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发酵法生产腺苷的培养基优化

作　者: 毛利斯
导　师: 陈长华
学　校: 华东理工大学
专　业: 发酵工程
关键词: 腺苷枯草芽孢杆菌响应面法发酵优化
分类号: TQ920
类　型: 硕士论文
年　份: 2010年
下　载: 147次
引　用: 0次
阅　读: 论文下载

内容摘要

采用实验设计法对Bacillus subtilis ATCC 21616生产腺苷的培养基中两个重要成分(酵母粉和玉米浆)进行优化和探讨。通过响应面法(中心组合设计)建立了影响因素与响应值(腺苷产量)之间的函数关系。根据回归方程与典型相关法寻优得出,当酵母粉为7.27g/l,玉米浆为51.84 ml／l时,预测腺苷产量可达9.84 mg/ml.在最优条件下,实际产苷量为9.62 mg/ml,与原培养基比较产苷量提高58.22%。针对响应曲面深入探讨,利用典型相关分析法进一步了解稳定点之数学特征,发现为极大点。此外,还发现发酵培养基中添加抗生素以及表面活性剂有利于腺苷积累。结果表明,在发酵56小时添加1% Tween 80,腺苷产量可达12.04 mg/ml,与其对照相比较产苷量提高28.08%。研究了磷酸镁,EDTA,柠檬酸钠,硫酸镁等物质对腺苷发酵的影响。发现在基础料中添加浓度为0.1%的柠檬酸钠或0.12%硫酸镁,腺苷产量分别提高了14.88%和29.02%。EDTA与磷酸镁的作用不佳。其浓度与产苷量成反比。

全文目录

摘要  5-6
Abstract  6-11
Chapter 1 General Introduction and objectives of this thesis  11-25
  1.1 Physiological properties and clinical application of adenosine  11-13
    1.1.1 Role of adenosine in the heart  11-12
    1.1.2 Role of adenosine in the nervous system  12
    1.1.3 Role of adenosine in the skeletal muscle  12-13
  1.2 Importance of nucleosides, nucleotides and related compounds  13
  1.3 De novo biosynthesis of purine nucleotides and nucleosides  13-14
  1.4 Regulation of purine nucleotides production  14-16
  1.5 Methods for the production of nucleic acid-related substances  16-21
    1.5.1 Enzymatic hydrolysis of RNA  17-18
    1.5.2 Fermentation production of 5'-IMP  18
    1.5.3 Fermentation production of inosine  18-19
    1.5.4 Fermentation production of guanosine  19
    1.5.5 Fermentation production of adenosine and adenine  19-21
  1.6 Optimization procedures  21-23
    1.6.1 Empirical optimization  21
    1.6.2 Statistically-based optimization  21-22
    1.6.3 Analysis of quadratic models  22-23
  1.7 Objectives of this thesis  23-25
Chapter 2 Materials and Methods  25-31
  2.1 Equipments  25
  2.2 Chemical reagents  25-26
  2.3 Bacterial strain  26
  2.4 Culture media  26
  2.5 Preparation of seed culture  26
  2.6 Fermentation conditions  26-27
  2.7 Cell growth measurement  27
  2.8 Residual sugar determination  27-28
  2.9 Quantitative determination of bases and nucleosides  28-29
  3.0 HPLC assay of adenosine  29-31
Chapter 3 Results and Discussions  31-62
  3.1 Evaluation of factors affecting adenosine accumulation  31-33
  3.2 Effect of corn steep liquor and yeast extract on growth and adenosine production  33-41
    3.2.1 Culture conditions  34
    3.2.2 Experimental design  34-35
    3.2.3 Data analysis for cell growth  35-37
    3.2.4 Data analysis for adenosine accumulation  37-41
  3.3 Model validation  41-43
  3.4 Strain preservation  43-44
  3.5 Action of antibiotics and surface active agents on adenosine production  44-57
    3.5.1 Culture conditions  44
    3.5.2 Effect of antibiotics  44-49
    3.5.3 Effect of surface active agents  49-53
    3.5.4 Response surface representation of the effects of Tween 80 and time of addition  53-57
  3.6 Effects of additional medium components  57-62
    3.6.1 Magnesium phosphate  57-59
    3.6.2 EDTA  59
    3.6.3 Sodium citrate  59-60
    3.6.4 Effect of magnesium sulfate concentration  60-62
Chapter 4 Conclusions and suggestions for future work  62-64
References  64-70
Acknowledgments  70

发酵法生产腺苷的培养基优化

内容摘要

全文目录

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