研究报告
孙浩,于童,胡洪营.反渗透膜面细菌胞外多聚物提取方法研究[J].环境科学学报,2017,37(10):3629-3634
反渗透膜面细菌胞外多聚物提取方法研究
- Study on extraction method of extracellular polymeric substances from bacteria on reverse osmosis membrane
- 基金项目：国家重点研发计划（No.2016YFE0118800）
- 作者
- 单位
- 孙浩
- 清华大学环境学院, 环境模拟与污染控制国家重点联合实验室, 国家环境保护环境微生物利用与安全控制重点实验室, 北京 100084
- 于童
- 清华大学环境学院, 环境模拟与污染控制国家重点联合实验室, 国家环境保护环境微生物利用与安全控制重点实验室, 北京 100084
- 胡洪营
- 1. 清华大学环境学院, 环境模拟与污染控制国家重点联合实验室, 国家环境保护环境微生物利用与安全控制重点实验室, 北京 100084;2. 清华-伯克利深圳学院, 环境与新能源技术工程实验室, 深圳 518055
- 摘要：城市污水脱盐系统的反渗透（RO）膜面生物污堵问题严重影响了该技术的稳定运行，解析生物污堵机理的关键因素是微生物胞外多聚物（EPS）特征分析，然而目前细菌EPS提取方法多样，无统一标准，制约了EPS分析的可靠性和可对比性.基于此，本文以再生水厂RO膜面污堵物中分离的细菌为对象，采用水浴、超声、离子树脂、碱液4种方法提取EPS，以DOC和DNA含量表征提取的EPS总量和提取过程细胞破裂程度.结果表明，45℃水浴法不造成细胞破裂，提取总量多，是一种有效的提取方法.进一步对比不同提取时间条件下EPS溶液中多糖和蛋白含量变化，以及分子量、荧光特征等，综合认为提取1 h时，提取的EPS中多糖与蛋白相对含量稳定，不同分子量化合物均溶于水中，能客观地反映EPS特征.研究表明，45℃水浴1 h可以作为细菌EPS提取的适用方法应用.
- Abstract：The biofouling of reverse osmosis (RO) membrane seriously affects the operational stability of municipal reclaimed water desalination system. Microbial extracellular polymeric substance (EPS) analysis is the key factor to dissect the biofouling mechanism. Despite that there are many methods for the EPS extraction, there is still no standard method at the moment. This restricts the uniformity and comparison of biofouling analysis. This study uses the bacteria isolated from RO membrane as the research object, and EPS was extracted using 4 different methods, namely water bathing at 45℃, ultrasound, ion exchange resin, and NaOH. The concentrations of DOC and DNA were measured in EPS solution, to characterize the EPS amount and the degree of cells lysis during the extraction processes respectively. The results suggest that 45℃ water bathing can effectively extract bacterial EPS without cell lysis. Furthermore, comparing the total amount of EPS, polysaccharide and protein compositions, molecular weight, and fluorescence characteristics in EPS solutions with different extraction time under the condition of 45℃ water bathing, suggest that when heating the sample for 1 h, the concentration of polysaccharide and protein is relatively stable. It was also found that different molecular weight compounds are dissolved in water, which is objectively reflected the characteristics of EPS. In conclusion, 45℃ water bath for 1h can be used as an effective method for EPS extraction in pure bacteria system.
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