摘  要：苯酚是环境中水污染的重要来源之一，为了寻找高效降解苯酚的微生物，本文从连云港入海口采集污泥样品并进行微生物的分离，经过富集分离和纯化，最终筛选出一株具有苯酚降解能力的菌株，编号为 SDP-2。然后对该菌株进行形态特征观察，并通过菌株16S rRNA基因PCR扩增与测序及系统发育分析进行鉴定。同时对该菌株的降解特性进行了初步研究，包括菌株的生长盐浓度和pH范围，菌株降解苯酚的最适底物浓度、接种量和最适pH。结果表明，该菌株与棒杆菌属的有效种谷氨酸棒杆菌（Corynebacterium glutamicum）的16S rRNA基因相似性为100%，因此鉴定为谷氨酸棒杆菌的菌株。菌株 SDP-2的最适生长盐浓度为 1%-5%，最适生长 pH 为8-9；菌株降解苯酚的最适底物浓度为500 mg/L，最适接种量为 10%-15%，最适 pH 为8.0。论文为后续微生物降解苯酚的深入研究提供了前期基础和菌种资源。

该论文有图9幅，表1个，参考文献16篇。

关键词：海洋细菌  16S rRNA基因  生长特性  降解特性

Enrichment-isolation and Identification of a Phenol Degrading Bacterium Isolated from Marine

Abstract： Phenol is one of the most harmful sources of water pollution in environment. In order to search for microorganisms which can degrade phenol efficiently, in this paper, the sludge samples were collected from the Lianyungang estuary and used for bacteria isolation. After enrichment isolation and purification, a strain with phenol degrading ability was obtained and named as SDP-2. The morphological characteristics of the strain were observed and it was identified by PCR amplification, sequencing and phylogenetic analysis of 16S rRNA gene. The degradation characteristics of the strain were also studied, including the salt concentration and pH range, the optimal substrate concentration, inoculum size and optimal pH. The results showed that the similarity of 16S rRNA gene between the strain and the type species of Corynebacterium glutamicum was 100%, and the strain was identified as Corynebacterium sp. The optimum growth concentration of SDP-2 was 1%-5%, the optimum growth of pH was 8-9, and the optimum substrate concentration of phenol was 500 mg/L, the optimum inoculum was 10%-15%, and the optimum pH was 8.0. This paper provides the basis for the further study of microbial degradation of phenol.

Key Words: Marine bacteria  16S rRNA gene  growth characteristics  Degradation characteristics

目  录

摘  要 III

Abstract IV

目  录 V

图清单 VI

表清单 VI

变量注释表 VII

1 绪论 9

2 材料与方法 11

2.1材料 11

2.2 方法 12

2.3 苯酚降解菌 SDP-2 的初步鉴定 13

2.4 苯酚浓度的测定 13

2.5 苯酚降解菌 SDP-2的生长特性研究 14

2.6 菌株SDP-2 苯酚降解实验 15

3 结果与分析 16

3.1 菌株的分离与纯化 16

3.2 苯酚降解菌的 16S rRNA 序列分析 16

3.3 苯酚标准曲线 18

3.4 菌株SDP-2 的生长特性 19

3.5 菌株 SDP-2 的降解特性 21

4 结论 25

参考文献 26

致谢 27

图清单

图序号 图名称 页码

图3-1 菌株 SDP-2 在 LB 固体平板上的生长形态 16

图3-2 菌株 SDP-2 基于 16S rRNA 的系统发育树 18

图3-3 苯酚标准曲线 18

图3-4 菌株 SDP-2 的生长曲线 19

图3-5 盐浓度对 SDP-2 生长的影响 20

图3-6 pH 对 SDP-2 生长的影响 21

图3-7 底物浓度对菌株 SDP-2 降解苯酚的影响 22

图3-8 接种量对菌株 SDP-2 苯酚降解特性的影响 23

图3-9 pH 对菌株 SDP-2 苯酚降解特性的影响 24

表清单

表序号 表名称 页码

表2-1 实验仪器和设备 11

变量注释表

A 苯酚吸光度

LB 基础细菌培养基

rpm 摇床转速