Second Generation Sustainable Bacterial Biofuels

Completed

Renewable Energy Sources(Bio-Energy, Production of transport biofuels (incl. Production from wastes))
Renewable Energy Sources(Bio-Energy, Production of other biomass-derived fuels (incl. Production from wastes))

Basic and strategic applied research

BIOLOGICAL AND AGRICULTURAL SCIENCES (Biological Sciences)
PHYSICAL SCIENCES AND MATHEMATICS (Applied Mathematics)
ENGINEERING AND TECHNOLOGY (Electrical and Electronic Engineering)

Not Cross-cutting

Prof N Minton
Centre for Biomolecular Sciences
University of Nottingham

Research Grant

BBSRC

01 April 2009

30 September 2014

66 months

£2,127,704

Transport Systems and Vehicles

East Midlands

Bioenergy Initiative (BEN)

Prof N Minton, Centre for Biomolecular Sciences, University of Nottingham

Dr D Bolam, Institute for Cell and Molecular Biosciences (ICaMB), Newcastle University
Prof J Crowe, Electrical Systems and Optics, University of Nottingham
Dr J Heap, Centre for Synthetic Biology and Innovation, Imperial College London
Prof C Hodgman, School of Biosciences, University of Nottingham
Prof J King, Mathematical Sciences, University of Nottingham

Objectives not supplied

The generation of the butanol from lignocellulose (plant cell walls) has considerable BioEnergy potential. The major limitation to exploiting lignocellulose, however, is the rate at which these composite structures can be degraded by enzyme consortia. The most economic method of delivering these enzymes into the biomass conversion process is to engineer the fermenting organism to synthesise the plant cell wall degrading apparatus. In this project we will develop a consolidated bioprocessing system by introducing the plant cell wall degrading multienzyme complex from Clostridium cellulolyticum (cellulosome) into the butanol producing bacterium Clostridium acetobutylicum. Genes will be stably introduced into the genome using a newly developed, and patented, technological innovation which allows the construction of complex operons encoding the large number of catalytic components involved. Initially in vitro experiments will be used to develop an enzyme cocktail that is optimized for plant cell wall degradation. To assemble the enzymes into a cellulosome, to maximise the essential synergy between the catalytic components, the scaffolding protein will be inserted from Clostridium thermocellum, which will be tethered to the bacterial cell wall though a type II cohesin-dockerin interaction. By engineering promoter strengths the stoichiometries of the enzymes will be optimized for cell wall degradation. The genetic approach will also be used to identify genes that significantly enhance the degradation process by selecting for the activation of 'pro-genes'. The strains developed will be evaluated in butanol fermentation trials using plant biomass as the carbon and energy source. The readout from these initial experiments will inform modelling of cellulosome composition and action, which will inform further modification of the cellulosome through an iterative process. Finally, the influence of scale up will be evaluated through our industrial partner.

22/11/13