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論文名稱: | 探討 Aeromonas salmonicida MAEF108所產 Agarase AS-I 之基因轉殖與特性分析
| Studies on Gene Cloning and Characterization of Aeromonas salmonicida MAEF108 Agarase AS-I
研究生: | 馮家輝
指導教授: | 潘崇良博士 Chorng-Liang Pan, Ph.D.
學位類別: | 碩士
學校名稱: | 國立臺灣海洋大學
系所名稱: | 食品科學系
學號: | M95320042
畢業學年度: | 96
語文別: | 中文
關鍵字: | 洋菜酶 agarase
| 基因轉殖 gene cloning
| 無限制酶轉殖 restriction free cloning
全文影像連結: | | (本論文20110729對校內公開)
| 論文全文(pdf)

論文頁數: | 123
摘要: | 本研究之目的為分離純化 Aeromonas salmonicida MAEF108 所產 Agarase AS-I，探討其生化特性和分解產物；並先後嘗試從 MAEF108 的 基因體 (genomic) DNA 中選殖三個洋菜酶基因 (Aga-2769、Aga-2379、和 Aga-2400) 至大腸桿菌 (E. coli DH5) 表現。A. salmonicida MAEF108 的粗酵素液經過離心、超過濾、陰離子交換層析法和膠體過濾層析法得到純化的 Agarase AS-I，其純化倍率為53，分子質量為 86 kDa。Agarase AS-I 水解 0.2% 洋菜糖 (agarose) 的最適作用溫度及 pH 值分別為 45oC 和 6.0。對比於其他已測試的金屬離子，Mn2+ 顯示最高的相對活性，但 Fe3+ 和 Sn2+ 會使 Agarase AS-I 完全失活。Agarase AS-I 對 agarose 的活化能為 25.46 J K-1 mol-1 (6.09 cal K-1 mol-1)。以不同濃度的 agarose 做為基質測定 Agarase AS-I 動力學參數時，可得到酵素與基質的親和力 (Km) 為 0.49 mg/mL。Agarase AS-I 能降解紅藻類中的熱水萃取多醣如 (石花菜和龍鬚菜) 以及市售實驗試藥洋菜 (agar)，並可水解洋菜醣 (agarose) 產生以新洋菜六醣 (neoagarohexaose, DP6) 和新洋菜四醣 (neoagarotetraose, DP4) 為主要組成物的寡醣產物。但 Agarase AS-I 並不能將半乳醣 (galactose)合成其他醣類。另一方面，以無限制酶轉殖技術 [restriction-free (RF) cloning] 進行基因轉殖實驗時，洋菜酶基因Aga-2769、Aga-2379、和 Aga-2400 分別與載體 pET21b 經過三次聚合酶連鎖反應 (polymerase chain reaction, PCR) 的成功擴增後，再利用DpnI 剪切處理及嘗試轉殖至 E. coli DH5中，但均未觀察到轉殖株的出現。之後分別調整引子 (primer) 的黏合溫度 (annealing temperature) 和鎂離子 (Mg2+) 濃度，亦未發現任何轉殖株的生長。再以較傳統的限制酶剪接法進行基因轉殖，洋菜酶基因 Aga-2769 和 Aga-2379 與 載體 pET21b 先經過 NdeI 和 SacI 剪切，再利用 DNA 接合酶 (T4 ligase) 結合後轉殖至 E. coli DH5 中，亦尚未觀察到轉殖株。
| The aim of this study is to separate and purify Agarase AS-I, which produced by Aeromonas salmonicida MAEF108, and its biochemical properties and digested oligomeric products from algal polysaccharides were also being investigated.  Three agarase genes (Aga-2769, Aga-2379, and Aga-2400) were picked up from the genomic of strain MAEF108 and trials have been done for cloning them into E. coli DH5 host cell for expression of agarase activity.  A. salmonicida MAEF108 crude agarase solution was centrifuged and ultrafiltration, then applied to anion-exchange and gel premeation chromatography, to obtain Agarase AS-I which was 53 fold and estimated molecular mass as a 86 kDa.  The optimal temperature and pH to hydrolytic 0.2% agarose were 45oC and 6.0, respectively.  Mn2+ showed the highest relative activity a comparison other metal ions, Fe3+ and Sn2+ could make Agarase AS-I inactivity.  Activation energy and Km of Agarase AS-I were 25.46 J K-1 mol-1 (6.09 cal K-1 mol-1) and 0.49 mg/mL of hydrolytic agarose, respectively.  Agarase AS-I could degrade Rhodphyta extracted polysacchariedses (Gelidium sp. and Gracilaria sp.) and commercial agar.  While agarose was degraded by Agarase AS-I, hydrolysed products include mainly neoagarohexaose (DP6) and neoagarotetraose (DP4).  In addition, Agarase AS-I showed no effect to biosynthesize any product from galactose.  In restriction-free (RF) cloning experiment, agarase gene of Aga-2769, Aga-2379, and Aga-2400 used polymerase chain reaction (PCR) with pET21b, and then utilized DpnI digestion and transformation in E. coli DH5, none of transformant found in medium.  More trials have been done on adjust annealing temperature and concentration of magnesium ion (Mg2+), but there is no transformant observed in medium.  In tradition of restriction splicing experiment, at first, agarase gene of Aga-2769 and Aga-2379 as well as pET21b utilized NdeI and SacI digestion, and then used T4 ligase ligation and transformation in E. coli DH5, the result as same as RF cloning, none of transformant found on ampicillin selective agar plate.
論文目次: | 目錄    i
| 表目錄    v
| 圖目錄    vi
| 附錄目錄    ix
| 中文摘要    x
| 英文摘要 (Abstract)      xii
| 壹、前言    1
| 貳、文獻整理    3
| 1. 洋菜的構造與特性    3
| 2. 洋菜酶    3
| 2.1 洋菜酶生產菌株及其分離來源    4
| 2.2 洋菜酶之生化特性    4
| 2.2.1 溫度和pH值    4
| 2.2.2 分子質量    5
| 2.2.3 金屬離子    6
| 3. 海藻之來源及應用    7
| 3.1 石花菜    7
| 3.2 馬尾藻    8
| 3.3 海菜    8
| 4. 海藻多醣    9
| 5. 洋菜酶分解海藻多醣之產物    10
| 5.1 生理活性    10
| 6. 海藻寡醣組成成份之鑑定    12
| 6.1 薄層層析法 (thin-layer chromatography, TLC)    12
| 6.2 高效能液相層析法 (high-performance liquid
| chromatography, HPLC)    13
| 6.3 磁核共振光譜法 (nuclear magnetic resonance, NMR)    13
| 7. 洋菜酶轉殖    14
| 7.1 利用限制酶 (restriction enzyme, RE) 剪切進行基因轉殖    14
| 7.2 無限制酶剪切位置之基因轉殖 (restriction free cloning)      15
| 參、實驗設計    16
| 肆、實驗材料    17
| 1. 實驗菌株與質體    17
| 1.1 產洋菜酶菌株    17
| 1.2 表現洋菜酶基因之質體    17
| 1.3 表現洋菜酶基因之勝任細胞    17
| 2. 海藻多醣    18
| 3. 培養基    18
| 4. 試驗藥品    18
| 5. 儀器設備    21
| 6. 套組 (kit)      23
| 伍、實驗方法    24
| 1. Agarase AS-I之純化分離    24
| 1.1 MAEF108 粗酵素液之製備    24
| 1.1.1 洋菜酶活性測定    24
| 1.1.2 蛋白質的定量    24
| 1.2 MAEF108-agarases 之濃縮    25
| 1.3 Agarase AS-I 之分離    25
| 1.3.1 DE-52 離子交換樹脂層析    25
| 1.3.2 SephacrylR S-200 膠體過濾層析    25
| 1.4 Agarase AS-I 之 N 端胺基酸定序    26
| 1.5 Agarase AS-I之生化性質測試    26
| 1.5.1 分子質量    26
| 1.5.2 最適作用溫度    26
| 1.5.3 最適作用pH值    27
| 1.5.4 金屬離子的影響    27
| 1.5.5 基質特異性    27
| 1.5.6 活化能的決定    27
| 1.5.7 酵素動力學    28
| 1.6 Agarase AS-I 分解和合成產物之探討    28
| 1.6.1 薄層層析法 (TLC)    28
| 1.6.2 高效能液相層析法 (HPLC)    29
| 2. MAEF108 agarase 的基因選殖    29
| 2.1 MAEF108 genomic DNA 之萃取純化及定序    29
| 2.2 質體製備    30
| 2.2.1 DNA 電泳分析    30
| 2.3 Agarase 基因序列比對    31
| 2.4 引子 (primer) 設計    31
| 2.5 洋菜酶轉殖    32
| 2.5.1 以 RF cloning 進行基因轉殖    31
| 2.5.1-A 擴增 agarases 基因    31
| 2.5.1-B 以限制酶 Dpn I 剪切 RF cloning 後的產物    34
| 2.5.2 以限制酶 (RE) 剪切進行基因轉殖    35
| 2.5.2-A 擴增 agarases 基因    35
| 2.5.2-B 雙股 DNA 之限制酶剪切反應 (digestion)    35
| 2.5.2-C 雙股 DNA 之黏合反應 (ligation)    36
| 2.6 將agarases基因轉形至大腸桿菌    36
| 2.6.1 以 E. coli DH5 製做勝任細胞 (competent cell)    36
| 2.6.2 以熱休克 (heat shock) 進行轉形作用 (transformation)
| 36
| 3. 統計分析    37
| 陸、結果與討論    38
| 柒、結論    56
| 捌、參考文獻    58
|
參考文獻: | 田勝朗。1990。作用於心臟血管系統之水産食品成分。食品工業。30 (3): 29-34。
| 江仁淳。2001。臺灣重緣葉馬尾藻海藻多醣體之免疫調節功能評估。國立臺灣海洋大學食品科學系碩士學位論文，基隆。臺灣。
| 江永棉、王瑋龍、黃椒芳。1990。臺灣海藻簡介。臺灣省立博物館。臺北。臺灣。
| 李健瑋。2000。高溫洋菜分解酵素生產菌株之菌種鑑定及洋菜分解酵素之純化。私立東吳大學微生物學系碩士學位論文，臺北。臺灣。
| 邱雅凰。2007。海藻萃取物抑制日本腦炎病毒感染及免疫功能調節之研究。國立臺灣海洋大學食品科學系碩士學位論文，基隆。臺灣。
| 吳紹祺。1999。海洋細菌所産 agarases 生產條件與海藻多醣經 agarases 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。
| 吳紹祺。2005。海藻多醣洋菜酶水解物及其發酵產物之生理活性研究。國立臺灣海洋大學食品科學系博士學位論文，基隆。臺灣。
| 吳欣蕙。2007。Pseudomonas vesicularis MA103 菌株所產 agarases PV-1 與 PV-2 之純化與性質探討。國立臺灣海洋大學食品科學系碩士學位論文，基隆。臺灣。
| 施珊汶。2006。海帶多醣水解液乳酸發酵產物之生理活性探討。國立臺灣海洋大學食品科學系碩士學位論文，基隆。臺灣。
| 孫乃云。2006。石花菜 (Gelidium)水萃取液抗氧化及抗癌活性之探討。國立臺灣海洋大學水產養殖學系碩士學位論文，基隆。臺灣。
| 馬盈瑜。2003。龍鬚菜多醣及其水解物生理活性之探討。國立屏東科技大學食品科學系碩士學位論文，屏東。臺灣。
| 張玉瓏、除乃芝、許素菁。2005。生物技術。新文京開發。臺中。臺灣。
| 康新楷。2007。Aeromonas salmonicida MAEF108所產MAEF108 Agarase II 之純化與特性研究暨水解二種藻類多醣萃取所得寡醣之生理活性探討。國立臺灣海洋大學食品科學系碩士學位論文。基隆。臺灣。
| 陳姿元。2006。Aeromonas salmonicida MAEF108 菌株所產洋菜酶的純化與性質之探討。國立臺灣海洋大學食品科學系碩士學位論文，基隆。臺灣。
| 傅郁文。2005。飼料中添加龍鬚菜與石花菜熱水萃取物增強白蝦免疫力及抗Vibrio alginolyticus 感染之研究。國立臺灣海洋大學水產養殖學系碩士學位論文。基隆。臺灣。
| 黃淑芳。2002。妙用無窮的海藻。科學發展。360: 8-13。
| 黃霨禎。2005。龍鬚菜寡醣水解物之製飾及其抗氧化與抗致突變性之探討與對雙叉桿菌生長影響。國立臺灣海洋大學食品科學系碩士學位論文。基隆。臺灣。
| 黃穰、廖婉茹。2003。海藻，來自海洋的保健藥草。科學發展。364: 30-39。
| 德永隆久、日高秀昌。1991。寒天オリグ糖の生產と利用。バイオサイエンスとインダストリ。49: 28-32。
| 魏岑芸。2007。増加基質結合部位殘基與基質間氫鍵對於 Sulfolobus solfataricus ATCC33092 麥芽寡糖苷海藻糖生成酶轉糖苷及水解作用之影響。國立臺灣海洋大學食品科學系碩士學位論文，基隆。臺灣。
| 蕭銘瑞。2007。鹽橋對於 Brevibacterium helvolum ATCC 11822 麥芽寡糖苷海藻糖生成酶熱穩定性之影響。國立臺灣海洋大學食品科學系碩士學位論文，基隆。臺灣。
| Allouch, J., M. Jam, W. Helbert, T. Barbeyron, B. Kloareg, B. Henrissat, and M. Czjzek.  2003.  The three-dimensional structures of two β-agarases.  Journal of Biological Chemistry  278: 47171-47180.
| Aoki, T., T. Araki, and M. Kitamikado.  1990.  Purification and characterization of -agarase from Vibrio sp. AP-2.  Nippon Suisan Gakkaishi  56:  825-830.
| Araki, T., M. Hayakawa, Z. Lu, S. Karita, and T. Morishita.  1998. Purification and characterization of agarases from a marine bacterium, Vibrio sp. PO-303. Journal of Bioscience and Bioengineering  6: 260-265.
| Barsanti, L., and P. Gualtieri.  2006.  Algae: Anatomy, Biochemistry, and Biotechnology.  Taylor and Francis, Boca Raton, FL, USA  pp. 1-2.
| Belas, R., D. Barlent, and M. Silverman.  1988.  Cloning and gene replacement mutagensis of a Pseudomonas atlantica agarase gene.  Applied and Environmental Microbiology  54: 30-37.
| Bradford, M. M.  1976.  A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding.  Biochemistry  72: 248-254.
| Chevolot, L., A. Foucault, F. Chaubet, N, Kervarec, C. Sinquin, A. M. Fisher, and C. Boisson-Vidal.  1999.  Further data on the structure of brown seaweed fucans: Relationships with anticoagulant activity.  Carbohydrate Research  319: 154-165.
| Colak, A. and S. Guner.  2004.  Polyhydroxyalkanoate degrading hydrolase-like activities by Pseudomonas sp. isolated from soil.  International Biodeterioration and Biodegradation  53:  103-109.
| Dong, J., S. Hashikawa, T. Konishi, Y. Tamaru, and T. Araki.  2006.  Cloning of the novel gene encoding -agarase C from a marine bacterium, Vibrio sp. strain PO-303, and characterization of the gene product.  Applied and Environmental Microbiology  72: 6399-6401.
| Dong, J., Y. Tamaru, and T. Araki.  2007.  Molecular cloning, expression, and characterization of a -agarase gene, agaD, from a marine bacterium, Vibrio sp. strain PO-303.  Bioscience, Biotechnology, and Biochemistry  71: 38-46.
| Durate, M. E. R., M. A. Cardoso, M. Noseda, and A. S. Cerezo.  2001.  Structural studies on fucoidans from the brown seaweed Sargassum stenophyllum.  Carbohydrate Research  333: 281-293.
| Enzyme Nomenclature Database.  2008.  Agarase. http://www.expasy.org/cgi-bin/enzyme-search-de  Switzerland. (July 6, 2008)
| Errea, M. I. and M. C. Matulewicz.  2003.  Unusual structures in the polysaccharides from the red seaweed Pterocladiella capillacea (Gelidiacead, Gelidiales).  Carbohydrate Research  338:  943-953.
| Flament, D., T. Barbeyron, M. Jam, P. Potin, M. Czjzek, B. Kloareg, and G. Michel.  2007.  Alpha-agarases define a new family of glycoside hydrolases, distinct from beta-agarase families.  Applied and Environmental Microbiology  73: 4691-4694.
| Freile-Pelegrín, Y., D. R. Robledo, and G. García-Reina.  1995.  Seasonal agar yield and quality in Gelidium canariensis (Grunow) seoane-camba (Gelidiales, Rhodophyta) from Gran Canaria, Spain.  Journal of Applied Phycology  7: 141-144.
| Fu, X. T., H. Lin, and S. M. Kim.  2007a.  Purification and characterization of novel -agarase, AgaA34, from Agarivorans albus YKW-34.  Applied Microbiology and Biotechnology  78: 265-273.
| Fu, Y. W., W. Y. Hou, S. T. Yeh, C. H. Li, and J. C. Chen.  2007b.  The immunostimulatory effects of hot-water extract of Gelidium amansii via immersion, injection and dietary administrations on white shrimp Litopenaeus vannamei and its resistance against Vibrio alginolyticus.  Fish and Shellfish Immunology  22: 673-685.
| Ha, J. C., G. T. Kim, S. K. Kim, T. K. Oh, J. H. Yu, and I. S. Kong.  1997.  β-Agarase from Pseudomonas sp. W7: Purification of the recombinant enzyme from Escherichia coli and the effects of salt on its activity.  Applied Biochemistry and Biotechnology  26: 1-6.
| Hassairi, I., R. B. Amar, M. Nonus, and B. B. Gupta.  2001.  Production and separation of α-agarase from Alteromonas agarlyticus strain GJ1B.  Bioresource Technology  79: 47-51.
| Hatada, Y., Y. Otha, and K. Horikoshi.  2006.  Hyperproduction and application of a-agarase to enzymatic enhancement of antioxidant activity of porphyran.  Agricultural and Food Chemistry  54: 9895-9900.
| Hodgson, D. A., and K. F. Chater.  1981.  A chromosomal locus controlling extracellular agarase production by Streptomyces coelicolor A3(2), and its inactivation by chromosomal integration of plasmid SCP1.  Journal of General Microbiology  124: 339-348.
| Hosoda, A., and M. Sakai.  2006.  Isolation of Asticcacaulis sp. SA7, a novel agar-degrading alphaproteobacterium.  Bioscience, Biotechnology, and Biochemistry  70: 722-725.
| Hosoda, A., M. Sakai, and S. Kanazawa.  2003.  Isolation and characterization of agar-degrading Paenigacillus sp. associated with the rhizosphere of Spinach.  Bioscience, Biotechnology, and Biochemistry  67: 1048-1055.
| Hu, B., Q. Gong, Y. Wang, Y. Ma, J. Li, and W. Yu.  2006.  Prebiotic effects of neoagaro-oligosaccharides prepared by enzymatic hydrolysis of agarose.  Anaerobe  12: 260-266.
| Huang, X., H. Zhou, and H. Zhang.  2006.  The effect of Sargassum fusiforme polysaccharide extracts on vibriosis resistance and immune activity of the shrimp, Fenneropenaeus chinensis.  Fish and Shellfish Immunology  20: 750-757.
| Jam, M., D. Flament, J. Allouch, P. Potin, L. Thion, B. Kloageg, M. Czjzek, W. Helbert, G. Michel, and T. Barbeyron.  2005.  The endo--agarases AgaA and AgaB from the marine bacterium Zobellia galatanivorans: Two paralogue enzymes with different molecular organizations and catalytic behaviours.  The Biochermical Journal  385: 703-713.
| Jiménez-Escring, A., and F. J. Sánchez-Muniz.  2000.  Dietary fiber from edible seaweeds: Chemical structure, physicochemical properties and effects on cholesterol metabolism.  Nutrition Research  20: 585-598.
| Kang, N. Y., Y. L. Choi, Y. S. Cho, B. K. Kim, B. S. Jeon, J. Y. Cha, C. H. Kim, and Y. C. Lee.  2003.  Cloning, expression and characterization of a -agarase gene from a marine bacterium, Pseudomonas sp. SK 38.  Biotechnology Letters  25:  1165-1170.
| Kendall, K., and J. Cullum.  1984.  Cloning, and expression of an extracellular agarase from Streptomyces coelicolor A3(2) in Streptomyces lividans 66.  Gene  29: 315-321.
| Kim, B. J., H. J. Kim, S. D. Ha, S. H. Hwang, D. S. Byun, T. H. Lee, and J. Y. Kong.  1999.  Purification and characterization of -agarase from marine bacterium Bacillus cereus ASK202.  Biotechnology Letters  21: 1011-1015.
| Kirimura, K., N. Masuda, Y. Iwasaki, H. Nakagawa, R. Kobayashi, and S. Usami.  1999.  Purification and characterization of a novel -agarase from an alkalophilic bacterium, Alteromonas sp. E-1. Journal of Bioscience and Bioengineering  87: 436-441.
| Kobayashi, R., M. Takimasa, T. Suzuki, K. Kirimura, and S. Usami.  1997.  Neoagarobiose as a novel moisturizer with whitening effect.  Bioscience, Biotechnology and Biochemistry  61: 162-163.
| Kong, J. Y.  2001.  Production and functional properties of agarooligosaccharides.  Proceedings of Symposium on Scientific Study and Industrialization of Health Food, Health Food Society of Taiwan, Taipei.  pp. 60-81.
| Koyanagi, S., N. Tanigawa, H. Nakagawa, S. Soeda, and H. Shimeno.  2003.  Oversulfation of fucoidan enhances its anti-angiogenic and antitumor activities.  Biochemical Pharmacology  65: 173-179.
| Lee, D. G., G. T. Park, N. Y. Kim, E. J. Lee, M. K. Jang, Y. G. Shin, G. S. Park, T. M. Kim, J. H. Lee, J. H. Lee, S. J. Kim, and S. H. Lee.  2006.  Cloning, expression, and characterization of a glycoside hydrolase family 50 -agarase from a marine Agarivorans isolate.  Biotechnology Letters  28:  1925-1932.
| Lee, J. B., K. Hayashi, M. Maeda, and T. Hayashi.  2004.  Antiherpetic activities of sulfated polysaccharides from green algae.  Planta Medica  70:  813-817.
| Leon, O., L. Quintana, G. Peruzzp, and J. C. Slebe.  1992.  Purification and properties of an extracellular agarase from Alteromonas sp. strain C-1.  Applied and Environmental Microbiology  58: 4060-4063.
| Li, X., X. Dong, C. Zhao, Z. Chen, and F. Chen.  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.
| Li, J. B., F. Han, X. Z. Lu, X. Y. Fu, C. P. Ma, Y. Chu, and W. G. Yu.  2007.  A simple method of preparing diverse neoagaro-oligosaccharides with -agarase.  Carbohydrate Research  342: 1030-1033.
| Ludikhuyze, L. R., I. V. den Broeck, C. A. Weemaes, C. H. Herremans, J. F. I. Impe, M. E. Hendrickx, and P. P. Tobback.  1997.  Kinetics for isobaric-isothermal inactivation of Bacillus subtilis -amylase.  Biotechnology Progress  13:  532-538.
| Ma, C., X. Lu, C. Shi, J. Li, Y. Gu, Y. Ma, Y. Chu, F. Han, Q. Gong, and W. Yu.  2007.  Molecular cloning and characterization of a novel -agarase, agaB, from marine Pseudoalteromonas sp. CY24. Journal of Biological Chemistry  282: 3747-3754.
| Maeda, M., T. Uehara, N. Haoki, M. Sekiguchi, and A. Hiraoka.  1991.  Heparinoid-active sulfated polysaccharides from Monostroma nitidum and their distribution in the chlorophyta.  Phytochemistry  30:  3611-3616.
| Meena, R., K. Prasad, and A. K. Siddhanta.  2006.  Studies on “sugar-reactivity” of agar extracted from some Indian agarophytes. Food Hydrocolloids  20: 1206-1215.
| Merck KGaA.  1998  pET-21 a-d (+) Vector map.  http://www.emdbiosciences.com/docs/docs/PROT/TB036.pdf  (June 4, 2008, USA).
| Morrice, J. M., M. W. McLean, W. F. Long, and F. B. Williamson.  1983a.  Porphyran primary structure. An investigation using β-agarase I from Pseudomonas atlantica and 13C-NMR spectroscopy.  Europe Journal of Biochemistry  133: 673-684.
| Morrice, L. M., M. W. McLean, F. B. Williamson, and W. F. Long.  1983b.  β-agarase I and II from Pseudomonas atlantica. Purification and some properties.  Europe Journal of Biochemistry  135: 553-558.
| Noda, H., H. Amano, H. Arashima, S. Hashimoto, and K. Nisizawa.  1989.  Studies on the antitumour activity of marine algae.  Nippon Suisan Kogyo Gakkaishi  55: 1259-1264.
| Ohta, Y., Y. Nogi, M. Miyazaki, Z. Li, Y. Hayada, S. Ito, and K. Horikoshi.  2004.  Enzymatic properties and nucleotide and amino acid sequences of a thermostable β-agarase from the novel marine isolate, JAMB-A94.  Bioscience, Biotechnology, and Biochemistry  68: 1073-1081.
| Ohta, Y., Y. Hayada, M. Miyazaki, Z. Li, Y. Nogi, S. Ito, and K. Horikoshi.  2005.  Purification and characterization of a novel a -agarase from a Thalassomonas sp.  Current Microbiology  50: 212-216.
| Osumi, Y., M. Kawai, H. Amano, and H. Noda.  1998.  Effect of oligosaccharides from prophyran in vitro digestions, utilizations by various intestinal bacteria, and levels of serum lipids in mice.  Nippon Suisan Kogyo Gakkaishi  64: 98-104.
| Percial, E. G. V., and R. H. McDowell.  1967.  Chemistry and Enzymology of Marine Algae Polysaccharide.  Animal Press, London, U.K.  pp. 26-50.
| Potin, P., C. Richard, C. Rochas, and B. Kloareg.  1993.  Purification and characterization of the -agarase from Alteromonas agarlyticus (Cataldi) comb. nov., strain GJ1B.  Europe Journal of Biochemistry  214: 599-607.
| Sambrook, J., and D. W. Russell.  2006.  The Condensed Protocols from Molecular Cloning: A Laboratory Manual.  Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA.
| SAS.  1999.  SAS/GRAPH User’s Guide 6.03ed.  SAS Institute Inc., Cary, USA.
| Schroeder, D. C., M. A. Jaffer, and V. C. Coyne.  2003.  Investigation of the role of a β(1-4) agarase produced by Pseudoalteromonas gracilis B9 in eliciting disease symptoms in the red alga Gracilaria gracilis.  Microbiology  149: 2919-2929.
| Singh, T.  1992.  Agar and agar production.  INFOFISH, Luala Lumpur.  pp. 1-25.
| Sugano, Y., I. Terada, M. Arita, M. Noma, and T. Matsumoto.  1993a.  Purification and characterization of a new agarase from a marine bacterium, Vibrio sp. strain JT0107.  Applied and Environmental Microbiology  59: 1549-1554.
| Sugano, Y., T. Matsumoto, H. Kodama, and M. Noma.  1993b. Cloning and sequencing of agaA, a unique agarase 0107 gene from a marine bacterium, Vibrio sp. strain JT0107.  Applied and Environmental Microbiology  59: 3750-3756.
| Suzuki, H., Y. Sawai, T. Sawai, and K. Kawai.  2003.  Purification and characterization of an extracellular -agarases from Bacillus sp. MK03.  Journal of Bioscience and Bioengineering  95: 328-334.
| Swartz, M., and N. Gordon.  1959.  Agarase from an agar-digesting bacterium.  Journal of Bacteriology  77: 403-409.
| Usov, A. I.  1998.  Structural analysis of red seaweed galactans of agar and carrageenan groups.  Food Hydrocolloids  12: 301-308.
| van den Ent, F., and J. Lowe.  2006.  RF cloning: A restriction-free method for inserting target genes into plasmids.  The Journal of Biochemical and Biophysical Methods  67: 67-74.
| Vera, J., R. Alvarez, E. Murano, J. C. Slebe, and O. Leon.  1998.  Identification of a marine agarolytic Pseudoalteromonas isolate and characterization of its extracellular agarase.  Applied and Environmental Microbiology  64: 4378-4383.
| Wang, J., H. Mou, X. Jiang, and H. Guan.  2006.  Characterization of a novel -agarase from marine Alteromonas sp. SY37–12 and its degrading products.  Applied Microbiology and Biotechnology  71: 831-839.
| Wu, S. C., and C. L. Pan.  2004.  Preparation of algal-oligosaccharide mixtures by bacterial agarases and their antioxidative properties.  Fisheries Science  70: 1164-1173.
| Wu, S. C., T. N. Wen, and C. L. Pan.  2005.  Algal-oligosaccharide-lysates prepared by two bacterial agarases stepwise hydrolyzed and their anti-oxidative properties.  Fisheries Science  71: 1149-1159.
| Yamaura, I., T. Matsumoto, M. Funatsu, H. Shigeiri, and T. Shibata.  1991.  Purification and properties of agarase from Pseudomonas sp. PT-5.  Agricultural Biological Chemistry  55: 2531-2536.
| Yeh, S. T., C. S. Lee, and J. C. Chen.  2006.  Administration of hot-water extract of brown seaweed Sargassum duplicatum via immersion and injection enhances the immune resistance of white shrimp Litopenaeus vannamei.  Fish and Shellfish Immunology  20: 332-345.
| Yoshizawa, Y., A. Ametani, J. Tsunehiro, K. Nomura, M. Itoh, F. Fukui, and S. Kaminogawa.  1995.  Macrophage stimulation activity of the polysaccharide fraction of a marine alga (Porphyra yezoensis): Structure-function relationships and improved solubility. Bioscience, Biotechnology, and Biochemistry  59: 1933-1937.
| Yoshizawa, Y., J. Tsunehiro, and K. Nomura.  1996.  In vivo macrophage-stimulation activity of the enzyme-degraded water-soluble polysaccharide fraction from a marine alga (Gracilaria verrucosa).  Bioscience, Biotechnology, and Biochemistry  60: 1667-1671.
| Young, K. S., S. S. Battacharjee, and W. Yahpe.  1978.  Enzymic cleavage of the -linkages in agarose, to yield agaro-oligosaccharides.  Carbohydrate Research  66: 207-212.
| Zhang, W. W., and L. Sun.  2007  Cloning, characterization, and molecular application of a beta-agarase gene from Vibrio sp. strain V134.  Applied and Environmental Microbiology  73: 2825-2831.
| Zhu, W., L. C. M. Chiu, V. E. C. Ooi, P. K. S. Chan, and P. O. Ang Jr.  2004.  Antiviral property and mode of action of a sulphated polysaccharide from Sargassum patens against herpes simplex virus type 2.  International Journal of Antimicrobial Agents  24: 81-85.
|
| Zhu, W., L. C. M. Chiu, V. E. C. Ooi, P. K. S. Chan, and P. O. Ang Jr.  2006.  Antiviral property and mechanisms of a sulphated polysaccharide from the brown alga Sargassum patens against herpes simplex virus type 1.  Phytomedicine  13: 695-701.
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2010 年 3 月 8 日 22:56