Utilising Steel Mill 'Off-Gas' for Chemical Commodity Production using Synthetic Biology

Lead Research Organisation: University of Nottingham
Department Name: School of Life Sciences

Abstract

Abstracts are not currently available in GtR for all funded research. This is normally because the abstract was not required at the time of proposal submission, but may be because it included sensitive information such as personal details.
 
Description At the initial workshop held in Nottingham the partnership decided to focus on the acetogen Clostridium carboxidivorans (the chassis of the US company Coskata), while at the same time both countries would implement programmes of work designed to isolate new acetogens capable of growth on CO/CO2. The choice of the former was made on the basis that C. carboxidivorans is able to make both C4 (butyrate and butanol) and as well as C2 (acetate and ethanol) and on the fact that SIBs had been able to demonstrate gene transfer by electroporation, paving the way for metabolic engineering. On this basis, the exemplar product chosen was butanol. The fall-back position was acetone.

As transformation of C. carboxidivorans could not be replicated at Nottingham, two UK students visited Shanghai for 6 weeks immediately prior to the International conference 'Clostridium XIII' which was hosted by SIBs in September 2014. As no evidence was obtained for transformation during this visit, a Chinese researcher visited Nottingham (March - May 2015) to demonstrate DNA transfer. No transfer was obtained. Accordingly, both laboratories elected to investigate alternative acetogens. Nottingham chose to focus on Eubacterium limosum whereas the Chinese groups looked to isolate novel strains.

While these investigations were ongoing, collaborative work was undertaken on the design and implementation of CRISPR/ Cas9 genome editing tools for use in clostridia and acetogens. Accordingly, visits were made to Nottingham by three SIBs researchers (July-Oct 2015; Nov 2015- Jan 2016 and Apr - June 2016). Three publications arose from this work (ACS Synth Biol. 2016; 5(12):1355-1361; Biotechnol J. 2016; 11(7): 961-72; Metab Eng. 2017; 39:38-48). In the absence of a chosen strain, both groups investigated the production of acetone in model acetogens - in C. lungdahlii at SIBS and in Eubacterium limosum at Nottingham.

A midterm review meeting was held in Shanghai in December 2016, at which the partnership exchanged information on the status of tool development. At this meeting it was agreed that a recombineering strategy formulated at Nottingham should be continued to be pursued as a means of overcoming the recalcitrance of C. carboxidivorans to DNA transfer, while at the same time implementation of genetics in E. limosum should continue to be pursued, with a view to transferring the technology to Shanghai in 2017.

These efforts at Nottingham have led to the formulation of high frequency transformation procedures in Eubacterium limosum, the demonstration of gene knock-out by directed (allelic exchange) and random (mariner transposon) and the exemplification of CRISPR/Cas9 genome editing. These innovations have led to the award of the EraCoBio-Tech project, BIOMETCHEM (BB/R021503/1). Apr 2018-Mar 2021. In the meantime funding was received to explore a novel recombineering strategy to overcome the restriction barriers present in C. carboxidivorans as a 'Proof of Concept' project from the BBSRC NIBB, C1net.

Now that we have solved the problems of having a genetically amenable C4 acetogen, we wish to transfer the necessary technologies to our Chinese partner through secondment of appropriate students (Elizabeth Redfern and Francois Seys) to Shanghai in the New Year, and thereafter hold a workshop in the spring (May/June) in Beijing with our various Chinese collaborators, to plan the future exploitation of the chassis.
In particular, the workshop will focus on GCRF opportunities between the two countries that also involve other ODA countries in the region. We anticipate that out of this workshop will emerge potential projects that could be submitted to GCRF or other joint UK:China funding calls.
Exploitation Route It is too early for others to exploit our findings.
Sectors Chemicals,Energy,Manufacturing, including Industrial Biotechology

 
Description There has yet to be any significant use of the findings of this project to date.
First Year Of Impact 2014
Sector Chemicals,Manufacturing, including Industrial Biotechology
Impact Types Economic