论文总字数：19410字

摘 要

类胡萝卜素在生活中是一类重要的色素，在现实生活中动物和人类没有生成类胡萝卜素的能力，因此只能依靠外界进食摄取，并在体内进行一系列反应，从而达到对人体有利的效果。在人体中可利用的类胡萝卜素主要是α-胡萝卜素、β-胡萝卜素等。耐辐射奇球菌(简称DR)是一种可以在极端恶劣的环境下生存的微生物，它对辐射损伤的耐受能力极强，是目前已被发现的生物中最耐辐射的物种之一。它对可以引发DNA损伤致死和DNA突变致死的的各类致死因素都有较强的抵抗能力。本实验选用Deinococcus xibeiensis R13，它能够耐受辐照及紫外辐射，能够产生一种具有抗氧化作用的红色色素。本研究针对耐辐射菌株R13，对其产生类胡萝卜素的产量及生长情况进行研究，首先测定了R13的生长曲线，并对其产生的类胡萝卜素进行了提取和含量的测定。其次，对R13菌株在不同碳源，氮源，无机离子等培养基条件下的类胡萝卜素产量进行了测定。最后，对R13菌株在不同发酵条件如温度，转速等外部发酵条件下对类胡萝卜素产量的影响进行了测定。经实验研究发现，R13的最佳培养时间为48h，最适合的碳源是葡萄糖，最合适的氮源是胰蛋白胨，R13适合生活在无无机盐的环境中，培养温度和培养的接种量对R13的生长有显著影响。R13在优化后的培养基中，在30℃，200r/min的条件下，经过48h的发酵，类胡萝卜素产量达到了1358.42 ug/g，相比优化前产量提升了22.31%。

关键词：Deinococcus xibeiensis R13 耐辐射菌 类胡萝卜素 发酵条件优化

ABSTRACT

Carotenoids are also an important pigment in life. Carotenoids are generally found in nature. In real life, animals and humans do not have the ability to produce carotenoids. Therefore, they can only rely on outside food intake and in vivo. A series of reactions to achieve a beneficial effect on the human body. Carotenoids available in the human body are mainly α-carotene, β-carotene, lycopene, and the like. Radiation-resistant Deinococcus (abbreviated as DR) is a microorganism that can survive in extremely harsh environments. Its ability to withstand radiation damage is extremely strong. It is one of the most radiation-tolerant species in organisms that have been discovered so far. It has strong resistance to various types of lethal factors that can cause DNA damage killing and DNA mutation lethal. This experiment used Deinococcus xibeiensis R13, which can tolerate irradiation and ultraviolet radiation, can produce red pigment antioxidant. In this study, for radiation-resistant strain R13, the production and growth of carotenoids [the latter results were all carotenoid production, no growth OD] were studied. The growth curve of R13 was first determined [Growth curve in the results No.] and the carotenoids produced were extracted and assayed. Secondly, the carotenoid production of R13 strain was determined under different carbon sources, nitrogen sources, and inorganic ions. Finally, the carotenoid production of shrimp strain R13 was determined under different fermentation conditions such as temperature, rotational speed and other external fermentation conditions. The experimental study found that the best incubation time of R13 is 48h, the most suitable carbon source is glucose, the most suitable nitrogen source is tryptone, R13 is suitable for living in the environment without inorganic salts, the culture temperature and the amount of inoculum cultured The growth of R13 has a significant effect. After optimization, the carotenoids of R13 reached 1356.42ug/g, an increase of 22.31% compared to the output before optimization.

Keywords: Deinococcus xibeiensis R13 Radiation-resistant bacteria Carotenoids Optimization of fermentation conditions

目 录

摘要 I

ABSTRACT II

第一章 文 献 综 述 1

1.1 耐辐射奇异球菌的研究进展 1

1.1.1 耐辐射奇异球菌的简介 1

1.1.2 耐辐射奇异球菌的特征 1

1.1.3 耐辐射奇异球菌基因组的结构 2

1.1.4 耐辐射奇异球菌的抗辐射机制 3

1.2. 类胡萝卜素的研究进展 4

1.2.1类胡萝卜素的简介 4

1.2.2类胡萝卜素的功能 4

1.2.3类胡萝卜素的提取方法 5

1.3. 课题研究的意义与目的 6

第二章 实验材料和方法 7

2.1 实验材料 7

2.1.1 生产菌株 7

2.1.2 实验仪器 7

2.1.3 实验试剂 7

2.1.4 培养基及试剂配制 8

2.2 实验方法 9

2.2.1 类胡萝卜素的提取和测定 9

2.2.2 细胞干重的测定 9

2.2.3 耐辐射奇异球菌发酵培养基碳源种类优化 9

2.2.4 耐辐射奇异球菌发酵培养基氮源种类优化 10

2.2.5 耐辐射奇异球菌发酵培养基无机盐种类优化 10

2.2.6 Plackett-Burman设计筛选影响类胡萝卜素液体发酵的显著因素 10

第三章 实验结果和讨论 11

3.1 底盘细胞对类胡萝卜素的影响 11

3.2 R13的生长曲线 12

3.3 发酵时间对类胡萝卜素的影响 12

3.4碳源种类对类胡萝卜素的影响 13

3.5 氮源种类对类胡萝卜素的影响 14

3.6无机盐种类对类胡萝卜素的影响 15

3.7 Plackett-Burman设计筛选影响类胡萝卜素液体发酵的显著因素 15

3.8 HPLC检测R13类胡萝卜素 17

3.9 优化前后类胡萝卜素产量对比 17

第四章 实验结论 18

4.1 实验结论 18

参考文献 19

致谢 23

第一章 文 献 综 述

1耐辐射奇异球菌的研究进展

1.1.1 耐辐射奇异球菌的简介

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