Engineered Biofilm Catalysts

Lead Research Organisation: University of Birmingham
Department Name: Chemical Engineering

Abstract

Historically, biofilms are predominantly perceived as problematic; associated with infections, dental caries, marine and reactor fouling and reduction in heat transfer. In their natural environment individual planktonic cells have the tendency to cluster together at surfaces and interfaces. Within these clusters they protect themselves from environmental and chemical stress by secreting extracellular polymeric substances (EPS) a protective and adhesive polysaccharide matrix. This EPS gives the biofilm a degree of protection against extremes of pH, temperature or the introduction of organic solvents which cannot be tolerated by individual cells. In the current global climate of searching for increasingly green approaches to the synthesis of fine chemicals and pharmaceuticals there is a growing interest in employing biotransformations: using cells to produce useful products. Although such a strategy enables the generation of enantiomerically pure compounds, the reduction of steps in a synthetic route and the possibility of eliminating environmentally detrimental catalysts, solvents and reagents, their use is problematic due to the fact that individual cells cannot in general tolerate the extreme conditions required. In this research we exploit biofilms to produce highly active Engineered Biofilm Catalysts (EBC). This is a new area as the use of biofilms in biotransformations has been largely confined to wastewater treatment and bioremediation generally by mixed microbial communities referred to as consortia. Reports of the use of biofilms for synthesis of compounds are few and far between, industrially the list of compounds generated in such a manner is little more extensive than simple compounds such as acetic acid, ethanol, butanol, 2,3-butanediol, lactic acid, fumaric acid, and succinic acid. This interdisciplinary proposal presents a novel methodology for the development and exploitation of engineered biofilm catalysts (EBC) for biotransformations relevant to the fine chemicals and pharmaceutical industries. Recently we have demonstrated the generation and utilisation of an Engineered Biofilm Catalyst (EBC). The immobilised EBC not only demonstrated unprecedented stability but proved to be a strikingly better catalyst than the free cells. As the EBC is artificially generated there is considerable scope to engineer its microstructure. We propose to explore the general application of EBC to catalysis as well as to investigate how microstructure and gene expression relate to catalytic activity. We will employ EBC using flow chemistry to generate a prototype EBC reactor for biotransformations as an exemplar for future industrial exploitation.

Technical Summary

In the current global climate of searching for increasingly green approaches to the synthesis of fine chemicals and pharmaceuticals there is a growing interest in employing biotransformations. Although such a strategy enables generation of enantiomerically pure compounds, reduction of steps in a synthesis or the possibility of eliminating environmentally detrimental materials, their use is problematic as individual cells cannot in general tolerate the extreme conditions required. This interdisciplinary proposal presents a novel methodology to exploit the tolerance to extreme conditions of biofilms in highly active Engineered Biofilm Catalysts (EBC). This is a new area: the use of biofilms in biotransformations has been largely confined to wastewater treatment and bioremediation generally by consortia. Recently we have demonstrated the generation and utilisation of an EBC using Escherichia coli K-12 ompR234 pretransformed with pSTB7 a plasmid enabling the over expression of tryptophan synthase. This enzyme mediates generation of a series of L-halotryptophans from serine and haloindoles. The immobilised EBC not only demonstrated unprecedented stability but proved to be a strikingly better catalyst than the free cells. In this research we propose to -Investigate physical and chemical parameters governing the microstructure and activity of the K-12 E. coli EBC and make preliminary investigations as to the role of gene expression in specifying the structure and function of the engineered biofilm; -Apply the EBC concept to other biotransformations and exploring the possibility of generating an EBC with other organisms for example antibiotic producers such as Saccharopolyspora erythraea; -Explore the possibility of a multifunctional EBC (in order to produce multiple products) using gene promoter switching; -Provide a preliminary assessment of scalability and feasibility of an EBC bioreactor via construction of a lab-scale prototype for flow chemistry

Planned Impact

Through this research we have potential to make a demonstrable contribution to impact over a multiple year timeframe. The main focus of impact will be Scientific Advancement and Knowledge (0-3+ years). The fundamental research programme within this project will be disseminated via high quality scientific journals in the field (e.g. Biotechnology and Bioengineering, Nature Biotechnology, Angewandte Chemie). We intend to present our work at 2 leading international conferences as well as national meetings. Processes and Products (3-10 years). The potential beneficiaries of this research programme beyond the project timescale are industries involved in the manufacture of fine chemicals, pharmaceutical intermediates and pharmaceutical products. Whilst the research is currently fundamental in nature and at a low level of technology readiness, through this proposal we will engage with industry at the earliest opportunity via our Steering Group and seek to expand our industry portfolio via CIKTN and existing contacts. In the long term, as well as the cost and environmental savings our new technologies could bring, there may be a positive impact on public health via cheaper and more available products. We take the opportunity for engagement and leverage at the annual Formulation Engineering Conference (FEC) held at Birmingham, where we will hold a separate spin-out seminar in years 1 and 2 of the project to which all key stakeholders including the above contacts are invited. Through the Natural Products and Drug Discovery undergraduate and MSC module at UEA, which Dr Goss coordinates, there are existing interactions with GSK (Richard Hartley), Astra Zeneca (Duncan Gill and Andy Wells), Novartis (Brian Cox) Novacta (Mike Dawson, Antony Appleyard) and Biotica (Barrie Wilkinson). This network will provide a good initial point for dissemination of research and fine-tuning of the programme as the research progresses in order to meet the needs of industry and SMEs. We will hold a Research Showcase Event in the final year of the project to disseminate results of the project to Industry and promote exploitation (resource is requested to support this event). Training and Investment in People (Jobs) (0-3+ years). This research presents the opportunity for exchanges within the research team to develop novel research ideas and shared knowledge beyond the traditional discipline boundaries. Simmons and Goss have already performed exchanges via discipline hopping project and the development of the EBC concept is directly attributable to this. The continued exchange of personnel between laboratories is critical to the success of the project and builds a highly capable research workforce with excellent employment potential. Further knowledge exchange will be accomplished by -weekly 'sheep-dips' -month long exchanges 'marinades' Additional Communication Resources (0-3 years). We will develop a website to disseminate the work which will be linked from websites accessed by stakeholders including IChemE Biochemical Engineering and Pharma Subject Groups, and Royal Society of Chemistry interest group pages including those of the BioOrganic Group as well as inputting into Knowledge Transfer Networks such as CIKTN.
 
Description We have developed a technology which enables cells to immobilise themselves on the surface of a reactor. These cells are genetically engineered to produce enzymes (complex molecules) or other useful chemicals in an atom-efficient way, reducing wastage compared with conventional wet chemistry or catalysis. The system has the potential to be self- regenerating and is tunable, so that different families of molecules may be produced depending on how the reaction is initiated.
Exploitation Route In the pharmaceutical and healthcare sectors as a more efficient production method for complex molecules (proteins, enzymes, small molecule drugs)
Sectors Chemicals,Manufacturing, including Industrial Biotechology,Pharmaceuticals and Medical Biotechnology

 
Description The research carried out in this project was fundamental in nature. The primary impact of the work has been to prepare further grant applications on the implementation of the engineered biofilms principle to other reaction systems and other reactor designs, including monolith bioreactors.
First Year Of Impact 2015
Sector Chemicals,Manufacturing, including Industrial Biotechology,Pharmaceuticals and Medical Biotechnology
Impact Types Societal,Economic

 
Description EPSRC Follow on Fund
Amount £11,002 (GBP)
Funding ID EPSRC UOBFOF23 
Organisation Engineering and Physical Sciences Research Council (EPSRC) 
Sector Public
Country United Kingdom
Start 09/2013 
End 09/2015
 
Description Collaboration between Birmingham, St. Andrews and University College London 
Organisation University College London
Department Biochemical Engineering
Country United Kingdom 
Sector Academic/University 
PI Contribution A new collaboration was made between the investigators of this project and other investigators in St. Andrews and UCL. The group prepared and submitted two grant applications in 2016 to InnovateUK and BBSRC. The group continue to discuss collaborative working.
Collaborator Contribution As above - preparation of grant applications, exchange of materials and know-how.
Impact Two grant applications: InnovateUK IB Catalyst; and BBSRC sLOLA. Neither was successful. This is a multidisciplinary collaboration: Microbiology; fluid dynamics; organic chemistry; biocatalysis; biophysics.
Start Year 2015
 
Description Collaboration between Birmingham, St. Andrews and University College London 
Organisation University College London
Department Department of Chemistry
Country United Kingdom 
Sector Academic/University 
PI Contribution A new collaboration was made between the investigators of this project and other investigators in St. Andrews and UCL. The group prepared and submitted two grant applications in 2016 to InnovateUK and BBSRC. The group continue to discuss collaborative working.
Collaborator Contribution As above - preparation of grant applications, exchange of materials and know-how.
Impact Two grant applications: InnovateUK IB Catalyst; and BBSRC sLOLA. Neither was successful. This is a multidisciplinary collaboration: Microbiology; fluid dynamics; organic chemistry; biocatalysis; biophysics.
Start Year 2015
 
Description Collaboration between Birmingham, St. Andrews and University College London 
Organisation University of St Andrews
Department School of Biology
Country United Kingdom 
Sector Academic/University 
PI Contribution A new collaboration was made between the investigators of this project and other investigators in St. Andrews and UCL. The group prepared and submitted two grant applications in 2016 to InnovateUK and BBSRC. The group continue to discuss collaborative working.
Collaborator Contribution As above - preparation of grant applications, exchange of materials and know-how.
Impact Two grant applications: InnovateUK IB Catalyst; and BBSRC sLOLA. Neither was successful. This is a multidisciplinary collaboration: Microbiology; fluid dynamics; organic chemistry; biocatalysis; biophysics.
Start Year 2015
 
Description ITN Industrial Training Network 
Organisation University College Dublin
Country Ireland 
Sector Academic/University 
PI Contribution Development of an ITN proposal for H2020 around the industrial applications of biofilms
Collaborator Contribution Development of an ITN proposal for H2020 around the industrial applications of biofilms
Impact ITN application in progress.
Start Year 2013