BIOVALUE PROJECT 2 ANDERS PETER S. ADAMSEN AARHUS UNIVERSITY HEALTH 04 JUNE 2015

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1 BIOVALUE PROJECT 2 ANDERS PETER ADAMSEN

2 CONTENT Protein work KU Effekt of time lag between harvest and processing Flow sheeting Pilot plant

3 OLIGOSACCHARIDES - A Upgrading of C5 oligomers Structural composition of short arabinoxylooligosaccharides was semi-quantitatively approximated by combinatorial model, which provided the basis for any structural considerations in future experiments with prebiotic activity testing. Short linear xylooligosaccharides (2-4 degree of polymerization) were produced from hydrothermally pretreated wheat straw as a result of xylanase and cellulases treatment. Enzymes for increased protein extraction Identification, cloning and characterisation of Chaetomium thermophilum feruloyl esterase was performed. Putative feruloyl esterase gene was identified in Chaetomium thermophilum transcriptome; novel recombinant enzyme was produced in Pichia pastoris and characterized.

4 PROTEIN PLATFORM - KU Development of a fast protein hydrolyses platform for amino acid composition analysis with microwave assisted acid hydrolysis: Hydrolysis % recovery of Bovine Serum Albumin Running in on proteins from green biomass under development. Looks fine. Analysis of cell wall proteins etc. Separation of proteins from cell wall Complete N balances (Sol. AA, NH4+, NO3- & proteins) in green biomass Running in a new carbohydrate analyses methods (monosaccharides)

5 PROTEIN FEED FROM GREEN BIOMASS Fertilizer Biogas plant Manure Heat/ power Maceration Grass refinery Pressing Residuel juice Precipitation Fractionation Grass Refinery Protein concentrate Fibre Monogastric animal feed Ruminant animal feed Food 5

6 EXPERIMENTAL SETUP Biomasses: Rye grass & White clover Rye grass White clover Harvest: Layed / Chopped (11 November) Storage: 1m 3 box, simulating a large pile Harvest Layed in swaths Time: Processed after 0, 4, 24, & 48 hours Processed in triplicates Chopped Layed 6

7 Influence of harvest method and post-harvest time on protein extraction yield Temp. C Temperature increased during storage Rye grass Layed Rye grass Chopped Chopped Layed Hours after harvest BioBase 7

8 CONCLUSIONS SO FAR No trend of dry matter or crude protein yields changing over time Microbial growth is induced by chopping the biomass The difference will be in the quality Total amino acid content is needed to give an estimation of true protein. 8

9 SIMPLE PROCESS SIMULATION IN EXCEL Anders Peter Adamsen & Morten Ambye-Jensen Authors: Denotation: Input cells: Green number Calculation: Blue numbers Fibre product Protein kg/h Protein ,8% Sol. carbohydrates kg/h Sol. carbohydrates ,0% Hemicellulose kg/h Biomass Hemicellulose 458 7,0% Cellulose kg/h Biomass reception & storage system Maceration and separation fibre Cellulose ,6% Lignin kg/h Capacity 24 h Feeding Fractionation Lignin ,8% Residual 500 kg/h reception and Screw press Residual 332 5,1% P 30 kg/h P 19 0,3% system K 300 kg/h K 187 2,9% Ash (-P&K) 670 kg/h storage Ash (-P&K) 425 6,5% Necessary to Water kg/h Water Sum DM kg/h Sum DM Sum total kg/h Recycling Protein Juice % Sum total DM 18,0% 18,0% 70% Sol. carbohydrates ,7% % DM 19% Protein 25,0% total DM 25,0% Hemicellulose ,2% % Protein 20% Temp. 10 C 10 Cellulose % Enthalpy 0 GJ 0 Target DM Lignin % 12% Residual % Total stream needed P % Capacity t DM/yr 10 t DM/h K % Working days 3000 h Ratio of sidestream Ash (-P&K) % Input DM content 18% Separation efficiency 70% Water % Maceration etc. Protein separation Sum DM Steam Condensed or cooled water Protein 25,0% DM 50% into the juice 90% into Protein Conc Sum total Sol. carbohydrates 20,0% DM 70% into the juice 30% into Protein Conc DM 6% 6% - - Hemicellulose 10,0% DM 70% into the juice 30% into Protein Conc Protein 3% 3% - - Cellulose 20,0% DM 5% into the juice 50% into Protein Conc - precipitation - Protein Lignin 10,0% DM 5% into the juice 50% into Protein Conc - Temp. 70 C - Residual 5,0% DM 50% into the juice 30% into Protein Conc - Δh 6 GJ/h - P 0,3% DM 50% into the juice 40% into Protein Conc Heat exchanger precipitation K 3,0% DM 60% into the juice 15% into Protein Conc Ash (-P&K) 6,7% DM 50% into the juice 40% into Protein Conc - Δt 30 C - Total (check) 100,0% DM Water 455,6% DM 60% into the juice 15% into Protein Conc - - Unit (kg/h) Feed Fibre product Protein Concentrate Side stream Control Protein % % % 38 4% 0 0,0% Sol. carbohydrates % % % % Hemicellulose % 458 7% % % Cellulose % % 51 2% 15 2% 0 51 Lignin % % 25 1% 8 1% Residual 500 5% 332 5% 100 4% 69 7% - 19 P 30 0% 19 0% 8 0% 3 0% K 300 3% 187 3% 42 2% 71 7% Ash (-P&K) 670 7% 425 7% 170 7% 75 8% % Water Side stream Protein Concentrate Sum total Protein 38 4% 130 4% Protein ,2% Sum DM % % % % - Sol. carbohydrates % % Sol. carbohydrates ,4% DM 18,0% 18,7% 28,3% 8,1% Hemicellulose % % Hemicellulose ,7% Cellulose 15 2% waste water 51 2% Protein Separation Protein concentrate Cellulose 51 2,0% Liquid/Solid Amt. (t DM/yr) Lignin 8 1% 25 1% Lignin 25 1,0% 100% 65% 25% 9% Residual 69 7% 232 7% Decenting centrifuge Residual 100 3,9% P 3 0% 11 0% separation P 8 0,3% 25% 25% 20% 20% 46% 46% 4% 4% K 71 7% 239 7% K 42 1,7% Necessary to Ash (-P&K) 75 8% 255 8% Ash (-P&K) 170 6,7% Water Water Sum DM Sum DM Sum total Sum total DM 8,1% 8,1% DM 28,3% Protein 4,1% 4,1% Protein 46,2% BioBase 9

10 PILOT PLANT

11 Biomass Feeding System Mixing feeder, Macerator, Lobe pump 19m 3, mixing, load sensor Recirculation of residue juice Pumpable feed stream Cuts down to 10 mm BioBase 11

12 Fractionation step Screw press - Prototype m 3 per hour μm filter Thin filter cake Separate juice outputs Fibre DM of ca. 35% BioBase 12

13 Protein precipitation and separation Buffer tank, Pump, Heat exchange, Decanter centrifuge Temperature increase: 10 -> 80 C Temperature decrease: 80 -> 20 C Centrifuge capacity 1-5 m 3 /hr Prevents air intake to reduce foaming BioBase 13