Renewable Energy from Livestock Waste Valorization: Amyloid-Based Feather Keratin Fuel Cells

Renewable Energy from Livestock Waste Valorization: Amyloid-Based Feather Keratin Fuel Cells | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Physical Sciences - Article Renewable Energy from Livestock Waste Valorization: Amyloid-Based Feather Keratin Fuel Cells Ali Miserez, Wei Long Soon, Mohammad Peydayesh, Tym de Wild, Felix Donat, and 4 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-2816640/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Increasing carbon emissions have accelerated climate change, resulting in devastating effects that have become tangible on an everyday basis 1 . This is mirrored by a projected increase in global energy demand by ca. 50% in a single generation timespan 2,3 , urging a shift from fossil-fuel derived materials toward greener materials and more sustainable manufacturing processes 4 . Bio-based industrial byproducts, such as food industry side-streams 5 , are attractive alternatives with a strong potential for valorization owing to their large volume, low cost, renewability, biodegradability, and intrinsic material properties 6 . Here we demonstrate the reutilization of industrial chicken feathers waste into proton conductive membranes for fuel cells 7 , protonic transistors 8 , and water-splitting devices 9 . Keratin was isolated from chicken feathers via a fast and economical process, converted into amyloid fibrils upon heat treatment, and further processed into membranes with imparted proton conductivity through a simple oxidative method 10 . The functionality of the membranes is demonstrated by assembling them into a hydrogen fuel cell capable of powering several types of devices using hydrogen and air as fuel. Additionally, these membranes could be used to generate hydrogen by water splitting, and in protonic field-effect transistors as a thin-film modulating protonic conductivity via the electrostatic gating effect. We believe that by converting industrial waste into renewable energy materials at low cost and high scalability, our green manufacturing process can contribute to a fully circular economy with neutral carbon footprint. Physical sciences/Chemistry/Green chemistry/Sustainability Physical sciences/Materials science/Materials for energy and catalysis/Fuel cells Physical sciences/Chemistry/Polymer chemistry/Biopolymers Physical sciences/Materials science/Biomaterials/Bioinspired materials Figures Figure 1 Figure 2 Figure 3 Figure 4 Full Text Global energy demand has been on the continuous rise and is anticipated to further increase by an astonishing 47% in the near future 2 . Current primary energy sources, which are still derived from non-renewable fossil fuels, are unsustainable and detrimental to the environment due to the large emissions of greenhouse gases (GHG) into the atmosphere. Hence, to curb global warming, there is an urgent need for new technologies based on renewable and sustainable energy generation sources. Fuel cells are considered a promising sustainable technology for energy conversion to generate electricity without CO 2 emissions using electrochemical reactions 11 . Among the various types of fuel cells, hydrogen fuel cells are particularly attractive from a decarbonization point-of-view, with efforts focusing on the production of green hydrogen as the next renewable fuel 12 . The heart of a fuel cell lies in the membrane electrode assembly (MEA), which consists of a proton exchange membrane (PEM) that selectively allows only the passage of protons between the electrodes. Perfluorosulfonic acid (PFSA) membranes such as NafionÔ and Aquivion Ò remain the benchmark state-of-the-art PEMs used in fuel cells due to their high proton conductivity, chemical inertness, and mechanical stability. However, the production of Nafion is very expensive due to the use of fluorinated chemicals 13 and pose serious environmental concerns relating to poor biodegradability and environmental toxicity of perfluorinated materials 14 . Moreover, the functionalization of polymers through sulfonation usually involve highly corrosive reagents, reducing their environmental friendliness, 15-17 boosting the search for greener and more economical materials for large-scale applications. Inspired by the ubiquity of proton conduction in the living world 18 , 19 as well as their intrinsic biodegradability, studies have explored biopolymers (proteins and polysaccharides) as potential proton conductive materials 20,21 . The proton conductivity of naturally occurring proteins is generally too low to be useful in practical applications but has been improved with functional group modifications 8,22 . Alternatively, high proton conductivity can be achieved in biopolymers through de novo design of peptides utilizing nanostructure-aided conductivity 23 or metal oxide-peptide composites 24 , but these approaches still face challenges regarding cost and scalability. Chicken feathers are produced at a staggering rate of 40 million tons annually as a byproduct from the poultry industry 5 , but have been underutilized due to their unsuitable nutritional profile. Furthermore, they face disposal challenges due to the generation of toxic sulfur dioxide from incineration. Efforts to valorize chicken feathers include their use as hydrogels 25,26 , biosorbents for heavy metals 27,28 , functional films 29 and fibers 30 . These studies point out that feather keratin may be re-configured into highly functional materials, notably exploiting the structural properties and high cysteine (Cys) content of keratin proteins. However, no attempts have been made to date to revalue keratin into renewable energy devices. In this work, we chose chicken feathers as a starting material for fabricating proton conductive biomaterials. Keratin proteins were extracted from industrial chicken feathers using an environmentally friendly and scalable process and converted into amyloid fibrils that were subsequently processed into free-standing amyloid fibril membranes. Proton conductive properties were imparted through a post-oxidative treatment which converted Cys thiols into sulfonic acid groups, using a benign environmental process with harmless and inexpensive chemical compounds. The performance of the membrane was demonstrated in a fuel cell device capable to transform by electrochemical reaction H 2 and O 2 (the latter directly from air) into electrical power and mechanical work (with H 2 O as sole byproduct). To demonstrate the general potential of this approach, we highlight two additional applications of these materials, as protonic field-effect transistors, as well as in the generation of H 2 via water splitting. Protein extraction and isolation An overview of the production of feather keratin proton conductive membranes is depicted in Fig. 1. Keratin was extracted and isolated from chicken feathers and heat treated to produce keratin amyloid fibrils (Fig. 1a), after which they were mixed with a crosslinker and dopant and cast onto a substrate to produce a free-standing membrane. The membrane was then heat cured followed by an oxidative treatment to produce a modified membrane with imparted proton conductive properties (Fig. 1b), which was then assembled into a fuel cell (Fig. 1c) or other devices utilizing proton conductivity. Among protein-containing industrial waste byproducts, chicken feathers have the highest protein content ( ca. 90%) and are most abundant in Cys amino acid ( ca. 8%, Fig. 2a) 5 . However, the extraction and isolation procedure of feather keratin on a large scale has faced challenges, notably the use of harsh toxic solvents (e.g. sodium sulfide) and reducing agents such as b-mercaptoethanol and dithiothreitol, or long isolation dialysis treatments 31 . Here, we extracted feather keratin using a basic solvent consisting of urea and thioglycolate. Urea acted as a chaotropic solvent disrupting the hydrogen bonds within the compact structure of feathers, while thioglycolate served as a reducing agent to reduce intra- and intermolecular disulfide bonds, resulting in the separation of protein inter-chains and eventually dissolution. The supernatant was then precipitated and washed to obtain a crude keratin isolate. We re-extracted this isolate in ammoniacal solution and precipitated it to obtain a pure keratin isolate, which displayed only a single protein band around 10 kDa as analyzed by electrophoresis (Fig. 2b), demonstrating minimal protein degradation during the entire process. An amino acid analysis of the obtained keratin isolate revealed 7 mol.% of Cys (Supplementary Fig. S1), in agreement with the expected 8 mol.% from earlier reports 32 . Our process produced a feather keratin isolate almost fully soluble in dilute acid, in contrast to keratin isolates obtained from other extraction procedures, which require concentrated acids 33 . Furthermore, the process does not require dialysis, enabling scalability and fast production rates. Protein fibrillization Within the family of keratin proteins, hair and wool keratins belong to a-keratins composed of a-helix intermediate filaments, while keratin from feathers, claws, and beaks belong to b-keratins dominated by b-sheet secondary structures 34,35 . While early x-ray diffraction (XRD) studies on feather barbs suggested the presence of amyloid-like fibrils 36 , the self-assembly of regenerated feather keratin into amyloid fibrils has not yet been reported to date. Sequence analysis of feather keratin from the UniProt database (P02450) using a variety of protein structure prediction algorithms (Supplementary Fig. S2) indicated that feather keratin could be assembled into amyloid fibrils. The structure of feather keratin has been reported to consist of a central region with predominantly hydrophobic residues adopting a b-sheet conformation 32 , with Cys residues primarily located at the N- and C-termini 37 . The highest propensity for b-sheet aggregation was predicted to occur between residues Val 32 –Leu 43 , which lie within this central region and have also been identified as a hotspot for amyloidogenic aggregation. Thus, we postulated that feather keratin could be self-assembled into amyloid fibrils, which was verified by the facile self-assembly of keratin monomers into amyloid fibrils after heat treatment at 90°C under acidic conditions. Using thiazole orange as a molecular probe for amyloid fibrils 38 , the fibrillization process proceeded relatively quickly and reached a plateau after only 2h, exhibiting a faster kinetic than that of animal-derived b-lactoglobulin 39 or plant-derived proteins 40 (Fig. 2c). Solutions of feather keratin amyloid fibrils also exhibited birefringence under cross-polarized light, corroborating the presence of amyloid fibrils (inset of Fig. 2c), which were found to be several micrometers in length after 5h as imaged by Atomic Force Microscopy (AFM) and Transmission Electron Microscopy (TEM) (Fig. 2d and 2i). The nanofibrils exhibited an average height of about 7 nm (Fig. 2e), a semiflexible behavior with comparable contour (Fig. 2f) and persistence (Fig. 2g) lengths in the range, and periodic pitches of 90 nm along the fibril axis (Fig. 2h), as expected for most amyloid fibrils. Membrane fabrication We then mixed feather keratin amyloid fibrils with glyoxal and mercaptosuccinic acid and cast the solution to obtain a clear free-standing flexible membrane (Supplementary Fig. S3a), followed by thermal curing (Supplementary Fig. S3b). As we hypothesized that proton conduction could be imparted into the membrane upon conversion of thiol groups into sulfonic acids, we immersed the membranes in peracetic acid to oxidize thiols and disulfide bonds to sulfonic acids (Supplementary Fig. S3c) 10,41 . The final membrane is denoted as Keratin-M. Oxidation into sulfonic acid was corroborated by both Fourier Transform Infrared Spectroscopy (FTIR) and Raman Spectroscopy, with the appearance of the S=O peak at 1040 cm -1 42 (Fig. 3a) in the FTIR spectrum, and the disappearance of the Raman shift peak at 2550 cm -1 assigned to the thiol 43 (Fig. 3b). This was further supported by XPS measurements, which showed the major peaks of -SO 3 - 2p 3/2 and 2p 1/2 doublets at 168.2 and 169.6 eV, respectively, after deconvolution 44-47 . Peaks of sulfone and sulfate groups were also observed, possibly due to side reactions in the fabrication process (Fig. 3c). As the modification of thiols into sulfonic acids confers a negative charge, the difference in surface potential before and after modification was detected by Kelvin Probe Force Microscopy (KPFM). Before modification, the keratin casting solution deposited onto the Highly Oriented Pyrolytic Graphite (HOPG) substrate exhibited a relative positive potential, which likely arose from positively charged amino acids Arg and Lys. Upon oxidative treatment, the membrane exhibited a relative negative potential that may have resulted from the conversion of neutral thiols into negatively charged sulfonic acids (Fig. 3d and Supplementary Fig. S5). Throughout the fabrication process, the membrane maintained its shape and mechanical integrity while still possessing birefringence arising from amyloid fibrils under cross-polarized light (Supplementary Fig. S3d), suggesting that the amyloid fibrils were preserved without significant alteration. The XRD pattern of the membranes displayed two dominant peaks at 9.3° and 19.6°, corresponding to d -spacings of 4.5 Å and 9.5 Å, respectively, that reflect the signature cross-b structure of amyloid fibrils 48,49 (Fig. 3e). A smooth surface was observed under SEM for keratin-M membranes (Supplementary Fig. S6), whereas at higher magnification a fibrillar network morphology was identified on the surface with pore sizes of 7–20 nm (Fig. 3f). In contrast, neat keratin membranes showed a denser morphology with lower porosity (Supplementary Fig. S7). Membrane properties The modification of neutral thiols into negatively charged sulfonic acids was also indicated by the increase in ion exchange capacity (IEC) from 0.18 meq g -1 to 0.84 meq g -1 after oxidative treatment, while a further increase to 1.56 meq g -1 was achieved with the addition of MSA due to the contribution of additional thiol groups (Table 1). The boost in IEC also resulted in a higher water uptake due to enhanced hydration and charge repulsion from the anionic sulfonic acid groups, which could possibly explain the larger pore size from keratin-M due to higher swelling compared to neat keratin membranes. Keratin-M membranes maintained good barrier properties despite the presence of nanopores, demonstrating almost no permeability to dye molecules such as Rhodamine B (Supplementary Fig. S8). They also exhibited 3-fold less permeability to small molecules such as methanol compared to Nafion (Supplementary Fig. S9), demonstrating their potential in direct methanol fuel cells in which Nafion performs less efficiently due to its high methanol permeability 50 . The membranes were mechanically robust and could be readily manipulated without damaging them. Their thermo-mechanical characterization is reported in the supplementary information (Young’s Modulus and tensile strength in Supplementary Table S1 and Fig. S10; thermal behavior and glass transition in Fig. S11). Supercritical CO 2 -dried keratin membranes measured with TGA displayed a 10% decrease in weight until 100°C attributed to the evaporation of water, followed by thermal degradation and deamidation with drastic weight loss (~ 60%) between 300°C to 400°C 51 (Supplementary Fig. S12). Table 1 . Feather keratin membrane properties before and after treatment Membrane IEC (meq g -1 ) Water Uptake (%) Pre-treated 0.18 ± 0.03 55.3 ± 6.7 Neat keratin 0.84 ± 0.05 131.3 ± 17 Keratin-M 1.56 ± 0.03 215.9 ± 31 Nafion 117 1.10 ± 0.02 17.0 ± 1.2 The main parameter in a fuel cell is the proton conductivity (s), which is essential for any polymer electrolyte membrane. After the successful conversion of thiols into sulfonic acids, feather keratin membranes exhibited an enhanced proton conductivity that increased with the MSA content from 0.02 mS cm -1 to 6.3 mS cm -1 in water (Fig. 4a). The appearance of the semi-circle in the Nyquist plot of neat membranes indicate a more capacitive behavior (Fig. 4b), as also evidenced in the large phase angle from the Bode plot (Supplementary Fig. S13a). This characteristic shifted towards a membrane with a resistor-type behavior for keratin-M membranes, as seen from an almost absent semi-circle in the Nyquist plot (Fig. 4c) and a phase angle approaching 0° at high frequencies in the Bode plot (Supplementary Fig. S13b). Comparing across biomaterials reported to date, the proton conductivity of keratin-M is the highest among all biopolymers, with the exception of specifically engineered peptides (Supplementary Table S2 and S3). But in contrast to the latter, feather keratin membranes are processed from low-value waste materials with a strong potential for scaling-up. Remarkably, because this comes from a waste stream intended for incineration (and CO 2 emissions), this process takes place with an overall negative carbon footprint, adding value to both sustainability and environmental friendliness. Furthermore, the proton conductivity of the membranes could still be further improved by doping with acid electrolytes, notably with sulfuric acid which provided the most significant increment to 22.8 mS cm -1 , and which may find applications in vanadium redox flow batteries. Fuel cell performance Having demonstrated a suitable proton conductivity for the ensued amyloid keratin membranes, we then tested their in-situ performance as a polymer electrolyte membrane in a hydrogen fuel cell (Supplementary Fig. S14). The open circuit voltage (OCV) of a fuel cell has been reported to provide a useful indication of the integrity and degradative behavior of the membrane such as thinning and pinhole formation 52 . The cell assembled with keratin-M membrane displayed an OCV between 0.95 – 1 V –comparable to Nafion – and polarization curves were obtained at increasing temperature. With air at the cathode, the cell generated an increased power density with temperature, reaching up to 20 mW cm -2 at 65°C (Fig. 4d). Similarly, the peak power of 25 mW cm -2 was generated with pure oxygen at 55°C, after which it decreased to 21 mW cm -2 at 65°C (Fig. 4e). This could be due to an increase in reactive oxygen species generated with increased oxygen concentration, leading to the formation of free radicals such as OH×, H×, and HOO× that have been reported to initiate membrane degradation 53,54 . Further increase in temperature to 80°C resulted in a drop in OCV to 0.85 – 0.9 V along with a decreased power density lower than that at 65°C. To ensure a high OCV and optimal performance, a membrane of low hydrogen permeability is desired. Utilizing staircase voltammetry with N 2 at the cathode (Supplementary Fig. S16), the hydrogen permeability of the keratin membrane was assessed and calculated using the intercept, yielding 3.8 ± 0.3 ´ 10 -10 mol cm -2 s -1 , (compare to Nafion - 3.8 ´ 10 -9 mol cm -2 s -1 ) 55 while the electrical resistance obtained from the slope was approximately 3200 ± 140 Ohm cm 2 (Supplementary Fig. S17) 56,57 . To demonstrate the applicability of the keratin-M membrane in hydrogen fuel cells, we assembled the membrane into a commercial test fuel cell setup. With hydrogen and air as the respective fuels at the anode and cathode, the cell was able to generate power to turn on both red and white LED lamps (Supplementary Fig. S18). In addition, the cell was responsive to the presence of fuel, turning the LED lamp on and off with the introduction and absence of hydrogen (Supplementary Movie S1). Most importantly, the fuel cell could perform mechanical work, e.g. drive a fan setup driven by a motor (Supplementary Movie S2) as well as a fuel cell toy car ((Supplementary Movie S4 and Fig. 4f). Furthermore, the keratin-M PEM membranes could be further used as an electrolyzer for the production of hydrogen and oxygen from water when applying an electric bias, as observed by the formation of bubbles at the respective outlets (Supplementary Movie S4 & Supplementary Fig. S19). Transistor performance Finally, we demonstrate the application of keratin-M as a solid-state film for the fabrication of protonic field-effect transistors (H + -FETs). Devices were fabricated by casting keratin-M film between two gold/palladium hydride (PdH x ) electrodes pre-deposited using e-beam evaporation process on a 50 nm hafnium oxide (HfO 2 ) gate dielectric layer (Fig. 4g). The conductivity of the keratin-M film was observed to be dependent on relative humidity (RH), displaying a surge in current when the RH increased from 50% to 90% (Fig. 4h and Supplementary Fig. S20), as a result of the increase in proton conductivity with increased hydration, which is commonly observed in most proton conductive materials 20,22,58 . The transfer characteristics of the FET device (Fig. 4i) measured for negative and positive gate voltages ( V GS ) at a source to drain voltage ( V SD ) of 0.5 V exhibited a ON-to-OFF ratio of ~ 1.4 with a low leakage current (Supplementary Fig. 21). The output characteristics of the device displayed a similar trend whereby the channel conductivity increased upon application of negative V GS (Supplementary Fig. 23 and 24) . Biasing the device with negative V GS induced positive charges into the channel proportional to V GS as the keratin-M and Si gate form a capacitor with HfO 2 as the dielectric. This resulted in an increased positive charge carrier concentration and thereby channel conductivity. The electrostatic gating effect highlights the potential use of keratin-M in H + -FETs. Conclusions We have demonstrated the fabrication of proton conductive membranes isolated from chicken feathers keratin through a green and low-cost process with high scalability. Amyloid-based keratin membranes were successfully used in a hydrogen fuel cell to power several devices with only hydrogen and air as the fuel, and also demonstrated pronounced electrostatic gating effect in a transistor and hydrogen generation in an electrolyzer. While the ensued proton conductive membranes do not yet fully match the performance (particularly at high temperature) of commercial benchmarks such as NafionÔ and Aquivion Ò , the membranes pose a radically different environmental and sustainability footprint and at a potentially much lower cost: our current estimates on the total production from raw material to membrane give ca . USD 850/m 2 . This value is at least half that of NafionÔ (USD 1500 – 3000/m 2 ) and is anticipated to be much lower when scaled up and optimized, hence highlighting another long sought-after feature of these original materials. Our study unambiguously demonstrates that livestock industrial waste can be reprocessed into valuable materials for renewable energy devices using a fully sustainable process, and brings us one step closer towards a circular economy concept based on zero -or even negative- carbon emissions. Materials and Methods Full details on Materials and Methods are provided in the Supporting Sections A-D. Hereby a summary of the main methods used is provided. Protein extraction and isolation to membrane fabrication Protein extraction and isolation from chicken feathers obtained from a local farm are detailed in Supplementary Section AI. Feather keratin amyloid fibrils were prepared by heating keratin isolates (Supplementary Section AII) and fabricated into membranes as described in Supplementary Section AIII. Protein and membrane characterization are provided in Supplementary Section B and C, respectively. Electrochemical measurements of membranes are described in Supplementary Section D. Fuel cell measurements For the in-situ fuel cell measurements, membranes were placed inside a 1 cm 2 cell between two platinum-coated commercial GDE’s (JM ELE0244, Johnson Matthey, United Kingdom) with a nominal loading of 0.4 mg Pt/cm 2 . Fuel cell tests were performed using a custom fuel cell test bench with a constant flow rate of 0.4 ln/min for all gasses. Polarization curves were recorded at 100% RH and a pressure of 1.5 bar. Hydrogen crossover currents were measured using staircase voltammetry at 100% RH and a pressure of 1 bar. Additional details are provided in Supplementary Section IV. To demonstrate powering of the devices, membranes were placed between two platinum-coated carbon paper electrodes of 0.5 mg/cm 2 (Fuel Cell Store, USA) to form the membrane electrode assembly in the Flex-Stak Electrochemical cell (Fuel Cell Store, USA). Pure H 2 and Air were supplied at the anode and cathode, respectively. Metal tabs were connected to a mini DC/DC (LiPower, Sparkfun) converter with an output of 3.3 V. Transistor fabrication Field-effect transistor devices (channel length of 100 mm and width of 1 mm) were fabricated by depositing gold/palladium metal electrodes using electron-beam evaporation on top of hafnium oxide (50 nm) coated silicon substrate. Palladium electrodes were exposed to pure hydrogen gas for 5 mins in a closed chamber for their transformation to PdH x . 0.5 mL of amyloid fibril mixed with glyoxal and MSA were deposited onto the device and air-dried. The substrate was then cured at 150°C for 50 min and immersed in peracetic acid solution at 37°C for 5 h. The device was then immersed in 0.5 M sulfuric acid for 1 h and then rinsed thoroughly with water. H + -FET Characterization Output and transfer characteristic of the transistor devices were recorded using the Keysight B2901A precision source measure unit. Relative humidity level was kept at 95% in a custom-built dry box chamber using a humidifier (BioAire Lifestyle). Declarations Acknowledgements This research was supported by the strategic initiative on biomimetic and sustainable materials (IBSM), Nanyang Technological University (NTU), Singapore. W.L.S thanks NTU Singapore for the Nanyang President Graduate Scholarship (NPGS). The authors acknowledge the support of Dr. Anne Greet Bittermann and Stephan Handschin of ScopeM/ETH Zurich. The authors thank Prof. Nripan Matthews and Dr. Timothy Ng for the use of e-beam deposition system and Prof. Cesare Soci group for the use of source measurement unit for characterization of FETs. Conflict of Interest R.M., W.L.S., and A.M. are the inventors of a patent application jointly filed by NTU and ETHZ. Author Contributions W.L.S., R.M and A.M. designed the study. W.L.S. carried out the experiments from protein extraction and isolation to membrane fabrication and characterization. T.D.W and L.G. carried out the in-situ fuel cell measurements. F.D. carried out the TGA and XRD measurements. 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Pure keratin membrane and fibers from chicken feather. International Journal of Biological Macromolecules 89 , 614-621 (2016). https://doi.org:https://doi.org/10.1016/j.ijbiomac.2016.04.039 Shavandi, A., Silva, T. H., Bekhit, A. A. & Bekhit, A. E.-D. A. Keratin: dissolution, extraction and biomedical application. Biomaterials Science 5 , 1699-1735 (2017). https://doi.org:10.1039/C7BM00411G Arai, K. M., Takahashi, R., Yokote, Y. & Akahane, K. Amino-Acid Sequence of Feather Keratin from Fowl. European Journal of Biochemistry 132 , 501-507 (1983). https://doi.org:https://doi.org/10.1111/j.1432-1033.1983.tb07389.x Aluigi, A. et al. Study on the structure and properties of wool keratin regenerated from formic acid. International Journal of Biological Macromolecules 41 , 266-273 (2007). https://doi.org:https://doi.org/10.1016/j.ijbiomac.2007.03.002 Fraser, R. D. B. & Parry, D. A. D. The role of β-sheets in the structure and assembly of keratins. 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Journal of Power Sources 184 , 338-343 (2008). https://doi.org:https://doi.org/10.1016/j.jpowsour.2008.06.013 Dandurand, J. et al. Conformational and thermal characterization of a synthetic peptidic fragment inspired from human tropoelastin: Signature of the amyloid fibers. Pathologie Biologie 62 , 100-107 (2014). https://doi.org:https://doi.org/10.1016/j.patbio.2014.02.001 Vilekar, S. A. & Datta, R. The effect of hydrogen crossover on open-circuit voltage in polymer electrolyte membrane fuel cells. Journal of Power Sources 195 , 2241-2247 (2010). https://doi.org:https://doi.org/10.1016/j.jpowsour.2009.10.023 Ramaswamy, N., Hakim, N. & Mukerjee, S. Degradation mechanism study of perfluorinated proton exchange membrane under fuel cell operating conditions. Electrochimica Acta 53 , 3279-3295 (2008). https://doi.org:https://doi.org/10.1016/j.electacta.2007.11.010 Gubler, L., Dockheer, S. M. & Koppenol, W. H. Radical (HO•, H• and HOO•) Formation and Ionomer Degradation in Polymer Electrolyte Fuel Cells. Journal of The Electrochemical Society 158 , B755 (2011). https://doi.org:10.1149/1.3581040 Francia, C., Ijeri, V. S., Specchia, S. & Spinelli, P. Estimation of hydrogen crossover through Nafion® membranes in PEMFCs. Journal of Power Sources 196 , 1833-1839 (2011). https://doi.org:https://doi.org/10.1016/j.jpowsour.2010.09.058 Li, S., Wei, X., Dai, H., Yuan, H. & Ming, P. Voltammetric and galvanostatic methods for measuring hydrogen crossover in fuel cell. iScience 25 , 103576 (2022). https://doi.org:10.1016/j.isci.2021.103576 Schoemaker, M., Misz, U., Beckhaus, P. & Heinzel, A. Evaluation of Hydrogen Crossover through Fuel Cell Membranes. Fuel Cells 14 , 412-415 (2014). https://doi.org:https://doi.org/10.1002/fuce.201300215 Kim, Y., Ketpang, K., Jaritphun, S., Park, J. S. & Shanmugam, S. A polyoxometalate coupled graphene oxide–Nafion composite membrane for fuel cells operating at low relative humidity. Journal of Materials Chemistry A 3 , 8148-8155 (2015). https://doi.org:10.1039/C5TA00182J Additional Declarations Yes there is potential Competing Interest. R.M., W.L.S., and A.M. are the inventors of a patent application jointly filed by NTU and ETHZ. Supplementary Files SupplementaryMovie1.mp4 Supplementary Movie 1 SupplementaryMovie2.mp4 Supplementary Movie 2 SupplementaryMovie3.mp4 Supplementary Movie 3 SupplementaryMovie4.mp4 Supplementary Movie 1 SoonetalSIfinal.docx Supplementary Information Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-2816640","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Physical Sciences - Article","associatedPublications":[],"authors":[{"id":198164855,"identity":"f7ad8d92-5e54-49af-97b2-bd995d332e80","order_by":0,"name":"Ali 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Institut","correspondingAuthor":false,"prefix":"","firstName":"Lorenz","middleName":"","lastName":"Gubler","suffix":""},{"id":198164863,"identity":"b10e8a99-b775-49f1-94fc-db913a9b3a48","order_by":8,"name":"Raffaele Mezzenga","email":"","orcid":"https://orcid.org/0000-0002-5739-2610","institution":"ETH Zurich","correspondingAuthor":false,"prefix":"","firstName":"Raffaele","middleName":"","lastName":"Mezzenga","suffix":""}],"badges":[],"createdAt":"2023-04-14 10:26:40","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-2816640/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-2816640/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":38417051,"identity":"a41a0515-2e3f-4e97-bf87-220ad1007ef1","added_by":"auto","created_at":"2023-06-12 17:06:41","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":410652,"visible":true,"origin":"","legend":"\u003cp\u003eGeneral schematic of the valorization of chicken feather keratin for fuel cell applications. \u003cstrong\u003ea\u003c/strong\u003e, Extraction, isolation, and amyloid fibril formation of feather keratin. \u003cstrong\u003eb\u003c/strong\u003e, Membrane fabrication through solvent casting, thermal curing, and post treatment. \u003cstrong\u003ec\u003c/strong\u003e, Application of keratin membrane into a fuel cell assembly.\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-2816640/v1/5af0987e59dc250ec869b637.png"},{"id":38417052,"identity":"49432ab6-b386-4f02-bc51-e198ad1401d2","added_by":"auto","created_at":"2023-06-12 17:06:41","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":358469,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003ea\u003c/strong\u003e, Comparison of Cys content in several major proteins from industrial food waste. \u003cstrong\u003eb\u003c/strong\u003e, SDS-PAGE of isolated feather keratin with and without reducing agent, respectively. \u003cstrong\u003ec\u003c/strong\u003e, Fibrillization behavior of feather keratin amyloid fibrils with time, as assessed with thiazole orange. Inset: amyloid fibril solution under cross-polarized light. \u003cstrong\u003ed\u003c/strong\u003e, AFM image of feather keratin amyloid fibrils. \u003cstrong\u003ee-ih\u003c/strong\u003e, AFM statistical analysis of feather keratin amyloid fibrils showing average fibril height (e); contour length (f); persistence length L\u003csub\u003ep\u003c/sub\u003e (g) obtained through the 2D mean square end-to-end displacement (MSED) method; and fibril pitch (h). \u003cstrong\u003ei\u003c/strong\u003e, TEM images of feather keratin amyloid fibrils.\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-2816640/v1/eaabd74eaf0c85ee49c1ccab.png"},{"id":38417053,"identity":"de2ce6a0-73a2-4db0-9373-795f5673ce3f","added_by":"auto","created_at":"2023-06-12 17:06:41","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":195371,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003ea\u003c/strong\u003e, FTIR spectra of keratin membranes. \u003cstrong\u003eb\u003c/strong\u003e, Raman spectra of keratin membranes. \u003cstrong\u003ec\u003c/strong\u003e, XPS spectrum of the keratin-M membrane. \u003cstrong\u003ed\u003c/strong\u003e, Surface potential of the keratin-M membrane before and after treatment using KPFM. \u003cstrong\u003ee\u003c/strong\u003e, XRD patterns of keratin membranes. \u003cstrong\u003ef\u003c/strong\u003e, SEM image of the keratin-M membrane.\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-2816640/v1/ea6232a15501578b2d0d17f4.png"},{"id":38417054,"identity":"2aa84247-36db-4743-bc74-aaa202bbfdfa","added_by":"auto","created_at":"2023-06-12 17:06:41","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":292401,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003ea\u003c/strong\u003e, Proton conductivity of keratin membranes with increasing MSA content. Nyquist plot of \u003cstrong\u003eb\u003c/strong\u003e, neat keratin membrane and \u003cstrong\u003ec,\u003c/strong\u003e keratin-M membrane. In-situ performance of keratin-M membrane at different temperatures using hydrogen at the anode and \u003cstrong\u003ed\u003c/strong\u003e, Air and \u003cstrong\u003ee\u003c/strong\u003e, O\u003csub\u003e2\u003c/sub\u003e at the cathode. \u003cstrong\u003ef\u003c/strong\u003e, Keratin-M membranes assembled into a commercial fuel cell to power a commercial fuel cell toy car. \u003cstrong\u003eg\u003c/strong\u003e, Schematic illustration of protonic field-effect transistor using keratin-M membrane. \u003cstrong\u003eh\u003c/strong\u003e, I-V plot of keratin-M on PdH\u003csub\u003ex\u003c/sub\u003e electrodes with increasing relative humidity (I\u003csub\u003eSD\u003c/sub\u003e in log scale). \u003cstrong\u003ei,\u003c/strong\u003e Transfer characteristic of keratin-M bioprotonic FET device at 90% RH and V\u003csub\u003eSD\u003c/sub\u003e\u0026nbsp;=\u0026nbsp;0.5\u0026nbsp;V on PdH\u003csub\u003ex\u003c/sub\u003e electrodes.\u003c/p\u003e","description":"","filename":"4.png","url":"https://assets-eu.researchsquare.com/files/rs-2816640/v1/71748eb5e50d71e0930e6ee0.png"},{"id":38417312,"identity":"a83398b5-8d83-4aa8-90b5-60ef1e6762bc","added_by":"auto","created_at":"2023-06-12 17:14:47","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1500175,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-2816640/v1/a7bf4907-316f-4484-bfc1-b3ba4e0b0d12.pdf"},{"id":38417311,"identity":"721cd12c-b24c-4403-9a36-5b807373cc00","added_by":"auto","created_at":"2023-06-12 17:14:41","extension":"mp4","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":36518527,"visible":true,"origin":"","legend":"Supplementary Movie 1","description":"","filename":"SupplementaryMovie1.mp4","url":"https://assets-eu.researchsquare.com/files/rs-2816640/v1/d418112ef88c162e7e34f11e.mp4"},{"id":38417055,"identity":"90f29195-2541-4990-9ae5-44184e05607c","added_by":"auto","created_at":"2023-06-12 17:06:41","extension":"mp4","order_by":2,"title":"","display":"","copyAsset":false,"role":"supplement","size":9443035,"visible":true,"origin":"","legend":"\u003cp\u003eSupplementary Movie 2\u003c/p\u003e","description":"","filename":"SupplementaryMovie2.mp4","url":"https://assets-eu.researchsquare.com/files/rs-2816640/v1/f3a7369ebac682a1118f12be.mp4"},{"id":38417057,"identity":"96426dd6-82f4-43fc-b552-e99abc70046c","added_by":"auto","created_at":"2023-06-12 17:06:41","extension":"mp4","order_by":3,"title":"","display":"","copyAsset":false,"role":"supplement","size":28220122,"visible":true,"origin":"","legend":"\u003cp\u003eSupplementary Movie 3\u003c/p\u003e","description":"","filename":"SupplementaryMovie3.mp4","url":"https://assets-eu.researchsquare.com/files/rs-2816640/v1/82202e9c6b47e9b76bc4c748.mp4"},{"id":38417059,"identity":"5160e4e2-47b5-4bd3-b24f-d73497954f25","added_by":"auto","created_at":"2023-06-12 17:06:41","extension":"mp4","order_by":4,"title":"","display":"","copyAsset":false,"role":"supplement","size":7106063,"visible":true,"origin":"","legend":"\u003cp\u003eSupplementary Movie 1\u003c/p\u003e","description":"","filename":"SupplementaryMovie4.mp4","url":"https://assets-eu.researchsquare.com/files/rs-2816640/v1/249a505e06252b644c0ce8ea.mp4"},{"id":38417056,"identity":"275d1b13-972f-4af0-9bfa-ea54550900de","added_by":"auto","created_at":"2023-06-12 17:06:41","extension":"docx","order_by":5,"title":"","display":"","copyAsset":false,"role":"supplement","size":8652125,"visible":true,"origin":"","legend":"\u003cp\u003eSupplementary Information\u003c/p\u003e","description":"","filename":"SoonetalSIfinal.docx","url":"https://assets-eu.researchsquare.com/files/rs-2816640/v1/18de3627f8142409538515aa.docx"}],"financialInterests":"\u003cb\u003eYes\u003c/b\u003e there is potential Competing Interest.\nR.M., W.L.S., and A.M. are the inventors of a patent application jointly filed by NTU and ETHZ.","formattedTitle":"Renewable Energy from Livestock Waste Valorization:\r\nAmyloid-Based Feather Keratin Fuel Cells","fulltext":[{"header":"Full Text","content":"\u003cp\u003eGlobal energy demand has been on the continuous rise and is anticipated to further increase by an astonishing 47% in the near future\u003csup\u003e2\u003c/sup\u003e. Current primary energy sources, which are still derived from non-renewable fossil fuels, are unsustainable and detrimental to the environment due to the large emissions of greenhouse gases (GHG) into the atmosphere. Hence, to curb global warming, there is an urgent need for new technologies based on renewable and sustainable energy generation sources.\u003c/p\u003e\n\u003cp\u003eFuel cells are considered a promising sustainable technology for energy conversion to generate electricity without CO\u003csub\u003e2\u003c/sub\u003e emissions using electrochemical reactions\u003csup\u003e11\u003c/sup\u003e. Among the various types of fuel cells, hydrogen fuel cells are particularly attractive from a decarbonization point-of-view, with efforts focusing on the production of green hydrogen as the next renewable fuel\u003csup\u003e12\u003c/sup\u003e. The heart of a fuel cell lies in the membrane electrode assembly (MEA), which consists of a proton exchange membrane (PEM) that selectively allows only the passage of protons between the electrodes. Perfluorosulfonic acid (PFSA) membranes such as Nafion\u0026Ocirc;\u0026nbsp;and Aquivion\u003csup\u003e\u0026Ograve;\u003c/sup\u003e remain the benchmark state-of-the-art PEMs used in fuel cells due to their high proton conductivity, chemical inertness, and mechanical stability. However, the production of Nafion is very expensive due to the use of fluorinated chemicals\u003csup\u003e13\u003c/sup\u003e and pose serious environmental concerns relating to poor biodegradability and environmental toxicity of perfluorinated materials\u003csup\u003e14\u003c/sup\u003e. Moreover, the functionalization of polymers through sulfonation usually involve highly corrosive reagents, reducing their environmental friendliness,\u003csup\u003e15-17\u003c/sup\u003e boosting the search for greener and more economical materials for large-scale applications.\u003c/p\u003e\n\u003cp\u003eInspired by the ubiquity of proton conduction in the living world\u003csup\u003e18\u003c/sup\u003e\u003csup\u003e,\u003c/sup\u003e\u003csup\u003e19\u003c/sup\u003e as well as their intrinsic biodegradability, studies have explored biopolymers (proteins and polysaccharides) as potential proton conductive materials\u003csup\u003e20,21\u003c/sup\u003e. The proton conductivity of naturally occurring proteins is generally too low to be useful in practical applications but has been improved with functional group modifications\u003csup\u003e8,22\u003c/sup\u003e. Alternatively, high proton conductivity can be achieved in biopolymers through \u003cem\u003ede novo\u003c/em\u003e design of peptides utilizing nanostructure-aided conductivity\u003csup\u003e23\u003c/sup\u003e or metal oxide-peptide composites\u0026nbsp;\u003csup\u003e24\u003c/sup\u003e, but these approaches still face challenges regarding cost and scalability.\u003c/p\u003e\n\u003cp\u003eChicken feathers are produced at a staggering rate of 40 million tons annually as a byproduct from the poultry industry\u003csup\u003e5\u003c/sup\u003e, but have been underutilized due to their unsuitable nutritional profile. Furthermore, they face disposal challenges due to the generation of toxic sulfur dioxide from incineration. Efforts to valorize chicken feathers include their use as hydrogels\u003csup\u003e25,26\u003c/sup\u003e, biosorbents for heavy metals\u003csup\u003e27,28\u003c/sup\u003e, functional films\u003csup\u003e29\u003c/sup\u003e and fibers\u003csup\u003e30\u003c/sup\u003e. These studies point out that feather keratin may be re-configured into highly functional materials, notably exploiting the structural properties and high cysteine (Cys) content of keratin proteins. However, no attempts have been made to date to revalue keratin into renewable energy devices. In this work, we chose chicken feathers as a starting material for fabricating proton conductive biomaterials. Keratin proteins were extracted from industrial chicken feathers using an environmentally friendly and scalable process and converted into amyloid fibrils that were subsequently processed into free-standing amyloid fibril membranes. Proton conductive properties were imparted through a post-oxidative treatment which converted Cys thiols into sulfonic acid groups, using a benign environmental process with harmless and inexpensive chemical compounds. The performance of the membrane was demonstrated in a fuel cell device capable to transform by electrochemical reaction H\u003csub\u003e2\u003c/sub\u003e and O\u003csub\u003e2\u003c/sub\u003e (the latter directly from air) into electrical power and mechanical work (with H\u003csub\u003e2\u003c/sub\u003eO as sole byproduct). To demonstrate the general potential of this approach, we highlight two additional applications of these materials, as protonic field-effect transistors, as well as in the generation of H\u003csub\u003e2\u003c/sub\u003e via water splitting.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eProtein extraction and isolation\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAn overview of the production of feather keratin proton conductive membranes is depicted in Fig. 1. Keratin was extracted and isolated from chicken feathers and heat treated to produce keratin amyloid fibrils (Fig. 1a), after which they were mixed with a crosslinker and dopant and cast onto a substrate to produce a free-standing membrane. The membrane was then heat cured followed by an oxidative treatment to produce a modified membrane with imparted proton conductive properties (Fig. 1b), which was then assembled into a fuel cell (Fig. 1c) or other devices utilizing proton conductivity.\u003c/p\u003e\n\u003cp\u003eAmong protein-containing industrial waste byproducts, chicken feathers have the highest protein content (\u003cem\u003eca.\u003c/em\u003e 90%) and are most abundant in Cys amino acid (\u003cem\u003eca.\u003c/em\u003e 8%, Fig. 2a)\u003csup\u003e5\u003c/sup\u003e. However, the extraction and isolation procedure of feather keratin on a large scale has faced challenges, notably the use of harsh toxic solvents (e.g. sodium sulfide) and reducing agents such as\u0026nbsp;b-mercaptoethanol and dithiothreitol, or long isolation dialysis treatments\u003csup\u003e31\u003c/sup\u003e. Here, we extracted feather keratin using a basic solvent consisting of urea and thioglycolate. Urea acted as a chaotropic solvent disrupting the hydrogen bonds within the compact structure of feathers, while thioglycolate served as a reducing agent to reduce intra- and intermolecular disulfide bonds, resulting in the separation of protein inter-chains and eventually dissolution. The supernatant was then precipitated and washed to obtain a crude keratin isolate. We re-extracted this isolate in ammoniacal solution and precipitated it to obtain a pure keratin isolate, which displayed only a single protein band around 10\u0026nbsp;kDa as analyzed by electrophoresis (Fig. 2b), demonstrating minimal protein degradation during the entire process. An amino acid analysis of the obtained keratin isolate revealed 7\u0026nbsp;mol.% of Cys (Supplementary Fig. S1), in agreement with the expected 8\u0026nbsp;mol.% from earlier reports\u003csup\u003e32\u003c/sup\u003e. Our process produced a feather keratin isolate almost fully soluble in dilute acid, in contrast to keratin isolates obtained from other extraction procedures, which require concentrated acids\u003csup\u003e33\u003c/sup\u003e. Furthermore, the process does not require dialysis, enabling scalability and fast production rates.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eProtein fibrillization\u003c/strong\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eWithin the family of keratin proteins, hair and wool keratins belong to\u0026nbsp;a-keratins composed of\u0026nbsp;a-helix intermediate filaments, while keratin from feathers, claws, and beaks belong to\u0026nbsp;b-keratins dominated by\u0026nbsp;b-sheet secondary structures\u003csup\u003e34,35\u003c/sup\u003e. While early x-ray diffraction (XRD) studies on feather barbs suggested the presence of amyloid-like fibrils\u003csup\u003e36\u003c/sup\u003e, the self-assembly of regenerated feather keratin into amyloid fibrils has not yet been reported to date. Sequence analysis of feather keratin from the UniProt database (P02450) using a variety of protein structure prediction algorithms (Supplementary Fig. S2) indicated that feather keratin could be assembled into amyloid fibrils. The structure of feather keratin has been reported to consist of a central region with predominantly hydrophobic residues adopting a\u0026nbsp;b-sheet conformation\u003csup\u003e32\u003c/sup\u003e, with Cys residues primarily located at the N- and C-termini\u003csup\u003e37\u003c/sup\u003e. The highest propensity for\u0026nbsp;b-sheet aggregation was predicted to occur between residues Val\u003csup\u003e32\u003c/sup\u003e\u0026ndash;Leu\u003csup\u003e43\u003c/sup\u003e, which lie within this central region and have also been identified as a hotspot for amyloidogenic aggregation. Thus, we postulated that feather keratin could be self-assembled into amyloid fibrils, which was verified by the facile self-assembly of keratin monomers into amyloid fibrils after heat treatment at 90\u0026deg;C under acidic conditions. Using thiazole orange as a molecular probe for amyloid fibrils\u003csup\u003e38\u003c/sup\u003e, the fibrillization process proceeded relatively quickly and reached a plateau after only 2h, exhibiting a faster kinetic than that of animal-derived\u0026nbsp;b-lactoglobulin\u003csup\u003e39\u003c/sup\u003e or plant-derived proteins\u003csup\u003e40\u003c/sup\u003e (Fig. 2c). Solutions of feather keratin amyloid fibrils also exhibited birefringence under cross-polarized light, corroborating the presence of amyloid fibrils (inset of Fig. 2c), which were found to be several micrometers in length after 5h as imaged by Atomic Force Microscopy (AFM) and Transmission Electron Microscopy (TEM) (Fig. 2d and 2i). The nanofibrils exhibited an average height of about 7 nm (Fig. 2e), a semiflexible behavior with comparable contour (Fig. 2f) and persistence (Fig. 2g) lengths in the\u0026nbsp;\u0026nbsp;\u0026nbsp;range, and periodic pitches of 90 nm along the fibril axis (Fig. 2h), as expected for most amyloid fibrils.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMembrane fabrication\u003c/strong\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eWe then mixed feather keratin amyloid fibrils with glyoxal and mercaptosuccinic acid and cast the solution to obtain a clear free-standing flexible membrane (Supplementary Fig. S3a), followed by thermal curing (Supplementary Fig. S3b). As we hypothesized that proton conduction could be imparted into the membrane upon conversion of thiol groups into sulfonic acids, we immersed the membranes in peracetic acid to oxidize thiols and disulfide bonds to sulfonic acids (Supplementary Fig. S3c)\u003csup\u003e10,41\u003c/sup\u003e. The final membrane is denoted as Keratin-M.\u003c/p\u003e\n\u003cp\u003eOxidation into sulfonic acid was corroborated by both Fourier Transform Infrared Spectroscopy (FTIR) and Raman Spectroscopy, with the appearance of the S=O peak at 1040\u0026nbsp;cm\u003csup\u003e-1\u003c/sup\u003e \u003csup\u003e42\u003c/sup\u003e (Fig. 3a) in the FTIR spectrum, and the disappearance of the Raman shift peak at 2550 cm\u003csup\u003e-1\u003c/sup\u003e assigned to the thiol\u003csup\u003e43\u003c/sup\u003e (Fig. 3b). This was further supported by XPS measurements, which showed the major peaks of -SO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e-\u003c/sup\u003e 2p\u003csub\u003e3/2\u003c/sub\u003e and 2p\u003csub\u003e1/2\u003c/sub\u003e doublets at 168.2 and 169.6 eV, respectively, after deconvolution\u003csup\u003e44-47\u003c/sup\u003e. Peaks of sulfone and sulfate groups were also observed, possibly due to side reactions in the fabrication process (Fig. 3c). As the modification of thiols into sulfonic acids confers a negative charge, the difference in surface potential before and after modification was detected by Kelvin Probe Force Microscopy\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e(KPFM). Before modification, the keratin casting solution deposited onto the Highly Oriented Pyrolytic Graphite (HOPG) substrate exhibited a relative positive potential, which likely arose from positively charged amino acids Arg and Lys. Upon oxidative treatment, the membrane exhibited a relative negative potential that may have resulted from the conversion of neutral thiols into negatively charged sulfonic acids (Fig. 3d and Supplementary Fig. S5).\u003c/p\u003e\n\u003cp\u003eThroughout the fabrication process, the membrane maintained its shape and mechanical integrity while still possessing birefringence arising from amyloid fibrils under cross-polarized light (Supplementary Fig. S3d), suggesting that the amyloid fibrils were preserved without significant alteration. The XRD pattern of the membranes displayed two dominant peaks at 9.3\u0026deg; and 19.6\u0026deg;, corresponding to \u003cem\u003ed\u003c/em\u003e-spacings of 4.5\u0026nbsp;\u0026Aring;\u0026nbsp;and 9.5\u0026nbsp;\u0026Aring;, respectively, that reflect the signature cross-b\u0026nbsp;structure of amyloid fibrils\u003csup\u003e48,49\u003c/sup\u003e (Fig. 3e). A smooth surface was observed under SEM for keratin-M membranes (Supplementary Fig. S6), whereas at higher magnification a fibrillar network morphology was identified on the surface with pore sizes of 7\u0026ndash;20 nm (Fig. 3f). In contrast, neat keratin membranes showed a denser morphology with lower porosity (Supplementary Fig. S7).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMembrane properties\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe modification of neutral thiols into negatively charged sulfonic acids was also indicated by the increase in ion exchange capacity (IEC) from 0.18\u0026nbsp;meq g\u003csup\u003e-1\u003c/sup\u003e to 0.84 meq g\u003csup\u003e-1\u0026nbsp;\u003c/sup\u003eafter oxidative treatment, while a further increase to 1.56\u0026nbsp;meq g\u003csup\u003e-1\u003c/sup\u003e was achieved with the addition of MSA due to the contribution of additional thiol groups (Table 1). The boost in IEC also resulted in a higher water uptake due to enhanced hydration and charge repulsion from the anionic sulfonic acid groups, which could possibly explain the larger pore size from keratin-M due to higher swelling compared to neat keratin membranes. Keratin-M membranes maintained good barrier properties despite the presence of nanopores, demonstrating almost no permeability to dye molecules such as Rhodamine B (Supplementary Fig. S8). They also exhibited 3-fold less permeability to small molecules such as methanol compared to Nafion (Supplementary Fig. S9), demonstrating their potential in direct methanol fuel cells in which Nafion performs less efficiently due to its high methanol permeability\u003csup\u003e50\u003c/sup\u003e. The membranes were mechanically robust and could be readily manipulated without damaging them. Their thermo-mechanical characterization is reported in the supplementary information (Young\u0026rsquo;s Modulus and tensile strength in Supplementary Table S1 and Fig. S10; thermal behavior and glass transition in Fig. S11). Supercritical CO\u003csub\u003e2\u003c/sub\u003e-dried keratin membranes measured with TGA displayed a 10% decrease in weight until 100\u0026deg;C attributed to the evaporation of water, followed by thermal degradation and deamidation with drastic weight loss (~ 60%) between 300\u0026deg;C to 400\u0026deg;C\u003csup\u003e51\u003c/sup\u003e (Supplementary Fig. S12).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003e1\u003c/strong\u003e. Feather keratin membrane properties before and after treatment\u003c/p\u003e\n\u003cdiv align=\"Left\"\u003e\n \u003ctable border=\"0\" cellspacing=\"0\" cellpadding=\"0\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd width=\"33.333333333333336%\"\u003e\n \u003cp\u003e\u003cstrong\u003eMembrane\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"33.333333333333336%\"\u003e\n \u003cp\u003e\u003cstrong\u003eIEC (meq g\u003csup\u003e-1\u003c/sup\u003e)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"33.333333333333336%\"\u003e\n \u003cp\u003e\u003cstrong\u003eWater Uptake (%)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"33.333333333333336%\"\u003e\n \u003cp\u003ePre-treated\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"33.333333333333336%\"\u003e\n \u003cp\u003e0.18\u0026nbsp;\u0026plusmn;\u0026nbsp;0.03\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"33.333333333333336%\"\u003e\n \u003cp\u003e55.3\u0026nbsp;\u0026plusmn; 6.7\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"33.333333333333336%\"\u003e\n \u003cp\u003eNeat keratin\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"33.333333333333336%\"\u003e\n \u003cp\u003e0.84\u0026nbsp;\u0026plusmn; 0.05\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"33.333333333333336%\"\u003e\n \u003cp\u003e131.3\u0026nbsp;\u0026plusmn; 17\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"33.333333333333336%\"\u003e\n \u003cp\u003eKeratin-M\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"33.333333333333336%\"\u003e\n \u003cp\u003e1.56\u0026nbsp;\u0026plusmn; 0.03\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"33.333333333333336%\"\u003e\n \u003cp\u003e215.9\u0026nbsp;\u0026plusmn; 31\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"33.333333333333336%\"\u003e\n \u003cp\u003eNafion 117\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"33.333333333333336%\"\u003e\n \u003cp\u003e1.10\u0026nbsp;\u0026plusmn; 0.02\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"33.333333333333336%\"\u003e\n \u003cp\u003e17.0\u0026nbsp;\u0026plusmn; 1.2\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n\u003c/div\u003e\n\u003cp\u003eThe main parameter in a fuel cell is the proton conductivity (s), which is essential for any polymer electrolyte membrane. After the successful conversion of thiols into sulfonic acids, feather keratin membranes exhibited an enhanced proton conductivity that increased with the MSA content from 0.02 mS cm\u003csup\u003e-1\u003c/sup\u003e to 6.3 mS cm\u003csup\u003e-1\u003c/sup\u003e in water (Fig. 4a). The appearance of the semi-circle in the Nyquist plot of neat membranes indicate a more capacitive behavior (Fig. 4b), as also evidenced in the large phase angle from the Bode plot (Supplementary Fig. S13a). This characteristic shifted towards a membrane with a resistor-type behavior for keratin-M membranes, as seen from an almost absent semi-circle in the Nyquist plot (Fig. 4c) and a phase angle approaching 0\u0026deg; at high frequencies in the Bode plot (Supplementary Fig. S13b). Comparing across biomaterials reported to date, the proton conductivity of keratin-M is the highest among all biopolymers, with the exception of specifically engineered peptides (Supplementary Table S2 and S3). But in contrast to the latter, feather keratin membranes are processed from low-value waste materials with a strong potential for scaling-up. Remarkably, because this comes from a waste stream intended for incineration (and CO\u003csub\u003e2\u003c/sub\u003e emissions), this process takes place with an overall negative carbon footprint, adding value to both sustainability and environmental friendliness. Furthermore, the proton conductivity of the membranes could still be further improved by doping with acid electrolytes, notably with sulfuric acid which provided the most significant increment to 22.8 mS cm\u003csup\u003e-1\u003c/sup\u003e, and which may find applications in vanadium redox flow batteries.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFuel cell performance\u003c/strong\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eHaving demonstrated a suitable proton conductivity for the ensued amyloid keratin membranes, we then tested their \u003cem\u003ein-situ\u003c/em\u003e performance as a polymer electrolyte membrane in a hydrogen fuel cell (Supplementary Fig. S14). The open circuit voltage (OCV) of a fuel cell has been reported to provide a useful indication of the integrity and degradative behavior of the membrane such as thinning and pinhole formation\u003csup\u003e52\u003c/sup\u003e. The cell assembled with keratin-M membrane displayed an OCV between 0.95\u0026nbsp;\u0026ndash;\u0026nbsp;1\u0026nbsp;V \u0026ndash;comparable to Nafion \u0026ndash; and polarization curves were obtained at increasing temperature. With air at the cathode, the cell generated an increased power density with temperature, reaching up to 20\u0026nbsp;mW cm\u003csup\u003e-2\u003c/sup\u003e at 65\u0026deg;C (Fig. 4d). Similarly, the peak power of 25 mW cm\u003csup\u003e-2\u003c/sup\u003e was generated with pure oxygen at 55\u0026deg;C, after which it decreased to 21 mW cm\u003csup\u003e-2\u003c/sup\u003e at 65\u0026deg;C (Fig. 4e). This could be due to an increase in reactive oxygen species generated with increased oxygen concentration, leading to the formation of free radicals such as OH\u0026times;, H\u0026times;, and HOO\u0026times; that have been reported to initiate membrane degradation\u003csup\u003e53,54\u003c/sup\u003e. Further increase in temperature to 80\u0026deg;C resulted in a drop in OCV to 0.85\u0026nbsp;\u0026ndash;\u0026nbsp;0.9\u0026nbsp;V along with a decreased power density lower than that at 65\u0026deg;C. To ensure a high OCV and optimal performance, a membrane of low hydrogen permeability is desired. Utilizing staircase voltammetry with N\u003csub\u003e2\u003c/sub\u003e at the cathode (Supplementary Fig. S16), the hydrogen permeability of the keratin membrane was assessed and calculated using the intercept, yielding 3.8 \u0026plusmn; \u0026nbsp;0.3 \u0026acute; 10\u003csup\u003e-10\u003c/sup\u003e mol cm\u003csup\u003e-2\u003c/sup\u003e s\u003csup\u003e-1\u003c/sup\u003e, (compare to Nafion - 3.8\u0026nbsp;\u0026acute;\u0026nbsp;10\u003csup\u003e-9\u003c/sup\u003e mol cm\u003csup\u003e-2\u003c/sup\u003e s\u003csup\u003e-1\u003c/sup\u003e)\u003csup\u003e55\u003c/sup\u003e while the electrical resistance obtained from the slope was approximately 3200 \u0026plusmn; 140 Ohm cm\u003csup\u003e2\u003c/sup\u003e (Supplementary Fig. S17)\u003csup\u003e56,57\u003c/sup\u003e.\u003c/p\u003e\n\u003cp\u003eTo demonstrate the applicability of the keratin-M membrane in hydrogen fuel cells, we assembled the membrane into a commercial test fuel cell setup. With hydrogen and air as the respective fuels at the anode and cathode, the cell was able to generate power to turn on both red and white LED lamps (Supplementary Fig. S18). In addition, the cell was responsive to the presence of fuel, turning the LED lamp on and off with the introduction and absence of hydrogen\u003csub\u003e\u0026nbsp;\u003c/sub\u003e(Supplementary Movie S1). Most importantly, the fuel cell could perform mechanical work, \u003cem\u003ee.g.\u003c/em\u003e drive a fan setup driven by a motor (Supplementary Movie S2) as well as a fuel cell toy car ((Supplementary Movie S4 and Fig. 4f). Furthermore, the keratin-M PEM membranes could be further used as an electrolyzer for the production of hydrogen and oxygen from water when applying an electric bias, as observed by the formation of bubbles at the respective outlets (Supplementary Movie S4 \u0026amp; Supplementary Fig. S19).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTransistor performance\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eFinally,\u0026nbsp;we demonstrate the application of\u0026nbsp;keratin-M as a solid-state film\u0026nbsp;for the fabrication of protonic field-effect transistors (H\u003csup\u003e+\u003c/sup\u003e-FETs).\u0026nbsp;Devices were\u0026nbsp;fabricated by casting\u0026nbsp;keratin-M\u0026nbsp;film between two gold/palladium hydride (PdH\u003csub\u003ex\u003c/sub\u003e) electrodes pre-deposited using e-beam evaporation process on a 50\u0026nbsp;nm hafnium oxide (HfO\u003csub\u003e2\u003c/sub\u003e) gate dielectric layer (Fig. 4g).\u0026nbsp;The conductivity of the keratin-M film was observed to be dependent on relative humidity (RH), displaying a surge in current when the RH increased from 50% to 90% (Fig. 4h and Supplementary Fig. S20), as a result of the increase in proton conductivity with increased hydration, which is commonly observed in most proton conductive materials\u003csup\u003e20,22,58\u003c/sup\u003e.\u0026nbsp;The transfer characteristics of the FET device\u0026nbsp;(Fig. 4i)\u0026nbsp;measured for negative and positive gate voltages (\u003cem\u003eV\u003c/em\u003e\u003csub\u003eGS\u003c/sub\u003e) at a source to drain voltage (\u003cem\u003eV\u003c/em\u003e\u003csub\u003eSD\u003c/sub\u003e) of 0.5 V exhibited a ON-to-OFF ratio of ~ 1.4 with a low leakage current (Supplementary Fig. 21). The output characteristics of the device displayed a similar trend whereby the channel conductivity increased upon application of negative \u003cem\u003eV\u003c/em\u003e\u003csub\u003eGS\u003c/sub\u003e (Supplementary Fig. 23 and 24)\u003csub\u003e.\u0026nbsp;\u003c/sub\u003eBiasing the device with negative V\u003csub\u003eGS\u003c/sub\u003e induced positive charges into the channel proportional to \u003cem\u003eV\u003c/em\u003e\u003csub\u003eGS\u003c/sub\u003e as the keratin-M and Si gate form a capacitor with HfO\u003csub\u003e2\u003c/sub\u003e as the dielectric. This resulted in an increased positive charge carrier concentration and thereby channel conductivity. The electrostatic gating effect highlights the potential use of keratin-M in H\u003csup\u003e+\u003c/sup\u003e-FETs.\u003c/p\u003e"},{"header":"Conclusions","content":"\u003cp\u003eWe have demonstrated the fabrication of proton conductive membranes isolated from chicken feathers keratin through a green and low-cost process with high scalability. Amyloid-based keratin membranes were successfully used in a hydrogen fuel cell to power several devices with only hydrogen and air as the fuel, and also demonstrated pronounced electrostatic gating effect in a transistor and hydrogen generation in an electrolyzer. While the ensued proton conductive membranes do not yet fully match the performance (particularly at high temperature) of commercial benchmarks such as Nafion\u0026Ocirc;\u0026nbsp;and Aquivion\u003csup\u003e\u0026Ograve;\u003c/sup\u003e, the membranes pose a radically different environmental and sustainability footprint and at a potentially much lower cost: our current estimates on the total production from raw material to membrane give \u003cem\u003eca\u003c/em\u003e. USD 850/m\u003csup\u003e2\u003c/sup\u003e. This value is at least half that of Nafion\u0026Ocirc;\u0026nbsp;(USD 1500\u0026nbsp;\u0026ndash;\u0026nbsp;3000/m\u003csup\u003e2\u003c/sup\u003e) and is anticipated to be much lower when scaled up and optimized, hence highlighting another long sought-after feature of these original materials. Our study unambiguously demonstrates that livestock industrial waste can be reprocessed into valuable materials for renewable energy devices using a fully sustainable process, and brings us one step closer towards a circular economy concept based on zero -or even negative- carbon emissions.\u003c/p\u003e"},{"header":"Materials and Methods","content":"\u003cp\u003eFull details on Materials and Methods are provided in the Supporting Sections A-D. Hereby a summary of the main methods used is provided.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eProtein extraction and isolation to membrane fabrication\u003c/strong\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eProtein extraction and isolation from chicken feathers obtained from a local farm are detailed in Supplementary Section AI. Feather keratin amyloid fibrils were prepared by heating keratin isolates (Supplementary Section AII) and fabricated into membranes as described in Supplementary Section AIII. Protein and membrane characterization are provided in Supplementary Section B and C, respectively.\u0026nbsp;Electrochemical measurements of membranes are described in Supplementary Section D.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFuel cell measurements\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eFor the in-situ fuel cell measurements, membranes were placed inside a 1 cm\u003csup\u003e2\u003c/sup\u003e cell between two platinum-coated commercial GDE\u0026rsquo;s (JM ELE0244, Johnson Matthey, United Kingdom) with a nominal loading of 0.4 mg Pt/cm\u003csup\u003e2\u003c/sup\u003e. Fuel cell tests were performed using a custom fuel cell test bench with a constant flow rate of 0.4 ln/min for all gasses. Polarization curves were recorded at 100% RH and a pressure of 1.5 bar. Hydrogen crossover currents were measured using staircase voltammetry at 100% RH and a pressure of 1 bar. Additional details are provided in Supplementary Section IV.\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003eTo demonstrate powering of the devices, membranes were placed between two platinum-coated carbon paper electrodes of 0.5 mg/cm\u003csup\u003e2\u003c/sup\u003e (Fuel Cell Store, USA) to form the membrane electrode assembly in the Flex-Stak Electrochemical cell (Fuel Cell Store, USA). Pure H\u003csub\u003e2\u003c/sub\u003e and Air were supplied at the anode and cathode, respectively. Metal tabs were connected to a mini DC/DC (LiPower, Sparkfun) converter with an output of 3.3 V.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTransistor fabrication\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eField-effect transistor devices (channel length of 100\u0026nbsp;mm and width of 1 mm) were fabricated by depositing gold/palladium metal electrodes\u0026nbsp;using electron-beam evaporation\u0026nbsp;on top of hafnium oxide (50 nm) coated silicon substrate. Palladium electrodes were exposed to pure hydrogen gas for 5 mins in a closed chamber for their transformation to PdH\u003csub\u003ex\u003c/sub\u003e.\u0026nbsp;0.5\u0026nbsp;mL of amyloid fibril\u0026nbsp;mixed with glyoxal and MSA were deposited onto the device\u0026nbsp;and\u0026nbsp;air-dried. The substrate was then cured at 150\u0026deg;C\u0026nbsp;for 50 min and immersed in peracetic acid solution at 37\u0026deg;C\u0026nbsp;for 5 h. The device was then immersed in 0.5 M sulfuric acid for 1 h and then rinsed thoroughly with water.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eH\u003csup\u003e+\u003c/sup\u003e-FET Characterization\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eOutput and transfer characteristic of the transistor devices were recorded using the Keysight B2901A precision source measure unit. Relative humidity level was kept at 95% in a custom-built dry box chamber using a humidifier (BioAire Lifestyle).\u0026nbsp;\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eAcknowledgements\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis research was supported by the strategic initiative on biomimetic and sustainable materials (IBSM), Nanyang Technological University (NTU), Singapore. W.L.S thanks NTU Singapore for the Nanyang President Graduate Scholarship (NPGS). The authors acknowledge the support of Dr. Anne Greet Bittermann and Stephan Handschin of ScopeM/ETH Zurich. The authors thank Prof. Nripan Matthews and Dr. Timothy Ng for the use of e-beam deposition system and Prof. Cesare Soci group for the use of source measurement unit for characterization of FETs.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConflict of Interest\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eR.M., W.L.S., and A.M. are the inventors of a patent application jointly filed by NTU and ETHZ.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthor Contributions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eW.L.S., R.M and A.M. designed the study. W.L.S. carried out the experiments from protein extraction and isolation to membrane fabrication and characterization. T.D.W and L.G. carried out the in-situ fuel cell measurements. F.D. carried out the TGA and XRD measurements. W.L.S. and F.D. carried out the commercial fuel cell device demonstration. R.S. carried out the transistor performance measurements. A.M., R.M., and M.P. supervised the study. W.L.S., A.M., and R.M. wrote the manuscript. All the authors edited and approved the final manuscript.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eData Availability\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declare that the data supporting the findings of this study are available within the paper and its Supplementary Information files. Should any raw data files be needed in another format they are available from the corresponding author upon reasonable request.\u003c/p\u003e\n"},{"header":"References","content":"\u003col\u003e\n \u003cli\u003eRockstr\u0026ouml;m, J.\u003cem\u003e\u0026nbsp;et al.\u003c/em\u003e A safe operating space for humanity. \u003cem\u003eNature\u003c/em\u003e \u003cstrong\u003e461\u003c/strong\u003e, 472-475 (2009). https://doi.org:10.1038/461472a\u003c/li\u003e\n \u003cli\u003eMeghan, G. \u0026amp; Weber, M. Global energy demand to grow 47% by 2050, with oil still top source: US EIA. 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High strength nanostructured films based on well-preserved \u0026beta;-chitin nanofibrils. \u003cem\u003eNanoscale\u003c/em\u003e \u003cstrong\u003e11\u003c/strong\u003e, 11001-11011 (2019). https://doi.org:10.1039/C9NR02870F\u003c/li\u003e\n \u003cli\u003eKi, C. S., Kim, J. W., Oh, H. J., Lee, K. H. \u0026amp; Park, Y. H. The effect of residual silk sericin on the structure and mechanical property of regenerated silk filament. \u003cem\u003eInternational Journal of Biological Macromolecules\u003c/em\u003e \u003cstrong\u003e41\u003c/strong\u003e, 346-353 (2007). https://doi.org:https://doi.org/10.1016/j.ijbiomac.2007.05.005\u003c/li\u003e\n \u003cli\u003eHuang, Q. 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Voltammetric and galvanostatic methods for measuring hydrogen crossover in fuel cell. \u003cem\u003eiScience\u003c/em\u003e \u003cstrong\u003e25\u003c/strong\u003e, 103576 (2022). https://doi.org:10.1016/j.isci.2021.103576\u003c/li\u003e\n \u003cli\u003eSchoemaker, M., Misz, U., Beckhaus, P. \u0026amp; Heinzel, A. Evaluation of Hydrogen Crossover through Fuel Cell Membranes. \u003cem\u003eFuel Cells\u003c/em\u003e \u003cstrong\u003e14\u003c/strong\u003e, 412-415 (2014). https://doi.org:https://doi.org/10.1002/fuce.201300215\u003c/li\u003e\n \u003cli\u003eKim, Y., Ketpang, K., Jaritphun, S., Park, J. S. \u0026amp; Shanmugam, S. A polyoxometalate coupled graphene oxide\u0026ndash;Nafion composite membrane for fuel cells operating at low relative humidity. \u003cem\u003eJournal of Materials Chemistry A\u003c/em\u003e \u003cstrong\u003e3\u003c/strong\u003e, 8148-8155 (2015). https://doi.org:10.1039/C5TA00182J\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":true,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"","lastPublishedDoi":"10.21203/rs.3.rs-2816640/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-2816640/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eIncreasing carbon emissions have accelerated climate change, resulting in devastating effects that have become tangible on an everyday basis\u003csup\u003e1\u003c/sup\u003e. This is mirrored by a projected increase in global energy demand by \u003cem\u003eca.\u003c/em\u003e 50% in a single generation timespan\u003csup\u003e2,3\u003c/sup\u003e, urging a shift from fossil-fuel derived materials toward greener materials and more sustainable manufacturing processes\u003csup\u003e4\u003c/sup\u003e. Bio-based industrial byproducts, such as food industry side-streams\u003csup\u003e5\u003c/sup\u003e, are attractive alternatives with a strong potential for valorization owing to their large volume, low cost, renewability, biodegradability, and intrinsic material properties\u003csup\u003e6\u003c/sup\u003e. Here we demonstrate the reutilization of industrial chicken feathers waste into proton conductive membranes for fuel cells\u003csup\u003e7\u003c/sup\u003e, protonic transistors\u003csup\u003e8\u003c/sup\u003e, and water-splitting devices\u003csup\u003e9\u003c/sup\u003e. Keratin was isolated from chicken feathers via a fast and economical process, converted into amyloid fibrils upon heat treatment, and further processed into membranes with imparted proton conductivity through a simple oxidative method\u003csup\u003e10\u003c/sup\u003e. The functionality of the membranes is demonstrated by assembling them into a hydrogen fuel cell capable of powering several types of devices using hydrogen and air as fuel. Additionally, these membranes could be used to generate hydrogen by water splitting, and in protonic field-effect transistors as a thin-film modulating protonic conductivity via the electrostatic gating effect. We believe that by converting industrial waste into renewable energy materials at low cost and high scalability, our green manufacturing process can contribute to a fully circular economy with neutral carbon footprint.\u003c/p\u003e","manuscriptTitle":"Renewable Energy from Livestock Waste Valorization:\nAmyloid-Based Feather Keratin Fuel Cells","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2023-06-12 17:06:36","doi":"10.21203/rs.3.rs-2816640/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"2186f5d5-04e3-4595-a85d-701eee0e5f67","owner":[],"postedDate":"June 12th, 2023","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[{"id":21257524,"name":"Physical sciences/Chemistry/Green chemistry/Sustainability"},{"id":21257525,"name":"Physical sciences/Materials science/Materials for energy and catalysis/Fuel cells"},{"id":21257526,"name":"Physical sciences/Chemistry/Polymer chemistry/Biopolymers"},{"id":21257527,"name":"Physical sciences/Materials science/Biomaterials/Bioinspired materials"}],"tags":[],"updatedAt":"2023-06-12T17:06:36+00:00","versionOfRecord":[],"versionCreatedAt":"2023-06-12 17:06:36","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-2816640","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-2816640","identity":"rs-2816640","version":["v1"]},"buildId":"_2-kVJe1T_tPrBINL-cwx","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}