ERA-IB 5: Biological conversion of CO2 to the platform chemical 3-hydroxypropanoic acid

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

Abstract

Current energy and chemical needs are met by the extraction and processing of the fossil fuels.Such resources are finite and their use causes environmental pollution and greenhouse gas (GHG) emissions. The challenge facing humankind is, therefore, to identify new, sustainable and cleaner processes for chemical and energy generation. Biological routes represent a promising option, but strategies to date rely on the use of microbes to convert through fermentation the easily accessible carbohydrates (sugar and starch) of plants (such as sugar cane or corn) into chemicals and fuels. This has led to concerns over competition with the use of these carbohydrates as food, and a re-focussing of efforts on non-food, plant cell wall material (lignocellulose). However, lignocellulose is extremely resistant to being broken down into the sugar needed for fermentation. Overcoming this recalcitrance in a cost effective manner is proving extremely challenging.

There is, however, an exciting low-cost alternative, and that is to directly capture carbon, by harnessing the ability of certain bacteria to 'eat' single carbon GHG gases such as CO2. The gas is injected into the liquid medium of fermentation vessels where it is consumed by the bacteria and converted into the chemicals we need. Fortunately, such gases are an abundant resource, and may be derived from non-food sources such as waste gases from industry as well as 'synthesis gas' produced from the gasification (heating) of non-food biomass and domestic/ agricultural wastes. In this project, we will use this technology to make the platform chemical hydroxypropanoic acid. It has a multitude of uses, including the manufacture of plastics, coatings, adhesives, floor polishes and paints. By using non-food, waste gas as a feedstock, competition with food and land resources is avoided while at the same time providing benefits to the environment and society through a reduction in GHG emissions.

Technical Summary

One of the greatest challenges facing industry and society are the future sustainable production of chemicals and fuels from non-food feedstocks while at the same time reducing Green House Gas (GHG) emissions. To compete with existing petrochemical-based chemical manufacturing processes, low cost feedstocks for biological fermentation processes are essential, since the feedstock typically equates to over 60% of the overall production cost. To date, the focus has been to use lignocellulosic feedstocks. Their use is reliant on an energy intensive pre-treatment step, and thereafter, the addition of costly exogenous hydrolytic enzymes needed to convert the partially deconstructed biomass into the sugars needed by the fermentative process organisms. The costs involved are making the development of economic processes extremely challenging.

An innovative solution is to use waste gases as the feedstock. Gas fermenting microbes are able to grow on C1 compounds, such as carbon monoxide (CO), carbon dioxide (CO2) and methane (CH4) derived from non-food sources such as waste gases from industry (steel manufacture, oil refining, coal and natural/shale gas) as well as 'synthesis gas' (CO/CO2 and H2) produced from sustainable resources, such as biomass and domestic/agricultural wastes. This enables the production of fuels and chemicals in any industrialized geography without the consumption of valuable food or land resources. Moreover, by consuming gases otherwise destined for atmospheric release, a real contribution to the environment will be made through reductions in GHG emissions.

In this project, we will establish an alternative commercial process for the production of a major platform chemical via gas fermentation, from synthetic biology and genome modeling through fermentation process development and finally technoeconomic (TEA) and life cycle analysis (LCA) of a modelled integrated future commercial process.

Planned Impact

Successful completion of CO2CHEM will provide a novel route to the platform chemical 3-HP, based on the use of the cheap and abundant carbon source CO2. Multiple novel pathway enzymes will be characterized and their expression optimized in the production organism.

Primary outcomes will be a new platform microorganism, A. woodii, with a commercial process for fermentation of CO2 and H2. The project will develop general molecular biology methods needed for efficient engineering of future production organisms. Genome scale models will be developed and implemented for optimization of production organisms as well as media and fermentation conditions. These tools will advance the field of industrial biotechnology through future research collaborations and dissemination of project results.

The project will deliver key information needed for commercialization of CO2 + H2 fermentation: 1) target products; 2) optimized process conditions from lab-scale experiments, with integrated commercial process models; 3) TEA and LCA, showing benefits in GHG and pollution reductions; and 4) recommended commercial deployment scenarios. The novel combination of water electrolysis with gas fermentation can also be a means for large scale and long-term energy storage, important for load balancing in future energy systems with high shares of renewable power generation.

The CO2CHEM concept directly enhances the global competitiveness of biorefinery industries, which are expected to expand significantly in the coming decades. Successful proof-of-concept for CO2 + H2 fermentation will provide an offering to customers in CO2 intensive industries to monetize gaseous waste streams while lowering emissions. The project opens new markets for the industry partners. Post-project steps for commercial are: demonstration of the integrated process at pre-commercial scale, followed by commercial offering to LanzaTech's extensive current and future industrial customer base. In addition, Siemens will take individual technologies to the market, including water electrolyser, carbon capture processes, and gasifiers.

Dissemination Plan

The dissemination strategy has been defined to target different groups: researchers and students, equipment suppliers, and potential end-users of the technology (energy intensive industry, biorefineries and related industry). Every effort will be made to disseminate information about the goals of the project, on-going activities, progress, results and the potential benefits and opportunities afforded by the development of the project concepts. The goals of this strategy will be to:

- Enhance partners' visibility at national and international level
- Stimulate the interaction between industry partners and scientific community
- Attract the interest of related energy intensive industries
- Attract the interest of biorefineries and potential investors

During the project, the consortium will provide periodic reports including summaries that will be published in the public domain. It will include information on the expected final results and their wider societal implications, and the text will be understandable for a lay audience. The consortium will establish a website (open domain) for general dissemination, and a private (password protected) domain for dissemination among the consortium members. Each partner will ensure that the foreground generated is disseminated as swiftly as possible, once relevant IP has been protected.

The dissemination among stakeholders and potential end-users will be carried out through presentations and/or exhibition stands in industry relevant conferences. Moreover, the partners will seek to disseminate knowledge through the dedicated channels provided in European Technology Platforms, Cordis, other relevant EU websites and www.c1net.co.uk, the website of the BBSRC NIBB, C1net: Chemicals from Gas.
 
Description Gas eating bacteria have the potential to consume the waste gases responsible for global warming and convert them into the chemicals and materials a modern society needs. The bacterium being worked on here is called Acetobacterium woodii, and is particularly adept at growing of carbon dioxide. In this project we have set out to make a platform chemical 3-hydroxypropanoic acid (3-HP) which can be converted into many useful materials, such as nylon. As the organism does not naturally make 3-HP, it needs to be engineered to do so for which gene tools are required. Unfortunately, the available tools at the start of the project were rudimentary in nature. For competitive commercialisation, improved genetic toolsets are essential to successfully utilise a new bacterial chassis through stably inserting novel operons or deleting/disrupting competing pathways.

As Nottingham's part of the consortium, we have successfully applied the Clostridia Roadmap for gene tool development. Initially, plasmid transformation efficiencies were improved more than 3 orders of magnitude over existing published methodologies to 105 cfu/µg DNA. A defective Gram positive replicon was identified and used to construct a knockout plasmid that generated an auxotrophic pyrE mutant. Next, a pyrE complementation vector was designed and successfully tested. Through using pyrE as a counter selection marker, a knockout Allele-Coupled Exchange was developed that allows the reproducible generation of clean, in-frame deletions. The system was exemplified through disruption of A. woodii glucose, fructose and ethanol growth phenotypes via the deletion of a key enzyme in each of these three uptake pathways.

In addition to the development of tools for directed gene knock-outs, a random mutagenesis system was formulated. The alternate sigma factor TcdR was integrated into the pyrE locus allowing implementation of mariner transposition in the strain through suicide delivery of a vector carrying a mini transposon and the transposase under the control of the tcdB promoter. The system was used to generate a random mutant library pool comprising over 1 million individual mutants. Next generation sequencing of this pool following their growth on glucose has allowed the identification of all of those genes (ca. 400) that are essential for growth on glucose, characterised as genes which contain essentially no transposon insertions. Currently this analysis is being repeated on mutant pools prepared from cells that have been grown on carbon monoxide. This will allow the identification of all those genes/ enzymes needed for growth on CO2. In further experiments, cells will be grown in the presence of sub-inhibitory concentrations of 3-HP, thereby identifying genes/proteins important in tolerance to this product, laying the foundation for rational routes to increasing its production levels.
Exploitation Route The primary outcome of the project is the development of a new platform microorganism useful in the conversion of CO2 into useful chemicals. The project has put in place the general molecular biology methods needed for efficient engineering of future production organisms. It has also revealed those genes and enzymes that are important for CO2 utilisation, providing crucial information for future engineering. These tools have advanced the field of industrial biotechnology through future research collaborations and dissemination of project results, through the initial presentation of the results at the annual conferences organised by the Network in Industrial Biotechnology and Bioenergy (NIBB), C1net. Publications describing these advancements are shortly to be submitted.

The outcome has led to the award of a HORIZON 2020 project that seeks to convert the CO2 component of biogas derived from anaerobic digestion and landfill into the commercial solvent, acetone. Successful proof-of-concept for CO2 + H2 fermentation will provide an offering to customers in CO2 intensive industries to monetize gaseous waste streams while lowering emissions. The project opens new markets for the industry partners. The technology also forms the basis of a GCRF HUB bid that seeks to exploit Municipal Solid Waste (MSW) in India for conversion to value-added products.
Sectors Chemicals,Energy,Environment,Manufacturing, including Industrial Biotechology

 
Description BIOTEC-05-2017
Amount € 6,986,910 (EUR)
Funding ID 760994-2 
Organisation European Commission 
Department Horizon 2020
Sector Public
Country European Union (EU)
Start 01/2018 
End 12/2021