:::

| | | | | 第 | | 筆

詳目顯示區塊

第 1 筆
| (以作者名查詢海洋大學圖書館館藏系統)
| (以作者名查詢全國圖書書目資訊網)
| (以作者名查詢全國博碩士系統)
論文名稱: | Pseudomonas vesicularis MA103 所產 Alginate lyase pal III 之基因轉殖至大腸桿菌中表現之探討
| Studies on Gene Cloning and Expression of Pseudomonas vesicularis MA103 Alginate Lyase palIII in Escherichia coli
研究生: | 林慧傑 Hui-Jei Lin
指導教授: | 潘崇良 Chorng-Liang Pan
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: | M97320035
畢業學年度: | 98
語文別: | 中文
關鍵字: | 褐藻膠裂解酶 pal III
| 無限制酶基因選殖法
全文影像連結: | | (本論文20130730對校內公開)
| 論文全文(pdf)

論文頁數: | 120
摘要: | 本研究探討 Pseudomonas vesicularis MA103 經誘導產生褐藻膠  裂解酶 (Alginate lyase) 的純化，及藉 N 端胺基酸定序結果比對，  確認該 Alginate lyase 之完整基因序列；由已知的菌株 MA103 基因 體序列中挑選一 Alginate lyase pal III 基因，以無限制酶基因選殖法 (Restriction-free, RF cloning) 轉殖至大腸桿菌中，並觀察其 Alginate lyase rpal III 之酵素活性。以含有 0.2% Sodium alginate 的 Marine broth (MB) 和 Modified marine broth 2-alginate lyase (MMB2-Al)    培養基進行誘導 培養，可有效提高菌株 MA103 crude alginate lyases 的活性，經活性染色和 SDS-PAGE 確認誘導培養可產生兩個 Alginate lyase，由膠片結果推估其分子量大小分別約為 57 和 38 kDa。其粗酵素液以 DE-52 離子交換樹脂層析初步純化，可分離出兩個具有 Alginate lyase 活性的蛋白質波峰，個別收集後命名為 Pv-Al-1 及 Pv-Al-2，其比活性分別為 1142.22 U/mg 和 531.76 U/mg。以 12% SDS-PAGE 分析後得知，Pv-Al-1 與 Pv-Al-2 中仍有部分雜蛋白，需進一步以膠體過濾層析管柱純化；另一方面，從已完成定序的MA103 genomic DNA 中得知有 4 個可能產生 Alginate lyase 的基因片段， 挑選一段核苷酸序列全長為 1041 bp 之 Alginate lyase pal III，可轉譯出 348 個胺基酸，使用 pET 表現系統以及 RF cloning 方法設計兩組引子對，並以 PCR 製得插入 Alginate lyase pal III 基因之重組載體 pET-21b (+) 的 PCR 擴充產物，核苷酸序列全長約為 6500 bp，該  重組載體經電穿孔法轉殖至 E. coli BL21 (DE3) 後，將其載體上之  插入序列定序，其定序結果顯示 rpal III 與 pal III 除了因設計方便 後續純化而剔除的 stop codon 之外，其餘的序列皆相同。經 Isopropyl -D-1-thiogalactopyranoside (IPTG, 0.5 mM) 於 20oC 下 誘導 3 小時後，胞內粗酵素液可得到最高酵素活性 (21.06 U/mg protein)，該胞內粗酵素液經由鎳親和性管柱層析處理後，得到純化之 Alginate lyase rpal III (487.50 U/mg protein)，此純化酵素液之 SDS-PAGE 結果顯示，其分子量約為 40.5 kDa，與 rpal III 基因序列所推估的大小相似；在基質特異性方面 Alginate lyase rpal III 對於 Alginic acid 具有較高的酵素活性 12.41 U/mL。此外，除了對重緣葉馬尾藻熱萃液約略具有酵素活性 (0.43 U/mL) 外，對於其他的藻類熱萃液則無顯著的酵素活性產生。從 TLC 的分析中得知，Alginate lyase rpal III 在 25oC 與 pH 9.0 中將 Alginate 水解 24小時後所得到的褐藻膠寡醣，與 Zhang et al. (2005) 所發表的文獻數據相比對，推測其水解產物的聚合度 (Degree of polymerization, DP) 可能為 DP4 或 DP5 的褐藻膠寡醣； 而以 MA103 crude alginate lyase 在相同條件下將Alginate水解 24 小時，觀察到四種不同的褐藻膠寡醣產生，其聚合度可能在 DP4-DP8 之間。而從 HPLC 的分析結果得知 Alginate lyase rpal III 與 MA103 crude alginate lyase 在水解 Alginate 24 小時後，分別可能得到兩種和四種褐藻膠寡醣水解產物。未來會進一步將該褐藻膠寡醣分離純化後，再以 NMR 鑑定其褐藻膠寡醣的結構。
論文目次: | 目錄
|
|
| 目錄    i
| 表目錄    vi
| 圖目錄    vii
| 附錄目錄    x
| 中文摘要    xi
| 英文摘要 (Abstract)    xiii
| 壹、前言    1
| 貳、文獻整理    3
| 一、褐藻膠的來源    3
| 二、褐藻膠的構造與特性    3
| 三、褐藻膠裂解酶生產菌株及其分離來源    4
| 四、褐藻膠裂解酶對褐藻膠裂解的作用機制    5
| 五、褐藻膠裂解酶之生化特性    5
| 5-1. 溫度與 pH    5
| 5-2. 分子質量    6
| 5-3. 金屬離子    7
| 六、褐藻膠的應用    7
| 6-1. 微膠囊化固定    7
| 6-2. 金屬吸附    7
| 6-3. 褐藻膠培養系統    8
| 6-4. 在農業與生物防治上之應用    8
| 6-4-1. 對植物根部生長與種子萌芽的影響    8
| 6-4-2. 抗菌作用    9
| 七、褐藻膠裂解酶基因的選殖    10
| 7-1. 大腸桿菌或酵母菌做為宿主之轉殖系統    10
| 7-2. 無限制酶切位基因選殖 (Restriction-free cloning)    10
| 八、海藻寡醣組成成分鑑定    11
| 8-1. 薄層層析法 (Thin-layer chromatography, TLC)    11
| 8-2. 高效能液相層析法 (High-performance liquid chromatography, HPLC)    11
| 8-3. 磁核共振光譜法 (Nuclear magnetic resonance, NMR)    12
| 參、實驗設計    13
| 肆、實驗材料與方法    14
| 一、實驗材料    14
| 1-1. 實驗菌株    14
| 1-1-1. 菌種保存    14
| 1-1-2. 菌株活化    14
| 1-2 試驗藥品    15
| 1-2-1. 藥品    15
| 1-2-2. 載體    18
| 1-2-3. 引子    18
| 1-2-4. 聚合酶鏈鎖反應試劑    19
| 1-2-5. 限制劑    19
| 1-2-6. DNA 及蛋白質萃取純化套裝試劑    19
| 1-2-7. 電泳標準品    19
| 1-2-8. 膠體過濾標準品    20
| 1-2-9. 培養基組成    20
| 1-2-10.      褐藻膠裂解酶反應基質    21
| 1-2-11. 硫代巴比妥安酸法試劑      21
| 1-2-11-1. 0.3% TBA    21
| 1-2-11-2. 0.25 N H5IO6-0.125 N H2SO4    21
| 1-2-11-3. 2% NaAsO2-0.5 N HCl    21
| 1-2-12. 膠體過濾緩衝液    21
| 1-2-13. 電泳膠片製備    22
| 1-2-14. 電泳溶液    22
| 1-2-15. 活性染色法試劑    23
| 1-2-16. 無菌過濾    23
| 1-2-17. 離心式超過濾濃縮    23
| 1-2-18. 薄層層析矽膠片    24
| 1-3. 儀器設備    24
| 二、實驗方法    26
| 2-1. Alginate lyase 的純化分離    26
| 2-1-1. 褐藻膠裂解酶的產生    26
| 2-1-2. 褐藻膠裂解酶活性測定    26
| 2-1-3. 褐藻膠裂解酶活性定義    26
| 2-1-4. 蛋白質定量    27
| 2-1-5. 分子量的鑑定    27
| 2-1-6. 活性染色    27
| 2-1-7. 超過濾區分與濃縮    28
| 2-1-8.  DE-52 離子交換樹脂層析    28
| 2-1-9. Sepharcyl S-200 膠體過濾層析    28
| 2-1-10. Alginate lyase N 端胺基酸定序    29
| 2-2. Alginate lyase 的生化特性分析    29
| 2-2-1. 最適作用溫度    29
| 2-2-2. 熱安定性    29
| 2-2-3. 最適作用 pH 值    29
| 2-2-4.  pH 值的安定性    29
| 2-2-5. 金屬離子的影響    30
| 2-2-6. 基質特異性    30
| 2-3. Alginate lyase 之水解產物分析    30
| 2-3-1. TLC 分析    30
| 2-3-2. HPLC 分析    31
| 2-4. Alginate lyase 基因選殖    31
| 2-4-1.  MA103 染色體 DNA 的萃取純化    31
| 2-4-2. 引子設計    31
| 2-4-3. 無限制酶切位基因選殖 (RF cloning) 及 DpnI 限制酶剪切    32
| 2-4-4. 勝任細胞 (Competent cell) 之製作    34
| 2-4-5. 電穿孔 (Electroporation) 轉形    34
| 2-4-6. 利用PCR 確認插入之基因片段大小    35
| 2-4-7.  DNA 序列比對    35
| 2-5. Recombinant alginate lyase pal III的誘導表現    35
| 2-5-1. Recombinant alginate lyase pal III (rpal III) 的萃取  純化    36
| 2-5-2. 最適誘導時間    36
| 2-5-3. IPTG 最適誘導溫度與濃度    37
| 2-6. 褐藻膠裂解酶 rpal III 之生化性質測試    37
| 2-6-1. 最適作用溫度    37
| 2-6-2. 熱安定性    37
| 2-6-3. 最適作用 pH 值    37
| 2-6-4. pH 值的安定性    37
| 2-6-5. 金屬離子的影響    37
| 2-6-6. 基質特異性    38
| 2-7. Alginate lyase rpal III之水解產物分析    38
| 2-7-1. TLC 鑑定    38
| 2-7-2. HPLC 鑑定    38
| 2-8. 統計分析    38
| 伍、結果與討論    39
| 一、褐藻膠裂解酶的誘導與初步純化    39
| 1. 褐藻膠裂解酶的誘導    39
| 2.  MA103 crude alginate lyases 的生化特性    39
| 2-1. 最適作用溫度與熱安定性    39
| 2-2. 最適作用 pH 值與 pH 值安定性    40
| 2-3. 金屬離子之影響    40
| 2-4. 基質特異性    41
| 3.  DE-52陰離子交換層析部分純化    41
| 二、Alginate lyase palIII 基因大腸桿菌系統轉殖與表現    42
| 三、Recombinant alginate lyase pal III 基因誘導與表現    43
| 四、Recombinant alginate lyase pal III 之生化性質    45
| 4-1. 最適作用 pH 值與 pH 值安定性    45
| 4-2. 最適作用溫度及熱安定性    45
| 4-3. 金屬離子之影響    46
| 4.4. 基質特異性    47
| 五、褐藻膠水解產物之 TLC 與 HPLC 分析    47
| 5-1. TLC 分析    47
| 5-1-1. MA103 crude alginate lyases    47
| 5-1.2. Alginate lyase rpal III    47
| 5-2. TLC 分析    48
| 5-2-1. MA103 crude alginate lyases    48
| 5-2.2. Alginate lyase rpal III    48
| 陸、結論    49
| 柒、參考文獻    51
| 捌、附錄    96
|
| 表目錄
|
|
| 表一、  P. vesicularis MA103 所產褐藻膠裂解酶純化表    65
| 表二、  MA103 crude alginate lyases 對於9 種藻類多醣的基質            特異性    66
| 表三、  RF cloning 所使用之引子及其序列    67
| 表四、  不同培養時間對在1 mM IPTG 誘導之 E. coli BL21
| (DE3) alg 轉形株之胞內與胞外酵素活性表現的影響    68
| 表五、  在不同培養溫度及 IPTG 濃度下誘導 E. coli BL21
| (DE3) alg 轉形株之胞內與胞外酵素活性表現的影響    69
| 表六、  自 E. coli BL21 (DE3) alg 轉形株之胞內粗酵素液
| 純化 Alginate lyase rpal III    70
| 表七、  Alginate lyase rpal III 對於9 種藻類多醣的基質特異性    71
|
|
| 圖目錄
|
|
| 圖一、    P. vesicularis MA103 crude alginate lyases 之誘導生產    72
| 圖二、    P. vesicularis MA103 之生長曲線與所產 crude alginate lyases 之變化    73
| 圖三、    P. vesicularis MA103 crude alginate lyases 之
| SDS-PAGE  與活性染色圖    74
| 圖四、    P. vesicularis MA103 crude alginate lyases 之最適        反應溫度    75
| 圖五、    P. vesicularis MA103 crude alginate lyases 之熱安定性    76
| 圖六、    P. vesicularis MA103 crude alginate lyases 於不同      緩衝液中的最適作用 pH值    77
| 圖七、    P. vesicularis MA103 crude alginate lyases 於不同
| pH 值 緩衝液中的安定性    78
| 圖八、    金屬離子對 P. vesicularis MA103 crude alginate lyases    酵素活性的影響    79
| 圖九、    P. vesicularis MA103 粗酵素液經 30-100 kDa 超過濾  (UF) 區分的 alginate lyases 之 DE-52 陰離子交換    管柱層析圖    80
| 圖十、    P. vesicularis MA103 所產 alginate lyase Pv-Al-1 經  DE-52 離子交換樹脂層析管柱部份純化之 12% SDS-PAGE    81
| 圖十一、  P. vesicularis MA103 所產 alginate lyase Pv-Al-2 經 DE-52 離子交換樹脂層析管柱部份純化之 12% SDS-PAGE    82
| 圖十二、  Alginate lyase pal III 的完整 DNA 序列    83
| 圖十三、  RF cloning 實驗過程中 1st PCR 產物其 1% 洋菜醣
| 膠電泳分析    84
| 圖十四、  RF cloning 實驗過程中 2nd PCR 產物其 1% 洋菜醣
| 膠電泳分析    85
| 圖十五、  RF cloning 實驗過程中 3rd PCR 產物其 1% 洋菜醣    膠電泳分析    86
| 圖十六、  以 PCR 確認插入基因 rpal III 之片段大小    87
| 圖十七、  MA103 alginate lyase rpal III 轉殖株基因定序序列    88
| 圖十八、  Alginate lyase pal III 轉殖株 (alg clone) 與 MA103 alginate lyase pal III 基因序列比對    89
| 圖十九、  不同條件誘導下 alg clone 及帶有 vector pET-21b E. coli BL21 (DE3) 之胞內萃取蛋白質的 12%  SDS-PAGE 電泳分析圖    90
| 圖二十、  不同條件誘導下 alg clone 及帶有 vector pET-21b E. coli BL21 (DE3) 之胞內萃取蛋白質的 12% SDS-PAGE 電泳分析圖    91
| 圖二十一、 以鎳親和性管柱純化之重組 Alginate lyase pal III 其  12% SDS-PAGE 電泳分析圖    92
| 圖二十二、 MA103 algintae lyase pal III DNA 序列所演繹之胺基酸序列    93
| 圖二十三、 Alginate lyases rpal III之最適反應溫度    94
| 圖二十四、 Alginate lyase rpal III 之熱安定性    95
| 圖二十五、 Alginate lyases rpal III 於不同緩衝液中的最適作用    pH值    96
| 圖二十六、 Alginate lyases 於不同 pH 值緩衝液中的安定性    97
| 圖二十七、 金屬離子對 Alginate lyases rpal III 酵素活性之影響    98
| 圖二十八、 MA103 crude alginate lyases 水解 Sodium alginate
| 0-24 hr 之薄層色層分析圖譜    99
| 圖二十九、 Alginate lyase rpalIII 水解 Sodium alginate
| 0-24 hr 之薄層色層分析圖譜    100
| 圖三十 、  MA103 crude alginate lyases 水解 Sodium alginate
| 0-24 hr 之高效能液相層析圖譜    101
| 圖三十一、 Alginate lyase rpalIII 水解 Sodium alginate
| 0-24 hr 之高效能液相層析圖譜    102
|
|
| 附錄目錄
|
|
| 附錄一、      褐藻膠的基本組成單體構造    103
| 附錄二、      Alginate 與二價金屬洋離子的凝膠機制    104
| 附錄三之一、  來自藻類和海洋動物之褐藻膠裂解酶    105
| 附錄三之二、  來自海洋細菌之褐藻膠裂解酶    107
| 附錄三之三、  來自革蘭氏陰性菌之褐藻膠裂解酶    109
| 附錄三之四、  來自革蘭氏陽性菌之褐藻膠裂解酶    112
| 附錄三之五、  來自噬菌體、海洋真菌和病毒之褐藻膠裂解酶    113
| 附錄四、      P. vesicularis MA103 全基因體核苷酸序列中可能產生的alginate lyases    114
| 附錄五、      pET-21a(+) 的限制酶切位及基因圖譜    115
| 附錄六、      pET-21a-d (+) 上基因片段插入位區域的 DNA 序列    116
| 附錄七、      pET 重組蛋白表現系統原理圖    117
| 附錄八、      Alginate lyase pal III 基因大腸桿菌表現系統選殖反應結果彙整表    118
| 附錄九、      MA103 crude alginate lyases 與 Alginate lyase
| rpal III 之生化特性彙整表    119
| 附錄十、      RF cloning 實驗原理流程圖    120
|
|
|
參考文獻: | 陸、參考文獻
|
| 李盈誼。2005。褐藻膠培養系統對人類脂肪幹細胞軟骨新生之評估與及新型生物反應器動態培養技術之評估。國立中興大學化學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。
| 吳敬謙。2003。懸掛式固定化纖維酵素反應器之廢紙纖維素水解與醣類產物分析。中原大學化學系碩士學位論文，臺北，臺灣。
| 吳紹祺。2005。海洋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。國立台灣海洋大學食品科學系碩士學位論文，基隆，臺灣。
| 施姵岑。2009。石蓴寡醣之生產與其生理活性之探討。國立臺灣海洋大學食品科學系碩士學位論文，基隆，臺灣。
| 梁鳳鈺。1999。Pseudomonas vescicularis MA103 菌株所產洋菜酶的純化與性質。國立台灣海洋大學食品科學系碩士學位論文，基隆，臺灣。
| 梁敏婷。2009。Thermoanaerobacterium sp. NTOU2 氫氣生產相關酵素之基因選殖及葡萄糖 6-磷酸脫氫酶與 6-磷酸葡萄糖醛酸脫氫酶的特性探討。國立臺灣海洋大學食品科學系碩士論文，基隆，臺灣。
| 黃萬滄。2001。人類1-抗胰蛋白酶基因在哺乳類動物細胞及酵母菌中支表現。國立中興大學畜產學系碩士學位論文，臺中，臺灣。
| 郭熙志、羅耀光、羅達。2004。以可轉化人工染色體 (TAC) 載體為基礎的百脈根基因組文庫的構建及篩選。植物生理與分子生物學學報。30: 234-238。
| 張俊鴻。2006。以 EDTA、藻酸鈉對砷及鉻汙染土壤萃取效率之研究。國立中央大學環境工程研究所碩士在職專班論文，桃園，臺灣。
| 陳姿元。2006。Aeromonas salmonicida MAEF108 菌株所產洋菜酶純化與性質之探討。國立臺灣海洋大學食品科學系碩士學位論文，基隆，臺灣。
| 許志維。2008。 Aeromonas salmonicida MAEF108 所產 Agarase AS-IIIb 基因轉殖至大腸桿菌與酵母菌及其表現。國立臺灣海洋大學食品科學系碩士學位論文，基隆，臺灣。
| 臺灣海藻資訊網。2009。何謂海藻。http://www.ntm.gov.tw/seaweeds/a/a1_01.asp (August 20, 2009, Taiwan)。
| 賴宜甄。2006。藻類多醣體應用於化妝品之抗老化分析。嘉南藥理科技大學化妝品科技研究所碩士學位論文。嘉義。
| Abd El-Rehim, H. A.  2005.  Characterization and possible agricultural application of polyacrylamide/sodium alginate crosslinked hydrogels prepared by ionzing radiation.  National Center for Radiation and Technology, Atomic Energy Authority, Nasr City, Cairo, Egypt.
| An, Q. D., Zhang, G. L., Wu, H. T., Zhang, Z. C., Gong, W., Liu, Y., Li, X. Z. and Murata, Y.  2008.  Production and partial properties of alginase from newly isolated Flavobacterium sp. LXA.  Process Biochemistry  43: 842-847.
| An, Q. D., Zhang, G. L., Wu, H. T., Zhang, Z. C., Zheng, G. S., Luan, L. Murata, Y. and Li, X.  2009.  Alginate-deriving oligosaccharide production by alginase from newly isolated Flavobacterium sp. LXA and its potential application in protection against pathogens.  Journal of Applied Microbiology  106: 161-170.
| Bartell, P. F., Orr, T. E. and Lam, G. K. H.  1966.  Polysaccharide depolymerase associated with bacteriophage infection.  Journal of Bacteriology  92: 56-62.
| Bashan, Y., Hernadez, J. P., Leyva, L. A. and Bacilio, M.  2002.  Alginate microbeads as inoculant carriers for plant growth-promoting bacteria.  Biology and Fertility of Soils  35: 359-368.
| Boyen, C., Bertheau, Y., Barbeyron, T. and Kloare, B.  1990.  Preparation of guluronate from Pseudomonas alginovora for protoplast isolation in Laminaria.  Enzyme and Microbial Technology  12: 885-890.
| 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.
| Cao, L., Xie, L., Xue, X., Tan, H., Liu, Y. and Zhou, S.  2007.  Purification and characterization of alginate lyase from Streptomyces sp. strain A5 isolated from banana rhizosphere.  Journal of Agricultural and Food Chemistry  55: 5113-5117.
| Chavagnat, F., Duez, C., Guinand, M., Potin, P. and Barbeyron, T.  1996.  Cloning, sequencing and overexpression in Escherichia coli of the alginate-lyase-encoding aly gene of Pseudomonas alginovora: identification of three classes of alginate lyase.  Journal of Biochemistry  319: 575-583.
| Chavagnat, F., Heyraud, A., Colin-Morel, P., Guinand, M. and Wallach, J.  1998.  Catalytic properties and specificity of a recombinat, overexperssed D-mannuronate lyase.  Carbohydrate Research  108: 409-415.
| Chubinskaya, S., Huch, K., Schulze, M., Otten, L., Aydelotte, M. B. and Cole, A. A.  2001.  Gene expression by human articular chondrocytes cultured in alginate bead.  Journal of Histochemistry and Cytochemistry  49: 1211-1219.
| Clementi, F., Fantozzi, P., Mancini, F. and Moresi, M.  1995.  Optimal conditions for alginate production by Azotobacter vinelandii.  Enzyme and Micorbial Technology  17: 983-988.
| Davidson, I. W., Lawson, C. J. and Sutherland, I. W.  1977.  An alginate lyase from Azotobacter vinelandii phage.  Journal of General Microbiology  98: 223-229.
| Davis, T. A., Lianes, F., Volesky, B., Diaz-Pulido, G., McCook, L. and 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.
| Dong, T., Tamaru, Y. and Araki, T.  2007.  Molecular cloning, expression, and characterization of a-agarase gene, agaD, from marine bacterium, Vibrio sp. strain PO-303.  Bioscience Biotechnology and Biochemistry  71: 38-46.
| Favorov, V. V.  1973.  Purification of alginases by affinity chromatography on Bio-Gel alginate column.  International Journal of Biochemistry  4: 107-110.
| Fu, X. T., Lin, H. and Kim, S. M.  2007.  Purification and characterization of a Na+/K+ dependent alginate lyase from tuban shell Vibrio sp. YKW-34.  Enzyme and Microbial Technology  41: 828-834.
| Gacesa, P.  1992.  Enzyme degration of alginate.  International Journal of Biochemistry  24: 545-552.
| Gombotz, W. R. and Wee, S. F.  1998.  Protein release from alginate matrices.  Advance Drug Delivery Reviews  31: 267-285.
| Ha, J. C., Kim, G. T., Kim, S. K., Oh, T. K., Yu, J. H. and Kong, I. S.  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.
| Harris, P.  1990.  Food Gels. Elsevies Science Publishing Co., Inc. New York, U.S.A.
| Haug, A., Larsen, B. and Smidsrod, O.  1966.  A study of the constitution of alginic acid by partial acid hydrolysis.  Acta Chemica Scandinavica  20: 183-190.
| Hay, I. D., Gatland, K., Campisano, A., Jordens, J. Z. and Rehm, B. H. A.  2009.  Impact of alginate overproduction an attachment and biofilm architecture of a supermucoid Pseudomonas aeruginosa strain.  Applied and Environmental Microbiology  75: 6022-6025.
| Iwamoto, T., Araki, R., Iriyama, K. I., Oda, T., Fukuda, H., Hayashida, S. and 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.
| Iwasaki, K. and Matsubara, Y.  2000.  Purification of alginate oligosaccharides with root growth-promotion activity toward lettuce.  Bioscience Biotechnology and Biochemistry  64: 1067-1070.
| Kashiwabara, Y., Suzuki, H. and Nisizawa, K.  1969.  Alginate lyase of Pseudomonads.  Journal of Biochemistry  66: 503-512.
| Kenndey, L., McDowell, K. and Sutherland, I. W.  1992.  Alginases from Azotobacter species.  Journal of General Microbiology  138: 2465-2471.
| Kim, D. E., Lee, E. Y. and Kim, H. S.  2008.  Cloning and characterization of alginate lyase from a marine bacterium Streptomyces sp. ALG-5.  Marine Biotechnology  11: 10-16.
| Kirimura, K., Masuda, N., Iwasaki, Y., Nakagawa, H., Kobayashi, R. and Usami, S.  1999.  Purification and characterization of a novel agarase from an alkalophilic bacterium, Alteromonas sp. E-1.  Journal of Bioscience and Bioengineering  87: 436-441.
| Kitamikado, M., Yamaguchi, K., Tseng, C. H. and Okabe, B.  1990.  Method designed to detect alginate-degrading bacteria.  Applied and Environmental Microbiology  56: 2939-2940.
| Kitamikado, M., Tseng, C. H., Yamaguchi, K. and Nakamura, T.  1992.  Two types of bacterial alginate lyase.  Applied and Environmental Microbiology  58: 2472-2478.
| Koopmann, B., Rollwage, H., Nollenburg, M. and Rudolph, K.  2001.  Isolation and characterization of the algD gene of Pseudomonas syringae pv. phaseolicola and its distribution among other Pseudomonads and related organisms.  Journal of Phytopathology  149: 511-519.
| Larsen, B., Salem, D. M. S. A., Sallam, M. A. E., Mishrikey, M. M. and Beltagy, A. I.  2003.  Characterization of the alginates from algae harvested at the Egyptian red sea coast.  Carbohydrate Research  338: 2325-2336.
| Li, X., Dang, X., Zhao, C., Chen, Z. and 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.
| Madgwick, J., Haug, A. and Larsen, B.  1973.  Alginate lyase in the brown alga Laminaria digitata (Huds.) Lamour.  Acta chemica Scandinavica  27: 711-712.
| Madgwick, J., Haug, A. and Larsen, B.  1978.  Ionic requirments of alginate-modifying enzymes in the marine alga Pelvetia canaliculata (L) Dcne. et Thur. Botanica Marina  21: 1-3.
| Malissard, M., Duez, C., Guinand, M., Vacheron, M. J. and Michel, G.  1993.  Sequence of a gene encoding a (polyManA) alginate lyase active on Pseudomonas aeruginosa alginate.  FEMS Microbiolog Letters  110: 101-106.
| Malissard, M., Chavagnat, F., Duez, C., Vacheron, M. J. and Guinand, M.  1995.  Overproduction and properties of the mannuronate alginate lyase AlxMB.  FEMS Microbiology Letters  126: 105-112.
| Matsushima, R., Danno, H., Uchida, M, Ishihara, K., Suzuki, T., Kaneniwa, M., Ohtsubo, Y., Nagata, Y. and Tsuda, M.  2010.  Analysis of extracellular alginate lyase and its gene from a marine bacterial strain, Pseudoalteromonas atlantica AR06.  Application of Microbiology and Biotechnology  86: 567-576.
| Mikami, B., Hisano, T., Yamashita, T., Inose, T. and Takagi, M.  1994.  Crystallization and preliminary X-ray diffraction studies of bacterial alginate lyase.  Journal of Fermentation and Bioengineering  77: 691-692.
| Miyazaki, M., Obata, J., Iwamoto, Y., Oda, T. and 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.
| Mstsubara, Y., Kawada, R., Iwasaki, K., Oda, T. and Muramatsu, T.  1998.  Extracellular poly(-L-guluronate)lyase from Corynebacterium sp.: Purification, characterization, and conformation properties.  Journal of Protein Chemistry  17: 29-36.
| Muramatsu, T., Hirose, S. and Katayose, M.  1977.  Isolation and properties of alginate lyase from the mid-gut gland of wreath shell Turbo cornutus.  Agricultural and Biological Chemistry  41: 1939-1946.
| Alkawash, M. A., Soothill, J. S. and Schiller, N. L.  2006.  Alginate lyase enhances antibiotic killing of mucoid Pseudomonas aeruginosa in biofilms.  Acta Pathologica, Microbiologica et Immunologica Scandinavica  114: 131-138.
| Nakagawa, A., Qzaki, T., Chubachi, K., Hosoyama, T. and Okubo, T.  1998.  An effective method for isolated alginate lyase-pruducing Bacillus sp. ATB-1015 strain and purification and characterization of the lyase.  Journal of Applied Microbiology  84: 328-339.
| Nisizawa, K., Fujibayashi, S. and Kashiwabata, Y.  1968.  Alginate lyases of a marine mollusc, Dolabella auricula Solander.  Journal of Biochemistry  64: 25-37.
| Ott, C. M., Day, D. F., Koening, D. W. and Pierson, D. L.  2001.  The release of alginate lyase from growing Pseudomonas syringae pathovar phaseolicola.  Current Microbiology  42: 78-81.
| Panikkar, R. and Brasch, D. J.  1996.  Composition and block structure of alginates from New Zealand Brown Seaweeds.  Carbohydrate Research  293: 110-132.
| Pecina, A., Pascual, A. and Paneque, A.  1999.  Cloning and expression of algL gene, encoding the Azotobacter chroococcum alginate lyase: Purification and characterization of the enzyme.  Journal of Bacteriology  181: 1409-1414.
| Pitt, T. L. and Raisbeck, L. C.  1978.  Degradation of the mucoid polysaccharide of Pseudomonas aeruginosa by Beneckea pelagia.  Journol of Applied Biochemistry  45: 297-300.
| Preiss, J. and Ashwell, G.  1962.  Alginic acid metabolism in bacteria.  Journal of Biological Chemisrty  237: 309-316.
| Redenbath, M., Kieser, H. M., Denapaite, D., Eichner, A. and Cullum, J.  1996.  A set of ordered cosmids and a detailed genetic and physical map for the 8 Mb Streptomyces coelicolor A3(2) chromosome.  Molecular Microbiology  21: 77-96.
| Rioux, L. E., Turgeon, S. L. and Beaulieu, M.  2007.  Characterization of polysaccharides extracted from brown seaweeds.  Carbohydrate Polymers  69: 530-537.
| Sambrook, J. and D. W. Russell.  2006.  The Condensed Protocol from Molecular Cloning: A Laboratory Manual.  Cold Spring Harbor Lab. Press, Cold Spring Harbor, NY, U.S.A.  pp. 58-59, 177, 188-192, 243, 718-725, 730, 758-760.
| Sawabe, T., Ezura, Y. and Kimura, T.  1992.  Purification and characterization and of an alginate lyase from marine Alteromonas sp.  Nippon Suisan Gakkaishi  58: 521-527.
| Sawabe, T., Ohtsuka, M. and Ezura Y.  1997.  Novel alginate lyase from marine bacterium Alteromonas sp. strain H-4.  Carbohydrate Research  304: 69-76.
| Sawabe, T., Sawada, C., Suzuki, E. and Ezura, Y.  1998.  Intracellular alginate oligosaccharide degrading enzyme activity that is incapable of degrading intact sodium alginate from a marine bacterium Alteromonas sp.  Fisheries Science  64: 320-334.
| Sawabe, T., Takahashi, H., Ezura, Y. and Gacesa, P.  2001.  Cloning, sequence analysis and expreesion of Pseudoalteromonas elyakovii IAM 14594 gene (alyPEEC) encoding the extracellular alginate lyase.  Carbohydrate Research  335: 11-21.
| Sawabe, T., Takahasi, H., Saeki, H., Niwa, K. and Aono, H.  2007.  Enhanced expression of active recombinant alginate lyase AlyPEEC cloned from a marine bacterium Pseudoalteromonas elyakovii in Escherichia coli by calcium compounds.  Enzyme and Microbial Technology  40: 285-291.
| Schroeder, D. C., Jaffer, M. A. and Coyne, V. C.  2003.  Investigation of the role of a(1-4)agarase produced by Pseudoalteromonas gracilis B9 in eliciting disease symptoms in the red alga Gracilaria gracilis.  Microbiology  149: 2919-2929.
| Shiroma, R., Uechim S., Tavata, S. and Toka, M.  2006.  Isolation and characterization of alginate from Hizikia fusiformis and preparation of its oligosaccharides.  Journal of Applied Glycoscience  54: 85-90.
| Stevens, R. A. and Levin, R. E.  1977.  Purification and characterization of an alginase from Alginovibrio aquatilis.  Applied and Enviromental Microbiology  33: 1156-1161.
| Sutherland, I. W. and Keen, G. A.  1981.  Alginases from Beneckea pelagia and Pseudomonas spp.  Journal of Applied Biochemistry    3: 48-57.
| Suzuki, H., Suzuki, K. I., Inoue, A. and Ojima, T.  2006.  A novel oligoalginate lyase from abalone, Haliotis discus hannai, that releases disaccharide from alginate polymer in an exolytic manner.  Carbohydrate Research  341: 1809-1819.
| Takeshita, S., Sato, N., Igarashi, M. and Muramatsu, T.  1993.  A highly denaturant-durable alginate lyase from a marine bacterium: Purification and properties.  Bioscience, Biotechnology and Biochemistry  57: 1125-1128.
| Torres, M. R., Sousa, A. P. A., Silva Filho, E. A. T., Melo, D. F., Feitosa, J. P. A., de Paula R. C. M. and Lima, M. G. S.  2007.  Extraction and physicochenical characterization of Sargassum vulgare alginate from Brazil.  Carbohydrate Research  342: 2067-2074.
| Tseng, C. H., Yamagucho, K. and Kitamikado, M.  1992a.  Isolation and some properties of alginate lyase from a marine bacterium Vibrio sp. AL-128.  Nippon Suisan Gakkaishi  58: 743-749.
| Tseng, C. H., Yamagucho, K., Nishimura, M. and Kitamikado, M.  1992b.  Alginate lyase from Vibrio alginolyticus ATCC 17749.    58: 2063-2067.
| van den Ent F., and J. Lowe. 2006. RF cloning: A restriction-free method for inserting target genes into plasmids. Journal of Biochemical and  Biophysical Methods  67: 67-74.
| Wang, T. Y., Preston, L. A. and 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, Y. H., Yu, G. L., Wang, X. M., Lv, Z. H., Zhao, X., Wu, Z. H. and Ji, W. S.  2006.  Purification and characterization of alginate lyase from marine Vibrio sp. YWA.  Acta Biochimica et Biophysica Sinica  38: 633-638.
| Watanabe, T. and Nisizawa, K.  1982.  Enzymatic studies on alginat lyase from Undaria pinnatifida in relation to texture softening prevention by ash-treatment (Haiboshi).  Bulletin of the Japanese Society of Scientific Fisheries  48: 243-249.
| Weissbach, A. and Hurwitz, J.  1958.  The formation of 2-keto-3-deoxyheptonic acid in extracts of Escherichia coli B.  Journal of Biological Chemistry  234: 705-709.
| Williams, M. A. K., Marshall, A., Haverkamp, R. G. and Draget, K. I.  2007.  Stretching single polysaccharide molecules using AFM A potential method for the investigation of the intermolecular uronate distribution of alginate.  Food Hydrocolloids  22: 18-23.
| Xu, X., Iwamoto, Y., Kitamura, Y., Oda, T. and Muramatsu, T.  2003.  Root growth-promoting activity of unsaturated oligomeric urinates from alginate on carrot and rice plants.  Bioscience, Biotechnology and Biochemistry  67: 2022-2025.
| Yamamoto, S., Sahara, T., Sato, D., Kawasaki, K., Ohgiya, S., Inoue, A. and Ojima, T.  2008.  Catalytivally important amino acid residues of abalone alginate lyase HdAly assessed by site-directed mutagenesis.  Enzyme and Microbial Technology  43: 396-402.
| Yang, Z., Li, J. P. and Guan, H. S.  2004.  Preparation and characterization of oligomannuronates from alginate degrade by hydrogen peroxide.  Carbohydrate Polymers  58: 115-121.
| Ye, H., Zhou, C., Liu, J. and Zeng, X.  2008.  Purification, antitumor and antioxidant activities in vitro of polysaccharides from the brown seaweed Sargassum pallidum.  Food Chemistry  111: 428-432.
| Young, C. C., Rekha, P. D., Lai, W. A. and Arun, A. B.  2006.  Encapsulation of plant growth-promotion bacteria in alginate beads enriched with humic acid.  Biotechnology and Bioengineering  95: 76-83.
| Zhang, Z. Q., Yu, G. L., Z    hao, X., Li, Q. and Guan, H. S.  2005.  Thin layer chromatography analysis of several uronates and their oligosaccharides.  Chinese Journal of Analytical Chemistry  33: 1750-1752.
|
|

|

預覽及輸出檢索結果

記錄

欄位

| 本頁 1 筆
| 記錄 1 至 36
| 選擇之記錄

| 標題
| 全部欄位
| selected fields new window
| APA Style
| Chicago Style
| MLA Style
| CNS-13611 Style
| EndNote Style

e-mail 地址 | : |
e-mail 標題 | : |

| |

系統規劃：海洋大學圖書館
系統製作：飛資得資訊有限公司

2010 年 9 月 26 日 22:49