Biodiesel, high-value LC omega-3 oils & algal meal production from thraustochytrids in a biorefinery approach

Transcription

1 Biodiesel, high-value LC omega-3 oils & algal meal production from thraustochytrids in a biorefinery approach Kim Jye Lee Chang*, Maged P. Mansour, Peter D. Nichols, Geoff J. Dumsday, Carol Mancuso Nichols, Lesley Clementson, Susan I. Blackburn Algal Ecology and Resources CSIRO Oceans and Atmosphere, GPO Box 1538, Hobart TAS 7001 October 24, 2016 Algae Biomass Summit, Phoenix, Arizona

2 CSIRO Marine Laboratories, Hobart, Tasmania 2

3 Australian National Algae Culture Collection Algal Culture Laboratory

4 Why algal-derived biofuels? Area required to supply 50% of Australia s transportation fuel Does not compete with food supply Palm Can be harvested year round Use of non arable land Renewable Microalgae source; 2% global CO2 emissions Energy security; 1 Future supply issues (peak oil) 2 Jatropha Canola Wheat Crop (as reference) 4

5 Current commercial interest in algal biofuels Organic Biomass Carbohydrates Oils (TAG) Proteins Pigments Bioethanol Biodiesel Amino acids Antioxidants Transesterification Triacylglycerol (TAG) Methanol Glycerol Biodiesel (Fatty Acids Methyl Ester) Hydroprocessing Triacylglycerol Biobased jet (Hydrocarbon) ASTM Approved 5

6 20µm Heterotrophic thraustochytrids LC Omega-3 Oils: C20, two or more double bonds H 3 C COOH 20:5w3 Eicosapentaenoic - EPA Thraustochytrids SFA + MUFA Biofuels H 3 C Fish consume microalgae & COOH accumulate ω3 LC-PUFA 22:6w3 Docosahexaenoic - DHA Nutritional Anti-thrombotic supplements Anti-inflammatory Aquaculture feeds Other co-products EPS and carotenoid pigments 6

7 Research objectives Ø Isolation and characterisation of thraustochytrids - Endemic strains to protect Australia s biodiversity Ø Strain selection for production of biodiesel - High growth, high lipid production Ø Co-production of high value-added by-products - Long-chain (LC) omega-3 oils, pigments, EPS Ø Life Cycle Analysis - GHG emissions and ERoEI (Energy Returned on Energy Invested) Ø Waste stream utilization - Recycling crude glycerol from biodiesel production 7

8 Isolation and characterisation of thraustochytrids Acriflavine detection Positive Negative DNA extraction Lipid extraction Pigment extraction Amplification and sequencing of 18S ribosomal RNA genes Gas Chromatography (GC) & Mass Spectrometry (GC-MS) High Performance Liquid Chromatography (HPLC) Lee Chang et al. (2011) Phytochemistry Lee Chang et al. (2012) Appl. Microbiol. Biotechnol 8

9 Sample collection sites and habitats River mouth and estuary (Tas) Morphological characteristics of thraustochytrids Mangrove forest (Qld) 20µm 200µm Ectoplasmic net elements Zoospores (motile) 9

10 Genus Schizochytrium Thraustochytrium Ulkenia Aurantiochytrium Group A B C D E F G H Temperate Tropical + LC omega-3 oils :6ω3 DHA :5ω3 EPA SFA + MUFA Odd Chain-PUFA β,β-carotene Canthaxanthin + + Astaxanthin + + Cholesterol Tr Stigmasterol + + Tr + + Brassicasterol + + Tr Tr Tr Tr Lee Chang et al. (2011) Phytochemistry Lee Chang et al. (2012) Appl. Microbiol. Biotechnol Biodiesel 10

11 Potential Production of Biodiesel, LC Omega-3 Oils & EPS 1L Flask culture on shaker Centrifugation 85% saturated FA (biodiesel) LC omega-3 oils Biomass: lipid EPS- filtration supernatant 18.5 g cell dry weight per L (34% TFA) at 9 days of growth K. J. Lee Chang et al. (2014) Marine Biotechnology 11 (Schizochytrium sp. strain 300 mg EPS /L )

12 Lipid fractionation A. Soluble TAG fraction Response 16:0 47% 22:6w3 DHA 39% A 14:0 18:0 ARA EPA 20:4w3 22:5w3 B. Insoluble TAG fraction Response :0 71% 22:6w3 DHA 19% B 14:0 18:0 22:5w Retention time (min) 12

13 Fed-batch cultivation in bioreactors DCW" Total"FAME" 16:0" 22:6w3"DHA" Ammonia"added"(mL)" Glucose'(g/'L)' DCW'(g/L)' Total'FAME''(g/L)' 80" 70" 60" 50" 40" 30" 20" 10" 0" 0" 0" 10" 20" 30" 40" 50" 60" 70" 80" Time'(h)' Carbon source Biomass Lipid Time Lee Chang et al. (2013) Thraustochytrids Glycerol 70 g/l 52 % 69 h Appl. Microbiol. Biotechnol. Yan et al. (2011) Chlorella protothecoides 160" 140" 120" 100" 80" 60" 40" 20" Ammonia'added'(mL)' Molasses 71 g/l 58 % 178 h 13

14 Life Cycle Assessment: heterotrophic cultivation Thraustochytrids Bioreactor Centrifugation Homogenizers Aquaculture feeds Biofuels Transesterification Extraction Nutritional supplements 13

15 Preliminary LCA Results GHG emissions & ERoEI of the heterotrophic microalgae production system. GHG (gco2e/mj) Upstream Downstream Balance ERoEI Biodiesel (Glycerol) Diesel (Fossil) Biodiesel (Molasses) Lee Chang et al J Appl Phycol 15

16 Biomass yield with combination of the waste soybean meal, protein meal and porcine mucosa digest 4 % w/v crude glycerol from biodiesel plants Lee Chang et al. (2015) J Funct Foods 16

17 Lipid extraction & algal biomass Algal meal % Protein 30 Carbohydrate 50 Lipid 16 Ash 4 FAME % FA Pure Crude 16: :5ω6 DPA :6ω3 DHA SUM SFA SUM PUFA

18 Summary 36 new thraustochytrid strains - temperate & tropical environments Lipid profiles with potential biodiesel & LC omega-3 applications Aurantiochytrium sp. strains, Group E DHA ( 60%), Group G, H biodiesel Maximum non-optimized yield of EPS was observed for Schizochytrium sp. (299 mg/l) Scale-up in a fed-batch cultivation system Biomass & oil production improved 71 g/ L DCW (52% TFA) at 69h of fermentation LCA of heterotrophic system - comparable to fossil diesel 20 g/l DCW at 69h of fermentation in crude glycerol medium Future research further optimize biomass & oil production, scale-up, feeding trials 18

19 Thank you CSIRO Oceans and Atmosphere CSIRO National Research Collections CSIRO Food and Nutrition Intelligent Processing Transformational Capability Platform (IP TCP) Australian National Algae culture Collection (ANACC) Acknowledgements: Anthony Koutoulis, Graeme Dunstan, Dion Frampton, Ian Jameson, Cathy Johnston, Lucas Rye, Tim Grant, Helen Paul Kim Lee Chang phone: (+61)