摘要： β-苯乙醇是一种带有甜和轻柔的玫瑰香味的芳香醇。β-苯乙醇的应用十分广泛，主要包括食品、香料、新型燃料、化妆品和医药等行业。随着人们对天然产品需求的日益增长，微生物转化的方法生产天然β-苯乙醇越来越受到人们的重视。目前工业上生物转化法合成β-苯乙醇大都是利用Ehrlich途径。本课题以本实验室自主筛选的肠杆菌MF024为研究对象，采用外源性生物中间体加入的方法，初步探索此株肠杆菌合成β-苯乙醇的生化路径。实验结果表明，该菌株可以利用L-苯丙氨酸、苯丙酮酸、苯乙醛为底物生物转化合成β-苯乙醇，且以苯乙醛为底物的生物转化率最高，以苯丙酮酸为底物转化率次之，以L-苯丙氨酸为底物转化率最低。可以初步认为肠杆菌MF024是可以通过Ehrlich途径生物转化合成β-苯乙醇的。为进一步分子生物学研究代谢路径奠定基础，也为进一步构建高产β-苯乙醇工程菌提供理论指导。

关键词： β-苯乙醇；Ehrlich途径；生物转化

Preliminary study on the metabolic pathway of fermentation of natural flavor β-Phenylalcohol by Enterobacter MF024

Abstrcat:β-Phenylethanol is an aromatic alcohol with a sweet, gentle rose aroma. β-Phenylethanol is widely used, including spices, foods, new fuels, and cosmetics and pharmaceuticals.With the increasing demand for natural products, the production of natural β-phenylethanol by microbial transformation has received increasing attention. At present, most of the industrial synthesis of β-phenylethyl alcohol using the Ehrlich pathway. In this research project, Enterobacteriaceae MF024, which was independently selected by our laboratory, was used as a research object. The method of adding exogenous biological intermediates was used to initially explore the biochemical route of the synthesis of β-phenylethanol by this strain of Enterobacteriaceae. The experimental results show that the strain can be used for the bioconversion of β-phenylethanol using L-phenylalanine, phenylpyruvate and phenylacetaldehyde as substrates, and the bioconversion rate is highest with phenylacetaldehyde as the substrate. The conversion rate of acid was the second, and the conversion rate of L-phenylalanine was the lowest. It can be preliminarily assumed that Enterobacteriaceae MF024 can biosynthesize β-phenylethanol through the Ehrlich pathway. It lays the foundation for the further study of metabolic pathways in molecular biology and provides theoretical guidance for the further construction of high-yield beta-phenylethanol engineering bacteria.

Keyword：β-phenylethyl alcohol； the Ehrlich pathway；Biotransformation

目录

目录

1. 绪论 4

1.1β-苯乙醇的基本性质及应用 4

1.1.1β-苯乙醛的基本性质 4

1.1.1 β-苯乙醛的应用范围 5

1.2 β-苯乙醇的国内外研究现状 5

1.3 微生物转化法合成β-苯乙醇 6

1.4β-苯乙醇的相关代谢途径 7

1.4.1 β-苯乙醇从头合成途径 7

1.4.2  Ehrlich途径 8

1.4.3  苯乙胺途径 9

1.5本课题的研究目的和意义 10

2.材料与仪器设备 11

2.1 材料与培养基 11

2.1.1 材料和试剂 11

2.1.2 培养基 11

2.2 仪器与设备 11

3. 实验方法 12

3.1菌体制备 12

3.2生物转化 12

3.2.1葡萄糖为底物 12

3.2.2L-苯丙氨酸为底物 13

3.2.3苯丙酮酸为底物 13

3.2.4苯乙醛为底物 13

3.3产物鉴定的实验方法 13

3.3.1气相色谱法的原理 13

3.3.2质谱原理 14

3.3.3标准曲线的绘制 14

3.3.4样品的检测 14

4. 结果与分析 15

4.1标准曲线的绘制 15

4.2葡萄糖为底物 15

4.2L-苯丙氨酸为底物 16

4.3苯丙酮酸为底物 16

4.4苯乙醛为底物 17

5. 结论与展望 19

5.1 结论总结 19

5.2 课题展望 19

致谢 20