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引用本文:庞晓宇,段洪涛,张玉超,马荣华.富营养化湖泊水体中藻蓝蛋白提取方法的对比.湖泊科学,2014,26(5):799-806. DOI:10.18307/2014.0519
PANG Xiaoyu,DUAN Hongtao,ZHANG Yuchao,MA Ronghua.Comparison of the extraction methods of phycocyanin pigments in eutrophic lake waters. J. Lake Sci.2014,26(5):799-806. DOI:10.18307/2014.0519
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富营养化湖泊水体中藻蓝蛋白提取方法的对比
庞晓宇1,2, 段洪涛1, 张玉超1, 马荣华1
1.中国科学院南京地理与湖泊研究所湖泊与环境国家重点实验室, 南京 210008;2.西北大学城市与环境学院, 西安 710127
摘要:
藻蓝蛋白是蓝藻的指示型色素,目前还没有标准的提取方法,特别是萃取剂的选择品种较多,对测量结果影响较大.分别以室内培养的铜绿微囊藻和巢湖夏季野生蓝藻为提取对象,运用液氮反复冻融法破碎蓝藻细胞,分别以AsolctinCHAPS缓冲液(AC)、磷酸盐缓冲液(PBS)和Tris-HCl缓冲液来提取藻蓝蛋白,通过分光光度法分析提取液的吸收光谱、藻蓝蛋白浓度以及藻蓝蛋白浓度与叶绿素a浓度的相关性,比较3种缓冲液的提取效果.结果显示:3种缓冲液提取的藻蓝蛋白提取液在620 nm处都出现了藻蓝蛋白的特征吸收峰,但AC缓冲液和PBS缓冲液提取效率明显高于Tris-HCl缓冲液.另外,虽然AC缓冲液提取效率最高,但其成本昂贵、配制过程复杂且不易控制、难保存,不适宜大规模地推广应用;而PBS缓冲液的提取效率略低于AC缓冲液,但其配制方便快捷,更适合于大规模湖泊水质监测的要求,因此推荐PBS缓冲液作为常用的藻蓝蛋白提取液.
关键词:  藻蓝蛋白  Asolctin-CHAPS缓冲液  磷酸盐缓冲液  Tris-HCl缓冲液  蓝藻  色素提取  巢湖
DOI:10.18307/2014.0519
分类号:
基金项目:国家高技术研究发展计划"863"项目(2014AA06A509);国家自然科学基金项目(41171271,41171273,41101316);国家水体污染控制与治理科技重大专项项目(2012ZX07103-005);中国科学院南京地理与湖泊研究所"一三五"战略发展规划项目(NIGLAS2012135014,NIGLAS2012135010)联合资助
Comparison of the extraction methods of phycocyanin pigments in eutrophic lake waters
PANG Xiaoyu1,2, DUAN Hongtao1, ZHANG Yuchao1, MA Ronghua1
1.State Key Laboratory of Lake Science and Environment, Nanjing Institute of Geography and Limnology, Chinese Academy of Sciences, Nanjing 210008, P. R. China;2.College of Urban and Environmental Sciences, Northwestern University, Xi'an 710127, P. R. China
Abstract:
The paper compares the effects of three methods of phycocyanin extraction buffer and tries to find out which method is the best. Cultured Microcystis aeruginosa and cyanobacteria blooms water samples of Lake Chaohu were used as extract objects. By repeated freezing and thawing method, cyanobacterial cells were broken, and then applied with Asolctin-CHAPS buffer, phosphate buffer and Tris-HCl buffer solution as extractant agent for extracting phycocyanin. Finally, spectrophotometry was used to detect the content of phycocyanin. We analyzed the absorption spectrum of the phycocyanin extract, the concentration of phycocyanin and the relevance between phycocyanin concentration and chlorophyll-a concentration to compare the quality of three methods. Absorption spectra show that all the characteristic absorption peaks of phycocyanin are emerged at 620 nm. Experimental results show that AC buffer and PBS buffer are better than the Tris-HCl buffer in extraction efficiency. In comparison, AC buffer is more expensive and difficult to obtain and save than the other two buffers, which makes it not suitable for large-scale use. So we recommend PBS buffer as the normal buffer which can meet the requirements of large-scale water quality monitoring.
Key words:  Phycocyanin  Asolctin-CHAPS buffer  PBS buffer  Tris-HCl buffer  cyanobacteria  pigment extraction  Lake Chaohu
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