Citation: WU Sheng-Nan, ZHANG Hao, JIANG Ling, HUO Feng-Wei, TIAN Dan-Bi. A Fluorescence Sensor for Determination of Lipase Activity Based on Curcumin-Hg2+ Complex[J]. Chinese Journal of Analytical Chemistry, ;2018, 46(10): 1588-1594. doi: 10.11895/j.issn.0253-3820.181187
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A biosensor for detecting lipase activity was constructed by using the Tween 40-curcumin complex micelle as a fluorescence probe, Hg2+ as a quenching agent and methyl thioglycolate as the substrate of lipase. Hg2+ combined with the β-diketone structure of curcumin, resulting in quenching of Tween 40-curcumin complex micelle fluorescence. However, upon the addition of lipase, lipase could catalyze the hydrolysis of methyl thioglycolate to produce mercaptoacetic acid which could capture Hg2+ released from the curcumin-Hg2+ complex, resulting in the recovery of fluorescence of the Tween 40-curcumin complex micelle. The intensity of fluorescence recovery was related to lipase activity. The fluorescence sensing strategy was applied to quantitatively detect lipase activity. Under optimal experimental conditions, the change of fluorescence intensity was linear with the concentration of lipase in the range of 2-80 U/mL, and the detection limit was 0.01 U/mL. This method was employed to detect the activity of four other kinds of commercial lipases. The results were in agreement with those determined by potentiometric titration. The results showed that the method had the advantages of low cost, simple operation, high sensitivity, good practicability, and could be used for high throughput detection of lipase activity.
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