Technologies for utilization of plant biomass as Fermentation Media
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Workshop/Webinar
Moscow, Russia, December 5th 2014
“Fostering R&D cooperation
between German and
Russian stakeholders in the
bioeconomy, especially in
the industrial biotech area“
Technologies for utilization of plant biomass
as Fermentation Media
Petra Schönicke
Research Institute Bioactive Polymer Systems e.V., Teltow-Seehof, Germany
schoenicke@biopos.de
Introduction Biorefinery Systems
Biorefinery
Systems
LCF- Green
Biorefinery Biorefinery
• ligno-cellulosic feedstocks (LCF) • green ‘nature wet’ raw materials
• ‘nature dry’ biomass • green grass
• cellulosic biomass • alfalfa
• cellulosic waste • clover
• wood and fast growing trees • immature cereals
• straw
• reed
2
Kamm, B.; Kamm, M.; Principles of Biorefineries. Appl. Microbiol, Biotechnol., (AMB), 64 (2004) 137–145
Primary refining process and its products
Fields/ harvest Green crop drying plant Pellets for
Alfalafa, grass, clover 151,2 m3/d Drying and pelletizing fodder
20.000 t/a (DM: 20 %) 165 working days per year Sale
(May to October)
Fractionation step Waste heat
Screw extruder Steam / hot water
Press juice Press cake Drying and Pelletizing
76,8 m3/d 74,4 m3/d
(DM:5 %) (DM:35 %)
3
Secondary refining process and its products
Press juice Waste heat / steam
76,8 m3/d (DM 5%) hot water
75,8 m3/d (DM 5 %) 1 m3/d (DM 5 %)
Waste heat, hot water
Heat exchanger Fermentation reactor Protein coagulation
74,7 m3/d (DM 3 %) (60°C)
Waste heat steam Protein
coagulation (75°C)
Protein phase Decantation
Fermentation Decantation (Green protein) Phase separation
Media Phase separation 200 kg/d (DS 15%)
Ultrafiltration Fermentation Media
720 kg/d (DM 1,4%)
Protein Phase Protein phase
(Green protein) (White protein) Fermentation reactor
Waste heat hot water
Fluidized bed drying Spray drying
Press juice-line I Green proteins White proteins Press juice-line II
(Thermo-section) 1704 kg/d (DM 90 %) 11 kg/d (DM 90 %) (Membrane-section)
4
Product properties White proteins High functional potential • for foams, foam stabilizer, films (cosmetic application) Green proteins (high-quality feed) • Amino acids (Asp, Glu, Ser, His, Gly, Thre, Arg, Ala, Tyr, Val, Phe, Ile, Leu, Lys, Pro, Hydroxypro, Met, Cys, Trp) • Carotene • Xantophyll • Fats Brown Juice (for biotechnological processes) • Glucose, Fructose, Sucrose (~ 5-7 g/l) • Proteins, amino acids (N-total 3% TS) • Fats • Minerals (Na+, K+, Ca2+, Mg2+, P, NO3-, SO42-, Cl-) 5
Introduction Biorefinery Systems
Biorefinery
Systems
LCF- Green
Biorefinery Biorefinery
• ligno-cellulosic feedstocks (LCF) • green ‘nature wet’ raw materials
• ‘nature dry’ biomass • green grass
• cellulosic biomass • alfalfa
• cellulosic waste • clover
• wood and fast growing trees • immature cereals
• straw
• reed
6
Kamm, B.; Kamm, M.; Principles of Biorefineries. Appl. Microbiol, Biotechnol., (AMB), 64 (2004) 137–145
Feedstock composition – Wheat straw
Content in original material (as 100% dry matter)[2]
Parameter Value Parameter Value
Cellulose 36.7 % Ash (T=550°C) 7.46 %
Hemicellulose 23.9 % Ash (T=815°C) 7.15 %
Lignin 21.7 % C 42.2 %
Extractives 10.9 % H 5.51 %
Uronic acids 1.92 % N 0.23 %
Harvest of wheat straw [1]
O 42.00 %
Minerals [g/kg] other Minerals [g/kg]
Ca 2.9
(Al, B, Ba, Cd, Cl,
K 2.1 Co, Cr, Cu, Fe, Li,
Mg 0.5 Mn, Mo, Na, Ni, P, 0.9
S 0.5 Pb, Sb, Se, S, Sr,
Ti, V, Zn)
Si 26.9
Straw after grinding, particle size ~1- 5mm
[1] With permission of www.geograph.org.uk 7
[2] NREL methods, Kjeldahl method: EU project BioSynergy No. 038994
Technology of Biorefining lignocellulosic Biomass
Harvested in Brandenburg,
Germany, dry matter: 92 %
Hammer mill
Reactor with heating devices and
recovery of the auxiliary
Protein extraction with hot water
Separation by filtration and wash
water recovery
Solids for enzymatic hydrolysis,
Extractives are the first product
8
Technology of Biorefining lignocellulosic Biomass
Solids come from wheat
straw after pretreatment
Enzymatic Saccharification of celluloses
Water Enzyme coctail
Hydrolysis
at 50°C for 48-72h
Filtration and Solids
Two filtration steps to
Water recovery separate substances which
fine filtration (Lignin)
may disturb fermentation
Enhance sugar concentration
Evaporation Water
with reduced pressure
Final glucose content of
Hydrolysate hydrolysate is approximately
500 g/l
9
Technology of Biorefining lignocellulosic biomass
Enzymatic Hydrolysis
gluc + xyl + ara
100
80
Content of 60
[g/L]
monomeric 40
sugar
20
0
0 24 48 72
time [h]
Conditions of Pretreatment: T=200°C, NH3(25%):BM 5:1; τ =5 min. ;p=38 bar; with protein extraction 10
(BM:H2O = 1:4, 80 °C, 60min.)Utilization of the nonfood sugar solution for fermentation
Hydrolysates from 200 L scale:
Sugar g sugar / kg straw
Glucose 348
Xylose 161
Burkholderia sacchari DSM 17165 [7]
Arabinose 13
ÆDelivered for PHB fermentation[6]
ÆEthanol fermentation tests with yeasts
(Saccharomyces sp.)
ÆABE fermentation tests with bacteria
(Clostridia sp.)
[6] EU project bugworkers No. 246449 (till 2014)
PHB fermentation at IST Portugal [8]
[7] Brämer, C.O., et al.; Int. J. Syst. Evol. Microbiol. 51, 1709-1713 (2001).
[8] Cesario, M.T., Raposo, R. S., Almeida, M.C., Ferreira, B. S., van Keulen, F., da Fonseca, M. M.,„Bioconversion of wheat
straw hydrolysates into polyhydroxyalkanoates“, 6th ESBP, Dublin, Ireland, Sept. 26th-28th 2011
11Technology of Biorefining lignocellulosic biomass
Up-scaling 10 L reactor
1 L reactor
200 mL
Sugar 1L 10 L
Glucose 60.5 g/L 50.8 g/L After scale up: 18 L
Xylose 25.0 g/L 23.6 g/L Glucose 971 g
Arabinose 2.0 g/L 1.9 g/L Xylose 465 g
Arabinose 37 gTechnology of Biorefining lignocellulosic biomass
• Scale-up to
200L scale
“The research leading to these results has received funding from the European Union
13
Seventh Framework Programme (FP/2007-2013) under grant agreement number 246449”Summary
• Brown Juice as Product of a Green Biorefinery
is suitable as Fermentation Medium (Biogas so far)
- but less carbohydrates are available
• Hydrolysate as product of a LCF Biorefinery
is suitable as substrate supplement
(PHB, Ethanol, ABE)
• Utilization of Brown Juice together with Hydrolysate
as Fermentation Medium would be a new R&D issue Picture: biorefinery.de GmbH
• Partners for biomass R&D and biorefining
technology are very welcome
14Biorefinery-Activities FI BIOPOS e.V.
B. Kamm (Ed.) Microorganisms in
Biorefineries, In: Microbiology Monographs,
Series Editor A. Steinbüchel, 2015
http://www.springer.com/life+sciences/
microbiology/book/978-3-662-45208-0
Topics covered include: new metabolic pathways of
microbes living on green plants and in silage; using
lignocellulosic hydrolysates for the production of
polyhydroxyalkanoates; fungi such as Penicillium as
host for the production of heterologous proteins
and enzymes; bioconversion of sugar hydrolysates
into lipids; production of succinic acid, lactones,
lactic acid and organic lactates using different
bacteria species; cellulose hydrolyzing bacteria in
the production of biogas from plant biomass; and
isoprenoid compounds in engineered microbes.
15Contact Dipl.-Ing. Biotech. Petra Schönicke
Research Centre Teltow-Seehof
Prof. Dr. Birgit Kamm Kantstraße 55, D-14513 Teltow
Director of the Institute Biopos e.V. www.biopos.de
and Prof. at BTU Cottbus Fon: +49 (0)33 28-33 22-18
Research Centre Teltow-Seehof schoenicke@biopos.de
Kantstraße 55, D-14513 Teltow
www.biopos.de
Fon: +49 (0)33 28-33 22-10
Fax: +49 (0)33 28-33 22-11
kamm@biopos.de
Dr. Jörg Beckmann
biorefinery.de GmbH
Kantstraße 55, D-14513 Potsdam
Email:office@biorefinery.de
www.biorefinery.de
www.biorefinica.de
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