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毕业论文网 > 毕业论文 > 化学化工与生命科学类 > 生物工程 > 正文

利用季也蒙毕赤酵母生物转化合成2-苯乙醇发酵优化的研究毕业论文

 2022-01-22 11:01  

论文总字数:19096字

摘 要

本论文研究发现了在未经优化的培养基中,季也蒙毕赤酵母菌株的耐受性大致是 4 g/L。以上述耐受性的菌株开展单批生长发酵,得到2-苯乙醇的产量为 3 g/L。所以这种菌株作为生物法合成2-苯乙醇是较为适宜的。

为达到提高2-苯乙醇的产量的目标,我们采取控制单一变量法进行发酵培养。开始,以温度为单一变量进行温度梯度培养,温度梯度为:25℃,30℃,33℃,37℃。最终由实验结果得出菌株在30 ℃时,生长状况最好,产量最高,最高产量为1.55g/L;其次,进行葡萄糖浓度梯度的发酵培养,而葡萄糖浓度梯度为:20g/L。20g/L,60g/L,80g/L,100g/L。最终由实验所得,当菌株在葡萄糖浓度为40g/L时,其生长情况最优,最高产量为2.17g/L;然后,进行底物浓度梯度的发酵培养,并由实验结果表明当底物浓度为7g/L时,其产量最高为2.28 g/L;接下来,采用响应面原理设计上述3种实验条件,使其成为响应面体系,根据实验得到:24.735℃,63.545g/L Glu,10.705g/L L-phe为最适条件。

最后利用发酵罐放大的方法提高2-苯乙醇的产量,使其产量最大值达到了2.68g/L,极大的提高了2-苯乙醇的产量。

关键词: 2-苯乙醇 季也蒙毕赤酵母 发酵条件优化 控制单一变量法

Optimization of 2-Phenylethanol Fermentation Synthesis by Meyerozyma guilliermondii Biotransformation

Abstract
   This paper found that the tolerance of the Pichia pastoris strain was approximately 4 g/L in the unoptimized medium. A single batch of growth fermentation was carried out with the above-tolerant strain, and the yield of 2-phenylethanol was 3 g/L. Therefore, this strain is more suitable for the biosynthesis of 2-phenylethyl alcohol.
In order to achieve the goal of increasing the yield of 2-phenylethanol, we adopted a single variable method for fermentation culture. Initially, temperature gradient culture was carried out with temperature as a single variable, and the temperature gradient was: 25 ° C, 30 ° C, 33 ° C, 37 ° C. Finally, the experimental results showed that the strain had the best growth at 30 °C, the highest yield, the highest yield was 1.55 g/L; secondly, the glucose concentration gradient fermentation culture, and the glucose concentration gradient: 20 g/L. 20 g/L, 60 g/L, 80 g/L, 100 g/L. Finally, the experimental results showed that when the strain had a glucose concentration of 40g/L, the growth was optimal, and the highest yield was 2.17g/L. Then, the substrate concentration gradient fermentation culture was carried out, and the experimental results showed that the substrate concentration When it is 7g/L, its yield is up to2.28 g/L. Next, the above three experimental conditions are designed by the response surface principle to make it a response surface system. According to the experiment, it is obtained: 24.735 ° C, 63.545 g/L Glu, 10.705 g/L L-phe is the most suitable condition.
Finally, the yield of 2-phenylethanol was increased by the method of magnifying the fermenter, and the maximum yield was 2.68 g/L, which greatly increased the yield of 2-phenylethanol.
Key words: 2-phenylethanol, Pichia pastoris, fermentation condition optimization, single variable method

目录

第一章 文献综述 2

1.1关于2-苯乙醇的简述 2

1.1.1 2-苯乙醇的应用和市场 2

1.1.2 如何合成2-苯乙醇 2

1.2利用生物转化法合成2-苯乙醇 4

1.2.1制备与生产2-苯乙醇的微生物 4

1.2.2 酵母菌合成2-苯乙醇的代谢途径 5

1.2.3 2-苯乙醇浓度对微生物的促进与抑制作用 6

1.3 本论文的研究内容以及拟采用的手段 7

1.4 本论文的研究意义 7

1.5 提高2-苯乙醇产量的方法 7

第二章 材料与方法 9

2.1 仪器与试剂 9

2.1.1实验仪器 9

2.1.2 试剂 10

2.1.3 酵母菌株 10

2.1.4 发酵培养基 10

2.1.5液相高效色谱仪: 11

第三章 实验内容 12

3.1 菌株的2-PE耐受性实验 12

3.1.1耐受性实验一实验操作 12

3.1.2实验结果及分析讨论 12

3.2最适碳源 13

3.2.1实验操作 13

3.2.2 实验结果与分析 14

3.3温度梯度实验 14

3.3.1实验操作: 14

3.3.2实验结果及分析讨论 15

3.4菌株的葡萄糖浓度梯度实验 16

3.4.1 实验操作 16

3.4.2 实验结果与分析讨论 16

3.5 菌株的底物梯度实验 17

3.5.1 菌株的底物梯度实验一操作 17

3.5.2 实验结果与分析讨论 18

3.6发酵优化-响应面实验 19

R2=0.9710 R2 (Adj)=0.9337 R2 (Pred)=0.5362 19

3.6.1 响应面实验(1) 19

3.6.2 响应面实验(2) 22

3.7 发酵罐放大实验 26

3.7.1 发酵罐放大实验操作 26

3.7.2 实验结果与分析讨论 26

第四章 结语 28

参考文献 29

致谢 32

第一章 文献综述

1.1关于2-苯乙醇的简述

1.1.1 2-苯乙醇的应用和市场

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