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[1]李崇华 马贵党 尹 斌 焦小轩 季荣博 张春华 葛 滢**.莱茵衣藻胞外聚合物的提取和红外光谱表征*[J].应用与环境生物学报,2019,25(01):1-14.[doi:10.19675/j.cnki.1006-687x.2018.04020]
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莱茵衣藻胞外聚合物的提取和红外光谱表征*()
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《应用与环境生物学报》[ISSN:1006-687X/CN:51-1482/Q]

卷:
25卷
期数:
2019年01期
页码:
1-14
栏目:
研究论文
出版日期:
2019-02-25

文章信息/Info

Title:

Extraction and Infrared Spectrum Characterization of Extracellular Polymeric Substances (EPS) from Chlamydomonas reinhardtii*
文章编号:
201804020
作者:
李崇华1 马贵党1 尹 斌1 焦小轩1 季荣博1 张春华2 葛 滢1**
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
Author(s):
LI Chonghua1, MA Guidang1, YIN Bin1, JIAO Xiaoxuan1, JI Rongbo1, ZHANG Chunhua2 & GE Ying 1** 1College of Resources and Environmental Sciences, Jiangsu Provincial Key Laboratory of Marine Biology, Nanjing Agricultural University, Nanjing 210095, China?
2Demonstration Laboratory of Elements and Life Science Research, Laboratory Centre of Life Science, Nanjing Agricultural University, Nanjing 210095, China
关键词:
莱茵衣藻胞外聚合物提取方法多糖蛋白质
Keywords:
Chlamydomonas reinhardtii EPS extraction method polysaccharides protein
DOI:
10.19675/j.cnki.1006-687x.2018.04020
摘要:
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。本文以模式微藻--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。总之,加热法是莱茵衣藻EPS的最适宜提取方法,EPS的官能团信息为后续研究莱茵衣藻EPS与重金属之间的相互作用奠定了基础。
Abstract:
Extracellular polymeric substances are macromolecules produced in the metabolic process of microbial cells. Many functional groups from EPS may change speciation and activities of elements in the surrounding environment by complexation and transformation. These processes indicate that EPS have potential in the application in bioremediation of pollution. In our study, the EPS of Chlamydomonas reinhardtii was extracted by heating, NaOH, EDTA, cation exchange resin (CER) and high speed centrifugation method. To develop an suitable method for EPS extraction from Chlamydomonas reinhardtii, the contents and composition of 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 extraced 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 C. reinhardtii cells were only slightly deactivated by heating compared with NaOH. 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 study on the interactions between EPS and heavy metals in this microalga.

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备注/Memo

备注/Memo:
收稿日期: 2018-04-17 接受日期 Accepted: 2018-05-23
*国家自然科学基金项目(31770548)、南京农业大学本科生科研训练项目(1713C12)资助?
**通讯作者(E-mail: yingge711@njau.edu.cn)
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更新日期/Last Update: 2018-07-13