A NOVEL GREEN BIOREFINERY CONCEPT. María Santamaría-Fernández // Section for Sustainable Biotechnology // Aalborg University

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1 A NOVEL GREEN BIOREFINERY CONCEPT María Santamaría-Fernández // Section for Sustainable Biotechnology // Aalborg University

2 THE ORGANOFINERY CONCEPT Protein feed, biogas and fertilizer 2 MARIA SANTAMARIA-FERNANDEZ // Green Biorefining Seminar // JUNE 29, 2018

3 PROGRESS ALONG THE PROJECT 1 PROTEIN EXTRACTION YIELDS AMINO ACID COMPOSITION PROTEIN BIOREFINING LAB-SCALE 3 MARIA SANTAMARIA-FERNANDEZ // Green Biorefining Seminar // JUNE 29, 2018

4 4 Mech an ical fractionation of the fresh crops into green juice and press cake Red clover Clover grass Alfalfa Oilseed radish Screw pressing 40-50% ww PRESS CAKE 50-60% ww GREEN JUICE 4 MARIA SANTAMARIA-FERNANDEZ // Green Biorefining Seminar // JUNE 29, 2018

5 5 Lactic acid fermentation of green juices and production of protein concentrates GREEN JUICE Lactic acid fermentation Centrifugation PROTEIN CONCENTRATE Lactobacillus salivarius BC 1OO1 37 C, overnight Final ph % ww Overall, between 3-8% of crops wet weight was extracted into the protein concentrates BROWN JUICE 5 MARIA SANTAMARIA-FERNANDEZ // Green Biorefining Seminar // JUNE 29, 2018

6 Up to 23% of plant proteins were recovered in the protein concentrates PROTEIN CONCENTRATES Crude protein recovery (%) 25 23, ,1 15,1 12, Red clover Clover grass Alfalfa Oilseed radish 6 MARIA SANTAMARIA-FERNANDEZ // Green Biorefining Seminar // JUNE 29, 2018

7 The essential amino acids represented 53% of the true protein content Essential AA Non-essential AA Red clover 53% 397 Clover grass 53% 349 Alfalfa 53% 346 Oilseed radish 53% 374 Soybeans* 48% Amino acid composition (g kg -1 DM) Essential AA for poultry: ARG, HIS, ILE; LEU; LYS, MET, PHE, THR, VAL *Steenfeldt and Hammershøj, MARIA SANTAMARIA-FERNANDEZ // Green Biorefining Seminar // JUNE 29, 2018

8 Methionine is usually first-limiting amino acid in most poultry feeds Methionine Essential AA Red clover 4% 211 Clover grass 4% 185 Alfalfa 4% 183 Oilseed radish 5% 197 Soybeans* 3% Amino acid composition (g kg -1 DM) *Steenfeldt and Hammershøj, MARIA SANTAMARIA-FERNANDEZ // Green Biorefining Seminar // JUNE 29, 2018

9 PROGRESS ALONG THE PROJECT 1 PROTEIN EXTRACTION YIELDS AMINO ACID COMPOSITION 2 SCALABILITY AND PRACTICALITY PRODUCE PROTEIN CONCENTRATE FOR FEEDING TRIALS PROTEIN BIOREFINING LAB-SCALE PROTEIN BIOREFINING DEMO-SCALE 9 MARIA SANTAMARIA-FERNANDEZ // Green Biorefining Seminar // JUNE 29, 2018

10 Fermentation tank Grass clover feeding system Tank trailer Press cake, screw conveyor Screw press

11 Filter sieve 200µm Sand sedimentation tank 10 m 3 fermentation tank

12 Around 10 tons of wet protein concentrate were produced from 444 tons of clover grass 50% ww PRESS CAKE 223 tons SILAGE BALES 2% ww FRESH CLOVER GRASS 444 tons 40% ww GREEN JUICE 177 tons WET PROTEIN CONCENTRATE 7 tons DRY PROTEIN CONCENTRATE 1 tons 12 MARIA SANTAMARIA-FERNANDEZ // Green Biorefining Seminar // JUNE 29, 2018

13 The overall protein recovery in the protein concentrate was 7% Protein concentrate Press cake Brown juice Loss Crude protein recovery (%) Grass clover - Demo scale 17 Grass clover - Lab scale* 13 MARIA SANTAMARIA-FERNANDEZ // Green Biorefining Seminar // JUNE 29, 2018 *Santamaria-Fernandez et al., 2017

14 Content of amino acids was lower than in protein concentrates produced at lab scale Grass clover - Demo scale Essential AA 52% Non-essential AA 294 Grass clover - Lab scale* 53% 349 Red clover - Lab scale* 53% 397 Soybeans** 48% Amino acid composition (g kg -1 DM) *Santamaria-Fernandez et al., 2017 **Steenfeldt and Hammershøj, MARIA SANTAMARIA-FERNANDEZ // Green Biorefining Seminar // JUNE 29, 2018

15 PROGRESS ALONG THE PROJECT 1 PROTEIN EXTRACTION YIELDS AMINO ACID COMPOSITION 2 SCALABILITY AND PRACTICALITY PRODUCE PROTEIN CONCENTRATE FOR FEEDING TRIALS 3 PLANT DEVELOPMENT STAGE ON PROCESS YIELDS OPTIMAL HARVESTING TIME PROTEIN BIOREFINING LAB-SCALE PROTEIN BIOREFINING DEMO-SCALE PROTEIN BIOREFINING OPTIMIZATION 15 MARIA SANTAMARIA-FERNANDEZ // Green Biorefining Seminar // JUNE 29, 2018

16 Plants were harvested at d ifferen t development stages (D S) ensuring morphological changes Chicory Cichorium intybus Red clover Trifolium pratense Timothy Phleum pratense DS1 ELONGATION Main shoot ELONGATION 40% full height ELONGATION Second internode DS2 ELONGATION 5 to 9 internodes INFLORESCENCE EMERGENCE Some buds visible EARING 70% earing DS3 INFLORESCENCE EMERGENCE First individual flowers (closed) FLOWERING Half way to full flowering 16 MARIA SANTAMARIA-FERNANDEZ // Green Biorefining Seminar // JUNE 29, 2018

17 Crude protein content in the plants significantly decreased with maturity FRESH PLANTS Crude protein content (%DM) DS1 DS2 DS Chicory Red clover Timothy 17 MARIA SANTAMARIA-FERNANDEZ // Green Biorefining Seminar // JUNE 29, 2018

18 Protein extractab ility in protein concentrates decreased significantly with maturity Protein extractability g CP in Prot conc kg DM plant DS1 DS2 DS Chicory Red clover Timothy 18 MARIA SANTAMARIA-FERNANDEZ // Green Biorefining Seminar // JUNE 29, 2018

19 Production of protein concentrates also decreased with maturity DS1 DS2 DS3 Prot conc. production g dry Prot conc kg DM plant Chicory Red clover Timothy 19 MARIA SANTAMARIA-FERNANDEZ // Green Biorefining Seminar // JUNE 29, 2018

20 CONCLUSIONS & FUTURE PERSPECTIVES The Organofinery process allowed producing up to 235 kg dry protein concentrate per ton of dried biomass. Laboratory-scale trials: Lactic acid fermentation-centrifugation method i.e. between 40-80% of proteins in green juice can be recovered. Up to 42% of proteins in fresh biomass recovered in the protein concentrate. Several challenges to overcome in large scale production: separation of proteins and drying of the protein concentrate. Protein concentrate good source of essestial amino acids for poultry.

21 FUTURE PERSPECTIVES Techonoeconomical analysis Fresh crop Alternative products e.g. lactic acid Drying Screw pressing OPTIMIZATION Nutritional value of protein concentrates Protein separation Fermentation Food-grade protein concentrates? 21 MARIA SANTAMARIA-FERNANDEZ // Green Biorefining Seminar // JUNE 29, 2018

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