Bioreactor system for Industrial Biotechnology process development.

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.

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.

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.