论文总字数：21398字

摘 要

Baeyer-Villiger（拜尔-维利格）氧化反应在有机化学中占有相当重要的地位，涉及到酮在过氧化物条件下生成酯，酯类或内酯类具有广泛有效的生物学活性包括抗病毒，抗生素活性，并且能够进一步合成类固醇类物质等， 是有机合成和药物化学中重要的中间体，这种反应的研究潜力很大，并且具有一定生物活性，因此受到了人们的广泛关注。

随着对Baeryer-Villiger反应的不断研究，人们对其反应机理以及催化体系也有了进一步的了解。在这之中，具有高效催化性能的拜尔维利格单加氧酶成为了目前研究的主流方向。本文以2-丁酮为反应底物，选择了CHMO和PAMO的几种突变体，以2-丁酮为底物测试他们的活性和区域选择性。

本课题旨在研究苯丙酮单加氧酶及其突变体催化的非常规Baeyer-Villiger反应并且提高产物的区域选择性，实验的原料便于取得，实验的操作步骤并不困难，降低实验的经济成本，在有机合成方面有一定的研究和发展前景。

关键词 Baeyer-Villiger氧化反应； 单加氧酶； 2-丁酮； 生物催化； 区域选择性；

Abstract

The Baeyer-Villiger reaction is an important type of oxidation reaction in organic chemistry involving the production of esters or lactones under the conditions of peroxide, which have a wide range of effective biological activities including antiviral and antibiotic activity, and it is an important intermediate in organic synthesis and pharmacochemistry, which has high research value and has certain biological activity.

With the continuous research on the reaction of Baeryer-Villiger, people have a better understanding of their reaction mechanism and catalytic system. Among them, the Bayerville cells with high catalytic performance has become the mainstream of the current research. In this paper, several mutants of CHMO and PAMO were selected for regioselective boicatalysis reaction using 2-butanone as model substrate. After the optimal reaction conditions were determined, the possible mechanism of the reaction was deduced by conditional control experiment and literature review.

The aim of this study was to investigate the unconventional Baeyer-Villiger reaction catalyzed by Baeyer-Villiger monooxygenase and their mutants and to improve the regioselectivity of the biocatalysis reaction. The experimental raw materials were easy to obtain and the experimental procedure was not difficult to reduce the economic cost, in the organic synthesis of a certain degree of research and development prospects.

Key words ：Baeyer-Villiger oxidation reaction；Monooxygenase；2-Butanone； Biocatalysis；Regioselectivity;

目 录

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

ABSTRACT...............................................................................................Ⅱ

1. 文献综述......................................................................................3

1.1 Baeyer-Villiger氧化反应...............................................................3

1.1.1 Baeyer-Villiger反应的概要.........................................................3

1.1.2 Baeyer-Villiger反应的机理.........................................................3

1.1.3 Baeyer-Villiger反应的应用.........................................................4

1.1.4 Baeyer-Villiger反应的催化剂......................................................5

1.2 Baeyer-Villiger单加氧酶...............................................................7

1.2.1 Baeyer-Villiger单加氧酶的概要....................................................7

1.2.2 Baeyer-Villiger单加氧酶的催化机理.............................................9

1.2.3 Baeyer-Villiger单加氧酶的选择性...............................................10

1.2.4 Baeyer-Villiger单加氧酶的展望..................................................11

1.3 课题研究的内容及意义...................................................................12

第二章 实验部分....................................................................................14

2.1 仪器与试剂....................................................................................14

2.1.1 实验仪器.................................................................................14

2.1.2 实验试剂.................................................................................15

2.2 实验操作.......................................................................................15

2.2.1 配置培养基与灭菌操作...............................................................15

2.2.2 单加氧酶基因的重组质粒的构建...................................................16

2.2.3 SDS-PAGE蛋白凝胶电泳................................................................17

2.2.4 测试Baeye-Villiger反应的活性..................................................18

2.2.5 气相色谱.................................................................................18

2.3 结果分析.......................................................................................19

2.3.1 SDS-PAGE电泳结果分析...............................................................19

2.3.2 气相色谱分析............................................................................20

第三章 总结............................................................................................23

参考文献..................................................................................................24

致谢......................................................................................................... 26

第一章 文献综述

1.1 Baeyer-Villiger氧化反应

1.1.1 Baeyer-Villiger反应的概要

Baeyer-Villiger（拜尔-维利格）反应是一种氧化反应，它指的是在酮的羰基和与羰基相邻的烃基之间增加一个氧原子,这样原来的酮就变成了酯，如果反应物是环酮就变成内酯，这种反应必须在有过氧化物(比如过氧化羧酸和过氧化氢等)存在时才能发生^[1]，我们常常在反应的时候在酮或者环酮之中加入一种能使溶液保持在中性的物质，比如说磷酸氢二纳，这样做可以防止生成的酯发生互相交换的反应，从而便于使反应顺利进行。

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