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硅藻粘附性EPS提取方法的比较研究
于兆伟,谢雅清,张珮璇,鲍金,孙陈炼,靳翠丽,周晓见
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(1.扬州大学环境科学与工程学院,江苏 扬州 225127; 2.扬州大学海洋科学与技术研究所,江苏 扬州 225127)
摘要:
底栖硅藻是可见光能照射到的水下表面生物膜中的主要污损生物,胞外多聚物(Extracellular Polymeric Substances, EPS)是硅藻生物膜形成和发育过程中的关键物质,可溶性EPS和粘附性EPS的提取方法不同,对粘附性EPS的分离提取仍没有统一的方法。本研究选用了4种方法提取底栖硅藻(Amphora sp.)粘附性EPS:30℃水浴处理、70℃水浴处理、福尔马林-NaOH处理和阳离子交换树脂处理,研究了不同分离方法条件下,Amphora sp.粘附性EPS中各主要成分的提取量。结果表明,粘附性EPS中各主要成分普遍受提取方法的影响,其中DNA含量变化幅度最大,其次是蛋白质、总糖和糖醛酸含量,硫酸基含量受提取方法的影响最小。低温水浴处理提取效率低,高温水浴处理可能影响EPS的后续分析,阳离子交换树脂提取粘附性EPS的效果较好,但对细胞的破坏最严重,福尔马林-NaOH处理提取的EPS总量最高,提取总糖、糖醛酸的效果最好,而且没有造成严重的细胞破裂。因此福尔马林-NaOH处理是相对均衡和高效的提取硅藻粘附性EPS的方法。
关键词:  海洋生物学  硅藻  粘附性EPS  提取  福尔马林-NaOH处理
DOI:
基金项目:国家自然科学基金资助项目(41776156、41271521);扬州大学大学生科技创新资助项目(x20180466)
Comparison of extraction methods for diatom bound EPS
YU Zhao-wei,XIE Ya-qing,ZHANG Pei-xuan,BAO Jin,SUN Chen-lian,JIN Cui-li,ZHOU Xiao-jian
(1.College of Environmental Science and Engineering, Yangzhou University, Yangzhou 225127, China;2. Marine Science and Technology Institute, Yangzhou University, Yangzhou 225127, China)
Abstract:
Diatoms are the biofouling organisms that live on the underwater biofilm in the light reachable depth. The extracellular polymeric substances (EPS) is an important factor for diatoms adhering on the surfaces and plays crucial roles in biofilm formation and development. The methods to extract the soluble EPS and the bound EPS are different and no accredited extraction methods available for the bound EPS until now. In this study, the components of the bound EPS of benthic diatom Amphora sp. were studied with four different extraction methods including water bath treatment at 30℃ and 70℃, formalin-sodium hydroxide treatment and cationic exchange resin treatment. The results showed that the main components of bound EPS varied generally with the extraction methods, among which the range of DNA content changed the most, the next were protein, sugar and uronic acid, the least was sulfonic acid. The extraction efficiency by low temperature water bath was relatively low while high temperature water bath might have negative influences on the subsequent EPS analysis. The cationic exchange resin treatment showed a good extraction efficiency but it caused serious damages to cells. The method of formalinsodium hydroxide treatment gave a balanced and efficient result as the extraction derived the highest total EPS especially for sugar and uronic acid and less serious damage to the cells.
Key words:  marine biology  diatom  bound EPS  extraction  formalin-sodium hydroxide treatment

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