A Microplate Fluorimetric Assay for Sacchariase Activity Measurement
ZHANG Li-li1, 2, WU Zhi-jie1*, CHEN Li-jun1, LI Dong-po1, MA Xing-zhu1, 2, SHI Yun-feng1, 2
1. Instituite of Applied Ecology, Chinese Academy of Sciences, Shenyang 110016, China 2. Graduate University of Chinese Academy of Sciences, Beijing 100039, China
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.
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