EnzymeCatalyzed Synthesis of Biodiesel ENZYME PRETREATMENT Pretreatment activates

Enzyme-Catalyzed Synthesis of Biodiesel

ENZYME PRETREATMENT �Pre-treatment activates immobilized enzyme. �Examples of pretreatment conditions (30°C, 30 min): - t-Butanol: immersion for 1 hour and then soybean oil for a further 1 hour (24. 5% yield biodiesel). - Isopropanol: immersion for 1 hour and then soybean oil for further 1 hour (16. 8% yield biodiesel). - Oil: immersion for 4 hours in soybean oil (8. 6% yield biodiesel). - Nonpolar compounds like hexane did not work.

OPTIMIZING CONDITIONS (Novozym 435) Method Pre-treatment Time (min) T (°C) % yield (w/w) % Conversion % Biodeisel Conventional with t-butanol None 60 50 55 70 39 Microwave with t-butanol None 1 50 84 39 33 Conventional 1 hr t-butanol, 1 hr oil 60 30 62 65 40 Microwave 1 hr t-butanol, 1 hr oil 1 50 78 66 52

OUR RESULTS (Novozym 435) Method Pretreatment Time (min) T (°C) Power (W) % yield (w/w) % Conversion % Biodeisel Conventional (2 X Me. OH) 1 hr t-butanol, 1 hr oil 60 50 25 77 19 15 Conventional (4 aliquots Me. OH) 1 hr t-butanol, 1 hr oil 60 50 25 49 54 27 Microwave (4 aliquots Me. OH) 1 hr t-butanol, 1 hr oil 1 50 25 47 40 19 Microwave 1 hr t-butanol, 1 hr oil 15 50 5 37 20 7 Microwave 1 hr t-butanol, 1 hr oil 1 30 25 40 7 3

OPTIMIZING CONDITIONS (Rhizomucor miehei) Method Pre-treatment Time (min) T (°C) % yield (w/w) % Conversion % Biodiesel Conventional 1 hr oil 60 50 72 50 36 Microwave 1 hr oil 1 50 84 41 34 Conventional (2 X Me. OH) 1 hr oil 60 50 72 63 45 Microwave (2 X Me. OH) 1 hr oil 1 50 85 23 20

OPTIMIZING CONDITIONS (Thermomyces lanuginosus) Method Pre-treatment Time (min) T (°C) % yield (w/w) % Conversion % Biodiesel Conventional 1 hr oil 60 50 66 76 50 Microwave 1 hr oil 1 50 ---- 31 --- Conventional (2 X Me. OH) 1 hr oil 60 50 54 80 43 Microwave (2 X Me. OH) 1 hr oil 1 50 80 49 39

CONCLUSION �Our best microwave results were with Novozym 435: - Microwave, 50°C, 1 min run time, pretreated with t-butanol and oil for 1 h each �This was even better than the conventional results under comparable conditions: - Conventional, 50°C, 1 h run time, pretreated with t-butanol and oil for 1 h each

1. 2. 3. 4. 5. References Department of Energy-Energy Information Administration, http: //ww. eia. doe. gov, accessed January, 2009. Principles of Green Chemistry, http: //www. epa. gov/greenchemistry/pubs/principles. html, accessed January, 2009. Schuchardt, Ulf; Sercheli, Ricardo; Vargas, Rogerio Matheus Vargas “Transesterification of Vegetable Oils: A Review, ” Journal of Brazilian Chemical Society 1998, 9, 199 -210. Chen, Jech-Wei; Wu, Wen-Teng “Regeneration of Immobilized Candida antarctica Lipase for Transesterification, ” Journal of Bioscience and Bioengineering 2003, 95, 466 -469. Leadbeater, N. E. ; Stencel L. M. Fast “Easy Preparation of Biodiesel Using Microwave Heating, ” Energy and Fuels 2006, 20, 2281 -2283.

Acknowledgements �NSF-STEP �Dr. Nicole Bennett �A. R. Smith Dept. of Chemistry �Dr. Eric Allain �Monique Conway �The Biofuels Center of North Carolina