Award details

Mycoprotein 2.0

Reference	BB/P020364/1
Principal Investigator / Supervisor	Dr Richard Harrison
Co-Investigators / Co-Supervisors
Institution	National Inst of Agricultural Botany
Department	Centre for Research
Funding type	Research
Value (£)	539,699
Status	Completed
Type	Research Grant
Start date	01/11/2017
End date	31/10/2020
Duration	36 months

Abstract

Work is proposed to elucidate the complex regulation of metabolism by different carbon sources and the effect upon the nutritional status of mycoprotein. In this proposal we will improve the ~1000 contig draft genome of Fusarium venenatum using Pacbio and sequencing followed by scaffolding with Bionano optical mapping. We will annotate the genome using orthology anlaysis, reading over information from the plant pathogen Fusarium graminearum. Using bioinformatics analysis of the improved genome, physically clustered and environmentally co-regulated genes that respond to differences in primary carbon source will be idenfied using both existing RNAseq data and new RNAseq data from further deletion lines. The construction of reporter lines using rapid modular cloning techniques, along with optimisation of CRIPSR-Cas9 mediated expression modulation, mutagenesis and deletion will allow a range of screens to be undertaken to identify global and specific signalling components modulating both deleterious secondary metabolite biosynthesis and the upregulation advantageous vitamin precursors which affect the end nutritional status of mycoprotein. Targeted knockout of pathway-specific regulators and screening for enhanced growth conditions will improve the metabolic profile of mycoprotein and advance our fundamental understanding of how this important microbe can be manipulated for enhanced industrial production. EMS mutagensis and subsequent TILLING by either Illumina or MinION sequencing will identify mutants that can be used for downstream commercial production.

Summary

As prosperity rises, demand for meat increases as it is a rich source of protein. This in turn places demand on water resources, changes land use (in a manner highly dependent upon how the animal is fed) and leads to an increase in anthropogenic GHG emissions. This has been determined to be unsustainable by a number of international bodies, with some estimates predicting a 70% rise from current levels of 11% of total GHG emissions by 2050. However, demand for protein can also be met by crop-based sources (e.g. soy and pulses) and by mycoprotein, produced by fermentation of crop-derived glucose into biomass, which is harvested and processed into high quality protein. Mycoprotein remains a relatively under-exploited resource worldwide but offers great promise for year-round production of high quality protein, a vital requirement for future food security and human nutrition. The most significant challenge to production is the reliance on a single carbon source, a wheat-derived glucose, which requires special processing before it is suitable for use. Our recent work has revealed that while the fungus used to produce mycoprotein is grown on this glucose substrate, production of a number of essential vitamins is inhibited. Our recent work has revealed that expression of vitamins in some other carbon sources, for example beet derived sucrose syrup is observed. In some, but not all cases, this is coupled to an increase in other deleterious secondary metabolites. This leads to the question, how is the fungus regulating secondary metabolism in relation to carbon source? To expand both the nutritional value of mycoprotein and the range of carbon sources that can be utilised (enabling production to move to other regions of the world) we will use the latest DNA sequencing techniques to reveal the structure of the genome of Fusarium venenatum and study the regions of the genome that contain secondary metabolite genes. From work carried out in other related fungi it is knownthat control of secondary metabolism (SM) is regulated by the position of SM cluster in the genome, and by specific regulatory factors. Utilising the latest sequencing techniques will allow us to positionally resolve SM location and determine the underlying mechanisms regulating responses to different carbon sources. Through a series of controlled batch and continuous culture experiments we will develop techniques to selectively induce vitamin biosynthesis across a range of carbon sources, without inducing the expression of deleterious SM genes, providing both an understanding of the control of SM and an enhanced product for future product development. Building on our existing work we will expand the toolbox of molecular techniques in order to edit the genome of F. venenatum to remove deleterious secondary metabolite gene clusters and their regulatory factors which are induced in response to different carbon sources. As a result of this work, mycoprotein will be able to be produced using a larger range of carbon sources drawing upon a wider range of UK agricultural sources (maize, barley, rice) and even shift to sucrose-based production of mycoprotein (a carbon source that has currently been completely inaccessible), utilising UK sources of sucrose such as sugar beet. Furthermore, the ability to enhance the complement of micronutrients in mycoprotein will broaden its utility as an important component of global diets and offers a more sustainable and flexible alternative to meat.

Impact Summary

The formation of this collaboration allows a company (Marlow Foods) that has previously not worked on research of this nature to, in time, realise new market opportunities as a result of developments in basic bioscience and ensures that the UK maintains its leading position in the production of high quality mycoprotein. MF will gain further competititive advantages by gaining additional buying power for raw materials and would have the capacity to broaden its horizons for production in areas where wheat glucose is not an economic source of production. This grant will have a global impact, both on the research field internationally and on the international industry, especially the UK industry. Through full engagement with industry stakeholders, maximum translation of this research will be ensured, driving forward the UK food industry in a globally competitive market. NIAB EMR and MF will develop an exploitation plan for any IP that arises as a result of the project. Direct beneficiaries: 1. Commercial private sector The impact of releasing the constraints on production of mycoprotein using only unrefined wheat-derived glucose (available from only 1 supplier) is enormous. Production costs could immediately be lowered and MF would be able to source carbon sources from a larger range of suppliers and from a wider range of sources (maize, barley, rice) and even shift to sucrose-based production of mycoprotein, utilising UK sources of sucrose such as sugar beet, benefiting primary agriculture. It is estimated that in the short term production costs for MF could be reduced by at least £250k per annum. Simplification of the production process would allow more rapid growth of production, growing market share in the UK and worldwide which at its current rate is already growing MF turnover by approximately £12M per annum. Widening the possibilities for mycoprotein production from a range of carbon sources, could lead to large benefits in the wider supply chain and for consumers. A low carbon, sustainable source of protein, could fulfil multiple roles in the national supply chain, as demand for meat increases and lead to longer term sustainable production of nutritionally balanced food. This could have particular application in developing 'giant economies' and has significant potential for further growth of a UK business. Indirect beneficiaries Companies focused on enhancing secondary metabolism will benefit through the knowledge gained in this project about the link between carbon source and the control of secondary metabolism. This could aid in development of enhanced strains for production of valuable biomaterials. Government, public and policy benefits There are opportunities to deploy novel technologies to enhance the nutritional value of Quorn, which would allow overall dietary consumption to be altered and potentially expand the range of applications for mycoprotein. Production improvements in mycoprotein will have benefit for the long term sustainabilty of mycoprotein and contribute to a resilient food supply for the consumer in the UK and beyond. Technologies deployed in this proposal will also be instrumental in improving the nutritional quality of Quorn, broadening its applicability. CO2e (equivalent CO2) emissions from Quorn production compare favourably with chicken production (the most efficient mass-consumed meat source defined by feed-conversion ratio), with 2.6t of CO2e per tonne of mycoprotein coming from primary production. There are significant opportunities to lower production costs, CO2e production and energy consumption through improved utilisation of a range of technologies e.g., utilisation of a broader range of substrates, longer production runs, more efficient biomass conversion, waste stream recovery etc.

Committee	Research Committee D (Molecules, cells and industrial biotechnology)
Research Topics	Microbiology
Research Priority	X – Research Priority information not available
Research Initiative	X - not in an Initiative
Funding Scheme	Industrial Partnership Award (IPA)

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