Algal oils by design: a new biotech platform for high-value lipids.

Lead Research Organisation: University College London
Department Name: Structural Molecular Biology

Abstract

Industrial biotechnology (IB) is the harnessing of biological processes and materials for the production of industrially useful products on a large scale. If offers novel solutions to product synthesis that avoid fossil fuels, but instead use plants, bacteria, fungi and algae as renewable sources. This bio-economy is currently worth trillions globally and new advances could bring major benefits to UK industry sectors including chemicals, renewable energy, materials, and health, and could create thousands of jobs in the coming years. A major area of IB is the intensive cultivation of specific microorganisms in industrial scale fermentors, where the microbes have been selected or engineered to produce high levels of a desired commercial product such as an antibiotic, a complex metabolite or a therapeutic protein. These fermentation processes generally use either bacterial or fungal species as the 'cell factory'. However, a very large and diverse group of microorganisms - the algae - represent a rich biological resource that has yet to be exploited in the biotech industry. These photosynthetic microorganisms not only offer a huge treasure trove of natural compounds that have potential application in drug discovery (e.g. as nutraceuticals and food additives, as natural colorants or stabilizers in food and cosmetics) but also offer the possibility of cultivation using sunlight and CO2 to drive biological production, rather than the expensive sugars needed for bacterial or fungal growth.

In this proposal we aim to develop algae as cell platforms for IB applications, and in doing so bring a new, disruptive technology to the fore. We will build on recent advances in our knowledge of the genetics, biochemistry and physiology of algae, together with developments in genetic engineering and high-throughput cell screening technologies. As a focus for this four year project we will develop the algal platform initially for the production of high value oils (=lipids) such as the edible omega-3 and omega-6 long-chain fatty acids that are an essential component of animal diets, and contribute, inter alia, to infant brain development and cognitive function in adults.

The multidisciplinary project has four integrated programmes of work and builds on the world-leading expertise of the five participating groups. We will combine our biological knowledge on the genomes, transcriptomes and lipid metabolomes of two chosen algal species with advanced metabolic modeling tools to build a detailed, robust and predictive model of lipid metabolism in the algal cell. Such as model will allow us to evaluate in silico the effects of manipulating the lipid biology (for example by adding new enzyme pathways). In a second programme, we will use our expertise in algal genetic engineering to develop synthetic biology tools and a design pipeline that will greatly accelerate the process of creating genetic engineered strains. This is critical since, even with a sophisticated model, a large number of different engineering parameters need to be evaluated in order to achieve a fully optimized strain. Such strain selection will also be facilitated by integrating the genetic engineering with a high-throughput cell evaluation technology developed by one of the groups. This system involves encapsulation of single cells in oil microdroplets and ultra-fast analysis and sorting using microfluidic technology coupled to analytical equipment. This will enable the rapid identification and recovery within large populations, of rare engineered or mutated cells that have the desired phenotype. Finally, the technologies will be brought together and tested in the final programme to confirm that we can design and create 'bespoke' strains producing high levels of a desired oil.

Technical Summary

There is considerable interest worldwide in the exploitation of microalgae for the production of a wide range of molecules, including biofuels, bulk chemicals and high value products. These photosynthetic organisms offer many advantages in an industrial context - they do not compete with food crops for arable land, growth rates and productivity is very high for many species, and they are amenable to cultivation in fermenters or closed photobioreactors, where conditions can be controlled. However, to exploit them fully as platforms for industrial biotechnology, it is essential that they can be manipulated easily and robustly, as is possible for other commonly used hosts, such as E. coli, yeast and certain mammalian cell cultures. In this proposal we aim to establish the workflow and molecular tools for two marine microalgae: Phaeodactylum tricornutum (Pt) and Nannochloropsis gaditana (Ng), both of which have sequenced genomes, and accumulate large amounts of lipids. Pt is a marine diatom that is readily transformable, and for which a number of molecular tools are being developed. Ng is a eustigmatophyte alga, for which transformation by homologous recombination has been reported; this will provide the opportunity for knock-outs and knock-ins of genes of interest.

We will use genome and transcriptome data to develop metabolic network models of lipid metabolism, and then employ the models to inform strategies for which lipid biosynthetic enzyme(s) should be manipulated to optimize production of particular lipid molecules, focusing initially on the omega-3 fatty acid DHA. The metabolic engineering itself will be facilitated by development of synthetic biology approaches, including multiplex DNA assembly, registry of DNA parts, and development of synthetic circuits for regulation of transgene expression. We will also design and fabricate microfluidic devices for high-throughput selection of desired strains of algae.

Planned Impact

There is considerable interest in the UK and indeed worldwide in developing an algal industrial biotechnology (IB) sector, both at the level of policy makers and research funders, but also industry. However, because exploitation is at such an early stage, there needs to be some form of proven platform technology before industry would feel confident about moving into the area.

In this application we aim to provide the basis of this knowledge. We will develop tools to enable much more rapid generation of constructs for metabolic engineering of industrially important marine microalgae. We will also develop mathematical models to define lipid metabolic pathways, thus working towards the goal of predictive design principles to engineer the algae to make any desired lipid molecule. Our focus in this grant will be on the important omega-3 fatty acid, DHA, but the principles we establish, together with the generic tools, will provide the means to do the same for many different lipid products. Finally, in an innovative cross-disciplinary collaboration, we will use a microdroplets platform to study the algae at the single cell level, and to sort populations with high throughput. Our links with industrial partners provide the means to take any promising strains we identify through to the next stage of technology readiness, with the prospect of commercialization within 5-10 years.

As the technological advances with microalgae proceeds, economic sustainability will be enhanced, and algal IB will have genuine societal impact. The following sectors will benefit: Food, Feed, Nutraceuticals, but also as the range of products that algae can be engineered to make expands, there will be impacts on the Speciality / platform chemicals, Energy, Fertiliser, and Cosmeceuticals, sectors.

Government and Public Bodies will also benefit, since the outputs of this project can have an impact on several pressing current issues: building the knowledge economy will lead to job creation, and contribute to sustainable economic growth, improve security of food and materials, and contribute to enhanced nutrition.

There will also be benefit to the general public, since the research may contribute to creation of jobs, as algal IB starts to become a reality. Adoption of research outcomes may result in nutraceuticals from algae (eg omega fatty acids, antioxidants and vitamins) becoming more affordable and widely available, leading to improved nutrition and public health. The outreach activities aim to increase awareness of the public on issues of energy, food and material security, and how the development of IB and a knowledge economy can benefit the UK. This will encourage pupils to adopt a career in science and technology. Outreach activities will also increase awareness of resource efficiency in the public, particularly energy awareness, and thus may contribute to lowering energy consumption and household bills.The benefit of these interactions on informed policy choices should start during the lifetime of the grant, and through introductions to algal lobbying bodies such as the European Algae Biomass Association, may have a lasting effect on decision makers in the public domain.

As well as commercial and academic sectors, the work we will do will have impact on our understanding of algal biology generally. There are over 300,000 different algal species, and marine species contribute up to half of all global CO2 fixation, so the study of these organisms has much wider implications than biotechnological exploitation. Our project will ensure that there are scientists with skills for studying algal biology, not just the PDRAs employed on the grant but also other members of our groups.

Publications

10 25 50
 
Description The aim of the project is to develop the tools and technologies for designing and creating transgenic algae that are engineered to make specific high-value oils. This involves a combination of metabolic modelling, synthetic biology, lipid analysis and single cell sorting and analysis using a microfluidic platform. The research is still on-going, but three key outcomes so far are:

1. The creation of transgenic strains of a diatom that accumulate increased levels of storage lipids, and which contain within these lipids significantly elevated levels of the valuable omega-3 long chain polyunsaturated fatty acid, DHA.

2. A droplets-based microfluidic platform has been designed and built to encapsulate, grow and observe single microalgal cells. Furthermore, this platform can be used for high-throughput sorting of these cells based on: i) expression of the fluorescent protein, GFP, or ii) levels of lipid that accumulate in the cell.

3. The development of standardized molecular tools and methodologies for efficient genetic engineering of the microalgae.
Exploitation Route The ultimate aim of the work is to develop a 'designer algae' platform that can be exploited by Industrial Biotechnology companies for the light-driven synthesis of speciality oils.
Sectors Manufacturing, including Industrial Biotechology

 
Description BBSRC IBBE Strategy Panel
Geographic Reach National 
Policy Influence Type Membership of a guideline committee
Impact The Strategic panel provided advice and guidance for the setting up of the Networks in Industrial Biotechnology & Bioenergy (NIBBs), which aim to foster increased awareness and interaction between academics and industrialists in the bioeconomy. It was also important for the thinking behind the IB Catalysts, joint with Innovate UK & EPSRC. Many of the grants and proof-of-concept awards are aimed at increased sustainability through the use of IB solutions
URL http://www.bbsrc.ac.uk/funding/filter/networks-in-industrial-biotechnology/
 
Description Member of BBSRC Bioenergy review panel
Geographic Reach National 
Policy Influence Type Membership of a guideline committee
 
Description Agri-Inno: Stimulating Innovation and Entrepreneurship in Egypt's Agricultural Sector
Amount £28,520 (GBP)
Organisation Biotechnology and Biological Sciences Research Council (BBSRC) 
Sector Public
Country United Kingdom
Start 01/2017 
End 01/2017
 
Description Algal Biorefinery: routes for energy and nutrient recovery.
Amount £9,966 (GBP)
Organisation Natural Environment Research Council 
Sector Public
Country United Kingdom
Start 06/2015 
End 12/2015
 
Description Business Interaction Voucher
Amount £10,000 (GBP)
Funding ID PHYCBIV-23 
Organisation Biotechnology and Biological Sciences Research Council (BBSRC) 
Department Networks in Industrial Biotechnology and Bioenergy (NIBB)
Sector Academic/University
Country United Kingdom
Start 04/2018 
End 10/2018
 
Description Implementing Algebra guidelines to risk assessments of scale-up of non-native species
Amount £9,500 (GBP)
Organisation Biotechnology and Biological Sciences Research Council (BBSRC) 
Sector Public
Country United Kingdom
Start 05/2017 
End 10/2017
 
Description Metabolic characteristics of terrestrial algae and plants in Antarctica - research visit to Antarctica
Amount £74,000 (GBP)
Organisation British Antarctic Survey 
Sector Academic/University
Country United Kingdom
Start 05/2014 
End 03/2015
 
Description PHYCONET Proof of Concept award
Amount £99,328 (GBP)
Organisation Biotechnology and Biological Sciences Research Council (BBSRC) 
Sector Public
Country United Kingdom
Start 12/2015 
End 07/2016
 
Description Playground to Polar - how do we measure Antarctic and UK biodiversity?
Amount £3,000 (GBP)
Organisation The Royal Society 
Sector Academic/University
Country United Kingdom
Start 11/2017 
End 06/2018
 
Description Regulation of polyphosphate metabolism in Chlamydomonas and potential for exploitation as phosphorus sink in nutrient recovery systems
Amount £65,772 (GBP)
Organisation Biotechnology and Biological Sciences Research Council (BBSRC) 
Sector Public
Country United Kingdom
Start 10/2016 
End 10/2019
 
Description Scoping a new collaborative and training networks in microbial and invertebrate biotechnology and 'omics' in Malaysia.
Amount £4,064 (GBP)
Organisation Natural Environment Research Council 
Sector Public
Country United Kingdom
Start 03/2017 
End 05/2017
 
Description Synthetic Biology Centres
Amount £12,000,000 (GBP)
Funding ID BB/L014130/1 
Organisation Biotechnology and Biological Sciences Research Council (BBSRC) 
Sector Public
Country United Kingdom
Start 06/2014 
End 05/2019
 
Description Undiscovered High Value Products from Polar Algae.
Amount £19,961 (GBP)
Organisation Unilever 
Sector Private
Country United Kingdom
Start 09/2014 
End 03/2015
 
Description Promoting algae for industrial biotechnology 
Organisation Rothamsted Research
Country United Kingdom 
Sector Academic/University 
PI Contribution I have been very active in promoting the idea of using algae as industrial biotechnology (IB) hosts. This has led to a successful BBSRC sLoLa award, and also contributed to the debate about how to cement the rather fragmented algal expertise in the UK. Ultimately the introduction of the BBSRC networks in IB (NIBBs) were borne out of this debate, and these include Phyconet, which is specifically algal biotechnology. In addition several other NIBBs consider using algae as source of novel products, pathways, genes (NPRONet, HVCfP), or as platforms for community based metabolism (ADNET)
Collaborator Contribution Research and development collaborations with colleagues in the algal biotechnology field, but also in metabolic engineering and SMEs in algal biotechnology
Impact sLoLa Several BBSRC NIBBs, with Phyconet as specific one on algal biotechnology IB Catalyst award
Start Year 2009
 
Description Promoting algae for industrial biotechnology 
Organisation University College London
Department Division of Biosciences
Country United Kingdom 
Sector Academic/University 
PI Contribution I have been very active in promoting the idea of using algae as industrial biotechnology (IB) hosts. This has led to a successful BBSRC sLoLa award, and also contributed to the debate about how to cement the rather fragmented algal expertise in the UK. Ultimately the introduction of the BBSRC networks in IB (NIBBs) were borne out of this debate, and these include Phyconet, which is specifically algal biotechnology. In addition several other NIBBs consider using algae as source of novel products, pathways, genes (NPRONet, HVCfP), or as platforms for community based metabolism (ADNET)
Collaborator Contribution Research and development collaborations with colleagues in the algal biotechnology field, but also in metabolic engineering and SMEs in algal biotechnology
Impact sLoLa Several BBSRC NIBBs, with Phyconet as specific one on algal biotechnology IB Catalyst award
Start Year 2009
 
Description Promoting algae for industrial biotechnology 
Organisation University of Aberdeen
Country United Kingdom 
Sector Academic/University 
PI Contribution I have been very active in promoting the idea of using algae as industrial biotechnology (IB) hosts. This has led to a successful BBSRC sLoLa award, and also contributed to the debate about how to cement the rather fragmented algal expertise in the UK. Ultimately the introduction of the BBSRC networks in IB (NIBBs) were borne out of this debate, and these include Phyconet, which is specifically algal biotechnology. In addition several other NIBBs consider using algae as source of novel products, pathways, genes (NPRONet, HVCfP), or as platforms for community based metabolism (ADNET)
Collaborator Contribution Research and development collaborations with colleagues in the algal biotechnology field, but also in metabolic engineering and SMEs in algal biotechnology
Impact sLoLa Several BBSRC NIBBs, with Phyconet as specific one on algal biotechnology IB Catalyst award
Start Year 2009
 
Description AlgaeEurope 2015 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Industry/Business
Results and Impact Presentation of research into algal synthetic biology to the principle conference in Europe aimed at supporting and developing the algal biotechnology sector. About half the participants were from industry (including large multisector, SMEs and startups)and 10% or so policy makers and officials eg from EU. The aim was to inform this group about the potential of synthetic biology in developing algal industrial biotechnology, providing the impetus to consider algae as production platforms for high value products as well as simply for biomass production. As a result, a consortium has been formed of academic colleagues who will share best practice and resources in SynBio.

I also described our recently opened Algal Innovation Centre (AIC), where it is possible to carry out pilot-scale algal cultivation, including of GM algae. As a result of the talk several contacts have been made with companies who are interested in the AIC.
Year(s) Of Engagement Activity 2015
 
Description Festival of Plants, Botanical Garden, Cambridge 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Public/other audiences
Results and Impact Exhibit at the Festival of Plants (was called Fascination of Plants in 2012) in Cambridge University Botanic Garden. It provided basic information about algae, showcased our scientific work, and then explained how we were using this in collaboration with companies to carry out remeidation of waste water. There were activities such as viewing microalgae with microscopes and observing algae phototaxis. Children could handcraft their own algae using pipe cleaners or take part in an interactive algae quiz to win prices. Several members of the public asked for additional information.

Interest from several visitors about how to incorporate algal growth in their activities, eg anaerobic digestion or green energy generally
Year(s) Of Engagement Activity 2012,2013,2014,2015
URL http://www.botanic.cam.ac.uk/Botanic/Event.aspx?p=27&ix=351&pid=2718&prcid=0&ppid=2718
 
Description High Value Products from Plants conference 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Industry/Business
Results and Impact Presentation of algal synthetic biology approaches to help support algal industrial biotechnology. This was at a Network in Industrial Biotechnology & Bioenergy (NIBB) meeting organised by the High Value Compounds from Plants NIBB. Interest was sparked amongst researchers to consider using algae, as well as plants, and production platforms.
Year(s) Of Engagement Activity 2015
 
Description IntoBiology resource 
Form Of Engagement Activity Engagement focused website, blog or social media channel
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Schools
Results and Impact A final year undergraduate student in Plant Sciences produced a webpage describing work on algae in Plant Sciences and Biochemistry. The webpage is part of the programme established by IntoBiology, aimed at encouraging school children to consider studying plant biology.
Year(s) Of Engagement Activity 2015
URL http://intobiology.org.uk/unlocking-the-potential-of-algae-how-the-green-stuff-in-your-pond-might-go...
 
Description PHYCONET (BBSRC NIBB) Meeting: "UK Microalgal biotechnology, creating a unified vision" Cambridge February 2018 
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 A workshop of academia, funders, policy makers and industry, aimed at identifying ways to augment and support the UK microalgal business. As a result of the meeting, we made links with several companies and have obtained two Business Interaction Vouchers
Year(s) Of Engagement Activity 2018
URL http://www.phyconet.org.uk/document_category/uk-microalgal-biotechnology-creating-a-unified-vision
 
Description PHYCONET workshop 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Industry/Business
Results and Impact PHYCONET showcase workshop to highlight R&D opportunities fin industrial biotechnology relating to high value products from microalgae.
Year(s) Of Engagement Activity 2016
URL http://www.phyconet.org.uk/
 
Description Phyconet (BBSRC NIBB) Annual Meeting, Plenary speaker 
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 talk was aimed at showing that the potential exploitation of algae for industrial purposes needs to be underpinned by fundamental biological understanding. Several industrial colleagues agreed, saying they recognise the need for better algal taxonomy and physiology to support their activities and ensure their IP can be protected.
Year(s) Of Engagement Activity 2017
URL http://www.phyconet.org.uk/wp-content/uploads/2017/09/PAC17_programme.pdf
 
Description Pint of Science 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Public/other audiences
Results and Impact I presented a talk on the potential uses of algae for bioenergy production as part of the Pint of Science festival in Cambridge, where scientists engage with people having a drink in a local pub. This takes place in London, Cambridge, Bristol and other university towns each year. As result of my talk and questions afterwards, although most people were not aware initially what algae were, nor how they could be used to generate energy, their opinions became more positive and optimistic towards biofuels generally. Pint of Science was awarded "Points of Light" by the Prime Minister David Cameron in November 2015 and the founders were interviewed about it on London Live TV.
Year(s) Of Engagement Activity 2014,2015
URL https://en.wikipedia.org/wiki/Pint_of_Science
 
Description Science on Saturday, part of SET week in Cambridge 
Form Of Engagement Activity Participation in an open day or visit at my research institution
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Public/other audiences
Results and Impact We had a stand in the science tent outside Dept of Plant Sciences. We demonstrated what algae are, what we use for now and how they might be used in the future for energy production. There were activities such as viewing microalgae with microscopes, extracting different light-harvesting pigments, and observing algae phototaxis. Children could handcraft their own algae using pipe cleaners or take part in an interactive algae quiz to win prices.

School asked for visit to talk to years 10-13.

Established the material to mount a Royal Society Summer Science Exhibit
Year(s) Of Engagement Activity 2006,2007,2008,2015,2016
 
Description Science on Sundays 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Public/other audiences
Results and Impact Short talk and Q&A on the nature of algae and how their characteristics could be used for sustainable energy, water remediation, or other green tech solutions. Held in the Cambridge Botanic Garden, the audience were interested in plants, and so this was an opportunity to provide information on what algae are, and how they might be used in novel ways. The audience fed back that they felt much better informed.
Year(s) Of Engagement Activity 2015
URL http://www.botanic.cam.ac.uk/Botanic/NewsItem.aspx?p=27&ix=196
 
Description The Life Scientific on BBC Radio 4 
Form Of Engagement Activity A broadcast e.g. TV/radio/film/podcast (other than news/press)
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Public/other audiences
Results and Impact I was interviewed by Jim Al-Khalili about my scientific career and interest in algae and vitamins. The programme "The Life Scientific" was broadcast on BBC Radio 4 and was also available on BBC iPlayer and as a podcast. As a result I have had numerous contacts about possible scientific and industrial collaborations, as well as enquiries from the general public about the various topics.
Year(s) Of Engagement Activity 2017
URL http://www.bbc.co.uk/programmes/b08bzl8y