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Projects: Projects for Investigator
Reference Number	BB/M000265/1
Title	Engineering new capacities for solar energy utilisation in bacteria
Status	Completed
Energy Categories	Renewable Energy Sources(Bio-Energy, Other bio-energy) 10%; Not Energy Related 90%;
Research Types	Basic and strategic applied research 100%
Science and Technology Fields	BIOLOGICAL AND AGRICULTURAL SCIENCES (Biological Sciences) 100%
UKERC Cross Cutting Characterisation	Not Cross-cutting 100%
Principal Investigator	Professor N Hunter No email address given Molecular Biology and Biotechnolog University of Sheffield
Award Type	Research Grant
Funding Source	BBSRC
Start Date	02 February 2015
End Date	01 February 2020
Duration	60 months
Total Grant Value	£3,380,116
Industrial Sectors	Healthcare
Region	Yorkshire & Humberside
Programme	Longer and Larger Grants (LoLas)
Investigators	Principal Investigator	Professor N Hunter , Molecular Biology and Biotechnolog, University of Sheffield (99.994%)
	Other Investigator	Dr W Huang , Engineering Science, University of Oxford (0.001%) Dr J Reid , Chemistry, University of Sheffield (0.001%) Dr AJ Cadby , Physics and Astronomy, University of Sheffield (0.001%) Dr M Dickman , Chemical and Biological Engineering, University of Sheffield (0.001%) Dr M (Matt ) Johnson , Department of Molecular Biology and Biotechnology, University of Sheffield (0.001%) Professor PA Bullough , Department of Molecular Biology and Biotechnology, University of Sheffield (0.001%)
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Objectives	Economic impacts Nanotechnology: Development of nanotechnological tools for imaging and functional measurements of biological samples. AFM, including affinity-mapping, and super-resolution optical microscopy, as well as picosecond lifetime and spectral imaging, will be developed and applied to membranes from plants and bacteria, and to enzymes and surface-attached nanoarrays of membrane protein complexes. AJC will test new optical microscope technologies and develop new applications and data analysis approaches for industrial partner Andor Technology. Bruker Nano Surfaces Division test new developments in probe manufacture in the CNH laboratory. Separation technologies: We collaborate with Thermofisher on innovative methodologies for separating peptides prior to MS analysis, and with Bruker for mass spectrometry. Synthetic biology. Dr Pin Yang, CEO of InCelliGEN, a biotechnology company specialising in genome and pathway synthesis, has expressed interest in our programme to construct photosynthesis modules for expression in a variety of bacteria. High-Throughput Techniques. Development of novel high-throughput screening methods of single colonies of mutant bacterial strains (CNH/WH) will be undertaken in collaboration with BMG LabTech. Utilising next generation fluorescent plate readers which can produce fluorescent spectra will decrease the time taken to screen for new fluorescent pigments or knockouts. Biofuels, CO2 remediation and Biosensors: WH and CNH will work with Tata Steel, who are looking to cut CO2 emissions by 50% by 2015. Waste CO2 could be used as a carbon source for electrobiosynthesis in R. eutropha or R. sphaeroides. Societal Impacts The sLoLa team encompasses molecular genetics and biosynthetic pathway engineering (Huang, Hunter), biochemistry, spectroscopy, structural studies using electron microscopy and atomic force microscopy (Bullough, Hunter, Johnson), mass spectrometry (Dickman), and super-resolution/spectral imaging/lifetime microscopy (Cadby). The multidisciplinary nature of this research project will train young researchers in a wide variety of practical skills, providing an outstanding training environment for PhD students and postdoctoral scientists, working to meet ambitious technical goals and targets. Communication and engagement with potential beneficiaries. Synthetic biology, food and energy security and the nano-scale world are highly topical in the media, and the work in this application will be communicated by, for example, the Krebs Festival, to be held in November 2015. This three-month, large-scale public engagement and outreach activity will be supported by this sLoLa; Matt Johnson and named PDRA Dr Nathan Adams are part of the steering committee. Ongoing University outreach activities for younger audiences (Festival of the Mind, Mobile University, Sheffield Science and Engineering Festival, Researchers Night) w ill be supported by PIs (Reid/Cadby/Johnson), and PDRAs and PhD students will be encouraged to present their research at these events over the course of the sLola. Nathan Adams is a presenter at national science festivals; in 2013 he undertook 28 science presenter/public engagement activities at the British Science Festival, Bang!, Cheltenham Science Festival, CBBC Live, Edinburgh Festival, TV and Radio with the support of the University and professional bodies, reaching over 2 million people through media appearances and demonstrations of science. NA will continue his science presenting on TV and radio during the sLoLa. PIs and PDRAs will disseminate important findings via press releases and interviews with the broadcast and print media, in collaboration with the office for public engagement.
Abstract	This sLoLa programme is organised into four interdependent sections. In the first, we will combine chlorophyll (Chl), bacteriochlorophyll (BChl) and carotenoid biosynthesis genes from bacteria and plants to create new pigment pathways tailored that extend the spectral range for absorbing solar energy. The morphology, organisation and performance of altered photosynthetic complexes and membranes comprising these novel pigments will be assessed in the second section using functional, proteomic and state-of-the-art imaging approaches, including electron microscopy, affinity mapping AFM, and super-resolution optical imaging. By applying selection pressure to grow under defined, restricted light regimes we will select engineered strains with improved photosynthetic performance. In the third part of the programme we will compile and then test multigene constructs for biosynthesis of proteorhodopsins, pigments, lipids, assembly factors and photosystem apoproteins. Following co-expression of these gene assemblies in heterotrophic bacteria such as E. coli and Ralstonia eutropha the outcomes will be assessed using simple growth tests, mass spectrometry, atomic force and electron microscopy, and spectroscopy. The fourth part of the programme, to be conducted in parallel with the other sections, proceeds initially by optimising the light-driven production of alkanes in Rhodobacter sphaeroides and Synechocystis, and by establishing an electro-biosynthesis platform for metabolite production in Ralstonia eutropha and Rhodobacter sphaeroides. Then this section builds on the design and construction of new photosynthetic systems in the other sections; proteorhodopsins, engineered pigment biosynthesis and photosystem pathways, augmented by hitherto undiscovered genes, will be used to enhance solar-powered production of metabolites in photosynthetic bacteria and in heterotrophs converted for absorbing and using light.
Publications	(none)
Final Report	(none)
Added to Database	08/04/16