臺灣博碩士論文加值系統

本研究的目的是利用結合糖化菌與發酵菌之三菌共固定化系統將農業廢棄物中所含之大量纖維素與半纖維素，透過一好厭氧結合之重力沉降流式反應器，生產可作為替代能源之生質丁醇。實驗中使用 Trichoderma reesei、Aspergillus niger 為糖化水解菌種，以及 Clostridium beijerinckii 為丁醇發酵菌種。
本研究中所使用之丁醇菌，其發酵的過程會造成培養基酸鹼值大幅度之變化，且發酵產物為丙酮、丁醇以及乙醇，可能對糖化水解菌及其酵素造成影響，故分別進行測試。實驗結果顯示酸鹼值並不影響酵素之穩定性，且在酸鹼值4到5之間具有較佳之酵素活性；在發酵產物之抑制實驗結果顯示，單一抑制物之濃度在500 ppm以下對於T. reesei、A. niger的生長及酵素活性皆無顯著影響，但其混合濃度提升至1000 ppm以上時，則對菌種及酵素活性產生抑制，且隨混合濃度提高，影響越劇烈。反應器之相關實驗成果顯示，預培養有控制酸鹼值在4以上其酵素活性及產量(141 ppm)皆比沒有控制酸鹼值(113 ppm)來的佳；基質比較中發現，經稀硫酸處理過之前處理稻稈產量(242 ppm)高於基質使用CMC(173 ppm)及纖維素(233 ppm)之結果；於稻稈比較中發現基轉稻稈產量(169 ppm)高於野生稻稈(111 ppm)；最後於溶氧度探討中，我們發現雖然改良式的重力沉降流式反應器在好氧區的溶氧度有達到提升的效果，但仍然需要進行改善，以達到好氧區更好氧，厭氧區更厭氧的目標。

The purpose of this study is to produce biobutanol by using three microorganism co-immobilization systems that combine saccharifying and fermentative microorganisms. This study also uses large quantities of cellulose and hemicellulose in agricultural waste as a carbon source through an anaerobic Vertical Mass-flow type Bioreactor to produce biobutanol. In this study, Trichoderma reesei and Aspergillus niger are saccharified hydrolytic microorganisms, and Clostridium beijerinckii is a butanol fermentative bacterium.
The bacteria used for butanol production in this study lead to significant changes in pH values of the medium in the fermentation process. Fermentation products are acetone, butanol and ethanol, and they may affect saccharifying and hydrolyzing fungi and their enzymes, so they were tested separately. The results showed that the pH did not affect the stability of the enzyme, and had better enzyme activity from pH 4 to 5. The results of the inhibition experiments on the fermentation products showed that the concentration of single inhibitor below 500 ppm had no significant effect on the growth and enzyme activity of T. reesei and A. niger. However, when the mixture concentration is increased to more than 1000 ppm, the activities of the bacteria and enzymes were inhibited, and the more severe was followed by the mixed increasing concentration. The results of bioreactor show that pre-cultured with control of pH above 4, enzyme activity and yield (141 ppm) were better than without control of pH value (113 ppm). The results showed that the yield of butanol in rice straw treated with dilute sulfuric acid (242 ppm) was higher than that of CMC (173 ppm) and cellulose (233 ppm). In rice straw comparison, it was found that the yield of gene transfered rice straw (169 ppm) was higher than that of wild rice straw (111 ppm). Finally, in the discussion of dissolved oxygen, which level in the aerobic zone of the modified Vertical Mass-Flow type Bioreactor was required to further improve, so the bioreactor still need to further engineered to achieve the goal that aerobic zone is more aerobic, and anaerobic area is more anaerobic than the current bioreactor.

長庚大學碩(博)士學位論文指導教授推薦書
長庚大學碩(博)士學位論文口試委員審定書
致謝 iii
中文摘要 iv
英文摘要 vi
目錄 viii
圖目錄 xiv
表目錄 xvi
第一章 緒論 1
1-1 前言 3
1-2 研究動機與目的 5
第二章 文獻回顧 5
2-1 環境問題 5
2-2 再生能源 6
2-2-1 生質能 7
2-2-1-1 生質醇類 9
2-3 木質纖維素 11
2-3-1 木質纖維素的組成 12
2-3-2 纖維素的組成及結構 13
2-3-3 半纖維素的組成及結構 14
2-3-4 木質素組成及結構 15
2-3-5 纖維素的來源及利用 15
2-3-6 木質纖維材料前處理 18
2-3-7 纖維素分解酵素種類之介紹 24
2-3-7-1內切型纖維素分解酵素（Endo-β-gluconase, E.C.3.2.1.4） 24
2-3-7-2外切型纖維素分解酵素（Cellobiohydrolase, E.C.3.2.1.91） 25
2-3-7-3 纖維二醣酵素（Cellobiase, E.C.3.2.1.21） 25
2-3-8 纖維分解酵素的作用機制 錯誤! 尚未定義書籤。26
2-3-8-1 C1-Cx假說 26
2-3-8-2 修正假說 26
2-3-9 纖維素分解酵素之產物反饋抑制作用 27
2-3-10 分解纖維素的微生物種類 28
2-3-10-1 真菌 29
2-3-10-2 細菌 29
2-3-10-3 放射菌 29
2-4 醣質發酵 30
2-4-1 丙酮-丁醇-乙醇 (ABE) 發酵 30
2-4-2 丁醇生產菌株 32
2-4-2-1 Clostridia 的代謝 35
2-5 固定化 38
2-6 菌種共培養 40
2-6-1 直接共培養 43
2-6-1-1 聯和共培養 43
2-6-1-2 序列共培養 43
2-6-2 共固定化細胞混菌培養 44
2-7 實驗架構 45
第三章 實驗材料與方法 46
3-1 實驗設備 46
3-2 實驗藥品 47
3-3 菌株來源及種類 48
3-4 菌種保存方式 49
3-5 菌種培養 49
3-5-1 糖化水解菌 49
3-5-2 丁醇發酵菌 50
3-6 菌體細胞計數 50
3-7 培養基成分 50
3-7-1 T. reesei、A. niger 培養專用 51
3-7-2 C. beijerinckii培養基組成 51
3-7-2-1 C.beijerinckii 培養專用 51
3-7-2-2 C.beijerinckii發酵用培養基 51
3-8 DNS 試劑 55
3-9 實驗方法與步驟 56
3-9-1 菌體固定化方法 56
3-9-1-1 海藻酸鈣包埋法 56
3-9-1-2 Polyurethane (PU) foam 57
3-9-2 稻稈前處理 58
3-9-2-1 稻稈酸水解 58
3-9-3 酵素活性測定法 59
3-9-4 還原糖測定 ( DNS 測定法) 60
3-9-5 丙酮、丁醇、乙醇及糖類之定量分析 60
3-9-6 反應器氣體分析 61
3-9-7 生化反應器設計 61
第四章 結果與討論 64
4-1 DNS檢量線確立 64
4-2 糖化水解菌培養條件優化探討 65
4-2-1 糖化水解菌最適化探討 65
4-2-1-1 商業酵素穩定性探討-pH 66
4-2-1-2 糖化水解菌酵素穩定性探討-pH 67
4-2-1-3 酵素穩定性探討-D.O.值 69
4-2-2 ABE對糖化水解菌的抑制探討 70
4-2-2-1 單一抑制物 71
4-2-2-2 混合抑制物 74
4-2-3 培養基選擇探討 76
4-2-3-1 主發酵培養基選擇探討-糖化水解菌 76
4-2-3-2 主發酵培養基選擇探討-丁醇發酵菌 78
4-2-3-3 培養基質選擇探討-糖化水解菌 81
4-2-3-4 主發酵培養基質選擇探討-發酵菌 84
4-3 反應器探討 85
4-3-1 反應器探討-酸鹼調控 86
4-3-2 反應器探討-基質比較 89
4-3-3 反應器探討-稻稈比較 92
4-3-4 直立反應器溶氧度探討 95
4-3-4-1 第二版與第三版直立反應器溶氧度探討比較 95
4-3-4-2 反應器溶氧度提升之探討 100
第五章 結論 103
參考文獻 105

 
圖目錄
圖2-1 纖維素的結構組成 16
圖2-2 纖維素分解酵素吸附在纖維素上之後再進行分解作用 16
圖2-3 木質素立體結構式 17
圖2-4 木質素組成之三種醇類結構 17
圖2-5 早期纖維素分解酵素作用機制假說 27
圗2-6 纖維素水解酵素作用示意圖 27
圖2-7 Clostridia 的產醇代謝路徑 37
圖2-8 實驗架構 45
圖3-1 海藻酸鈉包埋裝置 57
圖3-2 Polyurethane foam 之規格 58
圖3-3 VMFB 反應器示意圖 63
圖4-1 DNS檢量線- 65
圖4-2 不同酸鹼值對商業酵素之影響 67
圖4-3 不同酸鹼值對糖化水解菌之影響 68
圖4-4 酵素穩定性探討-D.O.值 70
圖4-5 不同濃度之ABE對糖化水解菌之影響-單一抑制物 73
圖4-6 不同濃度之ABE對糖化水解菌之影響-混和抑制物 75
圖4-7 不同培養基對糖化水解菌之酵素活性差異 78
圖4-8 主發酵培養基選擇-丁醇發酵菌 80
圖4-9培養基質選擇-糖化水解菌 83
圖4-10 主發酵培養基質選擇探討-發酵菌 85
圖4-11反應器探討-酸鹼調控比較 88
圖4-12反應器探討-基質比較 91
圖4-13反應器探討-稻稈比較 94
圖4-14在第二版/第三版直立反應器預培養及主培養之好氧/厭氧區D.O.值變化 98
圖4-15 反應器溶氧度提升之架設圖 100
圖4-16 反應器打氣測試 102

 
表目錄
表2-1 再生能源的優缺點比較 7
表2-2 各種前處理方式比較 20
表2-3 具產生纖維素分解酵素能力的微生物 28
表2-4 野生型丁醇 34
表2-5 基因修飾型丁醇生產菌株整理 34
表3-1 Bushnell-Haas selection medium組成 52
表3-2 Bushnell-Haas selection medium-CMC & glucose (BHSD)成分 52
表3-3 Bushnell-Haas selection medium-CMC (BHSC)成分 53
表3-5 Clostridium beijerinckii pre-culture medium成分 54
表3-6 P2 stock solution 54
表3-7 DNS試劑成分 55
表4-1 改良式反應器預培養好氧區空氣層之空氣成分 99

Abdehagh, Niloofar, F. Handan Tezel, and Jules Thibault. "Separation techniques in butanol production: challenges and developments. " Biomass and Bioenergy 60 (2014): 222-246.

Akhtar, Nadeem, et al. "Recent advances in pretreatment technologies for efficient hydrolysis of lignocellulosic biomass." Environmental Progress & Sustainable Energy 35.2 (2016): 489-511.

Al-Hamamre, Zayed, et al. "Wastes and biomass materials as sustainable-renewable energy resources for Jordan." Renewable and Sustainable Energy Reviews 67 (2017): 295-314.

Álvarez, Consolación, Francisco Manuel Reyes‐Sosa, and Bruno Díez. "Enzymatic hydrolysis of biomass from wood." Microbial biotechnology 9.2 (2016): 149-156.

Bellido, Carolina, et al. "Acetone–butanol–ethanol (ABE) production by Clostridium beijerinckii from wheat straw hydrolysates: Efficient use of penta and hexa carbohydrates." Bioresource technology 167: 198-205. (2014)

Berezina, Oksana V., et al. "Reconstructing the clostridial n-butanol metabolic pathway in Lactobacillus brevis." Applied microbiology and biotechnology 87.2 (2010): 635-646.

Boisson, Anne-Marie, et al. "A simple and efficient method for the long-term preservation of plant cell suspension cultures." Plant methods 8.1 (2012): 4.

Brandon B. and Ezike R. Ethanol and butanol: Symbiotic partners for a modern fuel. Biofuel (2015).

Brodeur, Gary, et al. "Chemical and physicochemical pretreatment of lignocellulosic biomass: a review." Enzyme research 2011 (2011).

Bhutto, Abdul Waheed, et al. "Insight into progress in pre-treatment of lignocellulosic biomass." Energy 122 (2017): 724-745.

Dürre, Peter, ed. Handbook on clostridia. CRC press, (2005).

Dürre, Peter. "Fermentative butanol production." Annals of the New York Academy of Sciences 1125.1 (2008): 353-362.

Green, Edward M. "Fermentative production of butanol—the industrial perspective." Current opinion in biotechnology 22.3 (2011): 337-343.

Guilliams, Andrew, et al. "Physical and chemical differences between one-stage and two-stage hydrothermal pretreated hardwood substrates for use in cellulosic ethanol production." Biotechnology for biofuels 9.1 (2016): 30.

Veringa, H.J., Advanced techniques for generation of energy from biomass and waste, (2009).

International Energy Agency (IEA) World energy outlook. OECD/IEA, Paris, (2009).

Jones, David T., and David R. Woods. "Acetone-butanol fermentation revisited." Microbiological reviews 50.4 (1986): 484.

Kashket, Eva R., and Zhi-Yi Cao. "Isolation of a degeneration-resistant mutant of Clostridium acetobutylicum NCIMB 8052." Applied and environmental microbiology 59.12 (1993): 4198-4202.

Kettering C.F. The effect of the molecular structure of fuels and the power and efficiency of internal combustion engines. Ind Eng Chem 36(1944): 1079–1085.

Kim, Jun Seok, Y. Y. Lee, and Tae Hyun Kim. "A review on alkaline pretreatment technology for bioconversion of lignocellulosic biomass." Bioresource Technology 199 (2016): 42-48.

Köhler, Kirsten AK, et al. "Integration of biocatalyst and process engineering for sustainable and efficient n‐butanol production. " Engineering in Life Sciences 15.1 (2015): 4-19.

Kötting O, Santelia D, Edner C, Eicke S, Marthaler T, Gentry MS, Comparot-Moss S, Chen J, Smith AM, Steup M, Ritte G, Zeeman SC. STARCH-EXCESS4 is a laforin-like Phosphoglucan phosphatase required for starch degradation in Arabidopsis thaliana. (2009).

Kujawska, Anna, et al. "ABE fermentation products recovery methods—a review." Renewable and Sustainable Energy Reviews 48 (2015): 648-661.

Kumar, Manish, and Kalyan Gayen. "Developments in biobutanol production: new insights." Applied Energy 88.6 (2011): 1999-2012.

Kung, Chih-Chun, et al. "Bioenergy prospects in Taiwan using set-aside land–an economic evaluation." China Agricultural Economic Review 5.4 (2013): 489-511.

Kung, Chih-Chun, Liguo Zhang, and Meng-Shiuh Chang. "Promotion policies for renewable energy and their effects in Taiwan." Journal of Cleaner Production 142 (2017): 965-975.

Li, Lin, et al. "Enhanced butanol production by coculture of Clostridium beijerinckii and Clostridium tyrobutyricum." Bioresource technology 143 (2013): 397-404.

Li, Yu, et al. "Experimental comparative study on combustion, performance and emissions characteristics of methanol, ethanol and butanol in a spark ignition engine." Applied Thermal Engineering 115 (2017): 53-63.

Liu, Siqing, Nasib Qureshi, and Stephen R. Hughes. "Progress and perspectives on improving butanol tolerance." World Journal of Microbiology and Biotechnology 33.3 (2017): 51.

Lu, Y.,Gehan, J.P., and Sharkey, T.D. Daylength and circadian effects on starch degradation and maltose metabolism. Plant Physiol 138, (2005) :2280-2291..

Luo, Hongzhen, et al. "Enhancing butanol production under the stress environments of co-culturing Clostridium acetobutylicum/Saccharomyces cerevisiae integrated with exogenous butyrate addition." PloS one 10.10 (2015): e0141160.

Maiorella, B. "Comprehensive biotechnol. Young M (ed)" Pergamon Press, Oxford. Ethanol fermentation 3 (1985): 861-914.

Martin, Carlos, Helene B. Klinke, and Anne Belinda Thomsen. "Wet oxidation as a pretreatment method for enhancing the enzymatic convertibility of sugarcane bagasse." Enzyme and Microbial Technology 40.3 (2007): 426-432.

Mathioudakis, Vassias, et al. "The water footprint of second-generation bioenergy: a comparison of biomass feedstocks and conversion techniques." Journal of cleaner production 148 (2017): 571-582.

Matsumoto, David. "Culture and cultural worldviews: Do verbal descriptions about culture reflect anything other than verbal descriptions of culture?." Culture & Psychology 12.1 (2006): 33-62.

McGlade, Christophe, and Paul Ekins. "The geographical distribution of fossil fuels unused when limiting global warming to 2 [deg] C." Nature 517.7533 (2015): 187-190.

MicroMarket Monitor, Asia-Pacific n-butanol market by applications (butyl acrylate, butyl acetate, glycol ethers, and others) & geography – global trends & forecasts to 2019. (2015).

Niven R.K. Ethanol in gasoline: environmental impacts and sustainability review article. Renew Sustain Energy Rev 9(2005): 535–555.

Oudshoorn, Arjan, Luuk AM Van Der Wielen, and Adrie JJ Straathof. "Assessment of options for selective 1-butanol recovery from aqueous solution." Industrial & Engineering Chemistry Research 48.15 (2009): 7325-7336.

Pang, Feng, et al. "Effects of combination of steam explosion and microwave irradiation (SE–MI) pretreatment on enzymatic hydrolysis, sugar yields and structural properties of corn stover." Industrial Crops and Products 42 (2013): 402-408.

Park, Junyeong, et al. "Use of mechanical refining to improve the production of low-cost sugars from lignocellulosic biomass." Bioresource technology 199 (2016): 59-67.

Qureshi, Nasibuddin, and Hans P. Blaschek. "Production of acetone butanol ethanol (ABE) by a hyper‐producing mutant strain of Clostridium beijerinckii BA101 and recovery by pervaporation." Biotechnology progress 15.4 (1999): 594-602.

Ramos, Juan‐Luis, et al. "Benefits and perspectives on the use of biofuels." Microbial biotechnology 9.4 (2016): 436-440.

Rajan, Kalavathy, and Danielle Julie Carrier. "Effect of dilute acid pretreatment conditions and washing on the production of inhibitors and on recovery of sugars during wheat straw enzymatic hydrolysis." Biomass and Bioenergy 62 (2014): 222-227.

REN21. Renewable energy PolicyNetworkfor the 21st century. Renewables 2014. Global Status Report, (2016).

Serna, LV Daza, CE Orrego Alzate, and CA Cardona Alzate. "Supercritical fluids as a green technology for the pretreatment of lignocellulosic biomass." Bioresource technology 199 (2016): 113-120.

Shankar, Shiv. "Renewable and Nonrenewable Energy Resources: Bioenergy and Biofuels." Principles and Applications of Environmental Biotechnology for a Sustainable Future. Springer Singapore, 2017. 293-314.

da Silva Trindade, Wagner Roberto, and Rogério Gonçalves dos Santos. "Review on the characteristics of butanol, its production and use as fuel in internal combustion engines." Renewable and Sustainable Energy Reviews 69 (2017): 642-651.

Sindhu, Raveendran, Parameswaran Binod, and Ashok Pandey. "Biological pretreatment of lignocellulosic biomass–An overview." Bioresource technology 199 (2016): 76-82.

Singh, Renu, et al. "A review on delignification of lignocellulosic biomass for enhancement of ethanol production potential." Renewable and Sustainable Energy Reviews 32 (2014): 713-728.

Singer, Stephan, Jean-Philippe Denruyter, and Deniz Yener. "The energy report: 100% renewable energy by 2050." Towards 100% Renewable Energy. Springer, Cham, (2017). 379-383.

Suopajärvi, Hannu, et al. "Use of biomass in integrated steelmaking–Status quo, future needs and comparison to other low-CO 2 steel production technologies." Applied Energy 213 (2018): 384-407.

Sundaram, Vijay, Kasiviswanathan Muthukumarappan, and Srinivas Reddy Kamireddy. "Effect of ammonia fiber expansion (AFEX™) pretreatment on compression behavior of corn stover, prairie cord grass and switchgrass." Industrial Crops and Products 74 (2015): 45-54.

Swana, Jeffrey, et al. "An analysis of net energy production and feedstock availability for biobutanol and bioethanol." Bioresource technology 102.2 (2011): 2112-2117.

Terracciano, Joseph S., and Eva R. Kashket. "Intracellular conditions required for initiation of solvent production by Clostridium acetobutylicum." Applied and environmental microbiology 52.1 (1986): 86-91.

Travaini, Rodolfo, et al. "Ozonolysis: An advantageous pretreatment for lignocellulosic biomass revisited." Bioresource technology 199 (2016): 2-12.

Wang, Yue, et al. "Current advances on fermentative biobutanol production using third generation feedstock." Biotechnology advances (2017).

Wu, May, et al. Life-cycle assessment of corn-based butanol as a potential transportation fuel. No. ANL/ESD/07-10. Argonne National Laboratory (ANL), (2007).

Xu, D., L. Wang, and C. Du. "Progress in microbial co-culture--A review." Wei sheng wu xue bao= Acta microbiologica Sinica 55.9 (2015): 1089-1096.

Xue, Chuang, et al. "Integrated butanol recovery for an advanced biofuel: current state and prospects." Applied microbiology and biotechnology 98.8 (2014): 3463-3474.

Xue, Chuang, et al. "Recent advances and state-of-the-art strategies in strain and process engineering for biobutanol production by Clostridium acetobutylicum." Biotechnology advances 35.2 (2017): 310-322.

Yoshida M, Liu Y, Uchida S, Kawarada K, Ukagami Y, Ichinose H, et al. Effects of cellulose crystallinity, hemicellulose, and lignin on the enzymatic hydrolysis of Miscanthus sinensis to monosaccharides. Biosci Biotechnol Biochem. 72.3 (2008):805–10.

Zhang, Yi‐Heng Percival, and Lee R. Lynd. "Toward an aggregated understanding of enzymatic hydrolysis of cellulose: noncomplexed cellulase systems." Biotechnology and bioengineering 88.7 (2004): 797-824.

Zhang YH. Production of biofuels and biochemical by in vitro synthetic biosystems: opportunities and challenges. Biotechnol Adv.;33(2015):1467–83.

Zhang, Ke, Zhijian Pei, and Donghai Wang. "Organic solvent pretreatment of lignocellulosic biomass for biofuels and biochemicals: a review." Bioresource technology 199 (2016): 21-33.

Zhao, Zheng, et al. "N-methyl-2-pyrrolidonium-based Brönsted-Lewis acidic ionic liquids as catalysts for the hydrolysis of cellulose." Science China Chemistry 59.5 (2016): 564-570.

Zheng, Jin, et al. "Recent advances to improve fermentative butanol production: genetic engineering and fermentation technology." Journal of bioscience and bioengineering 119.1(2015): 1-9.

Zhuang, Xinshu, et al. "Liquid hot water pretreatment of lignocellulosic biomass for bioethanol production accompanying with high valuable products." Bioresource technology 199 (2016): 68-75.

Zickfeld, Kirsten, Susan Solomon, and Daniel M. Gilford. "Centuries of thermal sea-level rise due to anthropogenic emissions of short-lived greenhouse gases." Proceedings of the National Academy of Sciences 114.4 (2017): 657-662.

王秀華,木材化學及其應用(初版), 台北國立編譯館, (2002).

西澤俊一, 纖維素總論. 南山堂出版社, (1974).

行政院原能會, 新穎性纖維酒精技術及其量產廠成本效益評估, 委託研究計畫研究報告, (2012).

行政院原能會, 核研所生質能科技海外發酵, 纖維轉化酒精技術, (2015).

行政院環保署, 環保政策月刊, (2017).

林佩瑩, 不同 pH 環境下厭氧產氫菌 Clostridium 競爭情形之研究. 國立成功大學環境工程研究所, (2007).

余淑美, 植物生物科技在糧食與能源的應用, (2005)

吳耿東和李宏台, 生質能源化腐朽為能源, (2004).

吳權霖，紙質文物保護材料之研究。國立中興大學森林學系碩士論文, ( 2007 ).

胡柏泰, 開發以改良式 VMFB 生物反應器進行前處理稻稈生產丁醇之三菌共培養系統. 長庚大學生化與生醫工程研究所, (2016).

許淳鈞, 利用混合特定菌種生產氫氣之研究, 國立中央大學化學工程研究所, (2001).

張揚狀, 21世紀能源革命 –生質能, 台灣中油公司綠能科技研究所, (2016).

陳勁中, 林亞玄, 蔡昌廷, 高艾玲, 蘇宜欣, 蔡承佳, 以固定化丁醇生產菌 CPCNE 生產丁醇, 石油季刊, (2015)

曾秀月, 用鹼液萃取稻穀半纖維素. 中華生質能源學會會誌, (1992).

經濟部, 105 年度能源局報表. (2016).

楊翔如、康志堅、石蕙菱、陳志洋, 2017年第二季我國新興能源產業回顧與展望, 工研院, (2017)

綠色和平專題報導, 再生能源：世界各地的現在進行式, (2016).

藍士豪, 以絲瓜纖維做為固定化載體於改良式反應器中進行多澱粉稻桿之纖維酒精生產, 長庚大學生化與生醫工程研究所, (2014).

賴惠瑜, 以 Pichia stipitis 進行玉米桿水解液之酒精醱酵. 國立高雄應用科技大學化學工程與材料工程所, (2009).