论文总字数：22974字

摘 要

随着人们生活方式和环境的变化及饮食习惯的多元化，人们面对的健康问题也越来越多。其中因为病原微生物而导致的感染性疾病是最为常见和频发的。D-对羟基苯甘氨酸作为一种合成β-内酰胺类半合成抗生素的一种非常重要的医药中间体，通常应用于头孢类广谱抗生素的制备。

通过对D-对羟基苯甘氨酸相关性质以及现代医药发展方向的分析，最终决定在本论文中介绍由恶臭假单胞菌—一菌两酶法生产D-对羟基苯甘氨酸。本文详细阐述了恶臭假单胞菌菌株的活化，筛选以及相关条件的调整以达到D-对羟基苯甘氨酸菌种的优化。主要内容是将冻干粉保存的菌种进行种子液活化，再接种到平板和斜面上进行一代，二代和三代菌种的传代，待菌种经过活化传代后，进行挑单菌落，筛选出更加优秀的菌株。对筛选后的菌株进行进一步的种子液活化和平板传代。用筛选传代后的三代菌种进行最优生长时间，诱导剂用量以及微量元素的考察。最后甘油冷冻保存菌种。

关键词：D-对羟基苯甘氨酸 恶臭假单胞菌 一菌两酶法

Selection and optimization of the preparation of D-hydroxy phenylyci

-ne

Abstract

With the change of people’s lifestyle and environment and the diversification of t-

heir dietary habits,people are facing more and more heath problems. Infectious diseas-

es caused by pathogenic microorganisms are the most frequent and frequent.D-hydrox

-ypHenylglycine is a main pHarmaceutical intermediate for the synthesis of betalacta-

m semisynthetic antibiotics,and it is used to produce cepHalosporin broad-spectrum a-

ntibiotics

By analyzing the related properties of D-hydroxypHenylglycine and the develop-

ment direction of modern medicine, we finally decided to introduce the production of D- hydroxypHenylglycine by Pseudomonas maloda - one - bacteria two enzyme meth

-od in this paper. In this paper, the activation, screening and related conditions of Pse-

udomonas putida were discussed in detail, so as to achieve the optimization of D-hydr -oxypHenylglycine.The main content is to activate the seed liquid preserved by the fr

-ozen dry powder, and then inoculate it on the flat and inclined surface for a generatio

-n, the two generation and the three generation of the strains. After the bacteria are act

-ivated and passed on, the single colony is selected and the more excellent strains are screened. Further seed solution activation and plate subculture were carried out on the screened strain. The optimal growth time, dosage of inducers and trace elements were screened by screening the three generation of strains after passage. Finally, the glycer

-ol was frozen to preserve the bacteria.

Key Words: D-hydroxypHenylglycine;Pseudomonas putida;two enzyme method

目录

摘要.....................................................Ⅰ

Abstract.................................................Ⅱ

第一章 文献综述...........................................1

1.1 D-对羟基苯苷氨酸的概略........................................1

1.1.1 D-对羟基苯甘氨酸的相关介绍..............................1

1.1.2 D-对羟基苯苷氨酸的多样用途..............................1

1.1.3 D-对羟基苯甘氨酸的应用前景..............................2

1.2 制备D-对羟基苯苷氨酸菌种的筛选和优化.........................3

1.2.1 生物酶法................................................4

1.2.2 化学合成法..............................................5

1.2.3 拆分法..................................................5

1.3 课题研究目的,意义和内容.......................................5

1.3.1 课题研究目的,意义.......................................5

1.3.2 课题研究内容............................................6

第二章 实验的仪器,试剂和培养基.................................7

2.1 实验的仪器,试剂...............................................7

2.2 培养基的配制..................................................8

第三章 实验步骤...........................................11

3.1 菌种的复壮...................................................11

3.2 菌种的筛选优化...............................................11

3.3 菌种的传代培养及保存.........................................12

3.4 菌种的发酵...................................................13

3.5 底物的转化...................................................14

3.6 实验分析方法.................................................14

3.6.1 菌种生长情况的观察检测...................................14

3.6.2 菌种生物量的测定分析.....................................15

3.6.3 菌种发酵转化后底物、中间体及产物的检测分析...............16

3.6.4 高效液相的检测分析.......................................16

3.6.5 红外光谱检测.............................................17

第四章 菌种的相关考察.........................................18

4.1 冻干粉种子液活化前后生物量...................................18

4.2 菌种培养时间的考察...........................................18

4.3 诱导剂用量对菌种的影响.......................................20

4.4 发酵液中CoCl₂用量对的影菌种生物量的影响......................23

4.5 底物转化结论分析.............................................25

第五章 结论与展望...............................................29

5.1 结论...........................................................29

5.2 展望...........................................................29

参考文献....................................................31

致谢..............................................................33

请支付后下载全文，论文总字数：22974字