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Microplate fluorimetric assay of soil leucine aminopeptidase activity: alkalization is not needed before fluorescence reading

文献类型：期刊论文

作者Wang, Jipeng2,3; Huang, Shunyu3; He, Qingqing1,5; Bing, Haijian5; Chen, Xiao4; Zhang, Xifeng5; Tian, Xin1,5; Zhou, Jun5; Wilcke, Wolfgang2; Wu, Yanhong5
刊名BIOLOGY AND FERTILITY OF SOILS
出版日期2020-02-01
卷号56期号:2页码:281-285
ISSN号0178-2762
关键词7-Amino-4-methyl-coumarin L-Leucine-7-amido-4-methylcoumarin Leucine aminopeptidase Enzyme assay protocols Alkalization procedure Soil enzyme
DOI10.1007/s00374-019-01419-x
通讯作者Bing, Haijian(hjbing@imde.ac.cn) ; Wu, Yanhong(yhwu@imde.ac.cn)
产权排序3
文献子类Article
英文摘要The fluorescence responses of 7-amino-4-methylcoumarin (AMC) and L-leucine-7-amido-4-methylcoumarin (L-leucine-AMC) (the fluorescent dye and substrate used in the soil leucine aminopeptidase assay, respectively) to alkalization by NaOH and tris(hydroxymethyl)aminomethane (THAM) buffer were analyzed. The leucine aminopeptidase activities were also measured without and with the alkalization procedure using five soils with contrasting properties. The pH optimum of leucine aminopeptidase activity was 8 +/- 1 for the five studied soils. In this pH range, the alkalization procedure cannot increase the fluorescence of AMC to improve the assay sensitivity. Excess alkalization (final medium pH of 12 or higher) led to decreasing fluorescence of AMC with time and non-enzymatic hydrolysis of L-leucine-AMC, which caused irreproducible values of the calculated enzyme activities. Moreover, a medium pH of similar to 11 cannot terminate the enzymatic reaction, and thus it seems impractical to stop the enzyme reaction by alkalization without causing the unstable behavior of AMC and L-leucine-AMC. We recommend performing the soil leucine aminopeptidase assay at the optimal pH without alkalization and reading the fluorescence immediately after incubation. This approach may help to simplify and standardize the leucine aminopeptidase assay.
电子版国际标准刊号1432-0789
WOS关键词ENZYME-ACTIVITY ; STOICHIOMETRY ; NITROGEN
资助项目National Natural Science Foundation of China[41701288] ; National Natural Science Foundation of China[41630751] ; Youth Innovation Promotion Association, Chinese Academy of Sciences[2017424] ; Science & Technology Department of Sichuan Province[2018JY0163] ; China Scholarship Council[201708515106]
WOS研究方向Agriculture
语种英语
出版者SPRINGER
WOS记录号WOS:000515928600013
资助机构National Natural Science Foundation of China ; Youth Innovation Promotion Association, Chinese Academy of Sciences ; Science & Technology Department of Sichuan Province ; China Scholarship Council
源URL[http://ir.imde.ac.cn/handle/131551/34072]  
专题中国科学院水利部成都山地灾害与环境研究所
通讯作者Bing, Haijian; Wu, Yanhong
作者单位1.Univ Chinese Acad Sci, Beijing 100049, Peoples R China;
2.KIT, Inst Geog & Geoecol, D-76131 Karlsruhe, Germany;
3.Chengdu Univ Technol, Coll Ecol & Environm, Chengdu 610059, Peoples R China;
4.Karlsruhe Inst Technol, Inst Inorgan Chem, D-76131 Karlsruhe, Germany
5.Chinese Acad Sci, Inst Mt Hazards & Environm, Key Lab Mt Surface Proc & Ecol Regulat, 9,Block 4,Renminnanlu Rd, Chengdu, Peoples R China;
推荐引用方式
GB/T 7714		 Wang, Jipeng,Huang, Shunyu,He, Qingqing,et al. Microplate fluorimetric assay of soil leucine aminopeptidase activity: alkalization is not needed before fluorescence reading[J]. BIOLOGY AND FERTILITY OF SOILS,2020,56(2):281-285.
APA Wang, Jipeng.,Huang, Shunyu.,He, Qingqing.,Bing, Haijian.,Chen, Xiao.,...&Wu, Yanhong.(2020).Microplate fluorimetric assay of soil leucine aminopeptidase activity: alkalization is not needed before fluorescence reading.BIOLOGY AND FERTILITY OF SOILS,56(2),281-285.
MLA Wang, Jipeng,et al."Microplate fluorimetric assay of soil leucine aminopeptidase activity: alkalization is not needed before fluorescence reading".BIOLOGY AND FERTILITY OF SOILS 56.2(2020):281-285.

入库方式： OAI收割

来源：成都山地灾害与环境研究所

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