1. Investigating and Modeling Natural Biodegradation System in Soil; Application for Designing an Efficient Biological Pretreatment Technology for Biofuel Production. Mythreyi Chandoor, Deepak Singh and Shulin Chen Bioprocessing and Bioproduct Engineering Laboratory, Department of Biological Systems Engineering Washington State University.

2. outline Aim and importance of the project Background – Hypothesis of the project Experimental:  Microbiology  Chemical analysis of lignocellulose degradation in soil. Modeling  Lignocellulose degradation in soil  Application Acknowledgements

3. Aim and importance of the project Demand for an Alternative Fuel – The U.S. ethanol consumption is forecast to increase from 5.6 billion gallons last year to 13.5 billion gallons in 2012, (Thomson Reuters, 2009). What are the challenges ? One of the challenges lies in the deconstruction of lignin part of the biomass to release sugars. Need for novel pretreatment Technology !! Demand for an Alternative Fuel – The U.S. ethanol consumption is forecast to increase from 5.6 billion gallons last year to 13.5 billion gallons in 2012, (Thomson Reuters, 2009). What are the challenges ? One of the challenges lies in the deconstruction of lignin part of the biomass to release sugars. Need for novel pretreatment Technology !!

4. Delignification, repolymerization Humus formation Proteins in soil Lignocellulosic system in soi l

5. To understand the biodegradation of lignocellulose in soil To model the biodegradation of lignocellulose in soil Design the pretreatment system Aim of the Project

6. Methodology

7. Scanning Electron Microscopy (SEM)

9. Aromatic carbons attached to methoxy groups in syringol unit Guiacyl moities C2,C3,C5 of cellulose C4 of amorphous and crystalline cellulose Phenolmethoxyl of coniferyl and sinapyl moities 4 weeks 8 weeks 12 weeks 16 weeks Solid State NMR Analysis

10. The Oxidation of syringyl and guaicyl units of lignin will give rise to syringol and guaicol units.

11. Quantitatively, syringyl and guaicyl units have decreased where as the syringol and guaicol amounts have increased which shows that there is change in the chemical nature of lignin structure Solid State NMR Analysis

12. Batch samples for every four weeks % Concentration of the total compound Py-GC/MS Analysis

13. The increase in the lignin content is attributed to the kind of subunits taken into consideration ; Syringol,Guaicol, Ethanone and others were considered which are formed as a result of oxidation or modification of lignin.

14. Cellulose and Hemicellulose are proportionately decreasing while the lignin concentration is stable and increased after a period of 12 weeks Py-GC/MS Analysis

15. TG Analysis Cellulose and hemicellulose Lignin

16. Modeling

17. d(S1) / d(t) = -Vb1*S1*X1/(Ks1+S1) #Cellulose Balance S1(0) = 0.71 # weight in gm/gm d(S2) / d(t) = -Vb2*S2*X2/(Ks2+S2) #Hemicellulose Balance S2(0) = 0.48 # d(S3) / d(t) = -Vb3*S3*X3/(Ks3+S3) #Lignin Balance S3(0) = 0.28 # Modeling Considering the values as follows ; µmax1=0.08 ∆ 1=0.001 µmax2=0.05 ∆ 2=0.001 μmax3=0.03 ∆ 3=0.001 We derived an relation using polymath which defines the degradation pattern in the soil system.

18. Modeling Time (in hours ) Initial Substrate concentration in gm / gm

19. Application of the model The model developed is a relation drawn between the total initial concentrations of the cellulose, hemicellulose and lignin defined in a specific proportion at any point of time. Further,the model would correlate the various factors involved parallel to the degradation rates of each component respectively. The model developed is a relation drawn between the total initial concentrations of the cellulose, hemicellulose and lignin defined in a specific proportion at any point of time. Further,the model would correlate the various factors involved parallel to the degradation rates of each component respectively.

20. Conclusion The determination of the exact relation between these factors would be helpful in developing a model which would predict the specific ratio of cellulose, hemicellulose and lignin apart from other factors involved such as pH, temperature and other organic compounds. Thus providing a suitable mechanism for the pretreatment technology !! The determination of the exact relation between these factors would be helpful in developing a model which would predict the specific ratio of cellulose, hemicellulose and lignin apart from other factors involved such as pH, temperature and other organic compounds. Thus providing a suitable mechanism for the pretreatment technology !!

21. I would like to thank Dr. Ann Kennedy USDA-ARS Soil Scientist/ Adj. Prof. Crop and Soil Sciences,WSU. Dr. Greg Helms, NMR Center Director,WSU. Dr. Manuel Garcia-Perez. Assistant Professor / Scientist. Biological Systems Engineering,WSU. Dr. Bill, Assistant manager,NMR Center,WSU. And my Advisor … Dr. Shulin Chen, Professor/Scientist. Department of Biological Systems Engineering,WSU.

22. Acknowledgements And My Team …

23. Thank you … Any Questions ?