【作者】 贾凌志；

【导师】 李君文；

【作者基本信息】 中国人民解放军军事医学科学院 ， 劳动卫生与环境卫生， 2004， 硕士

【摘要】 当今，环境内分泌干扰物对人类健康造成了危害。双酚A(bisphenol A，Bpa)是其中重要的一种。随着Bpa的大量使用，在世界各地各种水体中都有发现，对Bpa污染的治理势在必行。为此，我们开展了下面一些研究。 1．Bpa降解菌的筛选及鉴定。采用经典的细菌筛选技术，得到了Bpa高效降解菌(W2)，该菌可在饱和的Bpa溶液里以其为唯一碳源和能源生长。具有较强的Bpa降解能力。经过种属鉴定，该菌株属于假单胞菌属。 2．Pseudomonas sp.W2培养条件的优化。在实验室里探讨碳源、氮源、通气量、起始pH值、温度等因素对菌株生长及对Bpa降解能力的影响。菌体最佳生长条件为：pH7.0，250ml摇瓶装液量为50ml，温度30℃；Bpa最佳降解条件为：pH7.0，250ml摇瓶装液量为100ml，35℃。 3．Pseudomonas sp.W2降解Bpa机理初探。首先，利用分子生物学技术对该菌株降解Bpa起始关键基因与关键酶进行了初步研究，发现降解Bpa的起始关键基因定位在一个19kb左右的质粒上，起始关键酶是一种胞外酶。其次，利用现代分析化学技术研究Pseudonomas sp.W2对Bpa的代谢途径，发现对羟基苯甲酸、对羟基苯甲醛、对羟基苯乙酮为三种中间代谢产物。最后对该菌株的原儿茶酸双加氧酶基因pcaG进行了扩增测序。 本研究成果将为进一步开发Bpa治理微生物资源及发现Bpa降解关键基因研究奠定基础。更多还原

【Abstract】 Nowadays, endocrine disrupter chemicals are endangering human health. Of all EDCs, Bisphenol A (Bpa) is very important one. With Bpa used in great quantity,it is detected in all kinds of waters all over the world. Control of Bpa pollution is very necessary. So we did some correlative work.First of all, the isolation of strain able to degrade Bpa effectively and classify. A strain (W2) that was isolated ,which can degrade BPA very effectively and grow on saturation solution of Bpa as a sole carbon and energy sources. It was identified as pseudonomas sp.Secondly, study of the cultivation condition of pseudonomas sp. W2. In lab, we researched the effect of cabon source, nitrogen source, pH at beginning, temperature and aerate to the growth of strain W2 and degradation of Bpa. The optimal growth condition: pH7, 30℃, 50ml culture medium in 250ml flask. The optimal Bpa degradation condition: pH7,35℃, 100ml culture medium in 250ml flask.Thirdly, study of Bpa-degrading mechanism of pseudonomas sp.W2. Through the moleculor biology technique, we acknowledged that the Bpa-degrading enzyme is out-cell and the key gene is located at a plasmit about 19kb. Then, we studied the pathway of bacterial metabolism of Bpa and identified three kinds of metabolon of p-Hydroxy benzoic acid , p-Hydroxy benzaldehyde and p-Hydroxy acetophenone.Finally,we amplified and sequenced the pcaG gene.Our study result laid a foundation for exploitation of environmental Bpa-remeding microorganism and for building a gene-engineering microorganism to promote bioremediation of Bpa contamination.更多还原