A synthetic biology approach to enhancing chemical production by anaerobic bacteria (SynBio-AnOx)

Completed

Not Energy Related
Renewable Energy Sources(Bio-Energy, Other bio-energy)

Basic and strategic applied research

BIOLOGICAL AND AGRICULTURAL SCIENCES (Biological Sciences)

Not Cross-cutting

Dr J Heap
Centre for Synthetic Biology and Innovation
Imperial College London

Research Grant

BBSRC

15 January 2015

14 January 2018

36 months

£351,532

Healthcare

London

Dr J Heap, Centre for Synthetic Biology and Innovation, Imperial College London

The overall aim of this programme is to develop, validate and apply novel and existing synthetic biology approaches to the metabolic engineering of obligate anaerobes, particularly for sustainable manufacturing of chemicals at high yields. The model obligate anaerobe Clostridium acetobutylicum will be engineered to achieve a homo-butanol fermentation from carbohydrate substrate. The anticipated outcomes of this programme will have impacts on a variety of beneficiaries. The new approaches developed in the research will benefit academic and industrial researchers in the related fields of Clostridium and other anaerobes, synthetic biology, metabolic engineering and industrial biotechnology. In these fields, academic and industrial researchers are closely linked, and academic research is often directly relevant to industrial research. The new approaches will improve metabolic engineering of obligate anaerobes, allowing progress not possible using previous techniques, thereby improving technology for sustainable manufacturing of chemicals. The high-yield biological manufacture of bulk chemicals using anaerobic fermentation is an industrial technology of great value and importance to numerous companies. There are several such companies worldwide, including some based in the UK, or otherwise active in the UK. The approaches developed in the research programme could directly benefit the existing manufacturing processes of such companies, and could be important in the development of new manufacturing processes, which might involve other organisms, feedstocks and products. These benefits could in turn lead to UK jobs and other contributions to the UK economy and UK wealth. Sustainable manufacturing is not only of economic benefit, but is also an important part of the necessary transition to sustainability and away from finite fossil carbon. This is crucial for the long-term energy security, food security, well-being and economic prosperity of the UK and nations worldwide. The research will benefit the expertise and development of those directly involved in performing the research, and those who are part of relevant research communities at Imperial College. These benefits contribute to the UK skill base in the important areas of synthetic biology and industrial biotechnology. The research is at the cutting-edge of synthetic biology and metabolic engineering, and will influence the advanced education of students at Imperial College by the principal investigator John Heap (JH) who lectures both undergraduate and postgraduate students on these topics.

The overall aim of this programme is to develop, validate and apply novel and existing synthetic biology approaches to the metabolic engineering of obligate anaerobes, particularly for sustainable manufacturing of chemicals at high yields. Previous research on improving anaerobic fermentation strains has made the best progress with facultative anaerobes such as E. coli. This programme aims to make similar progress in improving obligate anaerobes, which have various advantages, but also challenges for genetic and metabolic engineering that have not been overcome by conventional approaches. To overcome these challenges, more sophisticated synthetic biology approaches will be developed and applied. The model obligate anaerobe Clostridium acetobutylicum will be engineered to achieve a homo-butanol fermentation from carbohydrate substrate.

1. We will develop core synthetic biology tools for anaerobes, using oxygen-independent fluorescent reporters to develop promoter libraries for native and dedicated (orthogonal) expression systems and to validate computational RBS design in Clostridium acetobutylicum.
2. We will develop and implement a system for pyruvate node redox flux redistribution that is compatible with the biochemistry and metabolism of obligate anaerobes. This will allow reducing equivalents to be directed to butanol formation, resulting in a homo-butanol fermentation.
3. We will implement STABLE solutions to the key challenges of butanol pathway over-expression, megaplasmid stability, and resilience to acid crash; and combine these with known modifications and the novel redox flux partitioning system to maximise butanol yield.Improved butanol-producing strain(s) generated in this programme will be evaluated for industrial use by our industrial partner.

08/04/16