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Development of Nanoporous Gold Based Bioelectrodes

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Development of Nanoporous Gold Based Bioelectrodes Development of nanoporous gold based bioelectrodes Thesis presented for the award of Doctor of Philosophy (Ph.D.) by Xinxin Xiao Under the supervision of Professor Edmond Magner Submitted to the Faculty of Science and Engineering University of Limerick, Ireland August 2018 ii Submitted: August 2018 Declaration I declare that this thesis is entirely my own work carried at the University of Limerick and has not been previously submitted to this or any other university. Xinxin Xiao Abstract Dealloyed nanoprous gold (NPG) is a porous material that possesses three dimensional frameworks of bicontinuous pores and ligaments, fabricated by electro/- chemical dissolution of the less noble component from an Au alloy. In this thesis, bioelectrodes were prepared by immobilising enzymes onto NPG and characterised in detail. For redox enzymes, osmium polymers were used to host the enzyme and as electron transfer mediators. Enzymatic biofuel cells (EBFCs) were assembled with redox enzyme modified bioelectrodes and characterised for a number of applications. A glucose/O2 EBFC device has been developed that can harvest electricity in nonaqueous solvents, which may inspire new applications of EBFCs in bioelectrosynthesis. The EBFC was comprised of a NPG/[Os(4,4′-dimethyl-2,2′- +/2+ bipyridine)2(polyvinyl -imidazole)10Cl] (Os(dmbpy)2PVI)/glucose oxidase (GOx) +/2+ bioanode and a NPG/[Os(2,2′-bipyridine)2(polyvinylimidazole)10Cl] (Os(bpy)2PVI)/bilirubin oxidase (BOx) biocathode. The power output of the cell decreased with increasing solvent hydrophobicity in the alcohols examined and the response of each electrode was restored when the electrodes were placed in phosphate buffer solution after operation in organic solutions. To further expand the range of EBFC, a proof-of-concept “self-powered pulse generator” based on a supercapacitor/EBFC hybrid device has been developed. The device was prepared by immobilising redox enzymes with electrodeposited poly(3,4- ethylenedioxythiophene) (PEDOT) and Os(bpy)2PVI on NPG. Once charged by the internal EBFC, the device can be discharged as a supercapacitor at a current density of 2 mA cm-2 providing a maximum power density of 608.8 μW cm-2, an increase of a factor of 468 when compared to the power output from the EBFC itself. To address the constrained oxygen supply that occurs at the biocathode, an oxygen-independent and membrane-less glucose biobattery was prepared by replacing the BOx based biocathode, with a solid-state NPG/MnO2 cathode. The potential of the ii discharged MnO2 could be recovered, enabling the development of a proof-of-concept biobattery/supercapacitor hybrid device. The resulting device exhibited a stable performance for 50 cycles of self-recovery and galvanostatic discharge as a supercapacitor at 0.1 mA cm-2. Wearable EBFCs are emerging as potential power sources for wearable micro- electronic devices. A key requirement of such cells is the need for flexible electrodes. Mechanically stable and flexible NPG electrodes were prepared using an electrochemical dealloying method consisting of a pre-anodization process and a subsequent electrochemical cleaning step. A flexible lactate/O2 EBFC consisting of a lactate oxidase based bioanode using electrodeposited Os(bpy)2PVI, and a BOx biocathode was placed between two commercially available contact lenses to avoid direct contact with the eye. When tested in air-equilibrated artificial tear solutions (3 mM lactate), a maximum power density of 1.7±0.1 μW cm-2 and an open-circuit voltage of 380±28 mV was obtained, values slightly lower than in phosphate buffer solution (2.4±0.2 μW cm-2 and 455±21 mV, respectively). The decrease was mainly attributed to interference from ascorbate. After 5.5 h of operation, the EBFC retained 20% of its initial power output. Finally, the utilization of NPG in fluidic biocatalysis was investigated. An electrochemically triggered sol-gel process was used to generate a thin silica layer for the immobilisation of lipase onto dealloyed NPG. The catalytic response of the entrapped lipase was examined using the hydrolysis of 4-nitrophenyl butyrate (4- NPB) as a model reaction. A deposition time of 180 s and a lipase concentration of 3 mg/mL was used to prepare the optimised electrode. The operational stability of the silica immobilised enzyme was enhanced on NPG in comparison to that on planar gold, which may arise from confinement of the enzyme in the porous structure. The modified electrodes were incorporated into a 3D printed flow cell with conversion efficiencies of up to 100% after 8 cycles. iii ACKNOWLEDGEMENTS I would like to take this opportunity to thank my supervisor Prof. Edmond Magner for the great support on my IRC scholarship application and bring me to study abroad in Ireland. I am grateful for Edmond’s mentoring, encouragement, inspiration and never-ending patience on correcting my writings throughout the past four years. Most importantly, Edmond offers me the maximum freedom to conduct my research, making me an independent researcher. I am very grateful to be an Associated Fellow of the BIOENERGY Marie Curie ITN programme coordinated by Prof. Wolfgang Schuhmann at Ruhr University Bochum. I benefit from the useful seminars, workshop, knowledge transfer and discussion with all the PIs and fellows. Among them, Prof. Dónal Leech and Dr. Peter Ó Conghail at National University of Ireland Galway for providing Os polymers and Dr. Roland Ludwig from BOKU-University of Natural Resources and Life Sciences for supplying glucose dehydrogenase are greatly appreciated. I would like to thank my collaborators and friends for their kind support and hosting for my short-term visit: Assoc. Prof. Pengchao Si and Prof. Lijie Ci at Shandong University, Prof. Jens Ulstrup, Prof. Jingdong Zhang and Assoc. Prof. Qijin Chi at Technical University of Denmark, Prof. Yi Ding at Tianjin University of Technology, Prof. Aihua Liu at Qingdao University. My sincere thanks also goes to instrument scientists at Bernal Institute: Dr. Serguei Belochapkine for performing sputtering of Au/Ag alloy films, Dr. Yina Guo for training me to use TEM and SEM, Dr. Lekshmi Kailas for AFM and FTIR training, Dr. Fathima Laffir for Raman spectroscope training, Dr. Wynette Redington for TGA training. Internal collaborators, Dr Micheál D. Scanlon for nice discussion, Dr. Tadhg Kennedy for Autolab test and Robert Whelan for 3D printing of flow cells, are also acknowledged. iv Many thanks to members of “Edmond’s research warriors” for their significant help: Alessandro Serleti, Cristina Carucci, Fernando Otero Diez, Dr. Marcela Salazar Alvarez, Dr. Noreldeen Abdallah, Simon White, Dr. Till Siepenkoetter, Dr. Urszula Salaj-Kosla, Dr. Victoria Gascón Pérez. I would also like to thank my friends from the “Chinese Community” in Limerick for the homely type company: Dr. Feng Chen, Dr. Fengwei Sun, Haiyang Zhang, Huan Ren, Dr. Lei Shi, Dr. Xiaoming Ju, Dr. Yina Guo. Finally, I express my profound gratitude to my parents for their unfailing love and moral support. I thank Ms. Yan Li, who was my girlfriend when I just came to Ireland and is my wife now, for her long-distance accompany and continuous encouragement. Long, long had been my road and far, far was the journey; I would go up and down to seek my heart's desire. -Authored by Qu Yuan (c. 340-278 BC) -Translated by David Hawkes v Publications and conference presentations Publications 1. Xinxin Xiao, Peter Ó Conghaile, Dónal Leech, Edmond Magner; Use of polymer coatings to enhance the response of redox-polymer-mediated electrodes, ChemElectroChem 2018, DOI: 10.1002/celc.201800983 2. Xinxin Xiao, Edmond Magner; A quasi-solid-state and self-powered biosupercapacitor based on flexible nanoporous gold electrodes, Chemical Communications 2018, 54, 5823-5826. 3. Xinxin Xiao, Till Siepenkoetter, Peter Ó Conghaile, Dónal Leech, Edmond Magner; Nanoporous gold based biofuel cells on contact lenses, ACS Applied Materials & Interfaces 2018, 2018, 10 (8), 7107–7116 4. Ciaran Lyons, Pratibha Dev, Pathik Maji, Neelima Rathi, Praveen K. Surolia, Owen Byrne, Xinxin Xiao, Niall J. English, Edmond Magner, J. M. D. MacElroy, K. Ravindranathan Thampi; Silicon-bridged triphenylamine-based organic dyes for efficient dye-sensitised solar cells, Solar Energy 2018, 160, 64-75 5. Francesca Lopez, Till Siepenkoetter, Xinxin Xiao, Edmond Magner, Wolfgang Schuhmann, Urszula Salaj-Kosla; Potential pulse-assisted immobilization of Myrothecium verrucaria bilirubin oxidase at planar and nanoporous gold electrodes, Journal of Electroanalytical Chemistry 2018, 812, 194-198 6. Xinxin Xiao, Till Siepenkoetter, Robert Whelan, Urszula Salaj-Kosla, Edmond Magner; A continuous fluidic bioreactor utilising electrodeposited silica for lipase immobilisation onto nanoporous gold, Journal of Electroanalytical Chemistry 2018, 812, 180-185 7. Xinxin Xiao, Peter Ó Conghaile, Dónal Leech, Roland Ludwig, Edmond Magner; An oxygen-independent and membrane-less glucose vi biobattery/supercapacitor hybrid device, Biosensors and Bioelectronics 2017, 98, 421-427 8. Xinxin Xiao, Christian Engelbrekt, Minwei Zhang, Zheshen Li, Jens Ulstrup, Jingdong Zhang, Pengchao Si; A straight forward approach to electrodeposit tungsten disulfide/poly (3, 4-ethylenedioxythiophene) composites onto nanoporous gold for the hydrogen evolution reaction, Applied Surface Science 2017, 410, 308- 314 9. Xinxin Xiao, Peter Ó Conghaile, Dónal Leech, Roland Ludwig, Edmond Magner; A symmetric supercapacitor/biofuel cell hybrid device based on enzyme- modified nanoporous gold: an autonomous
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