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摘要对羟基苯甲酸（4-Hydroxybenzoate，4HBA）是自然界中普遍存在的芳香族化合物，是很多人工化合物的重要中间代谢产物，也是较常见的环境污染物之一。海洋环境中4HBA降解菌的研究，不仅可以更好地理解芳香族化合物的碳素循环，还有利于海洋环境污染的治理。本实验以4HBA作为唯一的碳源和能源，从浙江海底沉积物中分离和筛选出具有降解和利用4HBA能力的菌株M1。结合菌落特征、菌体形态、生理生化特征和16S rDNA序列分析，确定菌株M1为海杆菌属。通过单因素实验可知，菌株M1在20~35 ºC 均可生长，最适温度为35 ºC；在弱碱性条件下生长繁殖情况较好，最适pH为7.5。菌株M1具有较强的4HBA降解能力，48 h以内对4HBA降解率达到90%以上。有关海杆菌降解芳香族化合物的报道很少，菌株M1成功分离为利用海洋微生物治理环境污染问题的研究提供了一个新方向。42690
关键词：对羟基苯甲酸 芳香族化合物 降解 海洋细菌
Study on Degradation of 4-Hydroxybenzoate by Marine Microbial
Abstract
4-Hydroxybenzoate(4HBA) is a common aromatic compound in nature, and as a key intermediate metabolite for many artificial products. It is also one of the common environmental pollutants. Study on degradation of 4HBA by marine bacteria, not only can better understand the aromatic compounds, but also conducive to the management of marine environmental pollution. In this paper, we isolated strain M1 from marine sediment of Zhejiang sea, by using 4HBA as the sole carbon and energy source. Strain M1, capable of degrading 4HBA, was identified as Marinobacterium based on its morphologic, physiological characteristics and the analysis of 16S rDNA sequence. The single factor experiment showed that strain M1 could grow at 20 to 35 ºC and the optimum temperature was about 35 ºC. Alkalescent condition was suitable for the growth of strain M1 and the optimum pH was 7.5. The ability of strain M1 to degrade 4HBA was strong and could degrade over 90% of 4HBA within 72 h. So far, degradation of aromatic compounds by Marinobacterium was rarely reported. Therefore, the successful separation of strain M1 can provide a new research direction for the management of marine environmental pollution.
Key words: 4-hydroxybenzoate aromatic compound degradation marine bacteria
目录
摘要 I
Abstract II
表清单 IV
1绪论 1
2材料与方法 3
2.1材料与试剂 3
2.2仪器设备 4
2.3相关试剂溶液配制 5
2.4降解菌的筛选和纯化 6
2.6菌种保存 7
2.7革兰氏染色 7
2.8降解菌基因组总DNA提取 7
2.9 16S rDNA扩增 8
2.10 16S rDNA扩增产物检测及纯化 8
2.11 16S rDNA测序 9
2.12生理生化反应 9
2.13最适温度 9
2.14 最适pH 9
2.15 HPLC检测菌株对4HBA降解程度 10
3结果与分析 10
3.1海洋对羟基苯甲酸降解菌的富集培养、分离纯化和形态特征