1. Phalaris aquatica L. lignocellulosic biomass as second generation bioethanol feedstock I. Pappas, Z. Koukoura, C. Kyparissides, Ch. Goulas and Ch. Tananaki Aristotle University of Thessaloniki Faculty of Forestry and Natural Environment Range Science Laboratory The Centre for Research and Technology Hellas Chemical Process Engineering Research Institute EGF 2012, Lublin

2. Outline  Introduction  Objective  Study area  Material and Methods  Results and Discussion  Conclusions

3. The problem

4. The consequence

5. The sources a)Fossil fuel combustion b) Land use change-degradation

6. Rising oil prices Source: Energy Information Administration, Cushing OK Spot Price

7. Energy security

8. Renewable energy resources Biomass Wind Solar Geothermy Hydropower Figure 1: Production of primary energy, EU-27, 2008,% of total, based on tonnes of oil equivalent) (Source: Eurostat)

9. Herbaceous perennial energy crops criteria: 1)High biomass yield potential. 2)High lignocellulosic content of aboveground biomass. 3)Good suitability for no prime agricultural land. 4)Low establishment cost. 5)Positive environmental impacts ( erosion protection, soil carbon sequestration, nutrient recylce, biodiversity).

10. Phalaris aquatica L. species traits Perrenial C 3 temperate grass. Native species of the Mediterranean region. Yield stability. High lignocellulosic content of aboveground biomass. High drought tolerance. Abundant seeds. Sustainable biomass production (soil cover, carbon accumulation, nitrogen recylce).

11. Study area Figure 1. Chysopigi site Figure 2. Thermi site

12. Climatic conditions Figure 3. Climatic conditions of Thermi site Figure 4. Climatic conditions of Chysopigi

13. Material and Methods I. Biomass yield  Biomass samples were collected at the end of the growing season of 2009.  Biomass production was measured using 10 (1x1 m 2 ) plots.  Harvested biomass was dried at 60 0 C for 48h and milled to a size of 1 mm. II. Lignocellulosic content  Fiber analyses (NDF, ADF, ADL) was conducted using the Van Soest method (Van Soest et al., 1991).  Hemicellulose = NDF – ADF  Cellulose = ADF – ADL  Lignin = ADL

14. Results ThermiChrysopigi Biomass yield (Mg ha -1 DM) 6.26a4.90b NDF (g Kg -1 DM) 770a740b ADF (g Kg -1 DM) 503a441b ADL (g Kg -1 DM) 63a73b Soluble cell components (g Kg -1 DM) 230a260b Table 1. Mean biomass production and cell wall concentration of Phalaris aquatica at sites with different altitude and climatic conditions

15. Cell wall structure

16. Results Figure 3. Mean cell wall components concentration of Phalaris aquatica at sites with different altitude and climatic conditions

17. Biomass conversion path Lignocellulosic Biomass Thermochemical and Biochemical Process Bioethanol

18. Flowsheet of bioethanol production

19. Pretreatment Scope: Alter the structure of lignocellulosic biomass and make it accessible to enzymes for hydrolysis of carbohydrates to sugar monomers.

20. Experimental 1) Biomass loading = 10 % (w/v) 2) Temperature: 120 0 C 3) H 2 SO 4 concentration: 1.5% (w/w) 4) Time: 45 min Sugar monomers (g Kg -1 DM) Total sugars Xylose Arabi- nose Galactose, Glucose, mannose 134,329,148,6212 Figure 5. SEM photo of the pretreated biomass

21. Enzymatic hydrolysis Scope: Τhe hydrolysis of insoluble cellulose fraction of lignocellulosic biomass to fermentable sugar monomer (glucose). Factors: 1.Biomass characteristics (hemicellulose and lignin content, degree of polymerization, cellulose crystallinity) 2.Enzyme synergy (origin, composition) 3.Physicochemical characteristics (temperature, pH, buffer, agitation, reaction time). Schematic representation of the enzyme action on insoluble cellulose

22. Experimental conditions Biomass loading: 2% (w/v) Enzyme loading: 40 FPU/g biomass Time: 48 h Agitation speed = 150 rpm Citrate buffer, V = 50 ml, pH = 4.8 Pretreatment Enzymatic hydrolysis Monomeric sugars (g Kg -1 D.M ) 212424 Total sugars Released Cell wall carbohydrates Conversion efficiency (%) 63670789,9 Figure 5. Shaking incubator

23. Conclusions Phalaris aquatica L. is a potential second generation bioethanol feedstock at lowland sites of North Greece due to higher biomass yield and total structural carbohydrate concentration compared to upland ones. Acid pretreatment followed by enzymatic hydrolysis, is an effective biochemical method to release fermentable sugar monomers from Phalaris aquatica L. cell wall carbohydrates for bioethanol production.

24. Keep dreaming

25. Thank you very much!