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Projects: Projects for Investigator
Reference Number	BB/I018492/1
Title	Bilateral BBSRC-FAPESP / Targeted analysis of microbial lignocellulolytic secretomes - a new approach to enzyme discovery.
Status	Completed
Energy Categories	Renewable Energy Sources(Bio-Energy, Other bio-energy) 50%; Renewable Energy Sources(Bio-Energy, Production of transport biofuels (incl. Production from wastes)) 50%;
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	Prof N (Neil ) Bruce No email address given Biology University of York
Award Type	Research Grant
Funding Source	BBSRC
Start Date	31 March 2012
End Date	30 March 2015
Duration	36 months
Total Grant Value	£493,661
Industrial Sectors	Pharmaceuticals and Biotechnology
Region	Yorkshire & Humberside
Programme
Investigators	Principal Investigator	Prof N (Neil ) Bruce , Biology, University of York (99.998%)
	Other Investigator	Prof S (Simon ) McQueen-Mason , Biology, University of York (0.001%) Prof JPW (Peter ) Young , Biology, University of York (0.001%)
Web Site
Objectives	The strategic relevance of this programme of work is high and falls within the BBSRC strategic priority in Bioenergy and Industrial Biotechnology described in the strategic plan 2010-2015. Exploring the digestion of lignocellulose by microbial communities using an integrated proteomics and metatranscriptomics has the potential to identify new approaches to lignocellulose utilisation along with new enzymes and intellectual property of benefit to the biofuel and biorefinery industries. Combining this potential with the power of high-throughput protein expression and top class analytical facilities in York and Sao Paulo will place the UK and Brazil in a strong position in this internationally competitive area. Establishing an efficient platform for sugar production from biomass feedstocks will underpin the establishment of competitive and sustainable biorefinery industries particularly in the area of transportation fuels. Other beneficiaries will include the farmers and breeders of energy crops. Waste management is a significant problem in the UK and our work is also aimed at providing underpinning tools for the conversion of waste biomass materials into sugars and biofuels. The project management will use proven processes to protect IP and publish results in scientific journals and at conferences. We will also use existing UK networks (eg the Bioscience KTN, the NNFCC, the Biorenewables Network) to communicate progress through their events and web-based or printed media. A workshop will be held towards the end of year 2 to disseminate information to relevant industrial stakeholders and academic beneficiaries. The applicants have strong links with the University of York's Enterprise and Innovation office who will also ensure that any commercial opportunities are realised. The research will also be undertaken in the context of the Centre for Low Carbon Futures (CLCF), a translational research centre established by the four major Universities in Yorkshire and Humberside, supported by an investment of £5.7m by the Regional Development Agency. This centre has contractual responsibility to engage end-users with research conducted in the regional HEIs. When appropriate, discoveries will be disseminated by the University to the general media through press releases. To ensure professional management of intellectual property, CNAP operates regular IP reviews of all projects. This involves the PIs, the University business development officer and a local patent company with expertise in biotechnology. CNAP has an outstanding track record in commercialisation of strategic research. This includes ongoing collaborations with companies throughout the biorenewable supply chain from major international seed companies, through processors to end users such as global pharmaceutical, personal care and feed companies as well as SMEs within the UK and EU.
Abstract	From both a fundamental and industrial biotech viewpoint understanding the deconstruction of lignocellulose in soil and compost is of central importance. In the natural environments microbial communities can efficiently degrade or modify lignin to enable the effective enzymatic hydrolysis of the polysaccharides present in plant cell walls. The aim of this proposal is to use metatranscriptomics and proteomics to determine gene- and protein-centred details to determine new mechanisms and improved methods of lignocellulose deconstruction in mixed microbial communities from composting wheat straw and sugar cane bagasse. The novelty of our proteomic approach lies in the use of a biotin affinity tag to distinguish secreted proteins from intracelluar proteins that are released from lysed cells during the extraction process necessary to release the proteins that bind tightly to the decaying plant biomass. The secreted proteins will be tagged, affinity purified, digested with trypsin and the resulting peptide mixtures analyzed by LC-ESI-MS. In order to have a picture of the overall community dynamics in terms of species composition at the different stages in the composting process DNA will be extracted for SSU rRNA profiling. Saccharification of the lignocellulose will be monitored and the lignin content of the straw or bagasse analysed using FTIR spectroscopy and solid state NMR. Metatranscriptome analysis will be performed by preparing cDNA from samples taken at various time points from the lignocellulose enriched cultures, the cDNA will be bar coded pooled and sequenced using the Roche 454 GS FLX Titanium platform. The peptide sequences from the proteomics analysis will allow the identification of full and partial coding sequences in the library. These coding sequences will be cloned and expressed in established recombinant expression systems and the recombinant proteins screened for activity.
Publications	(none)
Final Report	(none)
Added to Database	30/09/13