Bioreactor system for Industrial Biotechnology process development.

Lead Research Organisation: Aberystwyth University
Department Name: IBERS

Abstract

Industrial biotechnology is a core research platform at the Institute of Biological Environmental and Rural Sciences (IBERS) at Aberystwyth University (AU). This platform will benefit greatly from an automated bioreactor system that will enable 16 simultaneous fermentations to be performed on a multi-millilitre scale (50-250ml) enabling rapid development of industrially scalable fermentation processes.

Data from this system will enable identification and testing of multi-factorial parameters critical to fermentation processes in order to optimise productivity and yield, prior to scale-up to an industrially relevant bioprocess. It will assess inhibitors produced as a result of pre-processing technologies and allow studies on microbe-microbe interactions and microbe communities. This equipment would result in a major reduction in time and expense in process optimisation and provide a step change increase in data acquisition and analysis.

Additionally this system will enable identification of pathways and metabolic networks by complementing the metabolomics and next generations sequencing data sets produced at IBERS and elsewhere. The advent of the omics era has bought about an improved understanding and modelling of microbial metabolism in traditional, academically characterised bacterial and yeast fermentation systems such as Escherichia coli and Saccharomyces cerevisiae. However, this level of understanding rarely exceeds the scale of shake flask experimentation and pertinently may not reflect IB conditions or industrial production strains. Although tightly controlled, environmental heterogeneity for pH, temperature, rheology product titre and substrate availability is experienced in fermentation vessels during scale-up to industrially relevant volumes (multiple hundreds of litres). There is a paucity of academic and industrial knowledge on the impact of this heterogeneity on cellular physiology, metabolic pools and alterations in global gene expression of both academic model and industrial biocatalysts during scale-up.

This equipment will complement existing fermentation systems at IBERS which includes 6x1L, 4x10L, 1x30L, 2x70L and 1x250L automated, precision controlled reaction vessels, allowing for similar control on a smaller scale. Au will provide technical support and make this equipment available to both the academic and industrial community and will highlight it on web sites, equipment databases and brochures.

Technical Summary

A multi-bioreactor system is requested which contains 16 reactor vessels each with accompanying ports, pumps, individual Peltier heating, condensation reduction, sensors for pH, dissolved oxygen, temperature, foaming and gas monitoring. This is controlled and monitored by a computer based control system. A 16 vessels system is required for an L16 Taguchi Design of Experiment orthogonal array. This allows for the simultaneous assessment of key process variables across the system. It also allows for replication after initial process development. We assessed other technologies including Biolector, but rejected this on the basis of vessel geometry and lack of scale up potential as well as non-standardised probes and sensors when compared to industrial units. The Applikon 250 ml MiniBio range of bioreactors is a true scale-down of the laboratory scale bioreactors in the 1 to 20 litre range. The MiniBio systems have the same flexibility as the laboratory scale and reflect pilot to industrial scale bioreactors. This means that the MiniBio systems can be customised to fit the demands of any process. The small volume reduces: 1) the production and costs of cultivation medium (e.g lignocellulosic hydrolysates); 2) the use of expensive bench space (approx. 1.8 m2) and 3) the time taken to generate large, continuous sets of scientific data. The Lucullus software communicates with the my-Control graphical interface and is compatible with Linux and Windows based operating systems with data export including both standard and processed formats.

The fermentation data will complement bioinformatics data generated on the response and stability of microbial cell physiology undergoing bio-processes development. The integration of these data with RNA seq data for both wild-type and improved strains designed through rational genome editing will provide detailed information on biocatalyst performance and provide essential key molecular performance indicators during scale-up.

Planned Impact

The Applikon MiniBio modular fermentation system will complement the pilot scale biorefining fermentation capability at IBERS and the expansion of this facility in the planned Aberystwyth Innovation and Enterprise Campus (AIEC). This equipment will add impact to the Universities aspirations in the area of biorefining and will have impact in the sourcing of additional funding and translational activities that will be supported by IBERS' embedded impact team and the AU Technology Transfer Team to deliver the strategy for knowledge exchange. The new capability will be made available to MSc and PhD research training programs which will enhance and improve the graduate programmes at IBERS.
Knowledge will be communicated and disseminated to the wider scientific, industrial and public audience through high impact publications, NiBB meetings and outreach activities with schools, Government and industrial bodies. The Applikon MiniBio system will be highlighted to academia and industry on a number of brochures and websites including: Biorefining and Bioconversion Group site (www.aber.ac.uk/en/ibers/research/research-groups/bioconversion_biorefining), the BEACON site (www.beaconwales.org), the NiBB P2P site with links to the equipment database (www.nibb-p2p.org/users/login), the new AIEC site (under construction) and the Biopilot UK site (http://biopilotsuk.com). The applicants will work with a range of companies to trouble shoot fermentation barriers and help in the translation of underpinning research and newly discovered microorganisms with industrial potential for applications in a range of industries. We will provide booking and costing structures and provide a technical feasibility assessment of potential project collaborations in association with academic and industrial sectors.
Building on the scientific excellence of IBERS, the Bioconversion & Biorefining Group have a portfolio of national and international industrially focused research programs that aim to stimulate bioeconomic growth and mitigate the impact of climate change. Selected examples of extended research programs that would generate impact from this equipment initiative include: 1) A recently awarded BBSRC/FAPESP project (JG Co-I) investigating the production of oligomeric sugars for conversion to bioethanol from lignocellulosic feedstocks. The bioethanol partner in this project is looking to integrate this new technology to replace existing processes; 2) A BBSRC ERA-Net project 'Oxypol' (JG UK PI) where major companies involved in this project, the Siam Cement Company and Norner are looking to develop this technology, to produce new polymers to replace those from the petro-chemical industry. The Siam Cement company are hoping to add on a processing facility to their existing kraft lignin processing facility in Thailand; 3) Optimised production of lactic acid from municipal solid waste (MSW) (JG PI). Successful demonstration of an economically viable process will encourage Fiberight to invest in the first UK plant, to be built in the UK, within the next 5 years.
Dedicated, experienced, trained research staff will be on hand to provide fermentation and design of experiment training to a range of users including but not limited to: Research scientists, students and employees of commercial organisations in need of up-skilling their work force in fermentation. We will use Knowledge Transfer Partnerships to facilitate training as part of continuing professional development in biotechnology. Access to this training will help build and expand the UKs excellence in industrial biotechnology research and delivering the next generation of academic and industrial fermentation scientists.

Publications

10 25 50
 
Description This funding enabled the purchase of 16, fully controllable, mini bioreactors which have been used in a number of research projects and for commercial research with both academic, including undergraduate and postgraduate students, and industry partners. These reactors has been used to optimize fermentation processes prior to scale up and to understand microbial growth, recombinant protein expression, morphology changes, metabolic systems as well as optimizing product production and inhibitor tolerance. The mini bioreactors has been used to optimize the production of; lactic acid by mesophiic and thermophilic microorganisms; proteins for use in novel glues; pre-biotics from grass juice using recombinant hydrolases; platform chemicals such as ethanol and xylitol from a range of different feedstocks including straws. They have also be used to optimize lignocellulosic hydrolysis from a range of feedstocks including municipal solid waste and apple pomace. Additionally there reactors have been used to help understand morphology shifts in Candida species and to assessment fermentation characteristics of genetically modified strains in comparison to the wild type.
Exploitation Route This mini bioreactor system allows researchers and industry to optimize both hydrolysis and fermentation processes, with replication, to develop an optimized system for scale up. This will increase speed to market or generate a better understanding of fundamental science questions.
Sectors Agriculture, Food and Drink,Chemicals,Energy,Manufacturing, including Industrial Biotechology

 
Description BEACON+ Biorefing Centre East Wales funded through ERDF via Welsh European Funding Office
Amount £443,888 (GBP)
Funding ID 82163 
Organisation Welsh Assembly 
Sector Public
Country United Kingdom
Start 01/2019 
End 12/2021
 
Description BEACON+ Biorefing Centre West Wales & the Valleys Extension funded through ERDF via Welsh European Funding Office
Amount £3,782,889 (GBP)
Funding ID 80851 
Organisation Welsh Assembly 
Sector Public
Country United Kingdom
Start 01/2019 
End 12/2021
 
Description Commercial
Amount £56,000 (GBP)
Organisation University of Bath 
Sector Academic/University
Country United Kingdom
Start 07/2018 
End 10/2018
 
Description European regional development fund
Amount € 2,884,534 (EUR)
Organisation Welsh Assembly 
Sector Public
Country United Kingdom
Start 04/2018 
End 03/2023
 
Description ICURE Follow on fund
Amount £296,628 (GBP)
Organisation Innovate UK 
Sector Public
Country United Kingdom
Start 01/2019 
End 03/2020
 
Description Industrial research
Amount £724,309 (GBP)
Funding ID TS/S003177/1 
Organisation Innovate UK 
Sector Public
Country United Kingdom
Start 12/2018 
End 05/2020
 
Description Innovate UK: EcoXyl scale up
Amount £107,000 (GBP)
Funding ID TS/S019456/1 
Organisation Innovate UK 
Sector Public
Country United Kingdom
Start 03/2019 
End 03/2020
 
Description KESS WEFO Optimising microbial production of lactic acid from municipal solid waste (MSW); Insights into genome stability for industrial biotechnology
Amount £71,000 (GBP)
Funding ID C80815 
Organisation European Social Fund (Welsh Government/ EU) 
Sector Public
Country United Kingdom
Start 08/2018 
End 07/2021
 
Description Materials and Manufacturing Round 3
Amount £398,801 (GBP)
Funding ID TS/R010544/1 
Organisation Innovate UK 
Sector Public
Country United Kingdom
Start 04/2018 
End 10/2020
 
Description Newton Bhabha Industrial Waste BIOREVIEW
Amount £1,001,109 (GBP)
Funding ID BB/S011994/1 
Organisation Biotechnology and Biological Sciences Research Council (BBSRC) 
Sector Public
Country United Kingdom
Start 09/2018 
End 08/2021
 
Description Open call
Amount £319,041 (GBP)
Funding ID TS/R010544/1 
Organisation Innovate UK 
Sector Public
Country United Kingdom
Start 05/2018 
End 12/2020
 
Description Phase 2 synthetic biology for transparent materials
Amount £149,999 (GBP)
Organisation Defence Science & Technology Laboratory (DSTL) 
Sector Public
Country United Kingdom
Start 07/2018 
End 06/2019
 
Description Value added materials from organic waste sugars VAMOS Horizon 20:20 BBI
Amount £11,872,746 (GBP)
Funding ID H2020-BBI-JTI-2018-837771 
Organisation European Commission 
Sector Public
Country European Union (EU)
Start 09/2019 
End 08/2022
 
Description AIEC opening 
Form Of Engagement Activity Participation in an open day or visit at my research institution
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Professional Practitioners
Results and Impact Opening of AU new Biorefining centre. Presentations were made on current biorefining projects along with equipment available to academics and industry.
Year(s) Of Engagement Activity 2019
 
Description Beer and cider special interest group 
Form Of Engagement Activity A formal working group, expert panel or dialogue
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Industry/Business
Results and Impact Presentation on capability and research projects
Year(s) Of Engagement Activity 2018
 
Description IBERS Centenary Celebrations 
Form Of Engagement Activity Participation in an open day or visit at my research institution
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Professional Practitioners
Results and Impact Celebration of IBERS 100 years in plant breeding. Presentations given on biorefining projects and available equipment.
Year(s) Of Engagement Activity 2019
 
Description Newton Bhabha UK-India industrial waste challenge 2nd to 3rd March 2020 Indian Institute of Technology Mumbai Mid-term review meeting 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Industry/Business
Results and Impact The objectives for the mid-term review were:
To review progress on each project covering each work-package, reviewing progress against the project plan and examining the outputs of the work so far.
To review the interaction between the national/ international partners, covering both academics and businesses and their interactions with their respective funding partners on each project to identify issues affecting progress and to elicit suggestions to facilitate progress in the second half of the programme.
To review progress on the programme and any added value for the programme as a whole from interactions between individual project staff that has taken place to date.
To provide assurance to the sponsors about the investment of funds and that value for money has (or will be) achieved by the end point of the programme.

During which the use of the BBSRC investment in the mini bioreactor system at AU was highlighted as a game changing advancement in bioprocess optimisation fo the IB production of xylitol from lignocellulosic hydrolysates.
Year(s) Of Engagement Activity 2019
 
Description Newtown rotary Club Visit 
Form Of Engagement Activity Participation in an open day or visit at my research institution
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Public/other audiences
Results and Impact Presented current and post research in the area of biorefining.
Year(s) Of Engagement Activity 2019
 
Description Presentation 4th LB conference 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Industry/Business
Results and Impact Oral presentation at the 4th LBNet International Conference, 20-22 March 2019 Cheshire UK. Presented on developments in the production of novel pre-biotics and xylitol.
Year(s) Of Engagement Activity 2019
 
Description Visit from UTM 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact Return Visit by UTM team for further discussion on potential collaborative research and studentships etc.
Year(s) Of Engagement Activity 2020
 
Description Visit to Universiti Teknologi Malaysia (UTM) 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact Major discussions with Staff at both of the UTM campuses on areas of collaboration in Industrial biotechnology, bio-circular economy, engagement with Business, possible studentships and available equipment.
Year(s) Of Engagement Activity 2019