Steve Skill. The Nottingham Microalgae Biorefinery

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1 The Nottingham Microalgae Biorefinery Steve Skill Sustainable Chemicals from Microalgae, Encompassing Biocrude through to Fine Chemicals Royal Society of Chemistry, The Chemistry Centre, London UK 19 th November 2013

2 Nostoc muscorum HETEROCYST VEGETATIVE CELL Steve Skill Background Lancaster University 1983 Biochemist- Restriction enzyme characterisation of cyanobacterial plasmids and cyanophage DNA. BBSRC 3 year project - Research assistant - Isolation and cloning of Heterocyst specific gene sequences. Enhanced H2 production N fixation in higher plants Carbon negative fertiliser production Haber Process No Molecular toolkit!!

3 Sustainable Algae Production Process Gas Store CH 4 /CO 2 Heat Exchange CHP Chicken Manure 10m 3 AD (Farm Gas) Raw liquor Belt/Filter Press Filtrate PBR 1000 L CH4/O2 Electricity Process Design: Steve Skill Microalgae: Spirulina platensis Anaerobic digestion seed: STW digester Company: Blue-Green Biotech Location: Sherwood Forest Water Solids (compost) Belt filter Spirulina Biomass Products Discharge PBR- Photobioreacto AD- Anaerobic Digester (mesophilic)

4 Biocoil Photobioreactor, Algae/bacteria consortium Tertiary Sewage Treatment Process Process Design: Steve Skill Microbes: Autoflocculating consortia of Chlorella & Scenedesmus Company: Biotechna Ltd Location: Stoke Bardolph STW, Severn Trent PLC, Nottingham Discharge from Activated Sludge Mix Tank 2000 L Settling Tank Low Nutrient Discharge Settled Algae Biocoil Photobioreactor 5000 L Settled Algae Algae Biomass

5 1992- Autoflocculating algae consortium (Chlorella/Scenedesmus) Animal feed? HTL feedstock? Direct use as Fuel?

6 1993 Algae Power Station? BBC Tomorrows World

7 Biotechna Graesser Ltd, Greencycle Ltd, Bioxtract Ltd. Escola Superior de Biotecnologia (ESB), Lisboa. (Vitor Verdhelo) Two stage Biocoil PBR system for the production of Beta Carotene from Dunalliella salina. The unit was constructed in the UK and transported to site near Sines in Portugal.

8 High Strength Wastewater Treatment Cationic Flocculent Photosynthetic Anaerobe Biomass Tapoica / Potato Starch Effluent Anaerobic PBR Whole Broth Settling Aerobic PBR Process Design: Steve Skill Microbes: Photosynthetic Anaerobe consortium Autoflocculating Chlorella Scenedesmus consortium Company: Biotechna Ltd Location: Dorking, UK. Algae Biomass PBR- Photobioreactor Settling Supernatant Discharge (Low N&P)

9 Digestate Enhancement CH 4 /CO 2 Solids Cationic Flocculent Photosynthetic Anaerobe Biomass Chicken Manure Anaerobic Digester Digestate Filter Press Filtrate Anaerobic PBR Whole Broth Settling Aerobic PBR Process Design: Steve Skill Microbes: Photosynthetic Anaerobe Consortium Autoflocculating Chlorella / Scenedesmus consortium Company: Biotechna Ltd Location: Poultry farm, Cork, Ireland Algae Biomass PBR- Photobioreactor Settling Supernatant Discharge (Low N&P)

10 Sustainable Aquaculture Carbohydrate Protein Feed Biofilm PBR Water Recycling Circuit Intensive fish grow-out tanks Filter screen Faeces & spent food AD Proprietor: Steve Skill Microbes: photosynthetic biofilm Company: Sherwood Forest Tilapia Location: Nottingham, UK. Tilapia Produce

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12 The Nottingham Microalgae Biorefinery

13 Microalgal biorefinery - Industry perspective What commercially viable biomolecules can we characterise and extract from the algae? Can we grow the algae and extract the biomolecules economically? Does the consumer want whatever we find? Whole supply chain consideration -Can we develop a carbon negative ingredient supply chain?

14 Potential Microalgae Products and Applications Anti-oxidants Anti-inflammatory Anti tumour Anti-microbial Sunscreens & Cosmetics Pharmaceuticals Nutraceuticals Flavours and fragrance High value bioactives Biodiesel Bioethanol Hydrogen Methane Biofuel CO 2 capture Bulk chemicals Building block chemicals MICROALGAE Bioremediation Waste water treatment Contamination remediation Bioplastics Materials science Ceramics Adhesives

15 CCIF Project 2.1m Steve Skill PI Optimising Yield Molecular Biology Biomass production and harvesting Health and Beauty Retailer Formulator and Manufacturer Healthcare provider Power Station Engineers and Consultants Chemistry Photophysiology Stress Response Modelling Biosynthesis Product Identification Product Extraction Downstream Processing Healthcare and Personalcare Ingredient Production

16 Which microalgal strain? Screening programme lead to the selection of a ROBUST cyanobacteria (CX68) Auto-flocculation Thermophilic Robust Monoalgal culture Storage Freshwater strain Wild type produces potentially valuable metabolites Di-azotrophe Prokaryote

17 Microalgal Biorefinery with Carbon Capture Nutrients N,P Low CO2 Flue gas Cosmetic Bioactive Extract Hot flue gases from Boots Power Station 16 m 3 PBR Harvesting (Passive) 14% Solids Ball Milling Tangential flow filtration Process Design: Steve Skill Microbes: Autoflocculating Thermophilic Cyanobacteria Company: Boots, Cognis (BASF) Location: Nottingham, UK. Protein (Hair care) PBR- Photobioreactor

18 Direct carbon capture PBR 15MW Power Station Flue gas duct 16,000 litre photobioreactor (PBR) Productivity: g/l/day Energy use: 20-70W/m 3

19 A B C D Basic mode of operation. A-transparent shell, B-rotor assembly, C- Fluid phase, D-liquid phase Contaction and gas exchange between C & D. Patent: Apparatus for the treatment of fluid streams and method for conducting the same. S. Skill 2009 Basic mode of operation. Phase flows, inlets and outlets Co-current or counter-current flow of phases. A B C D E F

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21 Product R&D Focus Antioxidants UV Protection (Suntan lotions) Anti inflammatory Anti ageing Protein Hydrolysates (hair care)

22 Multi-stage-stage biorefinery fractionation process using low energy ball mills. Harvest Growth Media Recycle Passive settling Ball Mill 1 Passive settling Ball Mill 2 Passive settling 14% solids Extra-cellular Extract Sunscreen Anti-inflammatory Anti-oxidants (Anti-ageing) Further Fractionation (Crossflow filtration) Lysate Anti-oxidants Carotenoids Natural colours Protein Hair Care

23 Efficacy testing A suite of bioassays and analytical protocols have been employed to validate the cosmetic efficacy of extracts recovered from the biomass including; Antioxidant, Lipid peroxidation, antiinflamatory, UVA & UVB protection and antimicrobial potential.

24 Developing a platform for sustainable chemical production Engineering microalgae for sustainable chemicals CX68 Prokaryote Genome Sequenced Transformation protocols

25 Pathways for manipulation? NDP-Glc Other polysaccharides/peptidoglycans Glycogen Glucose G6P/G1P 4,5 & 7 C sugars F6P Calvin- Glycerol-P G3P Benson RuBP + CO 2 cycle Shikimate pathway MAAs Phenylalanine Phenolic compounds Aromatic amino acids MEP isoprenoid pathway Isoprene Terpenoids Tocochromanols Carotenoids C 2 H 4 Amino acids RS 2-3-PGA 2-PGA PEP OAA Pyruvate Malate Fumarate Lipids Fatty acyl-acps Malonyl-CoA Acetyl-CoA PHA Citrate/isocitrate Succinate Fatty acids Fatty alcohols Alkanes Polyketides 2-OG NH 4 + Amino acids Glu/Gln Cyanophycin Purines Pyrimidines Tetrapyrroles SO 4 2- N 2 and NO 3 -

26 Towards the development of a platform for sustainable chemical production The Nottingham Microalgae Biorefinery Robust production and biorefinery system Downstream processing Molecular tool kit for Cx68 Metabolic engineering Synthetic pathways Sustainable chemical production

27 Thank you.. Steve Skill Carole Llewellyn microalgae metabolites bioactives Gary Farnham molecular biology Bangaru Balasundarm downstream processing