Optimizations of extraction and activity determination methods of nitric oxide reductase in activated sludge
YANG Rui1,2,3, YUAN Lin-jiang1,2,3, WANG Gang1,2,3, YUAN Lin-jie1,2,3, YOU Xiao-long1,2,3, NIU Wan-xia1,2,3, YU Li-ping1,2,3
1. School of Environment and Municipal Engineering, Xi'an University of Architecture and Technology, Xi'an 710055, China; 2. Key Laboratory of Environmental Engineering of Shaanxi Province, Xi'an 710055, China; 3. Key Laboratory of Northwest Water Resources and Environmental Ecology Ministry of Education, Xi'an 710055, China
Abstract：Orthogonal experiments were carried out to study the effects of different fragmentation methods (e.g. fragmentation by ultrasonic, fragmentation by low temperature and high-pressure) on the catalytic activity of nitric oxide reductase (nor). The release of intracellular soluble protein, nucleic acid (DNA) and the catalytic activity of nor were analyzed under different ultrasonic intensity and frequency, breaking pressure and frequency, and the amount of lysate addition. According to the catalytic characteristics of enzymes in activated sludge under different experimental conditions, the method for measuring the catalytic activity of nor was improved. According to the results, the parameters of nor extraction by ultrasonic fragmentation were suggested to set at 100times (ultrasonic times), 500W (ultrasonic intensity), and 0.1mL (amount of lysate addition). The parameters of nor extraction by low temperature and high-pressure fragmentation were suggested to set at 4times (breaking times), 50Mpa (breaking pressure), and 0.1mL (amount of lysate addition). The release of intracellular soluble protein, DNA and the maximum catalytic activity of nor by low temperature and high-pressure fragmentation were higher than that by ultrasonic fragmentation. Last but not least, the end time for activity determination of nor was suggested at 15min.
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