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Published on: July 28, 2016
Characterization of membrane-bound polyphenol oxidase from Granny Smith apple (Malus × domestica Borkh.)
Qianyun Han1, Fang Liu2, Yanling Hao3
1College of Food Science and Nutritional Engineering, China Agricultural University, No. 17 Qinghua East Road, Beijing 100083, China; National Engineering Research Center for Fruits and Vegetables Processing, Beijing 100083, China; Key Laboratory of Fruit and Vegetables Processing, Ministry of Agriculture, Beijing 100083, China.
Abstract:
Membrane-bound polyphenol oxidase (mPPO) from the Granny Smith apple was purified and characterized. The enzyme was purified by a factor of 20.53 with a recovery of 1.8%. The molecular weight of purified mPPO was determined to be 65 kDa by electrophoresis and nano-electrospray ionization mass spectrometry. mPPO exhibited its highest activity at a temperature of 35 °C and a pH of 7.0 and can be regarded as a diphenol oxidase. A low concentration of SDS (≤0.5 mM) enhanced the enzymatic activity, whereas mPPO was activated at high concentration EDTA (≥2 mM). The thermal transition temperature of mPPO was 76.98 °C. The circular dichroism spectrum showed that mPPO contains high α-helix content, the fluorescence spectroscopy indicated that the tryptophan residues of mPPO are partially buried. The particle size of mPPO was 5-10 nm with a complete structure. The structural characterization of mPPO provided better insights into the regions responsible for its activity.
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