A Synthetic Biology Approach to Engineering Living Photovoltaics.

The ability to electronically interface living cells with electron accepting scaffolds is crucial for the development of next-generation biophotovoltaic technologies. Although recent studies have focused on engineering synthetic interfaces that can maximize electronic communication between the cell and scaffold, the efficiency of such devices is limited by the low conductivity of the cell membrane. This review provides a materials science perspective on applying a complementary, synthetic biology approach to engineering membrane-electrode interfaces. It focuses on the technical challenges behind the introduction of foreign extracellular electron transfer pathways in bacterial host cells and the past and future efforts to engineer photosynthetic organisms with artificial electron-export capabilities for biophotovoltaic applications. The article highlights advances in engineering protein-based, electron-exporting conduits in a model host organism, E. coli, before reviewing state-of-the-art biophotovoltaic technologies that use both unmodified and bioengineered photosynthetic bacteria with improved electron transport capabilities. A thermodynamic analysis is used to propose an energetically feasible pathway for extracellular electron transport in engineered cyanobacteria and identify metabolic bottlenecks amenable to protein engineering techniques. Based on this analysis, an engineered photosynthetic organism expressing a foreign, protein-based electron conduit yields a maximum theoretical solar conversion efficiency of 6-10% without accounting for additional bioengineering optimizations for light-harvesting.

[1]  Kevin Barraclough,et al.  I and i , 2001, BMJ : British Medical Journal.

[2]  A. Krieger-Liszkay,et al.  Regulation of Photosynthetic Electron Transport and Photoinhibition , 2014, Current protein & peptide science.

[3]  C. Howe,et al.  Terminal oxidase mutants of the cyanobacterium Synechocystis sp. PCC 6803 show increased electrogenic activity in biological photo-voltaic systems. , 2013, Physical chemistry chemical physics : PCCP.

[4]  G. G. Stokes "J." , 1890, The New Yale Book of Quotations.

[5]  Byoung-In Sang,et al.  Extracellular electron transfer from cathode to microbes: application for biofuel production , 2016, Biotechnology for Biofuels.

[6]  V. Raghavan,et al.  Carbon neutral electricity production by Synechocystis sp. PCC6803 in a microbial fuel cell. , 2012, Bioresource technology.

[7]  A. Melis,et al.  Photosystem-II damage and repair cycle in chloroplasts: what modulates the rate of photodamage ? , 1999, Trends in plant science.

[8]  Tingyue Gu,et al.  A state of the art review on microbial fuel cells: A promising technology for wastewater treatment and bioenergy. , 2007, Biotechnology advances.

[9]  A. Torriero,et al.  Inquisition of Microcystis aeruginosa and Synechocystis nanowires: characterization and modelling , 2015, Antonie van Leeuwenhoek.

[10]  L. Thöny-Meyer,et al.  Light Harvesting Proteins for Solar Fuel Generation in Bioengineered Photoelectrochemical Cells , 2014, Current protein & peptide science.

[11]  Xiaocheng Jiang,et al.  Probing electron transfer mechanisms in Shewanella oneidensis MR-1 using a nanoelectrode platform and single-cell imaging , 2010, Proceedings of the National Academy of Sciences.

[12]  Peng Liang,et al.  A completely anoxic microbial fuel cell using a photo-biocathode for cathodic carbon dioxide reduction , 2009 .

[13]  Caroline M. Ajo-Franklin,et al.  The Mtr Pathway of Shewanella oneidensis MR‐1 Couples Substrate Utilization to Current Production in Escherichia coli , 2014 .

[14]  Baowei Chen,et al.  Isolation of a High-Affinity Functional Protein Complex between OmcA and MtrC: Two Outer Membrane Decaheme c-Type Cytochromes of Shewanella oneidensis MR-1 , 2006, Journal of bacteriology.

[15]  Yamini Jangir,et al.  Disentangling the roles of free and cytochrome-bound flavins in extracellular electron transport from Shewanella oneidensis MR-1 , 2016 .

[16]  P. Dobbin,et al.  Characterization of the Shewanella oneidensis MR-1 Decaheme Cytochrome MtrA , 2003, Journal of Biological Chemistry.

[17]  G. T. Feliciano,et al.  Thermally activated charge transport in microbial protein nanowires , 2016, Scientific Reports.

[18]  Michaela A. Teravest,et al.  CymA and Exogenous Flavins Improve Extracellular Electron Transfer and Couple It to Cell Growth in Mtr-Expressing Escherichia coli. , 2016, ACS synthetic biology.

[19]  M. C. Potter Electrical Effects Accompanying the Decomposition of Organic Compounds. II. Ionisation of the Gases Produced during Fermentation , 1911 .

[20]  K. Yunus,et al.  In situ fluorescence and electrochemical monitoring of a photosynthetic microbial fuel cell. , 2013, Physical chemistry chemical physics : PCCP.

[21]  T. Arakawa,et al.  Biochemical characterization of purified OmcS, a c-type cytochrome required for insoluble Fe(III) reduction in Geobacter sulfurreducens. , 2011, Biochimica et biophysica acta.

[22]  J. Bye,et al.  A functional description of CymA, an electron-transfer hub supporting anaerobic respiratory flexibility in Shewanella. , 2012, The Biochemical journal.

[23]  Byoung-Chan Kim,et al.  Tunable metallic-like conductivity in microbial nanowire networks. , 2011, Nature nanotechnology.

[24]  Sarah M. Strycharz-Glaven,et al.  Measuring conductivity of living Geobacter sulfurreducens biofilms. , 2016, Nature nanotechnology.

[25]  Alice Dohnalkova,et al.  Electrically conductive bacterial nanowires produced by Shewanella oneidensis strain MR-1 and other microorganisms. , 2006, Proceedings of the National Academy of Sciences of the United States of America.

[26]  S. Long,et al.  What is the maximum efficiency with which photosynthesis can convert solar energy into biomass? , 2008, Current opinion in biotechnology.

[27]  S. Golden,et al.  Proteins Found in a CikA Interaction Assay Link the Circadian Clock, Metabolism, and Cell Division in Synechococcus elongatus , 2008, Journal of bacteriology.

[28]  L. M. Markillie,et al.  Overexpression of multi-heme C-type cytochromes. , 2005, BioTechniques.

[29]  T. Wen,et al.  Current and voltage responses in instant photosynthetic microbial cells with Spirulina platensis , 2010 .

[30]  Kazuko Tanaka,et al.  Bioelectrochemical fuel‐cells operated by the cyanobacterium, Anabaena variabilis , 1985 .

[31]  E. Hall,et al.  DIAMINODURENE AS A MEDIATOR OF A PHOTOCURRENT USING INTACT CELLS OF CYANOBACTERIA , 1994 .

[32]  Michaela A. Teravest,et al.  Transforming exoelectrogens for biotechnology using synthetic biology , 2016, Biotechnology and bioengineering.

[33]  Ardemis A. Boghossian,et al.  Photoelectrochemical complexes for solar energy conversion that chemically and autonomously regenerate , 2010, Nature chemistry.

[34]  Jochen Blumberger,et al.  Multi-haem cytochromes in Shewanella oneidensis MR-1: structures, functions and opportunities , 2015, Journal of The Royal Society Interface.

[35]  Hanqing Yu,et al.  Extracellular electron transfer mechanisms between microorganisms and minerals , 2016, Nature Reviews Microbiology.

[36]  Aaas News,et al.  Book Reviews , 1893, Buffalo Medical and Surgical Journal.

[37]  C. Haigh,et al.  Organ , 1941, Definitions.

[38]  Christopher J. Howe,et al.  Photosynthetic biofilms in pure culture harness solar energy in a mediatorless bio-photovoltaic cell (BPV) system† , 2011 .

[39]  C. Mullineaux Electron transport and light-harvesting switches in cyanobacteria , 2014, Front. Plant Sci..

[40]  Tibor Fabian,et al.  Direct extraction of photosynthetic electrons from single algal cells by nanoprobing system. , 2010, Nano letters.

[41]  Stanford Schor,et al.  Directed evolution of Gloeobacter violaceus rhodopsin spectral properties. , 2015, Journal of molecular biology.

[42]  Jong Hyun Choi,et al.  Biomimetic strategies for solar energy conversion: a technical perspective , 2011 .

[43]  Yogeswaran Umasankar,et al.  Photocurrent generation by immobilized cyanobacteria via direct electron transport in photo-bioelectrochemical cells. , 2014, Physical chemistry chemical physics : PCCP.

[44]  Largus T Angenent,et al.  Cathodes as electron donors for microbial metabolism: which extracellular electron transfer mechanisms are involved? , 2011, Bioresource technology.

[45]  L. Gorton,et al.  Photo-electrochemical communication between cyanobacteria (Leptolyngbia sp.) and osmium redox polymer modified electrodes. , 2014, Physical chemistry chemical physics : PCCP.

[46]  Karsten Zengler,et al.  A new model for electron flow during anaerobic digestion: direct interspecies electron transfer to Methanosaeta for the reduction of carbon dioxide to methane , 2014 .

[47]  F. Wollman,et al.  A specific c-type cytochrome maturation system is required for oxygenic photosynthesis , 2007, Proceedings of the National Academy of Sciences.

[48]  Christopher J. Howe,et al.  Biophotovoltaics: oxygenic photosynthetic organisms in the world of bioelectrochemical systems , 2015 .

[49]  Christopher J Howe,et al.  Biological photovoltaics: intra- and extra-cellular electron transport by cyanobacteria. , 2012, Biochemical Society transactions.

[50]  James Barber,et al.  Comparing Photosynthetic and Photovoltaic Efficiencies and Recognizing the Potential for Improvement , 2011, Science.

[51]  J. Gralnick,et al.  Ecology and biotechnology of the genus Shewanella. , 2007, Annual review of microbiology.

[52]  Xin-Guang Zhu,et al.  Improving photosynthetic efficiency for greater yield. , 2010, Annual review of plant biology.

[53]  Michaela A. Teravest,et al.  Tuning promoter strengths for improved synthesis and function of electron conduits in Escherichia coli. , 2013, ACS synthetic biology.

[54]  E. Aro,et al.  Photoinhibition of Photosystem II. Inactivation, protein damage and turnover. , 1993, Biochimica et biophysica acta.

[55]  Stefano Freguia,et al.  Microbial fuel cells: methodology and technology. , 2006, Environmental science & technology.

[56]  J. M. Schuurmans,et al.  The Redox Potential of the Plastoquinone Pool of the Cyanobacterium Synechocystis Species Strain PCC 6803 Is under Strict Homeostatic Control1[C][W] , 2014, Plant Physiology.

[57]  T. Ogawa,et al.  Effects of light on the electrical output of bioelectrochemical fuel‐cells containing Anabaena variabilis M‐2: Mechanism of the post‐illumination burst , 2007 .

[58]  A. Spormann,et al.  Periplasmic Electron Transfer via the c-Type Cytochromes MtrA and FccA of Shewanella oneidensis MR-1 , 2009, Applied and Environmental Microbiology.

[59]  T. Hwa,et al.  Interdependence of Cell Growth and Gene Expression: Origins and Consequences , 2010, Science.

[60]  John M. Pisciotta,et al.  Light-Dependent Electrogenic Activity of Cyanobacteria , 2010, PloS one.

[61]  M. Madigan,et al.  Syntrophic anaerobic photosynthesis via direct interspecies electron transfer , 2017, Nature Communications.

[62]  V. Rotello,et al.  Reply to 'Measuring conductivity of living Geobacter sulfurreducens biofilms'. , 2016, Nature nanotechnology.

[63]  Deepak Pant,et al.  Nanotechnology to rescue bacterial bidirectional extracellular electron transfer in bioelectrochemical systems , 2016 .

[64]  D. Richardson,et al.  The c-Type Cytochrome OmcA Localizes to the Outer Membrane upon Heterologous Expression in Escherichia coli , 2008, Journal of bacteriology.

[65]  David E. Williams,et al.  A cost-effective microbial fuel cell to detect and select for photosynthetic electrogenic activity in algae and cyanobacteria , 2014, Journal of Applied Phycology.

[66]  L. Cronin,et al.  A Bioelectrochemical Approach to Characterize Extracellular Electron Transfer by Synechocystis sp. PCC6803 , 2014, PloS one.

[67]  D. R. Bond,et al.  Shewanella secretes flavins that mediate extracellular electron transfer , 2008, Proceedings of the National Academy of Sciences.

[68]  Bruce E Cohen,et al.  Engineering of a synthetic electron conduit in living cells , 2010, Proceedings of the National Academy of Sciences.

[69]  W. Marsden I and J , 2012 .

[70]  Yung-Chuan Liu,et al.  Characteristics of the photosynthesis microbial fuel cell with a Spirulina platensis biofilm. , 2013, Bioresource technology.

[71]  Christopher J. Howe,et al.  Quantitative analysis of the factors limiting solar power transduction by Synechocystis sp. PCC 6803 in biological photovoltaic devices , 2011 .

[72]  Andrew G. Glen,et al.  APPL , 2001 .

[73]  René H. Wijffels,et al.  Maximum Photosynthetic Yield of Green Microalgae in Photobioreactors , 2010, Marine Biotechnology.

[74]  Derek R. Lovley,et al.  Alignment of the c-Type Cytochrome OmcS along Pili of Geobacter sulfurreducens , 2010, Applied and Environmental Microbiology.

[75]  D. R. Bond,et al.  Electricity Production by Geobacter sulfurreducens Attached to Electrodes , 2003, Applied and Environmental Microbiology.

[76]  D. R. Bond,et al.  A trans-outer membrane porin-cytochrome protein complex for extracellular electron transfer by Geobacter sulfurreducens PCA , 2014, Environmental microbiology reports.

[77]  C. Wraight,et al.  The absolute quantum efficiency of bacteriochlorophyll photooxidation in reaction centres of Rhodopseudomonas spheroides. , 1974, Biochimica et biophysica acta.

[78]  Uwe Schröder,et al.  Photomicrobial Solar and Fuel Cells , 2010 .

[79]  D. Krogmann,et al.  Extinction coefficients and midpoint potentials of cytochrome c(6) from the cyanobacteria Arthrospira maxima, Microcystis aeruginosa, and Synechocystis 6803. , 1999, Biochimica et biophysica acta.

[80]  Kenji Kano,et al.  Photosynthetic bioelectrochemical cell utilizing cyanobacteria and water-generating oxidase , 2001 .

[81]  Hao Yan,et al.  Reengineering the optical absorption cross-section of photosynthetic reaction centers. , 2014, Journal of the American Chemical Society.

[82]  Jeffrey A. Gralnick,et al.  Towards Electrosynthesis in Shewanella: Energetics of Reversing the Mtr Pathway for Reductive Metabolism , 2011, PloS one.

[83]  A. Noy,et al.  Crossing Over: Nanostructures that Move Electrons and Ions across Cellular Membranes , 2015, Advanced materials.

[84]  S. Bhattacharya,et al.  Hydrogen production by Cyanobacteria , 2005, Microbial Cell Factories.

[85]  Heinrich Heide,et al.  A dynamic periplasmic electron transfer network enables respiratory flexibility beyond a thermodynamic regulatory regime , 2015, The ISME Journal.

[86]  A. Spormann,et al.  Dissimilatory iron reduction in Escherichia coli: identification of CymA of Shewanella oneidensis and NapC of E. coli as ferric reductases , 2008, Molecular microbiology.

[87]  P. Cameron,et al.  An investigation of anode and cathode materials in photomicrobial fuel cells , 2016, Philosophical Transactions of the Royal Society A: Mathematical, Physical and Engineering Sciences.

[88]  Christopher J Howe,et al.  A High Power-Density, Mediator-Free, Microfluidic Biophotovoltaic Device for Cyanobacterial Cells , 2014, Advanced energy materials.

[89]  U. Schröder Anodic electron transfer mechanisms in microbial fuel cells and their energy efficiency. , 2007, Physical chemistry chemical physics : PCCP.

[90]  T. Mehta,et al.  Outer Membrane c-Type Cytochromes Required for Fe(III) and Mn(IV) Oxide Reduction in Geobacter sulfurreducens , 2005, Applied and Environmental Microbiology.

[91]  M. Ratner,et al.  Redox equilibria in hydroxylamine oxidoreductase. Electrostatic control of electron redistribution in multielectron oxidative processes. , 2005, Biochemistry.

[92]  Zhen He,et al.  Applications and perspectives of phototrophic microorganisms for electricity generation from organic compounds in microbial fuel cells , 2014 .

[93]  John M. Pisciotta,et al.  Photosynthetic microbial fuel cells with positive light response , 2009, Biotechnology and bioengineering.

[94]  W. W. Adams,et al.  Photochemistry and xanthophyll cycle-dependent energy dissipation in differently oriented cladodes of Opuntia stricta during the winter , 1998 .

[95]  D. Sinton,et al.  A photosynthetic-plasmonic-voltaic cell: Excitation of photosynthetic bacteria and current collection through a plasmonic substrate , 2014 .

[96]  Graham R Fleming,et al.  Lessons from nature about solar light harvesting. , 2011, Nature chemistry.

[97]  A. Fisher,et al.  Porous ceramic anode materials for photo-microbial fuel cells , 2011 .

[98]  C. D. Vitry Cytochrome c maturation system on the negative side of bioenergetic membranes: CCB or System IV. , 2011 .

[99]  H. Lill,et al.  Expression of prokaryotic and eukaryotic cytochromes c in Escherichia coli. , 2000, Biochimica et biophysica acta.

[100]  Ching Leang,et al.  Direct Exchange of Electrons Within Aggregates of an Evolved Syntrophic Coculture of Anaerobic Bacteria , 2010, Science.

[101]  Daniel Picot,et al.  Spectral and redox characterization of the heme ci of the cytochrome b6f complex. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[102]  L. T. Angenent,et al.  Light energy to bioelectricity: photosynthetic microbial fuel cells. , 2010, Current opinion in biotechnology.

[103]  P. Girguis,et al.  Electron uptake by iron-oxidizing phototrophic bacteria , 2014, Nature Communications.

[104]  R. Ramasamy,et al.  Enhanced photo-bioelectrochemical energy conversion by genetically engineered cyanobacteria. , 2015, Biotechnology and bioengineering.

[105]  Kenji Kano,et al.  Electrochemical investigation of cyanobacteria Synechococcus sp. PCC7942-catalyzed photoreduction of exogenous quinones and photoelectrochemical oxidation of water , 2001 .

[106]  Akin Ali,et al.  Surface morphology and surface energy of anode materials influence power outputs in a multi-channel mediatorless bio-photovoltaic (BPV) system. , 2012, Physical chemistry chemical physics : PCCP.

[107]  Liang Shi,et al.  The ‘porin–cytochrome’ model for microbe‐to‐mineral electron transfer , 2012, Molecular microbiology.

[108]  S. Durell,et al.  The effect of ethylenediamine chemical modification of plastocyanin on the rate of cytochrome f oxidation and P-700+ reduction. , 1987, Biochimica et biophysica acta.

[109]  John M. Pisciotta,et al.  Nanostructured polypyrrole-coated anode for sun-powered microbial fuel cells. , 2010, Bioelectrochemistry.

[110]  H. Ochiai,et al.  Properties of semiconductor electrodes coated with living films of cyanobacteria , 1983 .

[111]  A. Melis,et al.  Solar energy conversion efficiencies in photosynthesis: Minimizing the chlorophyll antennae to maximize efficiency , 2009 .

[112]  Higinio Mora-Mora,et al.  μ-MAR: Multiplane 3D Marker based Registration for depth-sensing cameras , 2015, Expert Syst. Appl..

[113]  S. Elliott,et al.  Solution-Based Structural Analysis of the Decaheme Cytochrome, MtrA, by Small-Angle X-ray Scattering and Analytical Ultracentrifugation , 2011, The journal of physical chemistry. B.

[114]  Anna Obraztsova,et al.  Current Production and Metal Oxide Reduction by Shewanella oneidensis MR-1 Wild Type and Mutants , 2007, Applied and Environmental Microbiology.

[115]  N. Schuergers,et al.  Appendages of the Cyanobacterial Cell , 2015, Life.