臺灣博碩士論文加值系統

D-塔格糖 (D-tagatose) 為 D-半乳糖 (D-galactose) 之異構物，具有低熱量 (1.5 kcal/g) 、高甜度 (約為蔗糖甜度的 92%) 及低升糖指數的特性。藉由 L-阿拉伯糖異構酶 (L-arabinose isomerase, L-AI) 將 D-半乳糖轉換成 D-塔格糖，其具有副產物少、對環境友善、成本較低等優勢。Thermoanaerobacterium saccharolyticum NTOU1 來源之 L-AI (TsAI) 具有良好的熱穩定性，然而 TsAI 之反應速率與某些菌種來源之 L-AI 相比較差，因此本篇研究期望藉由蛋白質工程提昇 TsAI 的反應速率並搭配固定化技術進而提昇 D-塔格糖工業生產的可能性。以網路伺服器 SWISS-MODEL 模擬 TsAI 之蛋白質結構，再將模擬之 TsAI 結構與基質 D-半乳糖進行分子對接 (docking) 並分析可能與基質產生交互作用之胺基酸，之後透過定位點突變獲得突變型菌株，其中突變型 F270N 在 50℃ 下之 kcat 約為原生型酵素之 8 倍，於降低 D-塔格糖褐變與生產具有優勢。為了增加重複利用性與穩定性，將含有 TsAI 之 Escherichia coli ClearColi BL21 (DE3) 重組菌體以海藻酸鈉進行固定化。含 TsAI 之固定化菌體的最適固定化方式為利用含 0.05% Triton X-100 之緩衝溶液將菌體懸浮至 50 g/L 之菌體濃度，之後與等體積之 5% 海藻酸鈉混合再滴入 200 mM 氯化鈣溶液中；當以最適固定化條件將含 TsAI 之菌體進行固定化，含原生型 TsAI 之固定化菌體於 55℃ 下之活性為 16.1 U/g cell、固定化效率約為 20.5%；而含突變型 F270N 之固定化菌體於相同條件下之固定化菌體之活性約為 29.8 U/g cell、固定化效率約為33.2%，結果顯示含突變型 F270N 之固定化菌體於 55℃ 下具有較高之活性與固定化效率。後續藉由固定化菌體搭配填充床反應器生產 D-塔格糖，當以 10% D-半乳糖為基質時，含原生型 TsAI 之固定化菌體於 55℃ 下反應 36 小時後達平衡，轉換率約為 30%，而含突變型 F270N 之固定化菌體於相同條件下反應 6 小時後即達平衡，轉換率約為 40%。綜合上述結果，與原生型 TsAI 之固定化菌體相比，以含有突變型 F270N 之固定化菌體生產 D-塔格糖具有較高反應速率與轉換率之優勢。

D-Tagatose is an isomer of D-galactose and it has some properties such as low calories (1.5 kcal/g), high sweetness (92% as sweet as sucrose) and low glycemic index. With less by-products, eco-friendly and less costly advantages, production of D-tagatose from D-galactose using L-arabinose isomerase (L-AI) has been studied intensively in recent years. L-AI from Thermoanaerobacterium saccharolyticum NTOU1 (TsAI) has been characterized in a previous study and it has good thermostability. However, the reaction rate of TsAI was inferior to some L-AIs from other microorganisms. This study attempted to improve the reaction rate of TsAI by protein engineering and produce D-tagatose using immobilized cells for increasing its applicability to industry. The structure of TsAI was modeled by SWISS-MODEL and the modeling structure was further docked with D-galactose. Mutant residues were selected by analyzing the relative residues of binding site which might interact with substrate and mutations were made by site-directed mutagenesis. According to enzyme kinetics, the kcat of mutant F270N was eight times larger than that of wild-type TsAI at 50℃. It revealed that F270N was more suitable for producing D-tagatose with decreasing browning of D-tagatose. To improve reusability and stability, the recombinant Escherichia coli ClearColi BL21 (DE3) cells harboring TsAI were immobilized with alginate. The optimal conditions for immobilization were conducted with 5% alginate, 200 mM CaCl2, 0.05% Triton X-100 and 50 g/L cell mass. On the optimal conditions, the activity of immobilized cells harboring wild-type TsAI was 16.1 U/g cell and immobilized efficiency was 20.5% at 55℃; the activity of immobilized cells harboring F270N TsAI was 29.8 U/g cell and immobilized efficiency was 33.2% at the same temperature. It concluded that the immobilized cells harboring F270N was presented with higher activity and efficiency at 55℃. To further exploring the industrial application, production of D-tagatose from 10% D-galactose was performed at 55℃ by immobilized cells in packed-bed reactor. The immobilized cells harboring wild-type TsAI could reach reaction equilibrium in 36 h at about 30% conversion yield and the immobilized cells harboring F270N TsAI could reached reaction equilibrium in 6 h at about 40% conversion yield in the same condition. In summary, it had advantages to produce D-tagatoe by immobilized cells harboring F270N with higher reaction rate and higher conversion yield than those of wild-type TsAI.

摘要 I
Abstract II
目錄 III
圖目錄 VI
表目錄 VIII
壹、 研究背景與目的 1
一、 研究背景 1
二、 研究目的 1
貳、 文獻整理 2
一、 稀有醣類 2
1. 簡介 2
2. 應用 2
3. 生產 2
4. D-塔格糖 (D-tagatose) 3
二、 L-阿拉伯糖異構酶 (L-arabinose isomerase, L-AI) 4
1. 簡介 4
2. 來源 4
3. 立體結構 4
4. 源自 T. saccharolyticum NTOU1 之 L-阿拉伯糖異構酶 5
三、 電腦輔助之蛋白質工程 6
1. 同源模擬 (Homology modeling) 6
2. 分子對接 (Molecular docking) 6
四、 氫鍵對酵素催化中心之影響 7
五、 大腸桿菌表現菌株與質體 7
1. ClearColi BL21 (DE3) 7
2. CodonPlus-RIL 8
六、 固定化 8
1. 簡介 8
2. 固定化材料介紹 9
參、 實驗設計與流程 11
一、 突變型 TsAI 酵素製備及特性分析 11
1. 以電腦軟體模擬進行突變點選擇及定位點突變 11
2. 突變點酵素之表現及特性探討 11
二、 TsAI 於 ClearColi 宿主表現 11
三、 固定化菌體並生產 D-塔格糖 12
1. 探討含 TsAI 菌體之固定化最適條件 12
2. 以固定化菌體生產 D-塔格糖 13
肆、 實驗材料與方法 14
一、 實驗材料 14
1. 菌株與載體 14
2. 抗生素 14
3. 標準品 14
4. 市售套組 14
5. 酵素 14
6. 培養基 15
7. 化學藥品 15
8. 實驗設備 16
9. 軟體 17
二、 藥品配製 18
1. Terrific broth (TB) 培養液 18
2. Super optimal broth (SOB) 培養液 18
3. Super optimal broth with catabolite repression (SOC) 培養液 18
4. Ampicillin (100 mg/mL) 19
5. Chloramphenicol (34 mg/mL) 19
6. DNA 瓊脂膠電泳膠片製備 19
7. 定量蛋白質之相關試劑 19
8. SDS-PAGE 相關藥劑製備 19
9. 小量破菌相關試劑 21
10. Cysteine-carbazole 呈色法相關試劑 21
三、 實驗方法 22
1. 電腦軟體模擬 22
2. 製備質體 DNA 22
3. 定位突變 23
4. 轉形作用與重組質體之篩選 26
5. 篩選突變型酵素 27
6. 重組型 TsAI 之蛋白質表現與純化 30
7. 重組型 TsAI 之活性分析與特性探討 31
8. TsAI 於 ClearColi BL21 (DE3) 表現與最適菌體培養條件探討 34
9. 固定化菌體條件與特性探討 35
10. D-塔格糖生產 38
11. 統計分析 38
伍、 結果與討論 39
一、 TsAI 之定位突變 39
1. TsAI 結構模擬 39
2. TsAI 之突變點搜尋 39
3. TsAI 之定位點突變 40
二、 突變型 TsAI 之活性快篩 40
三、 原生型與突變型 TsAI 之特性探討 40
1. TsAI 之酵素表現與純化 41
2. TsAI 之酵素特性 41
四、 原生型 TsAI 於 ClearColi BL21 (DE3)-CodonPlus-RIL 之表現 42
1. 最適 IPTG 誘導濃度 42
2. 最適誘導溫度 43
3. 最適誘導時間 43
五、 固定化菌體之固定化條件探討 43
六、 含原生型及突變型 TsAI 固定化菌體之特性探討 44
1. 固定化菌體之最適作用條件 44
2. 固定化菌體於最適條件下之活性 44
3. 固定化菌體之熱穩定性 45
4. 固定化菌體之儲存穩定性 45
七、 利用含 TsAI 之固定化菌體生產 D-塔格糖 45
陸、 結論 47
柒、 參考文獻 48
捌、 圖表 58
玖、 附錄 99

方弘懿，2016，以蛋白質工程及固定化提昇 Agrobacterium sp. ATCC 31750 來源重組 D-阿洛酮糖表異構酶之熱穩定性，國立臺灣海洋大學食品科學系碩士學位論文，基隆。
陳于君，2014，利用蛋白質工程改變 Thermoanaerobacterium saccharolyticum NTOU1 L-鼠李糖異構酶的基質特異性，國立臺灣海洋大學食品科學系碩士學位論文，基隆。
許仲霆，2014，利用蛋白質工程提昇 D-阿洛酮糖表異構酶之活性回收及熱穩定性，國立台灣洋大學食品科學系碩士學位論文，基隆。
張哲源，2016a，源自 Thermoanaerobacterium saccharolyticum L-鼠李糖異構酶之突變、固定化條件探討及 D-阿洛糖的生產，國立臺灣海洋大學食品科學系碩士學位論文，基隆。
張雅如，2016b，以蛋白質工程提昇 Geodermatophilus obscurus DSM43160 重組L-核糖異構酶的酵素活性及熱穩定性，國立臺灣海洋大學食品科學系碩士學位論文，基隆。
黃幸光，2014，熱穩定性阿拉伯糖異構酶之表現、特性探討、稀有醣類生產及蛋白質工程，國立臺灣海洋大學食品科學系博士學位論文，基隆。
游銘遠，2012，Geodermatophilus obscurus DSM 43160 來源之 L-核糖異構酶之基因選殖、表現、純化及特性探討，國立臺灣海洋大學食品科學系碩士學位論文，基隆。
魏岑芸，2007，增加基質結合部位殘基與基質間氫鍵對於 Sulfolobus solfararicus ATCC35092 麥芽寡糖苷海藻糖生成酶轉糖苷及水解作用之影響，國立台灣海洋大學食品科學系碩士學位論文，基隆。
Abagyan R, Totrov M, and Kuznetsov D. (1994). ICM—A new method for protein modeling and design: applications to docking and structure prediction from the distorted native conformation. Journal of Computational Chemistry, 15: 488-506.
Baek SH, Kwon SY, Lee HG, and Baek HH. (2008). Maillard browning reaction of D-psicose as affected by reaction factors. Food Science and Biotechnology, 17: 1349-1351.
Beadle JR, Saunders JP, and Wajda Jr TJ. (1992). Process for manufacturing tagatose. In: US Patent 5,078,796.
Beerens K, Desmet T, and Soetaert W. (2012). Enzymes for the biocatalytic production of rare sugars. Journal of Industrial Microbiology and Biotechnology, 39: 823-834.
Benkert P, Schwede T, and Tosatto SC. (2009). QMEANclust: estimation of protein model quality by combining a composite scoring function with structural density information. BMC Structural Biology, 9: 1-17.
Benkert P, Tosatto SC, and Schomburg D. (2008). QMEAN: A comprehensive scoring function for model quality assessment. Proteins: Structure, Function, and Bioinformatics, 71: 261-277.
Bertelsen H, Jensen BB, and Buemann B. (1999). D-Tagatose a novel low-calorie bulk sweetener with prebiotic properties. World Rev Nutr Diet, 85: 98-109.
Bhairi SM, and Mohan C. (1997). A guide to the properties and uses of detergents in biology and biochemistry. Doc. No. CB0068-0892, Calbiochem-Novabiochem Corp, LaJolla, CA.
Biasini M, Bienert S, Waterhouse A, Arnold K, Studer G, Schmidt T, Kiefer F, Cassarino TG, Bertoni M, and Bordoli L. (2014). SWISS-MODEL: modelling protein tertiary and quaternary structure using evolutionary information. Nucleic Acids Research, DOI: 10.1093/nar/gku340.
Bordoli L, Kiefer F, Arnold K, Benkert P, Battey J, and Schwede T. (2009). Protein structure homology modeling using SWISS-MODEL workspace. Nature Protocols, 4: 1-13.
Bradford MM. (1976). A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry, 72: 248-254.
Bryan P, Pantoliano MW, Quill SG, Hsiao HY, and Poulos T. (1986). Site-directed mutagenesis and the role of the oxyanion hole in subtilisin. Proceedings of the National Academy of Sciences, 83: 3743-3745.
Carvalho GM, Alves TLM, and Freire DM. (2000). L-DOPA production by immobilized tyrosinase. Applied Biochemistry and Biotechnology, 84: 791-800.
Chang MW, Ayeni C, Breuer S, and Torbett BE. (2010). Virtual screening for HIV protease inhibitors: a comparison of AutoDock 4 and Vina. PloS One, 5: e11955.
Cheetham P, and Wootton A. (1993). Bioconversion of D-galactose into D-tagatose. Enzyme and Microbial Technology, 15: 105-108.
Cheng L, Mu W, Zhang T, and Jiang B. (2010). An L-arabinose isomerase from Acidothermus cellulolytics ATCC 43068: cloning, expression, purification, and characterization. Applied Microbiology and Biotechnology, 86: 1089-1097.
Choi JM, Lee YJ, Cao TP, Shin SM, Park MK, Lee HS, Di Luccio E, Kim SB, Lee SJ, and Lee SJ. (2016). Structure of the thermophilic L-arabinose isomerase from Geobacillus kaustophilus reveals metal-mediated intersubunit interactions for activity and thermostability. Archives of Biochemistry and Biophysics, 596: 51-62.
Chouayekh H, Bejar W, Rhimi M, Jelleli K, Mseddi M, and Bejar S. (2007). Characterization of an L-arabinose isomerase from the Lactobacillus plantarum NC8 strain showing pronounced stability at acidic pH. FEMS Microbiology Letters, 277: 260-267.
Cleland W, and Kreevoy MM. (1994). Low-barrier hydrogen bonds and enzymic catalysis. Science, 264: 1887-1887.
DiCosimo R, McAuliffe J, Poulose AJ, and Bohlmann G. (2013). Industrial use of immobilized enzymes. Chemical Society Reviews, 42: 6437-6474.
Dische Z, and Borenfreund E. (1951). A new spectrophotometric method for the detection and determination of keto sugars and trioses. Journal of Biological Chemistry, 192: 583-587.
Donner T, Wilber J, and Ostrowski D. (1999). D‐Tagatose, a novel hexose: acute effects on carbohydrate tolerance in subjects with and without type 2 diabetes. Diabetes, Obesity and Metabolism, 1: 285-291.
Ensor M, Banfield AB, Smith RR, Williams J, and Lodder RA. (2015). Safety and efficacy of D-tagatose in glycemic control in subjects with type 2 diabetes. Journal of Endocrinology, Diabetes and Obesity, 3: 1-24.
Ewing TJ, Makino S, Skillman AG, and Kuntz ID. (2001). DOCK 4.0: search strategies for automated molecular docking of flexible molecule databases. Journal of Computer-Aided Molecular Design, 15: 411-428.
Fan C, Liu K, Zhang T, Zhou L, Xue D, Jiang B, and Mu W. (2014). Biochemical characterization of a thermostable L-arabinose isomerase from a thermoacidophilic bacterium, Alicyclobacillus hesperidum URH17-3-68. Journal of Molecular Catalysis B: Enzymatic, 102: 120-126.
Felix H. (1982). Permeabilized cells. Analytical Biochemistry, 120: 211-234.
Fersht AR, Shi J, Knill J, Jones D, Lowe A, Wilkinson D, Blow PB, Carter P, Waye M, and Winter G. (1985). Hydrogen bonding and biological specificity analysed by protein engineering. Nature, 314: 235-238.
Forli S, Huey R, Pique ME, Sanner MF, Goodsell DS, and Olson AJ. (2016). Computational protein-ligand docking and virtual drug screening with the AutoDock suite. Nature Protocols, 11: 905-919.
Genisheva Z, Teixeira J, and Oliveira J. (2014). Immobilized cell systems for batch and continuous winemaking. Trends in Food Science & Technology, 40: 33-47.
George M, and Abraham TE. (2006). Polyionic hydrocolloids for the intestinal delivery of protein drugs: alginate and chitosan—a review. Journal of Controlled Release, 114: 1-14.
Goodsell DS, Morris GM, and Olson AJ. (1996). Automated docking of flexible ligands: applications of AutoDock. Journal of Molecular Recognition, 9: 1-5.
Gumina G, Song GY, and Chu CK. (2001). L-Nucleosides as chemotherapeutic agents. FEMS Microbiology Letters, 202: 9-15.
He YC, Zhang ZJ, Xu JH, and Liu YY. (2010). Biocatalytic synthesis of (R)-(−)-mandelic acid from racemic mandelonitrile by cetyltrimethylammonium bromide-permeabilized cells of Alcaligenes faecalis ECU0401. Journal of Industrial Microbiology and Biotechnology, 37: 741-750.
Hirst E, Hough L, and Jones J. (1949). Composition of the gum of Sterculia setigera: occurrence of D-tagatose in nature. Nature, 163: 177.
Hossain MA, Kitagaki S, Nakano D, Nishiyama A, Funamoto Y, Matsunaga T, Tsukamoto I, Yamaguchi F, Kamitori K, and Dong Y. (2011). Rare sugar D-psicose improves insulin sensitivity and glucose tolerance in type 2 diabetes Otsuka Long-Evans Tokushima Fatty (OLETF) rats. Biochemical and Biophysical Research Communications, 405: 7-12.
Huang T, Gao D, Hei Y, Zhang X, Chen X, and Fei Z. (2016). D-Allose protects the blood brain barrier through PPARγ-mediated anti-inflammatory pathway in the mice model of ischemia reperfusion injury. Brain Research, 1642: 478-486.
Hung XG, Tseng WC, Liu SM, Tzou WS, and Fang TY. (2014). Characterization of a thermophilic L-arabinose isomerase from Thermoanaerobacterium saccharolyticum NTOU1. Biochemical Engineering Journal, 83: 121-128.
Ikeda S, Furuta C, Fujita Y, and Gohtani S. (2014). Effects of D‐psicose on gelatinization and retrogradation of rice flour. Starch‐Stärke, 66: 773-779.
Ishihara Y, Katayama K, Sakabe M, Kitamura M, Aizawa M, Takara M, and Itoh K. (2011). Antioxidant properties of rare sugar D-allose: effects on mitochondrial reactive oxygen species production in Neuro2A Cells. Journal of Bioscience and Bioengineering, 112: 638-642.
Izumori K. (2006). Izumoring: a strategy for bioproduction of all hexoses. Journal of Biotechnology, 124: 717-722.
Izumori K, Miyoshi T, Tokuda S, and Yamabe K. (1984). Production of D-tagatose from dulcitol by Arthrobacter globiformis. Applied and Environmental Microbiology, 48: 1055-1057.
Izumori K, and Tsuzaki K. (1988). Production of D-tagatose from D-galactitol by Mycobacterium smegmatis. Journal of Fermentation Technology, 66: 225-227.
Jørgensen F, Hansen O, and Stougaard P. (2004). Enzymatic conversion of D-galactose to D-tagatose: heterologous expression and characterisation of a thermostable L-arabinose isomerase from Thermoanaerobacter mathranii. Applied Microbiology and Biotechnology, 64: 816-822.
Jayamuthunagai J, Gautam P, Srisowmeya G, and Chakravarthy M. (2016). Biocatalytic production of D-tagatose: a potential rare sugar with versatile applications. Critical Reviews in Food Science and Nutrition.
Kim BC, Lee YH, Lee HS, Lee DW, Choe EA, and Pyun YR. (2002). Cloning, expression and characterization of L-arabinose isomerase from Thermotoga neapolitana: bioconversion of D-galactose to D-tagatose using the enzyme. FEMS Microbiology Letters, 212: 121-126.
Kim BJ, Hong SH, Shin KC, Jo YS, and Oh DK. (2014). Characterization of a F280N variant of L-arabinose isomerase from Geobacillus thermodenitrificans identified as a D-galactose isomerase. Applied Microbiology and Biotechnology, 98: 9271-9281.
Kim H, Kim J, Oh H, and Oh D. (2006). Characterization of a mutated Geobacillus stearothermophilus L‐arabinose isomerase that increases the production rate of D‐tagatose. Journal of Applied Microbiology, 101: 213-221.
Kim HJ, and Oh DK. (2005). Purification and characterization of an L-arabinose isomerase from an isolated strain of Geobacillus thermodenitrificans producing D-tagatose. Journal of Biotechnology, 120: 162-173.
Kim JH, Prabhu P, Jeya M, Tiwari MK, Moon HJ, Singh RK, and Lee JK. (2010). Characterization of an L-arabinose isomerase from Bacillus subtilis. Applied Microbiology and Biotechnology, 85: 1839-1847.
Kim JW, Kim YW, Roh HJ, Kim HY, Cha JH, Park KH, and Park CS. (2003). Production of tagatose by a recombinant thermostable L-arabinose isomerase from Thermus sp. IM6501. Biotechnology Letters, 25: 963-967.
Kim P. (2004). Current studies on biological tagatose production using L-arabinose isomerase: a review and future perspective. Applied Microbiology and Biotechnology, 65: 243-249.
Kleber-Janke T, and Becker W-M. (2000). Use of modified BL21 (DE3) Escherichia coli cells for high-level expression of recombinant peanut allergens affected by poor codon usage. Protein Expression and Purification, 19: 419-424.
Klibanov AM. (1983). Immobilized enzymes and cells as practical catalysts. Science, 219: 722-727.
Kramer B, Rarey M, and Lengauer T. (1999). Evaluation of the FLEXX incremental construction algorithm for protein–ligand docking. Proteins: Structure, Function, and Bioinformatics, 37: 228-241.
Lee DW, Choe EA, Kim SB, Eom SH, Hong YH, Lee SJ, Lee HS, Lee DY, and Pyun YR. (2005). Distinct metal dependence for catalytic and structural functions in the L-arabinose isomerases from the Mesophilic Bacillus halodurans and the thermophilic Geobacillus stearothermophilus. Archives of Biochemistry and Biophysics, 434: 333-343.
Lee DW, Jang HJ, Choe EA, Kim BC, Lee SJ, Kim SB, Hong YH, and Pyun YR. (2004). Characterization of a thermostable L-arabinose (D-galactose) isomerase from the hyperthermophilic eubacterium Thermotoga maritima. Applied and Environmental Microbiology, 70: 1397-1404.
Lee KY, and Mooney DJ. (2012). Alginate: properties and biomedical applications. Progress in Polymer Science, 37: 106-126.
Lee SJ, Lee DW, Choe EA, Hong YH, Kim SB, Kim BC, and Pyun YR. (2005). Characterization of a thermoacidophilic L-arabinose isomerase from Alicyclobacillus acidocaldarius: role of Lys-269 in pH optimum. Applied and Environmental Microbiology, 71: 7888-7896.
Levin GV. (2002). Tagatose, the new GRAS sweetener and health product. Journal of Medicinal Food, 5: 23-36.
Li Y, Zhu Y, Liu A, and Sun Y. (2011). Identification and characterization of a novel L-arabinose isomerase from Anoxybacillus flavithermus useful in D-tagatose production. Extremophiles, 15: 441-450.
Linke D. (2009). Detergents: an overview. Methods in Enzymology, 463: 603-617.
Longo MA, and Combes D. (1999). Thermostability of modified enzymes: a detailed study. Journal of Chemical Technology and Biotechnology, 74: 25-32.
Lu Y. (2001). Humectancies of D‐tagatose and D‐sorbitol. International Journal of Cosmetic Science, 23: 175-181.
Lu Y, and Levin G. (2002). Removal and prevention of dental plaque with D‐tagatose. International Journal of Cosmetic Science, 24: 225-234.
Luecke KJ, and Bell LN. (2010). Thermal stability of tagatose in solution. Journal of Food Science, 75: 346-351.
Ma T, Pai SB, Zhu YL, Lin JS, Shanmuganathan K, Du J, Wang C, Kim H, Newton MG, and Cheng YC. (1996). Structure-activity relationships of 1-(2-deoxy-2-fluoro-β-l-arabino-furanosyl) pyrimidine nucleosides as anti-hepatitis B virus agents. Journal of Medicinal Chemistry, 39: 2835-2843.
Mamat U, Woodard RW, Wilke K, Souvignier C, Mead D, Steinmetz E, Terry K, Kovacich C, Zegers A, and Knox C. (2013). Endotoxin-free protein production - ClearColiTM technology. Nature Methods, 10.
Manjasetty BA, and Chance MR. (2006). Crystal structure of Escherichia coli L-arabinose isomerase (ECAI), the putative target of biological tagatose production. Journal of Molecular Biology, 360: 297-309.
Mei W, Wang L, Zang Y, Zheng Z, and Ouyang J. (2016). Characterization of an L-arabinose isomerase from Bacillus coagulans NL01 and its application for D-tagatose production. BMC Biotechnology, 16: 1-11.
Men Y, Zhu Y, Zhang L, Kang Z, Izumori K, Sun Y, and Ma Y. (2014). Enzymatic conversion of D-galactose to D-tagatose: cloning, overexpression and characterization of L-arabinose isomerase from Pediococcus pentosaceus PC-5. Microbiological Research, 169: 171-178.
Michaelis L, and Menten ML. (1913). Die kinetik der invertinwirkung. Biochem. Z, 49: 334-339.
Nagata Y, Kanasaki A, Tamaru S, and Tanaka K. (2015). D-Psicose, an epimer of D-fructose, favorably alters lipid metabolism in sprague–dawley rats. Journal of Agricultural and Food Chemistry, 63: 3168-3176.
Neumann E, Schaefer-Ridder M, Wang Y, and Hofschneider P. (1982). Gene transfer into mouse lyoma cells by electroporation in high electric field. The EMBO Journal, 1: 841-845.
Noguchi C, Kamitori K, Hossain A, Hoshikawa H, Katagi A, Dong Y, Sui L, Tokuda M, and Yamaguchi F. (2016). D-Allose inhibits cancer cell growth by reducing GLUT1 expression. The Tohoku Journal of Experimental Medicine, 238: 131-141.
Obón J, Maiquez JR, Canovas M, Kleber H-P, and Iborra J. (1997). L(−)-Carnitine production with immobilized Escherichia coli cells in continuous reactors. Enzyme and Microbial Technology, 21: 531-536.
Oh HJ, Kim HJ, and Oh DK. (2006). Increase in D-tagatose production rate by site-directed mutagenesis of L-arabinose isomerase from Geobacillus thermodenitrificans. Biotechnology Letters, 28: 145-149.
Orwick‐Rydmark M, Arnold T, and Linke D. (2016). The use of detergents to purify membrane proteins. Current Protocols in Protein Science: 4.8. 1-4.8. 35.
Paterna J, Boess F, Stäubli A, and Boelsterli U. (1998). Antioxidant and cytoprotective properties of D-tagatose in cultured murine hepatocytes. Toxicology and Applied Pharmacology, 148: 117-125.
Police SB, Harris JC, Lodder RA, and Cassis LA. (2009). Effect of diets containing sucrose vs. D‐tagatose in hypercholesterolemic mice. Obesity, 17: 269-275.
Prabhu P, Tiwari MK, Jeya M, Gunasekaran P, Kim I-W, and Lee J-K. (2008). Cloning and characterization of a novel L-arabinose isomerase from Bacillus licheniformis. Applied Microbiology and Biotechnology, 81: 283-290.
Rhimi M, Bajic G, Ilhammami R, Boudebbouze S, Maguin E, Haser R, and Aghajari N. (2011). The acid-tolerant L-arabinose isomerase from the mesophilic Shewanella sp. ANA-3 is highly active at low temperatures. Microbial Cell Factories, 10: 96-107.
Rhimi M, and Bejar S. (2006). Cloning, purification and biochemical characterization of metallic-ions independent and thermoactive L-arabinose isomerase from the Bacillus stearothermophilus US100 strain. Biochimica et Biophysica Acta (BBA)-General Subjects, 1760: 191-199.
Rhimi M, Ilhammami R, Bajic G, Boudebbouze S, Maguin E, Haser R, and Aghajari N. (2010). The acid tolerant L-arabinose isomerase from the food grade Lactobacillus sakei 23K is an attractive D-tagatose Producer. Bioresource Technology, 101: 9171-9177.
Rollini M, and Manzoni M. (2005). Bioconversion of D-galactitol to tagatose and dehydrogenase activity induction in Gluconobacter oxydans. Process Biochemistry, 40: 437-444.
Sadana A, and Henley JP. (1987). Single‐step unimolecular non‐first‐order enzyme deactivation kinetics. Biotechnology and Bioengineering, 30: 717-723.
Salonen N, Nyyssölä A, Salonen K, and Turunen O. (2012). Bifidobacterium longum L-arabinose isomerase—overexpression in Lactococcus lactis, purification, and characterization. Applied Biochemistry and Biotechnology, 168: 392-405.
Schägger H, and Von Jagow G. (1987). Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis for the separation of proteins in the range from 1 to 100 kDa. Analytical Biochemistry, 166: 368-379.
Schleif R. (2000). Regulation of the L-arabinose operon of Escherichia coli. Trends in Genetics, 16: 559-565.
Schwede T, Kopp J, Guex N, and Peitsch MC. (2003). SWISS-MODEL: an automated protein homology-modeling server. Nucleic Acids Research, 31: 3381-3385.
Seddon AM, Curnow P, and Booth PJ. (2004). Membrane proteins, lipids and detergents: not just a soap opera. Biochimica et Biophysica Acta (BBA)-Biomembranes, 1666: 105-117.
Sitton O, and Gaddy J. (1980). Ethanol production in an immobilized‐cell reactor. Biotechnology and Bioengineering, 22: 1735-1748.
Smidsrød O, and Skja G. (1990). Alginate as immobilization matrix for cells. Trends in Biotechnology, 8: 71-78.
Stano J, Nemec P, Weissová K, Kovács P, Kákoniová D, and Lisková D. (1995). Decarboxylation of L-tyrosine and L-DOPA by immobilized cells of Papaver somniferum. Phytochemistry, 38: 859-860.
Suárez L, Díez MA, García R, and Riera FA. (2012). Membrane technology for the recovery of detergent compounds: a review. Journal of Industrial and Engineering Chemistry, 18: 1859-1873.
Sui L, Nomura R, Dong Y, Yamaguchi F, Izumori K, and Tokuda M. (2007). Cryoprotective effects of D-allose on mammalian cells. Cryobiology, 55: 87-92.
Tønnesen HH, and Karlsen J. (2002). Alginate in drug delivery systems. Drug Development and Industrial Pharmacy, 28: 621-630.
Tseng WC, Lin JW, Wei TY, and Fang TY. (2008). A novel megaprimed and ligase-free, PCR-based, site-directed mutagenesis method. Analytical Biochemistry, 375: 376-378.
Valeri F, Boess F, Wolf A, Göldlin C, and Boelsterli UA. (1997). Fructose and tagatose protect against oxidative cell injury by iron chelation. Free Radical Biology and Medicine, 22: 257-268.
Van de Velde F, Lourenço ND, Pinheiro HM, and Bakker M. (2002). Carrageenan: a food-grade and biocompatible support for immobilisation techniques. Advanced Synthesis and Catalysis, 344: 815-836.
Wanarska M, and Kur J. (2012). A method for the production of D-tagatose using a recombinant Pichia pastoris strain secreting β-D-galactosidase from Arthrobacter chlorophenolicus and a recombinant L-arabinose isomerase from Arthrobacter sp. 22c. Microbial Cell Factories, 11: 1.
Xu Z, Li S, Feng X, Liang J, and Xu H. (2014). L-Arabinose isomerase and its use for biotechnological production of rare sugars. Applied Microbiology and Biotechnology, 98: 8869-8878.
Xu Z, Qing Y, Li S, Feng X, Xu H, and Ouyang P. (2011). A novel L-arabinose isomerase from Lactobacillus fermentum CGMCC2921 for D-tagatose production: gene cloning, purification and characterization. Journal of Molecular Catalysis B: Enzymatic, 70: 1-7.
Yamaguchi F, Kamitori K, Sanada K, Horii M, Dong Y, Sui L, and Tokuda M. (2008). Rare sugar D-allose enhances anti-tumor effect of 5-fluorouracil on the human hepatocellular carcinoma cell line HuH-7. Journal of Bioscience and Bioengineering, 106: 248-252.
Yoon SH, Kim P, and Oh DK. (2003). Properties of L-arabinose isomerase from Escherichia coli as biocatalyst for tagatose production. World Journal of Microbiology and Biotechnology, 19: 47-51.
Zhang YW, Prabhu P, and Lee JK. (2010). Alginate immobilization of recombinant Escherichia coli whole cells harboring L-arabinose isomerase for L-ribulose production. Bioprocess and Biosystems Engineering, 33: 741-748.
Zhao WR, Huang J, Peng CL, Hu S, Ke PY, Mei LH, and Yao SJ. (2014). Permeabilizing Escherichia coli for whole cell biocatalyst with enhanced biotransformation ability from L-glutamate to GABA. Journal of Molecular Catalysis B: Enzymatic, 107: 39-46.