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稀土离子对纤维酶活性的影响及其机理

作　者: 余兴莲
导　师: 王丽
学　校: 宁波大学
专　业: 无机化学
关键词: Rare earth ion Cellulase Enzyme activity Effect mechanism
分类号: Q55
类　型: 硕士论文
年　份: 2008年
下　载: 28次
引　用: 0次
阅　读: 论文下载

内容摘要

This paper was studied the influences of the different concertration of rare earth ions (lanthanum, cerium, praseodymium, neodymium, samarium, europium, gadolinium) on cellulase degradation cellulose three substrates (filter paper, carboxymethylcellulose sodium, cellobiose) systemically, as well as the effection of rare earth ion which was the effect remarkable rare earth on the cellulose complexes with citric acid on the cellulase enzyme activity, obtained the best activation rare earth ion and the best activation density. The results showed: The effection of the rare earth on enzyme activity decided to the rare earth density. the cellulase activity was activated by the low concentration rare earth. Contrarily, the high concentration had the inhibition action to the enzyme activity.The Nd3+, Eu3+ had the most remarkable activition among the extent of experiment concentration for 1.0×10-9-1.0×10-7 g·L-1. the free rare earth ion on the cellulase enzyme activity was better than the rare earth complexes of citrate.The effect mechanism of rare earth on the cellulase activity and its structure had primarily discussed by atom force microscope, polyacrylamide gel electrophoresis(PAGE), SDS-polyacryla- mide gel electrophoresis(SDS-PAGE). By atom force microscope showed: the high concentrated rare earth ion made cellulase outside rough and uneven in surface, the edge irregular, it was obvious accumulated, conglomerated. But the low concentration have no effect on cellulase surface topography; By PAGE and SDS-PAGE showed: the cellulase electrophoresis band of some component had shifted up, conglomerated when it edded the high concentrated rare earth ion; And the low concentration have no effect on cellulase electrophoresis band. So the high concentrated rare earth ion made the cellulase accumulated, affected its structure, inhibited cellulase activity so made the efficiency of the cellulase degeneration cellulose was reduced. Contrarily, the low concentratio- n rare earth ion does not have the influence on the cellulase structure nearly. Maybe it adsorbe in the cellulase CBD or CD linke among them, improve the cellulase and cellulose combining better each other for increasing the catalyze efficiency.Using the remarkable activition rare earth ion and the best density on cellulase activity to search after the influence of rare earth ion and the cellulase to degrades the native cellulose wild bamboo. It showed that Nd3+ was the most remarkable activation function. Through the orthogonal experiment, the best technological conditions of the rare earth ion and the cellulase to degrade the wild bamboo was: pH 4.5, temperature for 52°C, reaction time 75 min.This research results had revealed the influence of rare earth ion on cellulose enzyme activity, and had discussed the rare earth ion and the cellulase effect mechanism, for increasing the cellulose enzyme degradation cellulose hydrolyzation efficiency, resolving the energy crisis have important practical or immediate significance and theory value.

全文目录

Abstract  4-9
引言  9-10
1 绪论  10-27
  1.1 纤维素研究概况  10-14
    1.1.1 纤维素的分子结构  11-13
    1.1.2 纤维素的降解  13-14
  1.2 纤维素酶的研究概况  14-21
    1.2.1 纤维素酶的组成及物化性质  15-18
    1.2.2 纤维素酶的分子结构及降解机理  18-21
  1.3 稀土离子与酶相互作用的研究进展  21-22
    1.3.1 稀土金属离子对酶作用的类型及作用特征  21
    1.3.2 稀土离子对酶作用的位点  21-22
    1.3.3 稀土离子对酶作用的时效性  22
    1.3.4 酶活性与稀土离子半径的关系  22
  1.4 纤维素酶降解纤维素的影响因素  22-25
    1.4.1 纤维素结构对酶解的影响  22-23
    1.4.2 酶和酶用量对酶解的影响  23
    1.4.3 温度、pH 值、酶促作用时间和显色时间对酶解的影响  23-24
    1.4.4 抑制剂和激活剂对酶解的影响  24-25
  1.5 选题依据和意义  25-27
2 稀土离子及稀土柠檬酸配合物对纤维素酶酶活的影响规律  27-55
  2.1 实验试剂及仪器  27-28
    2.1.1 试剂与规格  27-28
    2.1.2 仪器  28
  2.2 实验方法  28-34
    2.2.1 溶液制备  28-30
    2.2.2 葡萄糖标准曲线测定  30-31
    2.2.3 稀土离子对滤纸酶活的影响  31-32
    2.2.4 稀土离子对羧甲基纤维素钠(CMC-Na)酶活的影响  32-33
    2.2.5 稀土离子对纤维二糖酶活的影响  33-34
    2.2.6 钕、铕柠檬酸配合物对滤纸酶活的影响  34
    2.2.7 钕、铕柠檬酸配合物对羧甲基纤维素钠酶活的影响  34
    2.2.8 钕、铕柠檬酸配合物对纤维二糖酶活的影响  34
  2.3 实验结果与讨论  34-54
    2.3.1 葡萄糖标准曲线的绘制及回归曲线方程  34-35
    2.3.2 纤维素酶降解三种底物的酶活  35-36
    2.3.3 稀土离子对纤维素酶降解三种底物酶活的影响  36-50
      2.3.3.1 镧离子对纤维素酶降解三种底物酶活的影响  36-38
      2.3.3.2 铈离子对纤维素酶降解三种底物酶活的影响  38-40
      2.3.3.3 镨离子对纤维素酶降解三种底物酶活的影响  40-42
      2.3.3.4 钕离子对纤维素酶降解三种底物酶活的影响  42-44
      2.3.3.5 钐离子对纤维素酶降解三种底物酶活的影响  44-46
      2.3.3.6 铕离子对纤维素酶降解三种底物酶活的影响  46-48
      2.3.3.7 钆离子对纤维素酶降解三种底物酶活的影响  48-50
    2.3.3 钕、铕柠檬酸配合物对纤维素酶降解三种底物酶活的影响  50-54
      2.3.3.1 钕柠檬酸配合物对纤维素酶降解三种底物酶活的影响  50-52
      2.3.3.2 铕柠檬酸配合物对纤维素酶降解三种底物酶活的影响  52-54
  2.4 小结  54-55
3 原子力显微镜(AFM)下稀土离子与纤维素酶作用形貌的观察  55-61
  3.1 实验试剂及仪器  55
    3.1.1 试剂与规格  55
    3.1.2 仪器  55
  3.2 实验方法  55-56
    3.2.1 样品制备  55
    3.2.2 制样  55-56
  3.3 实验结果与讨论  56-60
    3.3.1 纤维素酶的AFM 图成像分析  56
    3.3.2 稀土钕离子与纤维素酶的AFM 图成像分析  56-58
    3.3.3 稀土铕离子与纤维素酶的AFM 图成像分析  58-60
  3.4 小结  60-61
4 纤维素酶蛋白电泳  61-67
  4.1 实验试剂及仪器  61-62
    4.1.1 试剂与规格  61
    4.1.2 仪器  61-62
  4.2 实验方法  62-64
    4.2.1 溶液的配制  62-63
      4.2.1.1 SDS-聚丙烯酰胺垂直凝胶电泳溶液的配制  62
      4.2.1.2 聚丙烯酰胺垂直凝胶电泳溶液的配制  62-63
    4.2.2 SDS-聚丙烯酰胺垂直凝胶电泳实验过程  63
    4.2.3 聚丙烯酰胺垂直凝胶电泳实验过程  63-64
  4.3 实验结果与讨论  64-66
    4.3.1 纤维素酶SDS-PAGE 浓度梯度电泳图  64
    4.3.2 稀土离子与纤维素酶SDS-PAGE 电泳图  64-65
    4.3.3 稀土离子与纤维素酶PAGE 电泳图  65-66
  4.4 小结  66-67
5 稀土离子对纤维素酶降解天然纤维素的影响  67-72
  5.1 实验材料  67
    5.1.1 试剂及规格  67
    5.1.2 仪器  67
  5.2 实验方法  67-69
    5.2.1 纤维素的预处理  67
    5.2.2 纤维素材料中纤维素含量的测定  67
    5.2.3 稀土离子对纤维素酶降解纤维素原料的影响  67-68
    5.2.4 还原糖的分析方法  68
    5.2.5 正交实验  68-69
  5.3 实验结果与讨论  69-71
    5.3.1 原料的纤维素含量  69
    5.3.2 稀土离子对纤维素酶降解纤维素原料的影响  69-70
    5.3.3 正交实验结果  70-71
  5.4 小结  71-72
6 总结  72-73
参考文献  73-80
在学研究成果  80-81
致谢  81

稀土离子对纤维酶活性的影响及其机理

内容摘要

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