臺灣博碩士論文加值系統

Actinotalea fermentans ATCC 43279 含有一可能為 L-核糖異構酶之假設性蛋白，合成此蛋白之基因並構築至 pET-21b 載體後以
Escherichia coli 大量表現。此重組型 L-核糖異構酶 (Af-RI) 的最適作用溫度為 45ºC，最適作用 pH 值為 8.0 (Tricine-NaOH buffer)；不需要額外添加金屬離子即有活性，但汞離子會使酵素完全失活；於 50ºC 之活性半衰期約為 74 分鐘；於 45ºC 反應至平衡時 L-核酮糖對 L-核糖之比例約為 30:70；對 L-核糖之 KM 值為 31.3 mM，Vmax 值為 232 U/mg，kcat 值為 6700 min-1，催化效率 (kcat/KM) 為 214 min-1·mM-1。與各來源之 L-RI 相比，Af-RI 具有最高的 Vmax 值、kcat 值、催化效率及酵素活性表現量，顯示 Af-RI 有生產 L-核糖之工業應用潛力。使用 MAESTRO 伺服器預測可能增加穩定性之突變，並建構一連接 SUMO (small ubiquitin-related modifier) 蛋白之 Af-RI，以期提昇 Af-RI 之熱穩定性，初步活性篩選的結果顯示，G39F、E61W、A68V、A68W 及 A165R 之突變型 Af-RI 的粗酵素液皆無活性；仍具活性之酵素進行熱穩定性篩選，於 50ºC 保溫 20 分鐘後，原生型 Af-RI、G176W、A233W、G43C/A238W 之突變型 Af-RI 及 SUMO-Af-RI 的殘餘活性分別為 19.0%、0%、16.2%、12.7% 及 9.1%，顯示各突變型 Af-RI 和 SUMO-Af-RI 的熱穩定性皆不如原生型 Af-RI。以不產內毒素之 E. coli ClearColi BL21 (DE3) 作為表現宿主，並藉由海藻酸鈣之包埋法製備含 Af-RI 的固定化菌體，其最適條件為使用 0.05% Triton X-100；5% 海藻酸鈉；50 g/L 之菌體；200 mM 氯化鈣；於氯化鈣溶液中固化 4 小時。以最適條件製備之固定化菌體的最適作用溫度為 50ºC；最適作用 pH 值為 8.0 (Tricine-NaOH buffer)；於 50ºC 之活性半衰期約為 1386 分鐘，約為未固定酵素的 19 倍。

A putative L-ribose isomerase gene of A. fermentans ATCC 43279 was chemically synthesized, subcloned into pET-21b vector, and then overexpressed in Escherichia coli. The purified recombinant L-ribose isomerase (Af-RI) demonstrated its optimal activity at 45ºC and pH 8 (in Tricine-NaOH buffer). Metal ions are not required for L-ribose isomerase (L-RI) activity, but Hg2+ inhibits its activity completely. The enzyme has a half-life of 74 min at 50ºC and the equilibrium ratio of 30:70 between L-ribulose and L-ribose at 45ºC. The Vmax, kcat, KM, and catalytic efficiency of Af-RI against L-ribose are 232 U/mg, 6700 min-1, 31.3 mM, and 214 min-1·mM-1, respectively. The high expression yield of the Af-RI and its highest Vmax, kcat, and catalytic efficiency among the characterized recombinant L-RIs suggest that this recombinant enzyme shows a potential application to produce L-ribose in industry. To enhance the thermostability of Af-RI by mutagenesis, several mutations predicted by using the MAESTRO sever was constructed, and a recombinant Af-RI with a fused SUMO (small ubiquitin-related modifier) protein was also constructed. The crude extracts containing G39F, E61W, A68V, A68W and A165R mutant Af-RIs are no activity. The residual activities of the crude extracts containing wild-type, G176W, A233W, G43C/A238C Af-RIs and SUMO-Af-RI after incubating at 50ºC for 20 min are 19.0, 0, 16.2, 12.7 and 9.1%, respectively. It means that the thermostabilities of all mutant Af-RIs and SUMO-Af-RI aren’t better than that of wild-type Af-RI. Af-RI was overexpressed in strain E. coli ClearColi BL21 (DE3) which doesn’t produce endotoxin. The immobilized cells harboring Af-RI was prepared by entrapment with calcium alginate.The optimal conditions of immobilization are 0.05% Triton X-100, 5% alginate, 50 g/L cells, 200 mM CaCl2 solution, hardening in CaCl2 solution with 4 h. The immobilized cells harboring Af-RI showed its optimal activity at 50ºC and pH 8 (in Tricine-NaOH buffer) and have a half-life of 1386 min at 50ºC. The half-life of immobilized cells at 50ºC is about nineteen times longer than that of free enzyme.

壹、研究背景與目的 1
一、研究背景 1
二、研究目的 2
貳、文獻整理 3
一、稀有醣類 3
1. 稀有醣類簡介 3
2. 核糖 3
二、L-核糖的生產 3
1. 化學合成法 4
2. 酵素轉換法 4
三、生產 L-核糖之相關酵素 5
1. 甘露醇-1-脫氫酶 5
2. 核糖醇脫氫酶 5
3. L-阿拉伯糖異構酶 5
4. 甘露糖-6-磷酸異構酶 6
5. L-核糖異構酶 6
四、不同來源之 L-RI 的特性 6
1. Acinetobacter sp. DL-28來源之 L-RI 6
2. C. parahominis MB426 來源之 L-RI 6
3. G. obscurus DSM 43160 來源之 L-RI 7
五、L-RI 之晶體結構 7
1. AcRI 之結構 7
2. CeRI 之結構 8
六、A. fermentans ATCC 43279 之簡介 8
七、電腦軟體輔助之蛋白質工程 9
1. 蛋白質結構模擬 9
2. 電腦軟體預測蛋白質穩定性 9
八、SUMO 蛋白簡介 10
九、大腸桿菌表現宿主 10
1. E. coli BL21 StarTM (DE3) 10
2. ClearColiTM BL21 (DE3) 11
十、固定化菌體 11
1. 固定化簡介 11
2. 固定化方法 12
3. 固定化材料 12
參、實驗設計與流程 13
一、A. fermentans ATCC 43279 來源之 L-RI 的表現及特性探討 13
1. A. fermentans ATCC 43279 來源之 L-ri 基因構築 13
2. Af-RI 之表現、純化與特性探討 13
二、以突變改變 Af-RI 之熱穩定性 14
三、固定化菌體之方法與特性探討 14
肆、實驗材料與方法 15
一、 材料 15
1. 菌株與載體 15
2. 抗生素 15
3. 標準品 15
4. 酵素 15
5. 市售套組 16
6. 培養基 16
7. 化學藥品 16
8. 實驗設備 18
9. 軟體 19
二、 實驗步驟及方法 20
1. A. fermentans 來源之 L-ri 基因構築 20
1.1 基因合成 20
1.2 製備 Terrific broth 培養液 20
1.3 抽取質體 DNA 20
1.4 質體 DNA 之確認 21
1.5 限制酶剪切 22
1.6 膠純化回收 DNA 片段 22
1.7 載體 DNA 之 Clean up 22
1.8 接合作用 22
1.9 電穿孔轉形 23
1.10 Colony PCR 24
1.11 基因序列分析 25
1.12 菌種保存 25
1.13 質體轉形至表現宿主 25
1.14 菌種保存 26
2. Af-RI 之表現與純化 26
2.1 以 IPTG 誘導表現 Af-RI 26
2.2 SDS-PAGE 26
2.3 大量誘導表現原生型 Af-RI 29
2.4 Af-RI 酵素純化 29
2.5 MALDI-TOF 質譜儀分析 30
2.6 分子篩管柱層析 30
3. Af-RI 之特性探討 30
3.1 蛋白質定量 30
3.2 重組酵素之粗酵素液活性測定 31
3.3 最適作用溫度 31
3.4 最適作用 pH 值 31
3.5 熱穩定性 31
3.6 保溫於 50ºC 之活性半衰期 32
3.7 pH 穩定性 32
3.8 金屬離子之影響 32
3.9 基質特異性 33
3.10 酵素動力學 33
3.11 反應產物之平衡比率 33
4.酵素穩定性之蛋白質工程 34
4.1 電腦軟體模擬結構及預測蛋白質穩定性 34
4.2 定位點突變 34
4.3 電穿孔轉形作用 36
4.4 基因定序 37
4.5 菌種保存 37
4.6 突變型質體轉形至表現宿主 37
4.7 菌種保存 37
4.8 SUMO 蛋白之基因構築 37
4.9 粗酵素之篩選 37
5.固定化菌體 38
5.1 誘導表現 Af-RI 之最適 IPTG 濃度 38
5.2 Af-RI 之最適誘導溫度 38
5.3 Af-RI 之最適誘導時間 38
5.4 大量誘導表現 Af-RI 39
5.5 菌體固定化 39
5.6 固定化菌體條件探討 39
5.7 固定化菌體特性探討 41
5.8 統計分析 42
伍、結果與討論 43
陸、結論 50
柒、參考文獻 51
捌、圖表 59
玖、附錄 97

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。
張雅如，2016，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。
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。
鄭閎文，2015，重組 L-核糖異構酶及其固定化菌體之特性探討，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，國立台灣洋大學食品科學系碩士學位論文，基隆。
Adachi O, Fujii Y, Ano Y, MOONMANGMEE D, Toyama H, Shinagawa E, THEERAGOOL G, LOTONG N, and Matsushita K. (2001). Membrane-Bound Sugar Alcohol Dehydrogenase in Acetic Acid Bacteria Catalyzes L-Ribulose Formation and Nad-Dependent Ribitol Dehydrogenase Is Independent of the Oxidative Fermentation. Bioscience, Biotechnology, and Biochemistry, 65(1), 115-125.
Ahmed Z. (2001). Production of Natural and Rare Pentoses Using Microorganisms and Their Enzymes. Electronic Journal of Biotechnology, 4(2), 13-14.
Ahmed Z, Shimonishi T, Bhuiyan SH, Utamura M, Takada G, and Izumori K. (1999). Biochemical Preparation of L-Ribose and L-Arabinose from Ribitol: A New Approach. Journal of Bioscience and Bioengineering, 88(4), 444-448.
Altschul SF, Madden TL, Schäffer AA, Zhang J, Zhang Z, Miller W, and Lipman DJ. (1997). Gapped Blast and Psi-Blast: A New Generation of Protein Database Search Programs. Nucleic Acids Research, 25(17), 3389-3402.
Ashley GW. (1992). Modeling, Synthesis, and Hybridization Properties of L-Ribonucleic Acid. Journal of the American Chemical Society, 114(25), 9731-9736.
Bílik V, Anderle D, and Alföldi J. (1974). Reactions of Saccharides Catalyzed by Molybdate Ions. Xi. Preparation of L-Glycero-L-Galacto-and-L-Glycero-L-Taloheptose. Chemical Papers, 28(5), 668-672.
Bagnara C, Toci R, Gaudin C, and Belaich J. (1985). Isolation and Characterization of a Cellulolytic Microorganism, Cellulomonas Fermentans Sp. Nov. International Journal of Systematic and Evolutionary Microbiology, 35(4), 502-507.
Beerens K, Desmet T, and Soetaert W. (2012). Enzymes for the Biocatalytic Production of Rare Sugars. Journal of Industrial Microbiology & Biotechnology, 39(6), 823-834.
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, gku340.
Brena BM, and BatistaViera F. (2006). Immobilization of Enzymes. Immobilization of Enzymes and Cells, 15-30.
CárdenasFernández M, Neto W, López C, Álvaro G, Tufvesson P, and Woodley JM. (2012). Immobilization of Escherichia Coli Containing Ω‐Transaminase Activity in Lentikats®. Biotechnology Progress, 28(3), 693-698.
Capriotti E, Fariselli P, Rossi I, and Casadio R. (2008). A Three-State Prediction of Single Point Mutations on Protein Stability Changes. BMC Bioinformatics, 9(2), 1.
Cheng J, Randall A, and Baldi P. (2006). Prediction of Protein Stability Changes for Single‐Site Mutations Using Support Vector Machines. Proteins: Structure, Function, and Bioinformatics, 62(4), 1125-1132.
Chothia C, and Lesk AM. (1986). The Relation between the Divergence of Sequence and Structure in Proteins. The EMBO Journal, 5(4), 823.
Consortium U. (2008). The Universal Protein Resource (Uniprot). Nucleic Acids Research, 36(suppl 1), D190-D195.
Dehouck Y, Grosfils A, Folch B, Gilis D, Bogaerts P, and Rooman M. (2009). Fast and Accurate Predictions of Protein Stability Changes Upon Mutations Using Statistical Potentials and Neural Networks: Popmusic-2.0. Bioinformatics, 25(19), 2537-2543.
Dhanoa TS, and Housner JA. (2007). Ribose: More Than a Simple Sugar? Current Sports Medicine Reports, 6(4), 254-257.
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(2), 583-587.
Du J, Choi Y, Lee K, Chun BK, Hong JH, and Chu CK. (1999). A Practical Synthesis of L-Fmau from L-Arabinose. Nucleosides, Nucleotides & Nucleic Acids, 18(2), 187-195.
Du Poët PDT, Arcand Y, Bernier R, Barbotin J, and Thomas D. (1987). Plasmid Stability in Immobilized and Free Recombinant Escherichia Coli Jm105 (Pkk223-200): Importance of Oxygen Diffusion, Growth Rate, and Plasmid Copy Number. Applied and Environmental Microbiology, 53(7), 1548-1555.
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 & Obesity, 3(1).
Falbe J. (2012). Surfactants in Consumer Products: Theory, Technology and Application: Springer Science & Business Media.
Giollo M, Martin AJ, Walsh I, Ferrari C, and Tosatto SC. (2014). Neemo: A Method Using Residue Interaction Networks to Improve Prediction of Protein Stability Upon Mutation. BMC Genomics, 15(4), 1.
Granström TB, Takata G, Tokuda M, and Izumori K. (2004). Izumoring: A Novel and Complete Strategy for Bioproduction of Rare Sugars. Journal of Bioscience and Bioengineering, 97(2), 89-94.
GrunbergManago M. (1999). Messenger Rna Stability and Its Role in Control of Gene Expression in Bacteria and Phages. Annual Review of Genetics, 33(1), 193-227.
Guerois R, Nielsen JE, and Serrano L. (2002). Predicting Changes in the Stability of Proteins and Protein Complexes: A Study of More Than 1000 Mutations. Journal of Molecular Biology, 320(2), 369-387.
Gumina G, Song G, and Chu CK. (2001). L-Nucleosides as Chemotherapeutic Agents. FEMS Microbiology Letters, 202(1), 9-15.
Hanahan D. (1983). Studies on Transformation of Escherichia Coli with Plasmids. Journal of Molecular Biology, 166(4), 557-580.
Harlow E, and Lane D. (1988). A Laboratory Manual. Cold Spring Harbor Laboratory, 579.
Helanto M, Kiviharju K, Granström T, Leisola M, and Nyyssölä A. (2009). Biotechnological Production of L-Ribose from L-Arabinose. Applied Microbiology and Biotechnology, 83(1), 77-83.
Helanto M, Kiviharju K, Leisola M, and Nyyssölä A. (2007). Metabolic Engineering of Lactobacillus Plantarum for Production of L-Ribulose. Applied and Environmental Microbiology, 73(21), 7083-7091.
Hu C, Li L, Zheng Y, Rui L, and Hu C. (2011). Perspectives of Biotechnological Production of L-Ribose and Its Purification. Applied Microbiology and Biotechnology, 92(3), 449-455.
Huang L, Gromiha MM, and Ho S. (2007). Iptree-Stab: Interpretable Decision Tree Based Method for Predicting Protein Stability Changes Upon Mutations. Bioinformatics, 23(10), 1292-1293.
Hung XG, Yu MY, Chen YC, and Fang TY. (2015). Characterization of a Recombinant L-Ribose Isomerase from" Geodermatophilus Obscurus" Dsm 43160 and Application of This Enzyme to the Production of L-Ribose from L-Arabinose. Journal of Marine Science and Technology, 23(4), 558-566.
Jung ES, Kim HJ, and Oh DK. (2005). Tagatose Production by Immobilized Recombinant Escherichia Coli Cells Containing Geobacillus Stearothermophilusl‐Arabinose Isomerase Mutant in a Packed‐Bed Bioreactor. Biotechnology Progress, 21(4), 1335-1340.
Jung H, Pan J, Pyun Y, Cho E, Lee D, Cha Y, and Lee S. (2007). Characterization of a Novel D-Lyxose. J. Bacteriol, 189(5), 1655.
Karel SF, Libicki SB, and Robertson CR. (1985). The Immobilization of Whole Cells: Engineering Principles. Chemical Engineering Science, 40(8), 1321-1354.
Khan S, and Vihinen M. (2010). Performance of Protein Stability Predictors. Human Mutation, 31(6), 675-684.
Kido M, Yamanaka K, Mitani T, Niki H, Ogura T, and Hiraga S. (1996). Rnase E Polypeptides Lacking a Carboxyl-Terminal Half Suppress a Mukb Mutation in Escherichia Coli. Journal of Bacteriology, 178(13), 3917-3925.
Kim KR, Seo ES, and Oh DK. (2014). L-Ribose Production from L-Arabinose by Immobilized Recombinant Escherichia Coli Co-Expressing the L-Arabinose Isomerase and Mannose-6-Phosphate Isomerase Genes from Geobacillus Thermodenitrificans. Applied Biochemistry and Biotechnology, 172(1), 275-288.
Klibanov AM. (1983). Immobilized Enzymes and Cells as Practical Catalysts. Science, 219(4585), 722-727.
Kylmä A, Granström T, and Leisola M. (2004). Growth Characteristics and Oxidative Capacity of Acetobacter Aceti Ifo 3281: Implications for L-Ribulose Production. Applied Microbiology and Biotechnology, 63(5), 584-591.
Laidler KJ. (1987). Chemical Kinetics.
Laimer J, HieblFlach J, Lengauer D, and Lackner P. (2016). Maestroweb: A Web Server for Structure Based Protein Stability Prediction. Bioinformatics, btv769.
Laimer J, Hofer H, Fritz M, Wegenkittl S, and Lackner P. (2015). Maestro-Multi Agent Stability Prediction Upon Point Mutations. BMC Bioinformatics, 16(1), 1.
Lee KY, and Mooney DJ. (2012). Alginate: Properties and Biomedical Applications. Progress in Polymer Science, 37(1), 106-126.
Levin GV. (2002). Tagatose, the New Gras Sweetener and Health Product. Journal of Medicinal Food, 5(1), 23-36.
Lim Y, and Oh D. (2011). Microbial Metabolism and Biotechnological Production of D-Allose. Applied Microbiology and Biotechnology, 91(2), 229-235.
Longo MA, and Combes D. (1999). Thermostability of Modified Enzymes: A Detailed Study. Journal of Chemical Technology and Biotechnology, 74(1), 25-32.
Lopez PJ, Marchand I, Joyce SA, and Dreyfus M. (1999). The C‐Terminal Half of Rnase E, Which Organizes the Escherichia Coli Degradosome, Participates in Mrna Degradation but Not Rrna Processing in Vivo. Molecular Microbiology, 33(1), 188-199.
Luedemann GM. (1968). Geodermatophilus, a New Genus of the Dermatophilaceae (Actinomycetales). Journal of Bacteriology, 96(5), 1848-1858.
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-Clearcoli (Tm) Technology. In): Nature Publishing Group MACMILLAN BUILDING, 4 CRINAN ST, LONDON N1 9XW, ENGLAND.
Masso M, and Vaisman II. (2008). Accurate Prediction of Stability Changes in Protein Mutants by Combining Machine Learning with Structure Based Computational Mutagenesis. Bioinformatics, 24(18), 2002-2009.
Mateus D, Alves S, and Da Fonseca M. (1999). Diffusion in Cell‐Free and Cell Immobilising Κ‐Carrageenan Gel Beads with and without Chemical Reaction. Biotechnology and Bioengineering, 63(5), 625-631.
Mathé C, and Gosselin G. (2006). L-Nucleoside Enantiomers as Antivirals Drugs: A Mini-Review. Antiviral Research, 71(2), 276-281.
Michaelis L, and Menten ML. (1913). Die Kinetik Der Invertinwirkung. Biochem. z, 49(333-369), 352.
Mizanur RM, Takata G, and Izumori K. (2001). Cloning and Characterization of a Novel Gene Encoding L-Ribose Isomerase from Acinetobacter Sp. Strain Dl-28 in Escherichia Coli. Biochimica et Biophysica Acta (BBA)-Gene Structure and Expression, 1521(1), 141-145.
Moran EJ, Tellew JE, Zhao Z, and Armstrong RW. (1993). Dehydroamino Acid Derivatives from D-Arabinose and L-Serine: Synthesis of Models for the Azinomycin Antitumor Antibiotics. The Journal of Organic Chemistry, 58(27), 7848-7859.
Morimoto K, Terami Y, Maeda Y, Yoshihara A, Takata G, and Izumori K. (2013). Cloning and Characterization of the L-Ribose Isomerase Gene from Cellulomonas Parahominis Mb426. Journal of Bioscience and Bioengineering, 115(4), 377-381.
Moyroud E, and Strazewski P. (1999). L-Ribonucleosides from L-Xylose. Tetrahedron, 55(5), 1277-1284.
Neumann E, Schaefer Ridder M, Wang Y, and Hofschneider P. (1982). Gene Transfer into Mouse Lyoma Cells by Electroporation in High Electric Fields. The EMBO Journal, 1(7), 841.
Oh D. (2007). Tagatose: Properties, Applications, and Biotechnological Processes. Applied Microbiology and Biotechnology, 76(1), 1-8.
Okano K. (2009). Synthesis and Pharmaceutical Application of L-Ribose. Tetrahedron, 65(10), 1937-1949.
Parthiban V, Gromiha MM, and Schomburg D. (2006). Cupsat: Prediction of Protein Stability Upon Point Mutations. Nucleic Acids Research, 34(suppl 2), W239-W242.
Pastinen O, Visuri K, Schoemaker HE, and Leisola M. (1999). Novel Reactions of Xylose Isomerase from Streptomyces Rubiginosus. Enzyme and Microbial Technology, 25(8), 695-700.
Patel SN, Sharma M, Lata K, Singh U, Kumar V, Sangwan RS, and Singh SP. (2016). Improved Operational Stability of D-Psicose 3-Epimerase by a Novel Protein Engineering Strategy, and D-Psicose Production from Fruit and Vegetable Residues. Bioresource Technology, 216, 121-127.
Perali RS, Mandava S, and Bandi R. (2011). A Convenient Synthesis of L-Ribose from D-Fructose. Tetrahedron, 67(22), 4031-4035.
Pires DE, Ascher DB, and Blundell TL. (2014). Mcsm: Predicting the Effects of Mutations in Proteins Using Graph-Based Signatures. Bioinformatics, 30(3), 335-342.
Pitsch S. (1997). An Efficient Synthesis of Enantiomeric Ribonucleic Acids from D‐Glucose. Helvetica Chimica Acta, 80(8), 2286-2314.
Prabhu P, Tiwari MK, Jeya M, Gunasekaran P, Kim I, and Lee J. (2008). Cloning and Characterization of a Novel L-Arabinose Isomerase from Bacillus Licheniformis. Applied Microbiology and Biotechnology, 81(2), 283-290.
Schindelin J, Rueden CT, Hiner MC, and Eliceiri KW. (2015). The Imagej Ecosystem: An Open Platform for Biomedical Image Analysis. Molecular Reproduction and Development, 82(7-8), 518-529.
Schleif R. (2000). Regulation of the L-Arabinose Operon of Escherichia Coli. Trends in Genetics, 16(12), 559-565.
Schlieker M, and Vorlop K. (2006). A Novel Immobilization Method for Entrapment: Lentikats®. Immobilization of Enzymes and Cells, 333-343.
Shi Z, Yang B, and Wu Y. (2001). A Stereospecific Synthesis of L-Ribose and L-Ribosides from D-Galactose. Tetrahedron Letters, 42(43), 7651-7653.
Shimonishi T, and Izumori K. (1996). A New Enzyme, L-Ribose Isomerase from Acinetobacter Sp. Strain Dl-28. Journal of Fermentation and Bioengineering, 81(6), 493-497.
Smidsrød O, and Skja G. (1990). Alginate as Immobilization Matrix for Cells. Trends in Biotechnology, 8, 71-78.
SPSS I. (2011). Ibm Spss Statistics for Windows, Version 20.0. New York: IBM Corp.
Takahashi H, Iwai Y, Hitomi Y, and Ikegami S. (2002). Novel Synthesis of L-Ribose from D-Mannono-1, 4-Lactone. Organic Letters, 4(14), 2401-2403.
Teng S, Srivastava AK, and Wang L. (2010). Sequence Feature-Based Prediction of Protein Stability Changes Upon Amino Acid Substitutions. BMC Genomics, 11(2), 1.
Terami Y, Yoshida H, Uechi K, Morimoto K, Takata G, and Kamitori S. (2015). Essentiality of Tetramer Formation of Cellulomonas Parahominis L-Ribose Isomerase Involved in Novel L-Ribose Metabolic Pathway. Applied Microbiology and Biotechnology, 99(15), 6303-6313.
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(2), 376-378.
Woodyer RD, Wymer NJ, Racine FM, Khan SN, and Saha BC. (2008). Efficient Production of L-Ribose with a Recombinant Escherichia Coli Biocatalyst. Applied and Environmental Microbiology, 74(10), 2967-2975.
Worth CL, Preissner R, and Blundell TL. (2011). Sdm—a Server for Predicting Effects of Mutations on Protein Stability and Malfunction. Nucleic Acids Research, 39(suppl 2), W215-W222.
Wulff G, and Hansen A. (1987). Synthesis of Monosaccharides with the Aid of a New Synthetic Equivalent for the Glycolaldehyde Anion. Carbohydrate Research, 164, 123-140.
Yamaguchi M, and Mukaiyama T. (1981). The Stereoselective Synthesis of D-and L-Ribose. Chemistry Letters, 10(7), 1005-1008.
Yeom S, Ji J, Kim N, Park C, and Oh D. (2009). Substrate Specificity of a Mannose-6-Phosphate Isomerase from Bacillus Subtilis and Its Application in the Production of L-Ribose. Applied and Environmental Microbiology, 75(14), 4705-4710.
Yeom S, Ji J, Yoon R, and Oh D. (2008). L-Ribulose Production from L-Arabinose by an L-Arabinose Isomerase Mutant from Geobacillus Thermodenitrificans. Biotechnology Letters, 30(10), 1789-1793.
Yeom S, Kim N, Park C, and Oh D. (2009). L-Ribose Production from L-Arabinose by Using Purified L-Arabinose Isomerase and Mannose-6-Phosphate Isomerase from Geobacillus Thermodenitrificans. Applied and Environmental Microbiology, 75(21), 6941-6943.
Yeom SJ, Seo ES, Kim BN, Kim YS, and Oh DK. (2011). Characterization of a Mannose-6-Phosphate Isomerase from Thermus Thermophilus and Increased L-Ribose Production by Its R142n Mutant. Applied and Environmental Microbiology, 77(3), 762-767.
Yi H, Schumann P, and Chun J. (2007). Demequina Aestuarii Gen. Nov., Sp. Nov., a Novel Actinomycete of the Suborder Micrococcineae, and Reclassification of Cellulomonas Fermentans Bagnara Et Al. 1985 as Actinotalea Fermentans Gen. Nov., Comb. Nov. International Journal of Systematic and Evolutionary Microbiology, 57(1), 151-156.
Yin S, Ding F, and Dokholyan NV. (2007). Eris: An Automated Estimator of Protein Stability. Nature Methods, 4(6), 466-467.
Yoshida H, Yoshihara A, Teraoka M, Terami Y, Takata G, Izumori K, and Kamitori S. (2014). X‐Ray Structure of a Novel L‐Ribose Isomerase Acting on a Non‐Natural Sugar L‐Ribose as Its Ideal Substrate. FEBS Journal, 281(14), 3150-3164.
Young CL, Britton ZT, and Robinson AS. (2012). Recombinant Protein Expression and Purification: A Comprehensive Review of Affinity Tags and Microbial Applications. Biotechnology Journal, 7(5), 620-634.
Zajkoska P, Rebroš M, and Rosenberg M. (2013). Biocatalysis with Immobilized Escherichia Coli. Applied Microbiology and Biotechnology, 97(4), 1441-1455.
Zemek J, Bílik V, and Zákutná L. (1975). Effect of Some Aldoses on Growth of Saccharomyces Cerevisiae Inhibited with Molybdenum. Folia Microbiologica, 20(6), 467-469.
Zhang L, Jiang B, Mu W, and Zhang T. (2009). Bioproduction of D-Psicose Using Permeabilized Cells of Newly Isolated Rhodobacter Sphaeroides Sk011. Frontiers of Chemical Engineering in China, 3(4), 393-398.
Zhang YW, Prabhu P, and Lee JK. (2009). Immobilization of Bacillus Licheniformis L-Arabinose Isomerase for Semi-Continuous L-Ribulose Production. Bioscience, Biotechnology, and Biochemistry, 73(10), 2234-2239.
Zhou H, and Zhou Y. (2002). Distance‐Scaled, Finite Ideal‐Gas Reference State Improves Structure‐Derived Potentials of Mean Force for Structure Selection and Stability Prediction. Protein Science, 11(11), 2714-2726.