生質酒精為綠色能源發展的重點，利用纖維素作為生質酒精的原料來源是近年來的研究方向之一。由於生質物料的木質纖維素結構緻密、不易分解，間接阻礙了酵素對纖維素的水解效果及糖類發酵製程，利用有效的前處理方法可破壞生質物料結構並解決上述問題。本研究以狼尾草 (Napiergrass, Pennisetum purpureum)作為生質酒精製程的纖維素來源，以超臨界CO2前處理為主，另以超臨界CO2為介質配搭超聲波於高壓系統內進行物理性前處理，並嘗試利用超臨界CO2前處理再配搭過氧化氫進行化學性前處理。研究中首先利用美國國家再生能源實驗室 (National Renewable Energy Laboratory, NREL)標準方法水解未經處理之狼尾草，並將檢測值視為糖類總組成含量。經不同前處理後之狼尾草則透過酵素水解分析，將其值除以糖類總組成含量視為糖類糖化率，並利用糖類糖化率判定前處理效果。透過實驗設計可得狼尾草經超臨界CO2前處理之最佳操作條件，在溫度180 oC、壓力3500 psi、液固比為4以及處理時間為75分鐘時，可得葡聚糖糖化率為39%，並且可知溫度為最顯著影響因子。此外，研究中亦嘗試以超臨界CO2為介質配搭超聲波於高壓系統內進行物理性前處理，由於超聲波需要高密度之介質傳遞能量，因此實驗皆採低溫高壓之操作方式，然而其葡聚糖糖化率提升效果不明顯。另一方面，若以超臨界CO2前處理再配搭過氧化氫化學性前處理，其葡聚糖糖化率為未處理狼尾草原料的3倍，約46%。在表面結構分析方面，透過掃描式電子顯微鏡 (Scanning electron microscopy, SEM)可觀察出狼尾草表面經不同前處理後均有開放性破壞的現象。最後利用商業軟體Aspen Plus進行狼尾草經超臨界CO2前處理放大製程之穩態模擬，計算超臨界CO2前處理程序所需能源之消耗，可得總操作成本為26.4 $/tonne dry biomass及49.0 $/tonne dry biomass，與文獻中不同前處理總操作成本 (19~60 $/tonne dry biomass)相比較，超臨界CO2前處理程序是具競爭力。

Aiello, C.; Ferrer, A. and Ledesma, A. “Effect of alkaline treatments at various temperatures on cellulase and biomass production using submerged sugarcane bagasse fermentation with Trichoderma reesei QM 9414,” Bioresour. Technol. 57, 13-18, 1996.
Alinia, R.; Zabihi, S.; Esmaeilzadeh, F. and Kalajahi, J. F. “Pretreatment of wheat straw by supercritical CO2 and its enzymatic hydrolysis for sugar production” Biosys. Eng. 107, 61-66, 2010.
Alvira, P.; Tomás-Pejó, E.; Ballesteros, M. and Negro, M. J. “Pretreatment technologies for an efficient bioethanol production process based on enzymatic hydrolysis: A review” Bioresour. Technol. 101, 4851-4861, 2010.
Béguin, P. “Cloning of cellulase gene” Crit. Rev. Biotechnol. 6, 129-162, 1987.
Benazzi, T.; Calgaroto, S.; Astolfi, V.; Rosa, C. D.; Oliveira, J. V. and Mazutti, M. A. “Pretreatment of sugarcane bagasse using supercritical carbon dioxide combined with ultrasound to improve the enzymatic hydrolysis” Enzyme Microb. Technol. 52, 247-250, 2013.
Bhat, M. K. and Bhat, S. “Cellulose degrading enzymes and their potential industrial applications” Biotechnol. Adv. 15, 583-620, 1997.
Bisaria, V. S. and Ghose, T. K. “Biodegradation of Cellulosic Materials – Substrates” Microorganisms, Enzymes and Products, Enzyme Microb. Technol. 3, 90-104, 1981.
Bisaria, V. S. and Mishra, S. “Regulatory Aspects of Cellulase Biosynthesis and Secretion” Crit. Rev. Biotechnol. 9, 61-103, 1989.
Bobleter, O. “Hydrothermal degradation of polymers derived from plants” Polym. Sci. 19, 797-841, 1994.
Buchanan, B. B.; Gruissem, W. and Jones, R. L. “Biochemistry and Molecular Biology of Plants” American Society of Plant Biologists, Rockville, Maryland, 2000.
Burden, D.W. and Whitney, D.B. “Biotechnology: Proteins to PCR. A Course in
Strategy and Lab Techniques” Birkhauser, Boston, MA, 43-47, 1995.
Bussemaker, M. J. and Zhang, D. “Effect of ultrasound on lignocellulosic biomass as a pretreatment for biorefinery and biofuel applications” Ind. Eng. Chem. Res. 52, 3563-3580, 2013.
Chang, C.-W.; Yu, W.-C.; Chen, W.-J.; Chang, R.-F. and Kao, W.-S. “A study on the enzymatic hydrolysis of steam exploded Napiergrass with alkaline treatment using artificial neural networks and regression analysis” Journal of the Taiwan Institute of Chemical Engineers, 42, 889-894, 2011.
Chuang, M-H. and Johannsen, M. “Characterization of pH in aqueous CO2-systems” in: Proceedings of the 9th International Symposium on Supercritical Fluids, 2009.
Conde-Mejíaa, C.; Jiménez-Gutiérreza, A. and El-Halwagi, M. “A comparison of pretreatment methods for bioethanol production from lignocellulosic materials” Process Saf. Environ. Prot. 90, 189-202, 2012.
Correia, J. A. C.; Júnior, J. E. M.; Gonçalves, L. R. B. and Rocha, M. V. P. “Alkaline hydrogen peroxide pretreatment of cashew apple bagasse for ethanol production: Study of parameters” Bioresour. Technol. 139, 249-256, 2013.
Cosgrove, D. J. “Cell Walls: Structures, Biogenesis, and Expansion. In: Plant Physiology.” In L. Taiz and E. Zeiger, eds. Sunderland: Sinauer Associates, Inc. 1998.
Galbe, M. and Zacchi, G. “Pretreatment: The key to efficient utilization of lignocellulosic” Biomass Bioenergy, 46, 70-78, 2012.
Gao, M.; Xu, F.; Li, S.; Ji, X.; Chen, S. and Zhang, D. “Effect of SC-CO2 pretreatment in increasing rice straw biomass conversion” Biosys. Eng. 106, 470-475, 2010.
Ghose, T. K. “Cellulase biosynthesis and hydrolysis of cellulosic substances” Advances in biochemical engineering, Berlin: Springer-Verlag, 6, 39-74, 1976.
Gould, J. M. “Studies on the mechanism of alkaline peroxide delignification of agricultural residues” Biotechnol. Bioeng. 27, 225-231, 1985.
Gu, T. “Green biomass pretreatment for biofuels production” Springer, Berlin-New York, 2013.
Hendriks, A.T.W.M. and Zeeman, G. “Pretreatments to enhance the digestibility of lignocellulosic biomass” Bioresour. Technol. 100, 10-18, 2009.
Hsu, T. A.; Ladisch, M. R. and Tsao, G. T. Alcohol from cellulose, Chemical Technol. 10, 315-319, 1980.
Kim, K. H. and Hong, J. “Supercritical CO2 pretreatment of lignocellulose enhances enzymatic cellulose hydrolysis” Bioresour. Technol. 77, 139-144, 2001.
King, J. W.; Srinivas, K.; Guevara, O.; Lu, Y.-W.; Zhang, D. and Wang, Y.-J. “Reactive high pressure carbonated water pretreatment prior to enzymatic saccharification of biomass substrates” J. Supercrit. Fluids, 66, 221-231, 2012.
Kumar, P.; Barrett, D. M.; Delwiche, M. J. and Stroeve, P. “Methods for Pretreatment of Lignocellulosic Biomass for Efficient Hydrolysis and Biofuel Production” Ind. Eng. Chem. Res. 48, 3713-3729, 2009.
Limayem, A. and Ricke, S. C. “Lignocellulosic biomass for bioethanol production: Current perspectives, potential issues and future prospects” Prog. Energy Combust. Sci. 38, 449-467, 2012.
Mazza, G. and Fu, D. “Optimization of processing conditions for the pretreatment of wheat straw using aqueous ionic liquid” Bioresour. Technol. 102, 8003-8010, 2011.
Mishima, D.; Tateda, M.; Ike, M. and Fujita, M. “Comparative study on chemical pretreatments to accelerate enzymatic hydrolysis of aquatic macrophyte biomass used in water purification processes” Bioresour. Technol. 97, 2166-2172, 2006.
Moniruzzaman, M. “Saccharification and alcohol fermentation of steam-exploded rice straw” Bioresour. Technol. 55, 111-117, 1996.
Mosier, N.; Wyman, C.; Dale, B.; Elander, R.; Lee, Y. Y.; Holtzapple, M. and Ladisch, M. “Features of promising technologies for pretreatment of lignocellulosic biomass” Bioresour. Technol. 96, 673-686, 2005.
Murphy, L.; Borch, K.; McFarland, K.C.; Bohlin, C. and Westh, P. “A calorimetric assay for enzymatic saccharification of biomass” Enzyme Microb. Technol. 46, 141-146, 2010.
Narayanaswamy, N. “Supercritical carbon dioxide pretreatment of various lignocellulosic biomasses” Ohio university, department of chemical and biomolecular, 2010.
Narayanaswamy, N.; Faik, A.; Goetz D. J. and Gu, T. “Supercritical carbon dioxide pretreatment of corn stover and switchgrass for lignocellulosic ethanol production” Bioresour. Technol. 102, 6995-7000, 2011.
Nikolic´, S.; Mojovic´, L.; Rakin, M.; Pejin, D. and Pejin, J. “Ultrasound-assisted production of bioethanol by simultaneous saccharification and fermentation of corn meal” Food. Chem. 122, 216-222, 2010.
Parveen, K.; Barrett, D. M.; Delwiche, M. J. and Stroeve, P. “Methods for pretreatment of lignocellulosic biomass for efficient hydrolysis and biofuel production” Ind. Eng. Chem. Res. 48, 3713-3729, 2009.
Pejin, D. J.; Mojovi´c, L. V.; Pejin, J. D.; Gruji´c, O. S.; Markov, S. L.; Nikoli´c, S. B. and Markovi´c, M. N. “Increase in bioethanol production yield from triticale by simultaneous saccharification and fermentation with application of ultrasound” J. Chem. Technol. Biotechnol. 87, 170-176, 2012.
Saha, B. C. “Hemicellulose bioconversion” J. Ind. Microbiol. Biotechnol. 30, 279-291, 2003.
Santos, A. L. F.; Kawase, K. Y. F. and Coelho, G. L. V. “Enzymatic saccharification of lignocellulosic materials after treatment with supercritical carbon dioxide” J. Supercrit. Fluids, 56, 277-282, 2011.
Schacht, C.; Zetzl, C. and Brunner, G. “From plant materials to ethanol by means of supercritical fluid technology” J. Supercrit. Fluids. 46, 299-321, 2008.
Shen, G.; Tao, H.; Zhao, M.; Yang, B.; Wen, D.; Yuan, Q. and Rao, G. “Effect of hydrogen peroxide pretreatment on the enzymatic hydrolysis of cellulose” J. Food Process Eng. 34, 905-921, 2011.
Silverstein, R. A.; Chen, Y.; Sharma-Shivappa, R. R.; Boyette, M. D. and Osborne, J. “A comparison of chemical pretreatment methods for improving saccharification of cotton stalks” Bioresour. Technol. 98, 3000-3011, 2007.
Srinivasan, N. and Ju, L.-K. “Statistical optimization of operating conditions for supercritical carbon dioxide-based pretreatment of guayule bagasse” Biomass Bioenergy, 47, 451-458, 2012.
Srinivasan, N. and Ju, L.-K. “Pretreatment of guayule biomass using supercritical carbon dioxide-based method” Bioresour. Technol. 101, 9785-9791, 2010.
Sun, Y. and Cheng, J. J. “Hydrolysis of lignocellulosic materials for ethanol production: a review” Bioresour. Technol. 83, 1-11, 2002.
Suslick, K. S. “Sonoluminescence and Sonochemistry” In Encyclopedia of Physical Science and Technology, 3rd Ed, Academic Press 15, 363-376, 2002.
Toor, S. S.; Rosendahl, L. and Rudolf, A. “Hydrothermal liquefaction of biomass: A review of subcritical water technologies” Energy. 36, 2328-2342, 2011.
Wooley, R. J. and Putsche, V. “Development of an ASPEN PLUS physical property database for biofuels components” Nrel/Mp-425-50685. Golden, Colorado National Labaratory of the U.S. Deparment of Energy, pp. 1-38, 1996.
Yachmenev, V.; Condon, B. D.; Klasson, K. T. and Lambert, A. H. “Acceleration of the enzymatic hydrolysis of corn stover and sugar cane bagasse celluloses by low intensity uniform ultrasound” J. Biobased Mater. Bioenergy, 3, 25-31, 2009.
Yunus, R.; Salleh, S. F.; Abdullah, N. and Biak, D. R. A. “Effect of ultrasonic pre-treatment on low temperature acid hydrolysis of oil palm empty fruit bunch” Bioresour. Technol. 101, 9792-9796, 2010.
Zheng, Y.; Lin, H.-M. and Tsao, G.T. “Pretreatment for cellulose hydrolysis by carbon dioxide explosion” Biotechnol. Prog. 14, 890-896, 1998.
Zhu, J. Y.; Pan, X. J.; Wang, G. S. and Gleisner, R. “Sulfite pretreatment (SPORL) for robust enzymatic saccharification of spruce and red pine” Bioresour. Technol. 100, 2411-2418, 2009.
古森本，“生質能源作物之開發與潛力”，植物種苗生技，第13期，P.46-53，2008
行政院農業委員會畜產試驗所，http://www.tlri.gov.tw/
成游貴，“狼尾草新品種台畜草二號介紹”，畜產專訊，第16期，P.1，2003
沈明來，“試驗設計學”，九州書局，1997
周昌弘，“台灣芒萃植物生態研究的回顧與前瞻”，中華民國雜草學會簡訊，第2期，1995
林祐生，李文乾，“生質酒精”，科學發展，第433期，P.20-25，2009
林威廷，“利用超臨界二氧化碳與超音波前處理狼尾草與甘蔗渣”，碩士論文，國立清華大學化學工程研究所，2012
陳文恆，郭家倫，黃文松，王嘉寶，“纖維素酒精技術之發展”，農業生技產業季刊，第9期，P.62-69，2009
陳怡傑，“以厭氧流體化床進行廚餘過篩液及狼尾草之氫發酵程序研究”，碩士
論文，國立成功大學環境工程學系，2009
陳盧亮，“我國狼尾草屬牧草主栽品種特性介紹”，中國奶牛，2012
齊倍慶，“從堆肥中篩選纖維素分解酵素生產菌及其酵素性質研究”，碩士論文，國立清華大學生命科學系，2000
黎正中編譯，“實驗設計與分析”，高立圖書有限公司，2010