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Purification and Partial Properties of a Superoxide Dismutase from Agave Hybrid No.11648
Authors:Bing Chen  Liang Chen  Li‐Jun Gao  Su‐Yu Ye  Rui Xiao  Pei‐Lian Liu  Bi‐Lian Chen  Yun‐Xia Zhang  Jun Yao
Institution:1. Chemistry Science and Technology School, Zhanjiang Normal University, 29 Cunjin Road, Zhanjiang 524048, P. R. China;2. Development Centre for New Materials Engineering & Technology in Universities of Guangdong, 29 Cunjin Road, Zhanjiang 524048, P. R. China;3. The Department of Traditional Chinese Medicine, Central People's Hospital of Zhanjiang, 2 Cunjin Road, Zhanjiang 524037, P. R. China
Abstract:A superoxide dismutase enzyme was extracted from the leaves of Agave Hybrid No.11648 using homogenisation, ammonium sulphate fractional precipitation, diethylaminoethyl cellulose ion exchange and Sephadex gel filtration chromatography. A superoxide dismutase parallelogram‐shaped crystal was formed after packing at 4 °C for two days. The optimum pH was 8, and the optimum temperature was 40 °C. The crystal was relatively stable at temperatures below 60 °C and at pH values from 5 to 10. The subunit size was determined to be approximately 27.5 kDa by sodium dodecyl sulphate‐polyacrylamide gel electrophoresis. It was also demonstrated that the enzyme is an iron‐containing superoxide dismutase based on its sensitivity to inhibitors of potassium cyanide, hydrogen peroxide and chloroform‐ethanol and based on the results of ultraviolet absorption spectroscopy.
Keywords:Agave hybrid No  11648  Superoxide dismutase  Purification  Properties  Purification
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