Using synthetic biology to manipulate bacterial social behaviours to maximise the microbial degradation of environmental waste plastics.

Lead Research Organisation: Brunel University London
Department Name: Life Sciences

Abstract

Plastic pollution is a growing worldwide problem, with 12,000 million metric tonnes of plastic waste predicted to be in landfill or the environment by 2050. Whilst recycling can give a second life to some plastic, not all plastic types are easily recycled and cost can be a limiting factor, resulting in only 9% of plastic waste having ever been recycled. If plastic is not recycled, it is either sent to landfill where it pollutes the soil and run off pollutes the global water systems, or it is incinerated, releasing toxic fumes and adding to carbon emissions. Plastic pollution is damaging the environments of animals all the way down to microbes, ultimately leading to ecosystem collapse. An environmentally friendly alternative is biodegradation of plastic by microorganisms into non-toxic breakdown products, some of which can be valorised with downstream industrial uses, such as polyethylene glycol, which has many uses including acting as an anti-foaming agent in food and drinks.
Multiple species of bacteria have been found to degrade waste plastic, however many of these act at a slow rate resulting in only a small reduction in plastic weight over a period of months. An example of this is an environmental consortia of plastic degrading microorganisms which was able to decrease polystyrene weight by ~5% in 6 months. Recent research has focused on identifying the enzymes produced by these bacteria and fungi that are capable of breaking down waste plastic and exploring if these enzymes can be modified to increase their ability to degrade waste plastic. While these approaches have yielded some improvements, these enzymes are still a long way off being a viable solution to tackling the plastic waste problem.
The majority of research in the field is currently focused on optimising the plastic eating enzymes themselves to improve their activity. In this proposal we aim to take a novel approach, exploring the frontiers of activity optimisation by modifying how bacteria behave, to increase their ability to degrade plastic. Bacteria like to attach to surfaces in communities called biofilms because, just like people who live villages, towns or cities, bacteria in biofilms are better protected from the environment and can share resources and nutrients with each other. To build a biofilm, bacteria produce a slime called an exopolysaccharide which surrounds the community. A good example of a bacterial biofilm that everyone has encountered at some point is dental plaque. This is a community of bacteria who like to grow in the mouth, so attach to our teeth and form a biofilm to help them stay in this environment. In this proposal we plan to harness this behaviour of bacteria using genetic engineering to trick plastic degrading bacteria into forming large biofilms on the surface of waste plastic. Forming a biofilm on waste plastic has two major advantages. The first advantage is that it increases the concentration of plastic degrading enzyme around the waste plastic and the more enzyme means the more degradation. The other major advantage is that the exopolysaccharide slime being produced by the bacteria will stop the enzymes from being washed away. We have performed some preliminary tests in the lab to show that using genetic engineering we can increase the levels of bacteria biofilm formation on waste plastic. We have also tested our approach using well-known plastic eating enzymes and shown that increasing the levels of biofilm formation leads to a major increase in the levels of plastic degradation. In a high risk, high reward strategy we now want to test our approach against the most common and difficult to degrade waste plastics such as Polyethylene terephthalate (PET). We also want to test this approach in a bioreactor where bacteria are fed waste plastic. We believe in the future; every house could have their own microbial plastic degrading bioreactor and this research could be the first steps in making this a reality.

Publications

10 25 50
 
Description Thus far, Objective 1 and 2 have been successfully delivered for this project. These objectives focused on the identification and characterisation of novel plastic degrading enzymes and determining the impact that enhancing biofilm formation has on the activity of these enzymes. The outcomes of these two objectives have resulted in 2 publications (PMID: 37515381 and 37788986). These publications demonstrate that plastic associated biofilms are a potential source of novel plastic degrading enzymes. We also demonstrate that modulating cyclic-di-GMP signalling can be used to augment the plastic degrading efficiency of plastic degrading bacteria. The final objective, Objective 3 "Explore the capacity of these engineered bacterial strains to valorise mixed plastics in an enclosed batch reactor system" is still live.
Exploitation Route This research has the capacity to accelerate plastic degradation in a closed system by enhancing the efficiency of plastic degradation. The outcomes could be exploited academic or non-academic partners to improve the overall rates of plastic degradation by specific microbes.
Sectors Chemicals

Environment

 
Description Engineering Biology Missions Hubs and Mission Awards
Amount £13,211,207 (GBP)
Organisation Biotechnology and Biological Sciences Research Council (BBSRC) 
Sector Public
Country United Kingdom
Start 02/2024 
End 02/2029
 
Title Biofilm Modulating Genetic Circuits for Plastic Degradation 
Description We designed a developed a series of genetic circuits that can be integrated into a variety of host bacteria that can offer tuneable control over biofilm formation. 
Type Of Material Technology assay or reagent 
Year Produced 2023 
Provided To Others? Yes  
Impact We demonstrated that these circuits can be used to improve the efficiency of previously characterised and novel plastic degrading enzymes. The circuits are being made available to the members of the Environmental Biotechnology Innovation Centre as well as the wider research community. 
 
Title In vitro Plastic Associated Biofilm Enrichment 
Description We developed a validated a methodology to perform lab adapted evolution of plastic associated biofilm communities, validating that this approached enriched for species that could use plastic substrates as a carbon source and that the resulting species could be a rich source of novel plastic degrading enzymes. 
Type Of Material Technology assay or reagent 
Year Produced 2023 
Provided To Others? Yes  
Impact We have had discussions with various research groups about how to implement this methodology. 
URL https://enviromicro-journals.onlinelibrary.wiley.com/doi/epdf/10.1111/1462-2920.16466
 
Description Interview for BBC Radio 4 Inside Science Programme 
Form Of Engagement Activity A press release, press conference or response to a media enquiry/interview
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Media (as a channel to the public)
Results and Impact I was invited to speak on BBC Radio 4 Inside Science programme on several aspects of our work to inhibit growth and biofilm formation in bacteria such as Pseudomonas aeruginosa and Acinetobacter baumannii. This was broadcast in March 2023 but is available to listen to at any point on BBC iPlayer
Year(s) Of Engagement Activity 2023
URL https://www.bbc.co.uk/programmes/m001k11r
 
Description Invited Seminar at University of Texas at Austin 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Postgraduate students
Results and Impact Invited to speak about current research ongoing in group at a seminar in the University of Texas at Austin. In this seminar several projects were discussed and immediately after the seminar I met with several academics and postgraduate researchers one to one to discuss the projects, methodologies and areas for collaboration. I also had follow up online meetings to detail specific methodologies and collaboration opportunities.
Year(s) Of Engagement Activity 2023
 
Description New Scientist Live - presenter; outreach event 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Public/other audiences
Results and Impact New Scientist Live is the world's greatest festival of ideas and discoveries and happens every year in October in London and is attended by about 20,000 people each year. In 2022 there were 56 main stage talks and 80 different exhibitors presenting the latest innovations from the scientific world ranging from space to medicine and agriculture. Lyuboslava Harkova was an exhibitor, representing McCarthy lab group and Brunel University London as part of the National Biofilms Innovation Centre (NBIC) team. She spent the day talking to people from all walks of life and various ages about biofilms, my research at Brunel and why it is important to investigate these communities more. She presented different biofilms-related activities to promote better understanding and engagement with the topic.
Year(s) Of Engagement Activity 2023
 
Description Speaker at ASM Microbe 2023 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Postgraduate students
Results and Impact Invited to speak at the American Microbiology Society annual Microbe Conference in Houston Texas. This presentation sparked several discussions afterwards and potential collaboration opportunities that are still being explored.
Year(s) Of Engagement Activity 2023