臺灣博碩士論文加值系統

本研究之目的在利用馬尾藻經熱酸水解、纖維素酶、澱粉酶、及馬尾藻多醣誘導之 Aeromonas salmonicida MAEF108 與 Pseudomonas vesicularis MA103 粗酵素液依序水解後，以所得馬尾藻多醣水解液 (SPSH) 利用梭狀芽孢桿菌 (Clostridium spp.) 發酵生產丁醇反應條件之探討。不同濃度稀鹽酸與檸檬酸進行熱酸水解之實驗結果顯示使用 0.4 N 稀鹽酸進行熱酸水解所得還原醣產率為最高，達到 12.70%。由 15 g 乾燥馬尾藻經 0.4 N 稀鹽酸於 121oC 下水解 20 min 後，以 13,680 U 纖維素酶、22,000 U 澱粉酶、及 100U (Alginate lyase activity) 馬尾藻多醣誘導之 MA103 與 MAEF108 粗酵素液水解，所得馬尾藻多醣水解液之總醣量與還原醣量分別為 42.30 及 29.86 mg/mL，再以超過濾離心區分出分子量小於 3 kDa 之寡醣，利用 TLC 分析發現水解液中含有單醣及雙醣等簡單醣類，以 HPLC 分析發現隨著水解程序進行，水解液中葡萄糖量逐漸增加，所得馬尾藻多醣水解液之葡萄糖產率為 56.77% (g/g dry biomass)。在耐鹽性試驗中 Clostridium acetobutylicum BCRC10639 與 Clostridium beijerinckii BCRC17950 之生長量 (OD600nm) 皆隨鹽度上升 (0-3%) 而下降，表示 2 株產丁醇菌在鹽度 > 2% 之培養基中生長較緩慢，甚至無法生長。由醣類利用性試驗之實驗結果可知 C. acetobutylicum BCRC10639 可以阿拉伯糖、葡萄糖、以及乳糖做為主要碳源進行發酵，而 C. beijerinckii BCRC17950 無法利用阿拉伯糖及乳糖做為主要碳源。以 60 g/L 葡萄糖發酵液為基質接種 10% C. acetobutylicum BCRC10639 進行攪拌批式發酵，經過 168 hr 後消耗 41.14 g/L 葡萄糖，產生 4.32 g/L 丙酮 (Acetone)、10.95 g/L 丁醇 (Butanol)、及 1.41 g/L 乙醇 (Ethanol)，而 ABE 產率則分別為 10%、27%、與 3% (g/g glucose)。將 10% C. beijerinckii BCRC17950 接種至 60 g/L 葡萄糖發酵液中進行攪拌批式發酵，在 168 hr 時消耗 40.99 g/L 葡萄糖，丙酮、丁醇、與乙醇之產量分別為 4.11、10.38 以及 1.53 g/L，而三者之產率分別為 10%、25% 以及 4% (g/g glucose)。之後以 3 倍稀釋之馬尾藻多醣水解液做為發酵基質，接種 10% C. acetobutylicum BCRC10639 進行攪拌批式發酵，總醣量與還原醣量分別由 14.10 與 9.95 mg/mL 下降至 5.61 與 3.72 mg/mL，而丙酮濃度與丁醇濃度分別為 0.40 與 0.49 g/L，丙酮與丁醇之產率分別為 0.80% 與 0.98% (g/g dry biomass)。利用 3 倍稀釋之馬尾藻多醣水解液進行靜置發酵，接種 10% C. acetobutylicum BCRC10639 發酵 120 hr 後產生 0.01% 丙酮與 0.09% 丁醇，其產率分別為 0.06% 與 0.46% (g/g dry biomass)；接種 10% C. beijerinckii BCRC17950 發酵 96 hr 生成丙酮與丁醇之濃度分別為 0.01% 與 0.04%，二者之產率分別為 0.04% 與 0.21% (g/g dry biomass)。

The purpose of this study is using Sagassum siliquosum polysaccharide hydrolysates (SPSH) that produced from Sagassum siliquosum dry powder treated with 0.4 N HCl heat-acid extraction, cellulase hydrolysis, amylase hydrolysis, and Sagassum siliquosum polysaccharides induced MA103 and MAEF108 crude enzyme hydrolysis as fermentation substrate for biobutanol production by Clostridium spp. The results of heat-acid extraction with different concentrations of diluted HCl and citric acid indicated that the reducing sugars yield of 0.4 N HCl heat-acid extraction was 12.70%. 15 g Sagassum siliquosum dry powder sequentially treated with 121oC hot water extraction for 20 min with 0.4 N HCl, 13,680 U cellulase hydrolysis, 22,000 U amylase hydrolysis, and 100 U (Alginate lyase) MA103 and MAEF108-Sar crude enzymes hydrolysis, the Sagassum siliquosum polysaccharide hydrolysates (SPSH) content 42.30 mg/mL total sugars and 29.86 mg/mL reducing sugars. Then, distinguished the oligosaccharide (OS) which molecular weight less than 3 kDa by ultrafiltration. The TLC analysis showed that SPSH containing monosaccharides and disaccharides. In HPLC analysis, The monosaccharide content increased gradually with hydrolysis procedures, and the glucose yield of SPSH was 56.77% (g/g dry biomass). In salt tolerance test of Clostridium spp., the growth amount of Clostridium acetobutylicum BCRC10639 and Clostridium beijerinckii BCRC17950 decreased with increasing of salinity. This result indicated that 2 Clostridium spp. grew slowly in medium with salinity > 2%, even can not grow. According to the experimental results of carbohydrate utilization test, C. acetobutylicum BCRC10639 can use arabinose, glucose, and lactose as the main carbon source for fermentation, and C. beijerinckii BCRC17950 can not use arabinose and lactose as the main carbon source. Using 60 g/L glucose fermentation solution as substrate inoculated C. acetobutylicum BCRC10639 to stirring batch fermentation, after 168 hr fermentation the glucose consumption was 41.14 g/L and produced 4.32 g/L acetone, 10.95 g/L butanol, and 1.41 g/L ethanol. The ABE yield of C. acetobutylicum BCRC10639 stirring fermentation in 60 g/L glucose solution were 10%, 27%, and 3% (g/g glucose), respectively. Inoculating 10% C. beijerinckii BCRC17950 into 60 g/L glucose solution for stirring batch fermentation. After 168 hr, the consumption of glucose was 40.99 g/L,and produced 4.11 g/L acetone, 10.38 g/L butanol, and 1.53 g/L ethanol. The ABE yield of C. beijerinckii BCRC17950 were 10%, 25%, and 4% (g/g glucose), respectively. Then, using 3-fold dilution SPSH as substrate for stirring fermentation, inoculated with 10% C. acetobutylicum BCRC10639,after 72 hr the total sugars and reducing sugars decreased form 14.10 and 9.95 mg/mL to 5.61 and 3.72 mg/mL, respectively. The acetone and butanol concentrations in fermentation solution were 0.40 and 0.49 g/L.The yield of acetone and butanol were 0.80% and 0.98% (g/g dry biomass), respectively. The 3-fold dilution SPSH inoculated with 10% C. acetobutylicum BCRC10639 for standing fermentation at 120 hr obtained 0.01% acetone and 0.09% butanol, and the yield of acetone and butanol were 0.06% and 0.46% (g/g dry biomass). Inoculating 10% C. beijerinckii BCRC17950 into 3-fold dilution SPSH for standing fermentation, at 96 hr obtained 0.01% acetone and 0.04% butanol, and the yields were 0.04% and 0.21% (g/g dry biomass), respectively.

中文摘要 I
英文摘要 (Abstract) II
目錄 IV
圖目錄 VIII
表目錄 IX
附錄目錄 XI
壹、前 言 1
貳、文獻整理 3
一、生質能源 3
二、丁醇之介紹 4
2-1. 丁醇之特性與應用 4
2-2. 丁醇之生產方法 5
2-2-1. 化學合成法 5
2-2-2. 微生物發酵法 5
三、產丁醇菌株之特性 6
3-1. 梭狀芽孢桿菌 (Clostriidum spp.) 6
3-2. 其他產丁醇菌株 8
四、馬尾藻 (Sargassum sp.) 9
4-1. 馬尾藻之生態特性 9
4-2. 馬尾藻之成分組成 9
五、褐藻膠裂解酶 (Alginate lyase) 9
5-1. 褐藻膠之組成結構 9
5-2. 褐藻膠裂解酶之生產菌株 10
5-3. 褐藻膠裂解酶之生化特性 10
六、Acetone-butanol-ethanol (ABE) 發酵生產生質丁醇 11
6-1. 批式發酵 (Batch fermantation) 生產生質丁醇 11
6-2. 以藻類生物質 (Algal biomass) 為原料生產生質丁醇 11
參、實驗設計 13
肆、實驗材料與方法 13
一、實驗材料 14
1-1. 原料 14
1-2. 實驗菌株 14
1-2-1. 褐藻膠水解酵素生產菌株 14
1-2-2. 產丁醇菌株 14
1-3. 試驗藥品 14
1-3-1. 藥品 14
1-3-2. 酵素 16
1-3-3. 培養基組成 16
1-3-4. 褐藻膠裂解酶酵素活性反應基質 18
1-3-5. 還原醣量反應試劑 18
1-3-6. 總醣量反應試劑 18
1-4. 儀器設備 18
二、實驗方法 20
2-1. 馬尾藻之前處理 20
2-2. 馬尾藻之一般成分分析 20
2-2-1. 水分含量 20
2-2-2. 粗脂肪含量 20
2-2-3. 粗蛋白含量 20
2-2-4. 灰分含量 21
2-3. 馬尾藻多醣誘導 MAEF108 與 MA103 粗酵素液之生產 21
2-3-1. 菌株保存 21
2-3-2. 菌株活化 21
2-3-3. 馬尾藻多醣誘導之粗酵素液製備 21
2-3-4. 誘導粗酵素液中褐藻膠裂解酶 (Alginate lyase) 之酵素活性 22
2-4. 馬尾藻多醣水解液之製備 22
2-4-1. 以不同濃度稀鹽酸與檸檬酸進行熱酸水解 22
2-4-2. 酵素水解 22
2-5. 產丁醇菌株之特性 22
2-5-1. 菌株保存 22
2-5-2. 菌株活化 23
2-5-3. 革蘭氏染色 (Gram stain) 23
2-5-4. 耐鹽性 23
2-5-5. 醣類利用性 23
2-6. 60 g/L 葡萄糖液批式發酵 23
2-6-1. 攪拌批式發酵 (Agitating batch fermentation) 23
2-6-2. 靜置批式發酵 (Static batch fermentation) 24
2-7. 馬尾藻多醣水解液 (3 倍稀釋) 批式發酵 24
2-7-1. 攪拌批式發酵 24
2-7-2. 靜置批式發酵 24
2-8. 分析方法 24
2-8-1. 總醣量測定 24
2-8-2. 還原醣量測定 24
2-8-3. Clostridium spp. 菌量測定 25
2-8-4. 有機溶劑產物 (丙酮、丁醇、乙醇) 濃度測定 25
2-9. 馬尾藻多醣水解液與發酵期間之醣類組成分析 25
2-9-1. 薄層色層分析 (Thin layer chromatography, TLC) 25
2-9-2. 高效液相層析 (High performance liquid chromatography, HPLC) 25
三、統計分析 26
伍、結果與討論 27
一、乾燥莢托馬尾藻粉末之一般成分分析 27
二、以不同濃度稀鹽酸與檸檬酸進行熱酸水解 27
2-1. 以不同濃度稀鹽酸進行熱酸水解 27
2-2. 以不同濃度檸檬酸進行熱酸水解 27
三、馬尾藻多醣水解液之製備與醣類組成分析 28
3-1. 馬尾藻多醣水解液 (Sargassum polysaccharide hydrolysates, SPSH) 28
3-2. 薄層色層分析 (TLC) 28
3-3. 高效液相層析分析 (HPLC) 28
四、Clostridium spp. 之生長曲線 29
4-1. Clostridium acetobutylicum BCRC10639 之生長曲線 29
4-2. Clostridium beijerinckii BCRC17950 之生長曲線 29
五、Clostridium spp. 之耐鹽性試驗與醣類利用性試驗 29
5-1. 耐鹽性試驗 29
5-2. 醣類利用性試驗 30
六、以 60 g/L 葡萄糖發酵液批式發酵 30
6-1. 攪拌批式發酵 30
6-2. 靜置批式發酵 31
七、以馬尾藻多醣水解液 (SPSH) 批式發酵 32
7-1. SPSH 攪拌批式發酵 32
7-2. SPSH 靜置批式發酵 32
八、馬尾藻多醣水解液 (SPSH) 靜置批式發酵之殘醣組成分析 32
陸、結 論 33
陸、參考文獻 35

王智薇。2008。淺談新興能源科技產業-生質能源。產經資訊，65: 22-27。
王仁、劉瓊霦。2012。孟宗竹發展生質能源之潛能。林業研究專訊，19: 50-54。
古森本。2008。生質能源作物之開發與潛力。農業生技產業季刊，13: 46-53。
李瑞真。2011。固定化 Clostridium acetobutylicum 進行連續式發酵生產丁醇之研究。東海大學化學工程與材料工程研究所碩士學位論文，臺中，臺灣。
吳紹祺。1999。海洋菌所產 Agarase 生產條件與海洋多醣經 Agarase 分解所得寡醣類組成之探討。國立臺灣海洋大學食品科學系碩士學位論文，基隆，臺灣。
吳弈霈。2012。馬尾藻經酵母菌發酵生產乙醇之最適反應條件探討。國立臺灣海洋大學食品科學系碩士學位論文，基隆，臺灣。
林佩瑩。2007。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學系碩士學位論文，臺南，臺灣。
林慧傑。2010。Pseudomonas vesicularis MA103 所產 Alginate lyase palIII 之基因轉殖至大腸桿菌中表現之探討。國立臺灣海洋大學食品科學系碩士學位論文，基隆，臺灣。
周仕凱、許梅娟。2009。新能源-生物產丁醇。科學發展，433: 26-31。
范繼中、吳建威、藍惠玲、吳純衡。2007。以海藻產製生質酒精之優勢。水試專訊，19: 24-26。
陳幸臣。2003。食品微生物學實驗法 (第 4 版)。華香園出版社，臺北市，臺灣。 pp. 25-26。
陳維新。2011。能源概論。高立圖書有限公司，新北市，臺灣。
黃淑芳。2004。臺灣海藻資訊網-重緣葉馬尾藻。國立臺灣博物館，臺北，臺灣。
黃培安、吳純衡。2006。海藻在保健食品、化妝品之開發。農業生技產業季刊。7: 21-25。
蔡文慶。2010。生質酒精的生產與發展現況。朝陽科技大學應用化學系碩士學位論文，臺中，臺灣。
潘崇良、陳衍昌、蔡震壽、方翠筠、黃意真、鄒文雄、林富邦、劉秀美、唐世杰。2009。海洋功能性醣體研究：藻類多醣生質乙醇與丁醇及碳交易附加收入。第三屆海峽兩岸魚類生理與養殖研討會，2009 年 10 月 19-21 日，基隆，臺灣。
潘崇良。2010。利用藻類生產生質能源。科學發展。448: 26-32。
蕭斯欣。2012。我國海洋生質能源產業的機會與挑戰。我國海洋生質能源產業發展趨勢學術研討會，國立臺灣海洋大學主辦，2012 年 3 月 2 日，基隆，臺灣。http://meet.ntou.edu.tw/0302www/index.html (March 5, 2012, Taiwan)。
戴雯琪。2013。再生能源在臺灣之發展與現況-環境基本法十周年的省思。全國律師月刊。38: 25-39。
謝志強、殷正華。2008。全球生質能源產業與技術發展現況與趨勢。科技發展政策報導。5: 15-39。
龔冬梅、張翠君、張戟、陳淑鳳。2003。車用乙醇汽油分層問題的研究。河南化工。6: 11-13。
An, Q. D., Zhang, G. L., Wu, H. T., Zhang, Z. C., Gong, W., Liu, Y., Li, X. Z., Murata, Y. 2008. Production and partial properties of alginase from newly isolated Flavobacterium sp. LXA. Process Biochemistry 43: 842-847.
AOAC. 1997. Official methods of analysis of AOAC international (16th ed.). Association of Official Analytical Chemists. Washington, DC, U.S.A.
Atsumi, S., Cann, A. F., Connor, M. R., Shen, C. R., Smith, K. M., Brynildsen, M. P., Chou, K. J. Y., Hanai, T., Liao, J. C. 2008. Metabolic engineering of Escherichia coli for 1-butanol production. Metabolic Engineering 10: 305-311.
Bahl, H., Andersch, W., Braun, K., Gottschalk, G. 1982. Effect of pH and butyrate concentration on the production of acetone and butanol by Clostridium acetobutylicum grown in continuous culture. European Journal of Applied Microbiology and Biotechnology 15: 201-205.
Bankar, S. B., Survase, S. A., Singhal, R. S., Granström, T. 2012. Continuous two stage acetone-butanol-ethanol fermentation with integrated solvent removal using Clostridium acetobutylicum B 5313. Bioresource Technology 106: 110-116.
Baron, A. J., Wong, T. Y., Hicks, S. J., Gacesa, P., Willcock, D., McPherson, M. J. 1994. Alginate lyase from Klebsiella pneumoniae subsp. aerogenes: Gene cloning, sequence analysis and high-level production in Escherichia coli. Gene 143: 61-66.
BCRC (Bioresource Collection and Research Center). 2012. Clostridium acetobutylicum BCRC 10639-medium. Bioresource Collection and Research Center, Taiwan. Retrieved Oct 24, 2012, from https://catalog.bcrc.firdi.org.tw/BS
AS_cart/controller?event=SEARCH&;bcrc_no=10639&;type_id=2&;keyword=.
Beckwith, P. 2010. The development of Butamax: A case study of the DuPont-BP joint venture to develop advanced biobutanol. Butamax, Wilmington, DE, US. Retrieved May 23, 2013, from http://www.butamax.com/_assets/pdf/LLC_deve-lopment.pdf.
Bochman, M., Cotton, F. A., Murillo, C. A., Wilkinson, G. 1999. Advanced inorganic chemistry. John Wiley and Sons, Inc, New York, U.S.A.
BP (British petroleum). 2012. BP statistical review of world energy June 2012. British Petroleum, London, United Kingdom. Retrieved May 23, 2013, from http://www.bp.com/assets/bp_internet/globalbp/globalbp_uk_english/reports_and_publications/statistical_energy_review_2011/STAGING/local_assets/pdf/statistical_review_of_world_energy_full_report_2012.pdf.
Bowles, L. K., Ellefson, W. L. 1985. Effects of Butanol on Clostridium acetobutylicum. Applied and Environmental Microbiology 50: 1165-1170.
Bryant, D. L., Blaschek, H. P. 1988. Buffering as a means for increasing growth and butanol production by Clostridium acetobutylicum. Journal of Industrial Microbiology and Biotechnology 3: 49-55.
Cao, L., Xie, L., Xue, X., Tan, H., Liu, Y., Zhou, S. 2007. Purification and characterization of alginate lyase from Streptomyces sp. strain A5 isolated from banana rhizosphere. Agricultural and Food Chemistry 55: 5113-5117.
Cheng, C. L., Che, P. Y., Chen, B. Y., Lee, W. J., Chien, L. J., Chang, J. S. 2012. High yield bio-butanol production by solvent-producing bacterial microflora. Bioresource Technology 113: 58-64.
Christensen, C. 2012. Corrosion and stress corrosion cracking in fuel grade ethanol. 7th Scandinavian Tank and Refinery Conference, Dec 6, 2012. Retrieved Jane 20, 2013, from http://www.forcetechnology.com/NR/rdonlyres/7FF02A1E-A6E6-4A
9C-AB87-E7891A591BE7/5955/3ACorrosionandcrackinginethanolandbioethanoltanksb.pdf.
Davis, T. A., Lianes, F., Volesky, B., Diaz-Pulido, G., McCook, L., Mucci, A. 2003. 1H-MNR study of Na alginates extracted from Sargassum spp. in relation of metal biosorption. Applied Biochemistry and Biotechnology 100: 75-90.
Dawes, C. J. 1998. Marine Botany (2nd ed.). John Wiley and Sons, Inc., New York, U.S.A. pp. 371-387.
DOW (The Dow chemical company). 2006. Product Safety Assessment: n-Butanol. The Dow Chemical Company, US. Retrieved Aug 20, 2012, from http://www.dow.com/productsafety/finder/¬nbut.htm.
Dubois, M., Gillles, K. A., Hamilton, J. K., Rebers, P. A. and Smith, F. 1956. Colorimetric method for determination of sugars and related substances. Analytical Chemistry 28: 350-356.
Duckworth, M. and Yaphe, W. 1970. Thin-layer chromatographic analysis of enzymatic hydrolysates of agar. Chromatography A 49: 482-487.
Dürre P. 2007. Biobutanol: An attractive biofuel. Biotechnology Journal 2: 1525-1537.
Efremenko, E. N., Nikolskaya, A. B., Lyagin, I. V., Senko, O. V., Makhlis, T. A., Stepanov, N. A., Maslova, O. V., Mamedova, F., Varfolomeev, S. D. 2012. Production of biofuels from pretreated microalgae biomass by anaerobic fermentation with immobilized Clostridium acetobutylicum cells. Bioresource Technology 14: 342-348.
Ertesvag, H., Erlien, F., Skjak-Bræk, G., Rehm, B. H. A., Valla, S. 1998. Biochemical properties and substrate specificities of a recombinantly produced Azotobacter vinelandii alginate lyase. Bacteriology 180: 3779-3784.
Ezeji, T. C., Qureshi, N., Blaschek, H. P. 2004. Acetone butanol ethanol (ABE) production from concentrated substrate: Reduction in substrate inhibition by fed-batch technique and product inhibition by gas stripping. Applied Microbiology and Biotechnology 63: 653-658.
Ezeji, T. C., Qureshi, N., Blaschek, H. P. 2005a. Industrially relevant fermentations. In Dürre P (Ed.). Handbook on clostridia. Taylor and Francis, London, U.K. pp. 797-812.
Ezeji, T. C., Karcher, P. M., Qureshi, N., Blaschek, H. P. 2005b. Improving performance of a gas stripping-based recovery system to remove butanol from Clostridium beijerinckii fermentation. Bioprocess and Biosystems Engineering 27: 207-214.
Ezeji, T. C., Qureshi, N., Blaschek, H. P. 2007. Production of acetone butanol (AB) from liquefied corn starch, a commercial substrate, using Clostridium beijerinckii coupled with product recovery by gas stripping. Journal of Industrial Microbiology and Biotechnology 34: 771-777.
Ezeji, T., Blaschek, H. P. 2008. Fermentation of dried distillers’ grains and solubles (DDGS) hydrolysates to solvents and value-added products by solventogenic clostridia. Bioresource Technology 99: 5232-5242.
Falbe, J. 1970. Carbon Monoxide in Organic Synthesis. Berlin, Germany: Springer Verlag.
Gombotz, W. R., Wee, S. F. 1998. Protein release from alginate matrices. Advance Drug Delivery Reviews 31: 267-285.
Harrigan, W. F., McCance, M. E. 1979. Laboratory methods in food and dairy microbiology. Academic Press, London, U.K.
Harris, P. 1990. Food Gels. Elsevies Science Publishing Co., Inc., New York City, NY, U.S.A.
Henstra, A. M., Sipma, J., Rinzema, A., Stams, A. J. M. 2007. Microbiology of synthesis gas fermentation for biofuel production. Current Opinion in Biotechnology 18: 200-206.
Heyraud, A., Colin-Morel, P., Girond, S., Richard, C., Kloareg, B. 1996. HPLC analysis of saturated or unsaturated oligoguluronates and oligomannuronates. Application to the determination of the action pattern of Haliotis tuberculata alginate lyase. Carbohydrate Research 29: 115- 126.
Hou, Z. Q., Zhang, M., Liu, B. Yan, Q., Yuan, F., Xu, D., Gao, Y. 2012. Effect of chitosan molecular weight on the stability and rheological properties of -carotene emulsions stabilized by soybean soluble polysaccharides. Food Hydrocolloids 26: 205-211.
Huesemann, M. H., Kuo, L. J., Urquhart, L., Gill, G. A., Roesijadi, G. 2012. Acetone-butanol fermentation of marine macroalgae. Bioresource Technology 108: 305-309.
Inui, M., Suda, M., Kimura, S., Yasuda, K., Suzuki, H., Toda, H., Yamamoto, S., Okino, S., Suzuki, N., Yukawa, H. 2008. Expression of Clostridium acetobutylicum butanol synthetic genes in Escherichia coli. Applied Microbiology and Biotechnology 77: 1305-1316.
Iwamoto, T., Araki, R., Iriyama, K. I., Oda, T., Fukuda, H., Hayashida, S., Muramatsu, T. 2001. Purification and characterization of bifunctional alginate lyase from Alteromonas sp. strain No. 272 and its action on saturated oligomeric substrates. Bioscience Biotechnology and Biochemistry 65: 133-142.
Jang, Y. S., Lee, J., Malaviya, A., Seung, D. Y., Cho, J. H., Lee, S. Y. 2012. Butanol production from renewable biomass: Rediscovery of metabolic pathways and metabolic engineering. Biotechnology Journal 7: 186-198.
John, R. P., Anisha, G. S., Nampoothiri, K. M., Pandey, A. 2011. Micro and macroalgal biomass: A renewable source for bioethanol. Bioresource Technology 102: 186-193.
Jones, D. T., Woods, D. R. 1986. Acetone-butanol fermentation revisited. Microbiological Reviews 50: 494-524.
Kamm, B., Kamm, M. 2007. International biorefinery systems. Pure and Applied Chemistry 79: 1983-1997.
Kim, J., Bajpai, R., Iannotti, E. L. 1988. Redox potential in acetone-butanol fermentations. Applied Biochemistry and Biotechnology 18: 175-186.
Kim, N. J., Li, H., Jung, K., Chang, H. N., Lee, P. C. 2011. Ethanol production from marine algal hydrolysates using Escherichia coli KO11. Bioresource Technology 102: 7466-7469.
Kim, H. K., Chung, J. H., Wang, D., Lee, J., Woo, H. C., Choi, I. G., Kim, K. H. 2012. Depolymerization of alginate into a monomeric sugar acid using Alg17C, an exo-oligoalginate lyase cloned from Saccharophagus degradans 2-40. Applied Microbiology and Biotechnology 93: 2233-2239.
Lee, S. Y., Park, J. H., Jang, S. H., Nielsen, L. K., Kim, J., Jung, K. S. 2008a. Fermentative butanol production by clostridia. Biotechnology and Bioengineering 101: 209-228.
Lee, S. M., Cho, M. O., Park, C. H., Chung, Y. C., Kim, J. H., Sang, B. I., Um, Y. 2008b. Continuous butanol production using suspended and immobilized Clostridium beijerinckii NCIMB 8052 with supplementary butyrate. Energy and Fuels 22: 3459-3464.
Lee, S., Oh, Y., Kim, D., Kwon, D., Lee, C. and Lee, J. 2011. Converting carbohydrates extracted from marine algae into ethanol using various ethanolic Escherichia coli strains. Applied Biochemistry and Biotechnology 164: 878-888.
Li, X., Dang, X., Zhao, C., Chen, Z., Chen, F. 2003. Isolation and some properties of cellulose-degrading Vibrio sp. LX-3 with agar-liquefying ability from soil. World Journal of Microbiology and Biotechnology 19: 375-379.
Lu, C., Zhao, J., Yang, S. T., Wei, D. 2012. Fed-batch fermentation for n-butanol production from cassava bagasse hydrolysate in a fibrous bed bioreactor with continuous gas stripping. Bioresource Technology 104: 380-387.
Lu, J., You, L., Lin, Z., Zhao, M., Cui, C. 2013. The antioxidant capacity of polysaccharide from Laminaria japonica by citric acid extraction. International Journal of Food Science and Technology 48: 1352-1358.
Luers, F., Seyfried, M., Daniel, R., Gottschalk, G. 1997. Glycerol conversion to 1,3-propanediol by Clostridium pasteurianum: Cloning and expression of the gene encoding 1,3-propanediol dehydrogenase. FEMS Microbiology Letters 154:337-345.
Lütke-Eversloh, T., Bahl, H. 2011. Metabolic engineering of Clostridium acetobutylicum: Recent advances to improve butanol production. Current Opinion in Biotechnology 22: 634-647.
Matsubara, Y., Iwasaki, K., Muramatsu, T. 1998. Action of poly (-L-guluronate) lyase from Corynebacterium sp. ALY-1 strain on saturated oligoguluronates. Bioscience Biotechnology and Biochemistry 62: 1055-1060.
Matsubara, Y., Kawada, R., Iwasaki, K., Kimura, Y., Oda, T., Muramatsu, T. 2000. Cloning and sequence analysis of a gene (aly PG) encoding poly (α-L-guluronate) lyase from Corynebacterium sp. strain ALY-1. Bioscience and Bioengineering 89: 199-202.
McCoy, E., Fred, E. B., Peterson, W. H., Hastings, E. G. 1926. A cultural study of the acetone butyl alcohol organism. The Journal of Infectious Diseases 39: 457-483.
Miller, G. L. 1959. Use of dinitrosalicylic acid reagent for reagent for determination of reducing sugars. Analytical Chemistry 31: 426-428.
Miyazaki, M., Obata, J., Iwamoto, Y., Oda, T., Muramatsu, T. 2001. Calcium-sensitive extracellular poly (-L-guluronate) lyase from a marine bacterium Pseudomonas sp. strain F6: Purification and some properties. Fisheries Science 67: 956-964.
Muramatsu, T., Yamada, K., Date, M., Yoshioka, S. 1993. Action of poly (-D-mannuronate) lyase from Turbo cornutus on oligomeric substrates. Bioscience Biotechnology and Biochemistry 57: 1990-1994.
Myra, G. B., Rizalinda, L. L., Joel, L. C. 2013. Bioethanol production from the macroalgae Sargassum spp. Bioresource Technology 138: 22-29.
Nielsen, D. R., Leonard, E., Yoon, S. H, Tseng, H. C., Yuan, C., Prather, K. L. 2009. Engineering alternative butanol production platforms in heterologous bacteria. Metabolic Engineering 11: 262-273.
Okuda, K., Oka, K., Onda, A., Kajiyoshi, K., Hiraoka, M., Yanagisawa, K. 2008. Hydrothermal fractional pretreatment of sea algae and its enhanced enzymatic hydrolysis. Journal of Chemical Technology and Biotechnology 83: 836-841.
Porter, J. R. 1961. Louis Pasteur: Achievements and disappointments, 1861. Bacteriological Reviews 25: 389-403.
Posten, C. H., Cooney, C. L. 1993. Growth of microorganisms. In Rehm, H. J. and Reed, G. (Eds), Biotechnology. Wiley-VCH, NY, U.S.A. pp. 121-122
Preston, L. A., Bender, C. L., Schiller, N. L. 2001. Analysis and expression of algL, which encodes alginate lyase in Pseudomonas syringae pv. syringae. DNA Sequence 12: 455-461.
Qureshi, N., Blaschek, H. P. 1999. Production of acetone-butanol-ethanol (ABE) by a hyper-producing mutant strain of Clostridium beijerinckii BA101 and recovery by pervaporation. Biotechnology Progress 15: 594-602.
Qureshi, N., Blaschek, H. P. 2001. Recent advances in ABE fermentation: Hyper-butanol producing Clostridium beijerinckii BA101. Journal of Industrial Microbiology and Biotechnology 27: 287-291.
Qureshi, N., Saha, B. C., Cotta, M. A. 2007. Butanol production from wheat straw hydrolysate using Clostridium beijerinckii. Bioprocess and Biosystems Engineering 30: 419-427.
Qureshi, N., Saha, B. C., Dien, B., Hector, R. E., Cotta, M. A. 2010a. Production of butanol (a biofuel) from agricultural residues: Part I - Use of barley straw hydrolysate. Biomass and Bioenergy 34: 559-565.
Qureshi, N., Saha, B. C., Hector, R. E., Dien, B., Hughes, S., Liu, S., Iten, L., Bowman, M. J., Sarath, G., Cotta, M. A. 2010b. Production of butanol (a biofuel) from agricultural residues: Part II - Use of corn stover and switchgrass hydrolysates. Biomass and Bioenergy 34: 566-571.
Rahman, M. M., Inoue, A., Tanaka, H., Ojima, T. 2010. Isolation and characterization of two alginate lyase isozymes, AkAly28 and AkAly33, from the common sea hare Aplysia kurodai. Biochemistry and Molecular Biology 157: 317-325.
Ramey, D. E. 2007. Butanol: The other alternative fuel. National Agricultural Biotechnology Council Report 19: 137-147.
SAS (Statistical Analytical System). 1999. SAS Use’s: Basic Statistical Analysis. SAS Institute Inc., Cary, NC, U.S.A.
Steen, E. J., Chan, R., Prasad, N., Myers, S., Petzold, C. J., Redding, A., Ouellet, M., Keasling, J. D. 2008. Metabolic engineering of Saccharomyces cerevisiae for the production of n-butanol. Microbial Cell Factories 7: 361-368.
Sugano, Y., Terada, I., Arita, M., Mona, M. and Matsumoto, T. 1993. Purification and characterization of new agarase from marine bacterium, Vibrio sp. strain JT0107. Applied and Environmental Microbiology 59: 1549-1554.
Tang, J. C., Taniguchi, H., Chu, H., Zhou, Q., Nagata, S. 2009. Isolation and characterization of alginate-degrading bacteria for disposal of seaweed wastes. Letters in Applied Microbiology 48: 38-43.
Tomas, C. A., Welker, N. E., Papoutsakis, E. T. 2003. Overexpression of groESL in Clostridium acetobutylicum results in increased solvent production and tolerance, prolonged metabolism, and changes in the cell’s transcriptional program. Applied and Environmental Microbiology 69: 4951-4965.
Torres, M. R., Sousa, A. P. A., Silva Filho, E. A. T., Melo, D. F., Feitosa, J. P. A., de Paula R. C. M., Lima, M. G. S. 2007. Extraction and physicochenical characterization of Sargassum vulgare alginate from Brazil. Carbohydrate Research 342: 2067-2074.
UN-Energy. 2007. Sustainable bioenergy: A framework for decision makers. United Nation Publications, Rome, Italy.
Wang, T. Y., Preston, L. A., Schiller, N. L. 2000. Alginate lyase: Review of major sources and enzyme characterization, structure- function analysis, biological roles, and applications. Annual Review of Microbiology 54: 289-340.
Wang, M. L., Wang, J. T., Choong, Y. M. 2004. Simultaneous quantification of methanol and ethanol in alcoholic beverage using a rapid gas chromatographic method coupling with dual internal standards. Food Chemistry 86: 609-615.
Wang, Y. H., Yu, G. L., Wang, X. M., Lv, Z. H., Zhao, X., Wu, Z. H., Ji, W. S. 2006. Purification and characterization of alginate lyase from marine Vibrio sp. YWA. Acta Biochimica et Biophysica Sinica 38: 633-638.
Wijffels, R. H., Barbosa, M. J. 2010. An outlook on microalgal biofuels. Science 329: 796-799.
Woods, D. R. 1995. The genetic engineering of microbial solvent production. Trends Biotechnology 13: 259-264.
Yeon, J. H., Lee, S. E., Choi, W. Y., Kang, D. H., Lee, H. Y., Jung, K. H. 2011. Repeated-batch operation of surface-aerated fermentor for bioethanol production from the hydrolysate of seaweed Sargassum sagamianum. Journal of Microbiology and Biotechnology 21: 323-331.
Zhang, Z. Q., Yu, G. L., Zhao, X., Li, Q. and Guan, H. S. 2005. Thin layer chromatography analysis of several urinates and their oligosaccharides. Chinese Journal of Analytical Chemistry 33: 1750-1752.
Zheng, Y. N., Li, L. Z., Xian, M., Ma, Y. J., Yang, J. M., Xu, X., He, D. Z. 2009. Problems with the microbial production of butanol. Journal of Industrial Microbiology and Biotechnology 36: 1127-1138.