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Purification and Partial Properties of a Superoxide Dismutase from Agave Hybrid No.11648
Author(s) -
Chen Bing,
Chen Liang,
Gao LiJun,
Ye SuYu,
Xiao Rui,
Liu PeiLian,
Chen BiLian,
Zhang YunXia,
Yao Jun
Publication year - 2012
Publication title -
journal of the chinese chemical society
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.329
H-Index - 45
eISSN - 2192-6549
pISSN - 0009-4536
DOI - 10.1002/jccs.201100462
Subject(s) - chemistry , superoxide dismutase , sephadex , size exclusion chromatography , potassium cyanide , hydrogen peroxide , chromatography , chloroform , polyacrylamide gel electrophoresis , ammonium sulfate precipitation , sodium , nuclear chemistry , sodium dodecyl sulfate , cyanide , enzyme , biochemistry , inorganic chemistry , organic chemistry
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.