMTNGAICWGPCPTAFRQIGNCGHFKVRCCKIR (SEQ ID NO:34)influenza A virus H1N1,IINNPITCMT (SEQ ID NO:35)H3N2,ITCMTNGAIC (SEQ ID NO:36)H5N1,NGAICWGPCP (SEQ ID NO:37)H7N7,WGPCPTAFRQ (SEQ ID NO:38)H7N9,TAFRQIGNCG (SEQ ID NO:39)SARS-CoV and MERS-CoVIGNCGHFKVR (SEQ ID NO:40)HFKVRCCKIR (SEQ ID NO:41)TAFRQIGNCGHFKVRCCKIR (SEQ ID NO:42)NGAICWGPCPTAFRQIGNCGHFKVRCCKIR (SEQ ID NO:43)IINNPITCMTNGAICWGPC (SEQ ID NO:44)IINNPITCMTNGAICWGPCPTAFRQIGNCG (SEQ ID NO:45)NGAICWGPCPTAFRQIGNCGHFKVRCCKIRDED (SEQ ID NO:46)ARLPR (SEQ ID NO:47) (C5)H1N1CIEQSFTTLFACQTAAEIWRAFGYTVKIMVDNGNCRLHVC (SEQ ID NO:48) (C40)H1N1IEEQAKTFLDKFNHEAEDLFYQSSLASWNYNTNIT (SEQ ID NO:49)Sars-Cov-2WLVFFVIFYFFR (SEQ ID NO:50)H1N1WLVFFVIAYFAR (SEQ ID NO:51)WLVFFVIFYFFRRRKK (SEQ ID NO:52)RRKKWLVFFVIFYFFR (SEQ ID NO:53)RRKKIFYFFR (SEQ ID NO:54)WLVFFVRRKK (SEQ ID NO:55)FFVIFYRRKK (SEQ ID NO:56)QMRRKVELFTYMRFD (SEQ ID NO:57)Enterovirus 71NDFRSKT (SEQ ID NO:58)H9N2CNDFRSKTC (SEQ ID NO:59)H9N3GCKKYRRFRWKFKGKFWFWG (SEQ ID NO:60)H7N7GKKYRRFRWKFKGKWFWFG (SEQ ID NO:61)H3N2GFWFKGKWRFKKYRGGRYKKFRWKGKFWFG (SEQ ID NO:63)H1N1SSNKSTTGSGETTTA (SEQ ID NO:63)H1N1.

7. The material of claim 1, wherein the contaminant binding domain is an influenza virus hemagglutinin binding peptide selected from: ARLPR (SEQ ID NO:47) (C5) and CIEQSFTTLFACQTAAEIWRAFGYTVKIMVDNGNCRLHVC (SEQ ID NO:48) (C40).

8. The material of claim 1, wherein the amyloid domain is of TasA (B. subtilis), CsgA (E. coli), PSMs (S. aureus), RmbC (V. cholera), CsgA (Enterobacter cloacae), FapC (Pseudomonas spp.), CsgA (Salmonella spp.) or PAc (Streptococcus mutans).

9. The material of claim 1, wherein the amyloid domain is a CsgA monomer.

10. The material of claim 1, wherein the microbial cells are Bacillus spp. (e.g. B.subtilis), Pseudomonas spp. (e.g. P. aeruginosa), Staphylococcus spp. (e.g. S. aureus), Salmonella ssp. (e.g. S. enterica), Vibrio spp. (e.g. V. cholera), Streptococcus spp. (e.g. Streptococcus mutans), Enterobacter spp. (e.g. Enterobacter cloacae), Lactobacillus spp. (e.g. L. plantarum) or Escherichia spp. (e.g. E. coli).

11. The material of claim 8, wherein the microbial cells are Bacillus spp. (e.g. B.subtilis), Pseudomonas spp. (e.g. P. aeruginosa), Staphylococcus spp. (e.g. S. aureus), Salmonella ssp. (e.g. S. enterica), Vibrio spp. (e.g. V. cholera), Streptococcus spp. (e.g. Streptococcus mutans), Enterobacter spp. (e.g. Enterobacter cloacae), Lactobacillus spp. (e.g. L. plantarum) or Escherichia spp. (e.g. E. coli).

12. The material of claim 9, wherein the microbial cells are Bacillus spp. (e.g. B.subtilis), Pseudomonas spp. (e.g. P. aeruginosa), Staphylococcus spp. (e.g. S. aureus), Salmonella ssp. (e.g. S. enterica), Vibrio spp. (e.g. V. cholera), Streptococcus spp. (e.g. Streptococcus mutans), Enterobacter spp. (e.g. Enterobacter cloacae), Lactobacillus spp. (e.g. L. plantarum) or Escherichia spp. (e.g. E. coli).

13. The material of claim 1 configured in a water decontamination system comprising the living engineered biofilm material, the system further comprising a water source comprising the contaminant.

14. The material of claim 1 configured in a water decontamination system comprising an industrial filler material colonized by the living engineered biofilm material.

15. The material of claim 10 configured in a water decontamination system comprising an industrial filler material colonized by the living engineered biofilm material.

16. The material of claim 11 configured in a water decontamination system comprising an industrial filler material colonized by the living engineered biofilm material.

17. The material of claim 12 configured in a water decontamination system comprising an industrial filler material colonized by the living engineered biofilm material.

18. The material of claim 1 configured in a water decontamination system comprising an industrial filler material colonized by the living engineered biofilm material, the system further comprising a water source comprising the contaminant.

19. A water decontamination system comprising an industrial filler material colonized by the living engineered biofilm material of claim 1.

20. A method of using the living engineered biofilm material of claim 1, comprising the step of contacting a water source comprising the contaminant with the biofilm material under conditions wherein the fusion protein binds the contaminant.