期刊
ENZYME AND MICROBIAL TECHNOLOGY
卷 71, 期 -, 页码 8-12出版社
ELSEVIER SCIENCE INC
DOI: 10.1016/j.enzmictec.2015.01.005
关键词
Candida rugosa lipase; p-nitrophenyl palmitate; Lipase enzymatic activity measurement; Uniform aqueous-based solution
资金
- National Natural Science Foundation of China [21274126, 51473143]
- National Twelfth Five-Year Plan for Science & Technology Support of China [2012BAI08B01]
We proposed a convenient and accurate method for the measurement of lipase activity in a uniform aqueous-based substrate solution. In this work, lipase from Candida rugosa was used as the model lipase to test its catalytic ability toward p-nitrophenyl palmitate (p-NPP), which was suspended in a mixture of p-NPP ethanol solution and buffer. An ultraviolet-visible spectrophotometer was used to efficiently measure the liberated p-nitrophenol without extraction or centrifugation. Several factors that affected lipase activity were investigated, such as the ratio of p-NPP ethanol solution to buffer, the concentrations of p-NPP and lipase, as well as the temperature, reaction time, pH and agitation rate. Additionally, enzyme catalytic parameters such as K-m, V-m and activation energy were also assessed. We determined the optimal conditions for lipase in this homogeneous system and demonstrated lipase's catalytic performance in this condition followed Michealis-Menten kinetics. (C) 2015 Elsevier Inc. All rights reserved.
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