1. Fiorello B. Abenes, Ph.D. Visiting Professor, Engineering R and D for Technology, 2009 Fulbright Fellow, 2009 DOST Balik-Scientist, 2008 Visiting Scientist, Philippine Carabao Center at CLSU Visiting Professor, Central Luzon State University Emeritus Professor, CalPoly University Pomona The Philippine Carabao: A Paradigm for Biomass Ethanol Production All rights reserved

2. Philippine Biofuels Act E-5 in 2009 E-10 in 2011

3. R.A. 9637 Bioethanol Feedstocks

4. “ The savior of the sugar industry is the biofuels industry.” Sen. Juan Miguel Zubiri, quoted by Sun Star Cebu, Feb.24, 2008

5. Socio-economic Issues of Biofuels

8. Food or Fuel?

9. Food prices have risen as more land is used to produce biofuels Biofuels 'crime against humanity' By Grant Ferrett BBC News A United Nations expert has condemned the growing use of crops to produce biofuels as a replacement for petrol as a crime against humanity

10. Dr. Hartmut Michel – Nobel Laureate (Chemistry) Rethink biofuel - There’s not real energy gained in biofuel We don’t need to tap other lands … use areas already planted to sugar. Sugar is not a basic food source. It is an additive, and so won’t compete directly with the population’s food requirement “ DECELERATE BIOFUEL” - provide funds for the newly created Biofuels Oversight Committee, to ensure that food acreage will not be prejudiced by biofuel acreage

11. It took 10 years to produce the first billion liters of fuel ethanol; another 10 years for the second billion It took only 18 months to produce the next 10 billion liters

12. World map: Kyoto Protocol, participation ██ Signed and ratified. ██ Signed, ratification pending. ██ Signed, ratification declined. ██ No position. As of April 2008, 178 states have signed and ratified the Kyoto Protocol to the United Nations Framework Convention on Climate Change, aimed at combating global warming.

13. The Alternative: Biomass

14. Biomass Structure

16. PCC/CLSU Biomass Deconstruction Paradigm An example serving as a model

18. Rumen Microflora and Microfauna

20. Metabolism in Yeast under Aerobic and Anaerobic conditions

21. The Heterolactic Fermentation Pathway The Entner-Doudoroff Fermentation Pathway UNIQUE PATHWAYS OF GLUCOSE METABOLISM IN PROKARYOTES

22. PCC/CLSU Biomass to Ethanol Production System

24. A Carabao fitted with rumen cannula Romy: Our rumen fluid donor

25. BIOMASS RUMEN FLUID 1 ST STAGE FERMENTATION PROPAGATED YEAST DISTILLATION CELLULOSE ETHANOL 2 nd stage FERMENTATION PASTEURIZATION Chopped to <10 mm

26. Efficiency of Cellulose to Sugar Conversion - Rice Straw 84.5% Efficiency of conversion Cellulose to glucose 186 g Amount of glucose 1.2% Glucose content 15.5 L Hydrolysate Volume 220 g Amt of cellulose in 500 g substrate

27. Efficiency of Conversion to Ethanol 32 g Ethanol Equivalent Yield (51.5% X 62 g) 62% Efficiency of sugar to ethanol conversion (% of theoretical yield) 51.5 Stoichiometric Yield, sugar to ethanol g/100 62 g Amount of sugar fermented .4% X 186 .4% Sugars used by yeast 1.2% - 0.8% 186 g Initial sugar in the hydrolysate

28. Ethanol Yield of the PCC/CLSU System 117 kg (39 gal or 148 L) Ethanol yield Ethanol Specific gravity = 0.79 Ethanol yield from sugar cane, fresh basis, is 20 gal/ton; sweet sorghum is 10.5 gal/ton Source: www.icrisat.org/Media/2004/media13.htm X 0.62 2 nd Stage sugar to ethanol conversion efficiency X 0.51 % Ethanol stoichiometric yield X 0.84 1 st Stage cellulose to sugar conversion efficiency X 0.44 % Cellulose 1 ton (1000 kg) Rice straw

29. Characterization of Ethanol produced from the PCC/CLSU System Contaminants differentiated from rice straw cellulose ethanol (CH3CH2OH) by molecular weight using mass spectrophotometer: 1,4-dioxane-2-6-dione(CH4H4O4 ) 2-formylhistamine (C6H9N3O) chloro-ethyl ester (C4H7ClO2) 1-amino-2-butanol(C4H11NO) acetic acid (C3H4O3) ethanedial (C2H2O2) urea N-methyl-N-nitroso ( C2H5N3O2) Ethanol “Purity” Range: 46.77% to 76.44% Average: 58.62%

30. Cellulose to Sugar Conversion of Sweet Sorghum Bagasse 1.2% 12 1.0% 15 1.4% 9 1.8% 6 1.4% 3 % Glucose in hydrolysate Days of Incubation with 1% Rumen Fluid + urea

31. Production Efficiencies of the Ethanol Technology Using Carabao Rumen Fluid Hydrolysis 246.98 kg (312.63 L or 82.5 gal.) Ethanol yield Ethanol Specific gravity = 0.79 X 0.515 2 nd Stage Biological system - Bakers yeast fermentation conversion efficiency X 0.97 1 st Stage biological system - carabao rumen fluid fermentation conversion efficiency X 49.44 Structural carbohydrates 1 tonne (1000 kg) Sweet sorghum bagasse

32. Theoretical Yields of Selected Biomass Feedstocks Feedstock Theoretical Yield (gal/ton) Corn Grain 124.4 Corn Stover 113.0 Rice Straw 109.9 Forest Thinnings 81.5 Hardwood Sawdust 100.8 Bagasse 111.5 Mixed Paper 116.2 http://www1.eere.energy.gov/biomass/ethanol_yield_calculator.html

33. + Microbial production Current B2B Protocol PCC/CLSU Protocol Methane and CO 2 Acid chopping cellulase cellulase & hemicellulases hexose hexose & pentose

34. Thank you! Rumenol - Powered to the last drop! R

35. Acknowledgements Perla C. Florendo, Ph.D. (Cand) Rene Agulto, Ph.D., Dean, CLSU College of Engineering Marlon Ocampo, PhD, DVM Felomino V. Mamuad, PhD, Director PCC at CLSU Adel Valdez, PhD, Director, CLSU-RSTC Juvy Monserate, PhD (Cand) CLSU Bioethanol Team Undergraduate Students Aris Marilao and R.C. Caluya

36. PICTURES

37. Gas Production and Reuse 1 st Biological System Inflation Deflation