葡萄糖氧化酶产生菌的诱变及发酵条件的研究

葡萄糖氧化酶产生菌的诱变及发酵条件的研究(8900字)
摘  要：从土壤中分离到一株能生产葡萄糖氧化酶菌株，对该菌株的菌落形态和菌体形态观察，确定其属于黑曲霉GD-8。以该菌株为出发菌株，经过微波诱变、紫外线诱变和硫酸二乙酯诱变剂进行诱变，选育出一株葡萄糖氧化酶高产菌株。对突变株A6进行了发酵条件研究，结果表明：该突变株适宜的发酵条件为发酵温度26℃，初始pH值5.0，装量为70mL/250mL三角瓶，接种量为15%（v/v）,摇床转速为150r/min，培养时间为4d。在此条件下，葡萄糖氧化酶活力达到16.04IU/mL 。
关键词：葡萄糖氧化酶；筛选；诱变；发酵条件

Mutagenes’s of Glucose Oxidase Producing Strain and its Fermentation Condition
Abstract: One strain of glucose oxidase producing strain was obstained from soil sample.Acording to the characteristics of the morpha and the colonies,the screened strain belongs to Asperillus niger GD-8.The strain was used as starting strain and mutated by UV,microwave and DES.And a high-glucose oxidase producing strain A6 was bred.The formentation conditions such as cultural temperature,initial pH value,inoculation, medium volume, rotatory speed and cultural time were studied by single factor experiment.The results showed that the optimum fermentation conditions were as follows:cultural temperation was 26℃,the initial pH value was 5.0, the medium volume was 70mL in every of 250mL flask, inoculation was 15%（v/v）,rotatry speed was 150r/min and cultural time was 4d.Under these conditions,the activity of glucose oxidase reached 16.04 IU/mL.
Key words: glucose oxidase；screening；mutation；fermentation conditions
 

目   录
摘要…………………………………………………………………………………………1
关键词………………………………………………………………………………………1
1前言…………………………………………………………………………………………2
2材料与方法………………………………………………………………………………3
    2.1材料…………………………………………………………………………………3
      2.2.1菌种……………………………………………………………………………3
      2.2.2 试剂与仪器………………………………………………………………3
      2.2.3 培养基………………………………………………………………………3
2.2 方法………………………………………………………………………………4
  2.2.1 菌株筛选……………………………………………………………………4
  2.2.2 培养方法…………………………………………………………………4
  2.2.3 菌种诱变……………………………………………………………………4
  2.2.4 产酶条件研究………………………………………………………………5
      2.2.5 酶活的测定………………………………………………………………5
3 结果与分析 ………………………………………………………………………………6
3.1 筛选结果…………………………………………………………………………6
3.2 紫外线诱变…………………………………………………………………6
  3.2.1紫外线诱变对黑曲霉GD-8孢子致死率的影响……………………………6
  3.2.2 紫外线诱变效果…………………………………………………………7
3.3 微波诱变…………………………………………………………………………7
  3.3.1微波诱变黑曲霉孢子致死率的影响………………………………………7
  3.3.2 微波诱变效果………………………………………………………………7
3.4硫酸二乙酯（DES）诱变…………………………………………………………8
3.5发酵条件对产酶的影响………………………………………………………9
   3.5.1培养温度对产酶的影响………………………………………………9
   3.5.2 培养基初始pH值对产酶的影响……………………………………………10
 3.5.3 装料量对产酶的影响………………………………………………………10
 3.5.4接种量对产酶的影响………………………………………………………11
3.5.5摇床转速对产酶的影响………………………………………………11
3.5.6 培养时间对产酶的影响……………………………………………………12
4 结论………………………………………………………………………………………12
参考文献 …………………………………………………………………………………13
致谢…………………………………………………………………………………………14