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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