Sustainable bioenergy from microalgae: A systems perspective

Lead Research Organisation: University of Sheffield
Department Name: Chemical & Biological Engineering

Abstract

Global energy demand and the environmental consequences of fossil fuel combustion to satisfy this demand is proving to be unsustainable. Bio-energy has the potential to mitigate carbon emission concerns and provide a renewable option for energy generation. But, this requires development of sustainable processes that are economically viable. The concept of using photoautotrophic metabolism in microalgae to convert solar energy and carbon dioxide to organic fuel precursors is attractive for sustainable energy generation. However, development of efficient processes that use the concept to make a difference to the energy economy has so far proved elusive. We believe that a greater understanding of microalgae metabolism derived through a systems approach will make a difference to this scenario and enable development of sustainable processes for bio-energy generation, including approaches that use synthetic biology and metabolic engineering. The successful development of any such process will require characterisation of organisms that are relevant to field cultivation rather than just laboratory strains.
We aim to combine UK and Indian facilities and expertise in phycology, bioremediation, 'omic' characterisations and biochemical engineering to conduct a systems level investigation on selected microalgae isolates with potential to accumulate biofuel precursors and develop sustainable processes for bio-energy generation from microalgae in India and the UK. We will perform a systems level investigation in selected isolates to determine key metabolic steps involved in the synthesis of biofuel precursors (in particular, carbohydrates, triacylglycerols (TAGs) and hydrocarbons) in microalgae. This will lead to optimisation of conditions to maximize product yields, and the identification of gene targets for strain manipulations. We will also examine engineering approaches to enhance product recovery and investigate exemplar biorefinery cases to enable the development of sustainable processes for bio-energy generation.
Indian partners at BDU have established a repository of marine microalgae with DBT support that has over 500 isolates in its collection, from the rich biodiversity hotspots of tropical southern Indian coastal areas and psychrophilic strains from the Arctic. Some of these isolates have been assessed for biodiesel generation and will be available for the project. Freshwater microalgae characterised for hydrocarbon and TAG accumulation with prior DBT support is available at Madurai. In the UK, SAMS have extensive experience at characterisation and maintenance of microalgae with relevance to the development of UK-related processes. We will combine this expertise with algae systems biology expertise available at Sheffield and Cambridge to investigate how pathways involved in accumulation of storage compounds that are biofuel precursors and how establish its relationship with growth of the organism (Figure 1). A novel patented microbubble technology incorporated in an airlift loop bioreactor developed at Sheffield (WBJZ) will also be investigated for process improvements and product recovery.
The proposed work will result in the identification of key metabolic steps in relevant algal biotransformations, optimised conditions that maximise product yields and selected or novel strains for use in developing bioprocess for sustainable generation of fuel precursors that make a difference to the energy economy in India and the UK. We also propose to hold joint workshops in the two countries and exchange visits to enable transfer of knowledge and expertise between partners, with active industrial involvement.

Technical Summary

Current methods to meet the growing energy demand are unsustainable and environmentally damaging. Bio-energy has the potential to mitigate carbon emission concerns and provide a renewable option for energy generation. But, this requires development of sustainable processes that are economically viable. The concept of using photoautotrophic metabolism in microalgae to convert solar energy and carbon dioxide to organic fuel precursors is attractive for sustainable energy generation. However, development of efficient processes that use the concept to make a difference to the energy economy has so far proved elusive. We believe that a greater understanding of microalgae metabolism derived through a systems approach will make a difference to this scenario and enable development of sustainable processes for bio-energy generation, including approaches that use synthetic biology and metabolic engineering. The successful development of any such process will require characterisation of organisms that are relevant to field cultivation rather than just laboratory strains. We aim to combine UK and Indian facilities and expertise in phycology, bioremediation, 'omic' characterisations and biochemical engineering to conduct a systems level investigation on selected microalgae isolates with potential to accumulate biofuel precursors and develop sustainable processes for bio-energy generation from microalgae in India and the UK. The proposed work will result in the identification of key metabolic steps in relevant algal biotransformations, optimised conditions that maximise product yields and selected or novel strains for use in developing bioprocess for sustainable generation of fuel precursors that make a difference to the energy economy in India and the UK. We also propose to hold joint workshops in the two countries and exchange visits to enable transfer of knowledge and expertise between partners, with active industrial involvement.

Planned Impact

The two collaborating industrial partners (Pioneer Jellice Ltd in India and Celtic distilleries in Scotland) as their waste streams will be tested for improved microalgal bioenergy generation potential - leading to processes that they could adopt.

The proposed work will impact the algal research community, as the research conducted here will provide initial leads and background information for future developments of technology for bioenergy generation from microalgae

Both UK and India will stand to benefit from the proposed work as sustainable processes relevant to the geographical areas could be developed based on the results of the work proposed.

The work proposed will enhance our knowledgebase that will not only be applicable in India and the UK, but also globally, elsewhere.

There will be exchange of knowledge and awareness of the socio-economic needs for the development of sustainable processes in India and the UK, by investigators from both the countries.

The developed processes will contribute to environmental sustainability and attract public and private sector investments for R&D

there will be transnational skills training possible

Publications

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Butler T (2020) Phaeodactylum tricornutum: A Diatom Cell Factory. in Trends in biotechnology

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C Esther Elizabeth Grace (2020) Biomolecular transitions and lipid accumulation in green microalgae monitored by FTIR and Raman analysis. in Spectrochimica acta. Part A, Molecular and biomolecular spectroscopy

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Esther Elizabeth Grace C (2022) Response to nutrient variation on lipid productivity in green microalgae captured using second derivative FTIR and Raman spectroscopy. in Spectrochimica acta. Part A, Molecular and biomolecular spectroscopy

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Kapoore RV (2016) Towards quantitative mass spectrometry-based metabolomics in microbial and mammalian systems. in Philosophical transactions. Series A, Mathematical, physical, and engineering sciences

 
Description The results of the research are still being interpreted, but some of the findings directly from the funding that we have been able to publish are listed below: (a) different species of microalgae may respond differently to environmental conditions that can be used to increase lipid productivities, required for sustainable biodiesel production (http://rdcu.be/IGu0); (b) cryopreserved algal cultures, stored at -15 or -80 C may loose their functionality to produce lipids, and mey need to be stored at liquid nitrogen temperatures (-196 C)(https://www.nature.com/articles/s41598-019-38588-6); (c) optimal conditions for quenching metabolism in algal cultures found that minimise metabolite leakage and hence loss of relevant information - a methodology related finding (https://www.mdpi.com/2218-1989/8/4/72); (d) key delineating features suitable for lipid accumulation in marine microalgae that would help in selecting appropriate strains and process conditions identified; (e) key regulatory features relevant to lipid accumulation in the sequenced diatom, P. tricornutum, identified through a systems level characterisation that reveals exemplary regulatory mechanisms employed by microalgae during lipid accumulation; (f) differences in CO2 uptake and nutrient assimilation identified between two strains from the same species isolated from a similar but geographically different habitat, suggesting adaptive evolution. The project enabled generic protocols to be developed on algae characterisations, including omic level investigations that will become evident in the publications to follow, for use by the wider research community. The project generated transcriptomic data that is still being analysed and will be deposited for public use soon, including a publication resulting from it, which is under preparation. The project enabled us to develop a working relationship between our labs and partner labs in India, which is being taken forward with funding applications for further research in algae biotechnology towards better environmentally sustainable (bio)manufacturing solutions. The project also enabled development of specialist skills in omic characterisation of algae. It resulted in the training of at least 5 research staff directly involved in the project, some of whom are now employed in the same field, participating in research activities within the UK and abroad. The project has enabled exploration of International collaboration in Indonesia, Malaysia and African countries within the GCRF context.
Exploitation Route The results of the work are still being interpreted. We have been disseminating the findings in conferences and publications, and anticipate to continue dissemination in impact publications. This will enable practitioners to develop suitable strategies for using algae biotechnology in energy generation, We have been engaging with industries to take the work forward and create socio-economic impact. We've conducted an international conference in India and in the UK and conducted workshops in India to create awareness of the work among academic and wider circles. We anticipate the findings from the project to slowly find its way to academic and non-academic players in the field (algae biotechnology). Specifically, the project has enabled improving the capabilities of our Indian partners and in enabling them to interact with Industry. Our work has informed lipid accumulation strategy in other algae (e.g., Biotechnol. Biofuels, 12, 151, 2019; Algal Res., 43, 101646, 2019; PLOS One, 14, 11, e0224701, 2019), formed the basis for an International workshop conducted by ICGEB in India, (https://www.icgeb.org/course-new-frontiers-in-algal-omics/), informed cryopreservation protocols for microalgae (e.g., J. Phycol., 56, 334, 2020; J Appl. Phycol., 32, 2967, 2020), macroalgae (e.g., Biopresevation and Biobanking, 17, 5, 378, 2019) and plants (Biol. Conservation, 250, 108736, 2020).
Sectors Agriculture, Food and Drink,Chemicals,Energy,Environment,Manufacturing, including Industrial Biotechology,Transport

URL https://www.sheffield.ac.uk/cbe/people/academic-staff/raman-vaidyanathan
 
Description The work has provided the basis for interaction with industrial partners, in particular SMEs in the UK and in India in engaging them further in grant applications, for example to an InnovateUK call to fund research with direct socio-economic benefits. Two successful business interaction voucher applications were made based on the work from this award, contributing towards UK economic progressiveness. In terms of direct impact we are trying to take forward our work with the Indian partners and the Indian Industry involved in the work to see how it can add direct value to the industry involved. There has been a successful grant application to a Mission Innovation challenge 5 (2019-22) by our Indian partner in which we are the MI country partner.
First Year Of Impact 2017
Sector Education,Energy,Environment
Impact Types Economic

 
Description "Outdoor mass cultivation of marine microalgae for biodiesel production with improved solar conversion efficiencies"
Amount ₹7,933,143 (INR)
Funding ID BT/PR31336/PBD/26/726/2019 
Organisation Government of India 
Sector Public
Country India
Start 12/2019 
End 11/2022
 
Description Accelerated Knowledge Transfer Pilot 2i
Amount £12,000 (GBP)
Funding ID 144 
Organisation Innovate UK 
Sector Public
Country United Kingdom
Start 12/2022 
End 03/2023
 
Description BBSRC ALERT14
Amount £400,000 (GBP)
Funding ID BB/M012166/1 
Organisation Biotechnology and Biological Sciences Research Council (BBSRC) 
Sector Public
Country United Kingdom
Start 01/2015 
End 12/2015
 
Description Marie-Sklodowska Curie Fellowship
Amount € 195,000 (EUR)
Funding ID 661063 - EQUIP 
Organisation Marie Sklodowska-Curie Actions 
Sector Charity/Non Profit
Country Global
Start 05/2015 
End 04/2017
 
Title Carbon dioxide uptake measurement 
Description A dynamic method to enable determination of carbon uptake kinetics in microalgae cultivations was developed, along with an improved stable protocol for the determination of dissolved inorganic carbon in aqueous environments. 
Type Of Material Technology assay or reagent 
Year Produced 2020 
Provided To Others? No  
Impact The methodology is yet to reach the public domain to create impact. We are preparing a publication for submission to an impactful journal. 
 
Title Cryopreservation of cultures 
Description The study reports for the first time the requirement of liquid nitrogen storage of biobanks and loss of functionality in otherwise cryopreserved microalgal cells using a systematic physiological experimentation on differently cryopreserved cells. Loss in lipid accumulation capacity as applicable to biodiesel production is demonstrated. 
Type Of Material Biological samples 
Year Produced 2018 
Provided To Others? Yes  
Impact It has informed cryopreservation protocols for microalgae (e.g., J. Phycol., 56, 334, 2020; J Appl. Phycol., 32, 2967, 2020), macroalgae (e.g., Biopresevation and Biobanking, 17, 5, 378, 2019) and plants (Biol. Conservation, 250, 108736, 2020). 
 
Title Sample processinig for metabolic profiling 
Description We have identified the importance of establishing the level of metabolome leakage for specific cell lines, irrespective of the methadology employed. 
Type Of Material Biological samples 
Provided To Others? No  
Impact Notable impact in processing of cells for metabolite profiling within our group. Yet to notice changes in practice elsewhere. 
 
Description Industrial - Varicon 
Organisation Varicon Aqua Solutions Ltd
Country United Kingdom 
Sector Private 
PI Contribution We tested material produced by the company in our labs. In particular, we collected data relevant to using the diatom (Phaeodactylum tricornotum) for producing multiple products in a biorefinery approach, using the photobioreactor rig and medium components provided by the company as in-kind contributions
Collaborator Contribution The company provided the materials to be used in the characterisation, including the photobioreactor rig, as well as staff time as in-kind contributions. This also led to a PhD student to work on placement in their facility.
Impact The results from the work are being analysed towards publications, which should appear soon.
Start Year 2018
 
Description Industrial-Ctech 
Organisation C-Tech Innovation
Country United Kingdom 
Sector Private 
PI Contribution Expertise (both myself and my team's), data from research
Collaborator Contribution Access to specific equipment and associated expertise for usage and application in research.
Impact Not yet. The collaboration is multidisciplinary, the disciplines involved were biological, biotechnological, chemical, engineering, electrical.
Start Year 2017
 
Description Mission Innovation collaborative project (IC5) 
Organisation Bharathidasan University
Country India 
Sector Academic/University 
PI Contribution Construction of the proposal and contribution of expertise for collaborative project work led by Partner institute. Costing factored towards hosting and travel for interactions and a symposium organisation.
Collaborator Contribution The partner is the lead organisation in the proposal partnering with us as the MI (Mission Innovation) collaborator
Impact Project is in progress, but is slow due to the pandemic. No outputs yet.
Start Year 2019
 
Description 6th UK Algae conference 2016 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Other audiences
Results and Impact A symposium of algae biotechnology was organised - had participation from EU countries and from across the UK. There were Industrial participants as well as academic and postgraduate and undergraduate students. This led to several network and collaboration initiations. This was also advertised on the social media such as Twitter and Facebook resulting in dialogues with wider community.
Year(s) Of Engagement Activity 2016
URL https://www.sheffield.ac.uk/algae/previousevents/6thukalgae2016
 
Description INDO-UK WORKSHOP ON 'DATA ANALYSIS IN VIBRATIONAL SPECTROSCOPY AND MASS SPECTROMETRY BASED METABOLIC PROFILING, Madurai, India 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Postgraduate students
Results and Impact Thirty participants, comprising research scholars and faculty members of life science and physical science departments from various colleges and universities, attended the workshop. Awareness was created regarding multivariate analysis and its usage in data analysis. This enabled participants to see value in data they had generated in different areas of science, including medical, chemical, environmental and biological fields.
Year(s) Of Engagement Activity 2017
URL http://www.thehindu.com/todays-paper/tp-national/tp-tamilnadu/campus-connect/article19572964.ece
 
Description International conference on Microalgae and Cyanobacterial Biotechnology (MACB) 2016, Thiruchirapally 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Other audiences
Results and Impact An International conference and workshop held in India. There was media attention and collaborative networking activities in addition to interaction with policymakers.
Year(s) Of Engagement Activity 2016
URL http://timesofindia.indiatimes.com/city/trichy/Biofuel-can-be-alternative-to-fossil-fuel/articleshow...
 
Description Invited keynote - India 
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 Invited to a keynote address in an International workshop held in India, where work from this project was used to conduct the workshop and dissipate knowledge and skill set to an international audience from at least 5 other countries.
Year(s) Of Engagement Activity 2018
 
Description Invited talk and interaction at International algae conference 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Postgraduate students
Results and Impact One of a handful of invited International speakers in an online event. Others included from countries as diverse as Finland, USA, Italy, Sweden, South Korea, Japan, etc. Invited talk was followed by questions by audiences that included students and practitioners.
Year(s) Of Engagement Activity 2020
 
Description Radio interview 
Form Of Engagement Activity A press release, press conference or response to a media enquiry/interview
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Public/other audiences
Results and Impact member of the team gave a radio interview about Microalgae biofuel production in the programme "En Fase Experimental" of Radio X.
Year(s) Of Engagement Activity 2016
 
Description Researchers night 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Schools
Results and Impact Members of the team were involved in explaining how algae can be used to produce biofuels. They organised and participated in the showcase event of the research as part of the British Science Week.
Year(s) Of Engagement Activity 2016
URL https://www.sheffield.ac.uk/researchersnight/programme/alfred-denny-building#algae
 
Description Science in the Classroom 
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 Member of the research team participated giving a talk about her research career entitled "Does the sea hide the future of the green oil?"
Year(s) Of Engagement Activity 2016
 
Description Science in the pub 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Public/other audiences
Results and Impact member of the research team gave an outreach talk entitled: "Everything you always wanted to know about algae, but were afraid to ask"
Year(s) Of Engagement Activity 2016