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[一种用于测量蔗糖酶活性的微孔板荧光测定法]

[A microplate fluorimetric assay for sacchariase activity measurement].

作者信息

Zhang Li-li, Wu Zhi-jie, Chen Li-jun, Li Dong-po, Ma Xing-zhu, Shi Yun-feng

机构信息

Instituite of Applied Ecology, Chinese Academy of Sciences, Shenyang 110016, China.

出版信息

Guang Pu Xue Yu Guang Pu Fen Xi. 2009 May;29(5):1341-4.

Abstract

With the fluorescent compound conjugates substrates, soil xylanase and cellulose in a free-air carbon dioxide enrichment (FACE) experiment were measured on the base of 96 microplate and fluorescence detection, aiming at testing its feasibility in sacchariase activity measurement. The results show that sacchariase activity can be tested and the data exhibit better repeatability(coefficient of variability <= 4.879%). Compared with spectrophotometric assay, this method allows a large number of soil samples and/or enzymes to be analyzed in a short time accurately and conveniently. Soil xylanase activity tends to be greater at elevated CO2 which significantly increases in jointing, heading and ripening stages of wheat and in heading and ripening stages of rice (P < 0.05), and the crop metabolizes rapidly under FACE condition and soil microorganisms are affected, which causes elevation of xylanase acitivy. Compared with ambient CO2, soil cellulose activity decreased slightly under elevated CO2 but there was no significant difference between treatments, indicating the cellulose activity was not influenced intensively in a short time.

摘要

以荧光化合物偶联物为底物,基于96孔板和荧光检测对自由空气二氧化碳浓度增高(FACE)试验中的土壤木聚糖酶和纤维素进行测定,旨在测试其在蔗糖酶活性测定中的可行性。结果表明,可以对蔗糖酶活性进行检测,且数据具有较好的重复性(变异系数≤4.879%)。与分光光度法相比,该方法能够在短时间内准确、便捷地分析大量土壤样品和/或酶。土壤木聚糖酶活性在CO2浓度升高时往往更高,在小麦的拔节期、抽穗期和成熟期以及水稻的抽穗期和成熟期显著增加(P<0.05),并且在FACE条件下作物代谢迅速,土壤微生物受到影响,这导致木聚糖酶活性升高。与环境CO2相比,CO2浓度升高时土壤纤维素活性略有下降,但处理之间没有显著差异,表明纤维素活性在短时间内没有受到强烈影响。

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