|本期目录/Table of Contents|

 LI Chonghua,MA Guidang,YIN Bin,et al.Extraction and infrared spectrum characterization of extracellular polymeric substances (EPS) from Chlamydomonas reinhardtii[J].Chinese Journal of Applied & Environmental Biology,2019,25(01):176-183.[doi:10.19675/j.cnki.1006-687x.2018.04020]





Extraction and infrared spectrum characterization of extracellular polymeric substances (EPS) from Chlamydomonas reinhardtii
李崇华 马贵党 尹斌 焦小轩 季荣博 张春华 葛滢
1南京农业大学资源与环境科学学院,江苏省海洋生物学重点实验室 南京 210095 2南京农业大学生命科学实验中心,元素与生命科学研究示范实验室 南京 210095
LI Chonghua1 MA Guidang1 YIN Bin1 JIAO Xiaoxuan1 JI Rongbo1 ZHANG Chunhua2 & GE Ying1**
1 College of Resources and Environmental Sciences, Jiangsu Provincial Key Laboratory of Marine Biology, Nanjing Agricultural University, Nanjing 210095, China 2 Demonstration Laboratory of Elements and Life Science Research, Laboratory Centre of Life Science, Nanjing Agricultural University, Nanjing 210095, China
Chlamydomonas reinhardtii extracellular polymeric substances (EPS) extraction method polysaccharide protein
胞外聚合物(EPS)是微生物细胞代谢过程中产生的高分子物质,其中的多种官能团通过络合、转化等反应改变周围环境的元素形态和活性,在污染生物修复中具有应用前景. 以模式微藻——莱茵衣藻为研究材料,采用加热法、NaOH法、EDTA法、阳离子交换树脂(CER)法、高速离心法提取莱茵衣藻的EPS,比较分析每种方法得到的EPS的含量及其组成成分,并结合LIVE/DEAD BacLight染色和激光共聚焦显微镜观察细胞活性,以确定莱茵衣藻EPS最适宜的提取方法. 结果表明,不同提取方法得到的EPS各组分含量有显著差异,5种方法的提取总量依次为NaOH法>加热法>CER法>EDTA法>离心法. 荧光染色和激光共聚焦显微镜检测结果表明,相对于NaOH法,加热法对微藻细胞的破坏程度较小,且傅里叶红外光谱分析结果表明,加热法提取的EPS出现N-H、C=O或C-N(蛋白质)和C-H、C-O-C、RHC(OH)(OR)(糖类)等吸收峰. 本研究表明加热法是莱茵衣藻EPS的最适宜提取方法,EPS的官能团信息sk可为后续研究莱茵衣藻EPS与重金属之间的相互作用奠定基础. (图7 表4 参37)
Extracellular polymeric substances are macromolecules produced in the metabolic processes of microbial cells. Many functional groups from extracellular polymeric substances (EPS) might change the speciation and activities of elements in the surrounding environment via complexation and transformation. These processes indicate that EPS have potential application in the bioremediation of pollution. In this study, we extracted the EPS of Chlamydomonas reinhardtii using heating, NaOH, EDTA, cation exchange resin (CER), and high-speed centrifugation methods. To develop a suitable method for EPS extraction from C. reinhardtii, the contents and composition of the EPS obtained from each method were compared. The cell viabilities after EPS removal under different extractions were observed using LIVE/DEAD BacLight staining and laser confocal microscopy. The results showed that the contents of EPS components were significantly different among the extraction methods. The total amount of EPS extracted by different methods was ranked as follows: NaOH > heating > cation exchange resin (CER) > EDTA > centrifugation. LIVE/DEAD BacLight staining and laser confocal microscopy showed that the C. reinhardtii cells were only slightly deactivated by heating compared with the NaOH treatment. The Fourier-transform infrared spectroscopy (FTIR) analysis demonstrated that the EPS extracted by the heating method had functional groups, such as N-H, C=O, or C-N (protein) and C-H, C-O-C, RHC(OH)(OR) (carbohydrate). In summary, heating was the most suitable EPS extraction method for C. reinhardtii. The information of EPS functional groups provided a basis for further studies on the interactions between EPS and heavy metals in this microalga.


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更新日期/Last Update: 2019-02-25