Aromatic feedstock chemicals from degradation of lignin
Lead Research Organisation:
University of Warwick
Department Name: Chemistry
Abstract
One of the biggest problems facing society today is our dependence on dwindling fossil fuels, and the contribution to global warming of power stations, car transport and domestic heating that make use of fossil fuels. Not only is oil used to prepare fuel, but also an essential by-product of oil refining is the production of raw materials for chemical and pharmaceutical synthesis. In 50-100 years time, all of these chemicals will need to be produced from renewable, non-petroleum sources, but at present we have very limited methods to do this, so a lot of new technology needs to be developed in a relatively short space of time. The carbon content of plant lignocellulose, found in plant cell walls, represents an abundant source of renewable carbon. One component of lignocellulose is an aromatic polymer called lignin, that binds the cellulose cell walls together. Lignin is very hard to break down, so at present lignocellulose is broken down via a 'pre-treatment' step, usually involving heating with acid and steam to 200 oC, which consumes a lot of energy. Therefore, if we could use Nature to break down the lignin, we would improve the efficiency of lignocellulose breakdown, and liberate useful aromatic by-products. We have recently developed a new method for identifying strains of bacteria that are able to break down lignin, and we have already found several strains of bacteria that are able to do this. We will isolate the lignin-degrading enzymes from these strains, and use molecular genetics to produce large quantities of these enzymes for further studies. We will then use these strains to try to produce useful aromatic chemicals from breakdown of lignocellulose. One example is vanillin, which is used for food flavouring in products such as vanilla ice cream; other examples are phenols that are used in the manufacture of plastics.
Technical Summary
Lignin is an aromatic polymer that is a major component of plant lignocellulose. Lignin is extremely hard to degrade, but represents an important source of aromatic carbon compounds, that could potentially be converted into useful, high value aromatic feedstocks. We have recently developed novel methods for the identification of bacteria that are able to degrade lignin, and we have already identified two strains of bacteria with this capability. The aim of this programme is to find novel lignin-degrading bacterial strains and enzymes, and to use them to convert the lignin component of lignocellulose into useful aromatic chemicals. In Part A, we will clone the genes encoding lignin-degrading enzymes from lignin-degrading strains, and the encoded enzymes will be expressed, purified, and characterised biochemically. Lignin-degrading strains, and recombinant lignin-degrading enzymes, will be tested for their ability to break down lignin and lignocellulose in small- and large-scale trials. In Part B, we will apply the lignin-degrading enzymes to convert plant lignocelluose into renewable feedstocks. Lignin-degrading strains and recombinant enzymes will be tested for their ability to produce the following high-value aromatic chemicals, by monitoring their production using HPLC and LC-MS methods: i.) vanillin is a high value chemical that can be produced from oxidative cleavage of the beta-aryl ether component of lignin; ii.) phenol and guaicol could be produced via breakdown of lignin containing di- and mono-substituted aryl units respectively, followed by oxidation and enzymatic decarboxylation; iii.) ferulic and caffeic acid are valuable anti-oxidants that are potential lignin breakdown products; iv.) lignin-derived bio-materials can be generated from partial lignin breakdown, and substituted styrenes can be generated enzymatically, that are feedstocks for material synthesis.
Planned Impact
The development of novel routes to high-value aromatic chemicals from lignocellulose will be of considerable interest to industry. Aromatic chemicals are used widely for plastics manufacture (e.g. phenol). Vanillin is used widely in the food industry, and ferulic and caffeic acid are used in the personal care industry. There is a growing market for renewable plastics, that could be made from styrene bio-products or lignocellulose breakdown products. Generation of high-value chemicals from lignin waste would also add considerable value to 2nd generation biofuel production, and the paper/pulp industry. Choice of suitable plant feedstock for chemicals production will also be of interest to plant breeders and seed manufacturers. In the long term, this work could also be of significant commercial benefit to the UK, since alternative routes to feedstock chemicals from non-petroleum sources must be found over the next 50-100 years.
People |
ORCID iD |
Timothy Bugg (Principal Investigator) | |
Paul Norris (Co-Investigator) |
Publications
Ahmad M
(2011)
Identification of DypB from Rhodococcus jostii RHA1 as a lignin peroxidase.
in Biochemistry
Ahmad M
(2010)
Development of novel assays for lignin degradation: comparative analysis of bacterial and fungal lignin degraders
in Molecular BioSystems
Bugg T
(2011)
Pathways for degradation of lignin in bacteria and fungi
in Natural Product Reports
Bugg TD
(2011)
The emerging role for bacteria in lignin degradation and bio-product formation.
in Current opinion in biotechnology
Konstantopoulou M
(2017)
Variation in susceptibility to microbial lignin oxidation in a set of wheat straw cultivars: influence of genetic, seasonal and environmental factors - OPEN ACCESS
in Nordic Pulp and Paper Research Journal
Roberts JN
(2011)
Characterization of dye-decolorizing peroxidases from Rhodococcus jostii RHA1.
in Biochemistry
Sainsbury PD
(2013)
Breaking down lignin to high-value chemicals: the conversion of lignocellulose to vanillin in a gene deletion mutant of Rhodococcus jostii RHA1.
in ACS chemical biology
Taylor CR
(2012)
Isolation of bacterial strains able to metabolize lignin from screening of environmental samples.
in Journal of applied microbiology
Williamson J
(2020)
Production of Substituted Styrene Bioproducts from Lignin and Lignocellulose Using Engineered Pseudomonas putida KT2440
in Biotechnology Journal
Description | This project resulted in the following significant results: 1. A new colorimetric assay method was developed for monitoring lignin breakdown 2. Using this assay method, 12 new bacterial lignin-degrading bacteria were identified from soil samples, and their lignin degradation capability assessed 3. A bacterial lignin-degrading peroxidase enzyme DypB was identified in Rhodococcus jostii RHA1. This is the first bacterial lignin peroxidase to be well characterised. 4. Using gene deletion strains of Rhodococcus jostii, high levels of metabolites in lignin degradation were produced by fermentation of lignocellulose, including vanillin, a valuable chemical for the food/flavour industry |
Exploitation Route | After consultation with Warwick Ventures regarding intellectual property, the results were published in international scientific journals. |
Sectors | Chemicals |
Description | The project led to a collaboration with Biome Bioplastics (SME), that resulted in a project funded by Innovate UK, and a further project funded by the IB Catalyst scheme, to produce monomers for bioplastics production from lignin. This collaboration has then resulted in a patent application "New polymers and methods" (Inventors: P. Law, P. Mines, T.D.H. Bugg), disclosure P62672GB filed by Biome Bioplastics (via Kilburn & Strode, London) in June 2015, describing bio-based polymers from 2,4-PDCA and 2,5-PDCA derived from lignin. P62672WO/ECC filed with European Patent Office on 15th June 2016 (application number PCT/EP2016/063779). |
First Year Of Impact | 2013 |
Sector | Chemicals,Manufacturing, including Industrial Biotechology |
Impact Types | Economic |
Description | Solutions for Sustainable Energy: Exploring Industrial Biotechnology |
Geographic Reach | National |
Policy Influence Type | Contribution to a national consultation/review |
Description | Evaluation of a novel process to obtain a high value polyester intermediate from lignin |
Amount | £65,207 (GBP) |
Funding ID | 131141 |
Organisation | Innovate UK |
Sector | Public |
Country | United Kingdom |
Start | 05/2013 |
End | 04/2014 |
Description | Investigation of the chemocatalytic and biocatalytic valorisation of a range of different lignin preparations |
Organisation | Imperial College London |
Department | Department of Life Sciences |
Country | United Kingdom |
Sector | Academic/University |
PI Contribution | We tested samples of lignins supplied by these Universities against three biocatalytic methods for lignin degradation discovered in my research group, in a proof of concept study funded by the Lignocellulosic Biorefinery Network (LBNet). |
Collaborator Contribution | They supplied the lignin samples, and analysed the lignin structure. |
Impact | Journal publication "An investigation of the chemocatalytic and biocatalytic valorisation of a range of different lignin preparations: the importance of ß-O-4 content" C.S. Lancefield, G.M.M. Rashid, F. Bouxin, A. Wasak, W-C. Tu, J. Hallett, S. Zein, J. Rodríguez, S.D. Jackson, N.J. Westwood, & T.D.H. Bugg, ACS Sustainable Chemistry & Engineering, 4, 6921-6930 (2016). |
Start Year | 2015 |
Description | Investigation of the chemocatalytic and biocatalytic valorisation of a range of different lignin preparations |
Organisation | University of Glasgow |
Department | Department of History |
Country | United Kingdom |
Sector | Academic/University |
PI Contribution | We tested samples of lignins supplied by these Universities against three biocatalytic methods for lignin degradation discovered in my research group, in a proof of concept study funded by the Lignocellulosic Biorefinery Network (LBNet). |
Collaborator Contribution | They supplied the lignin samples, and analysed the lignin structure. |
Impact | Journal publication "An investigation of the chemocatalytic and biocatalytic valorisation of a range of different lignin preparations: the importance of ß-O-4 content" C.S. Lancefield, G.M.M. Rashid, F. Bouxin, A. Wasak, W-C. Tu, J. Hallett, S. Zein, J. Rodríguez, S.D. Jackson, N.J. Westwood, & T.D.H. Bugg, ACS Sustainable Chemistry & Engineering, 4, 6921-6930 (2016). |
Start Year | 2015 |
Description | Investigation of the chemocatalytic and biocatalytic valorisation of a range of different lignin preparations |
Organisation | University of St Andrews |
Department | School of Chemistry St Andrews |
Country | United Kingdom |
Sector | Academic/University |
PI Contribution | We tested samples of lignins supplied by these Universities against three biocatalytic methods for lignin degradation discovered in my research group, in a proof of concept study funded by the Lignocellulosic Biorefinery Network (LBNet). |
Collaborator Contribution | They supplied the lignin samples, and analysed the lignin structure. |
Impact | Journal publication "An investigation of the chemocatalytic and biocatalytic valorisation of a range of different lignin preparations: the importance of ß-O-4 content" C.S. Lancefield, G.M.M. Rashid, F. Bouxin, A. Wasak, W-C. Tu, J. Hallett, S. Zein, J. Rodríguez, S.D. Jackson, N.J. Westwood, & T.D.H. Bugg, ACS Sustainable Chemistry & Engineering, 4, 6921-6930 (2016). |
Start Year | 2015 |
Description | School visit (Abingdon School) |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | Regional |
Primary Audience | Schools |
Results and Impact | 19th September 2016 Visited Abingdon School, talked to 6th form students Talk: "Renewable chemicals from plant biomass: a challenge for the 21st century" |
Year(s) Of Engagement Activity | 2016 |
Description | Talk at ZELCOR Summer School. 3rd-4th September, Wageningen University, Netherlands "Potential of bacteria and synthetic biology for conversion of lignin" |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Professional Practitioners |
Results and Impact | Participation in summer school for ZELCOR EU project |
Year(s) Of Engagement Activity | 2018 |