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Purification and characterization of peroxidase from avocado ( Persea americana Mill, cv. Hass)
Author(s) -
RojasReyes José O,
RoblesOlvera Victor,
CarvajalZarrabal Octavio,
Castro Matinez Claudia,
Waliszewski Krzysztof N,
AguilarUscanga María Guadalupe
Publication year - 2014
Publication title -
journal of the science of food and agriculture
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.782
H-Index - 142
eISSN - 1097-0010
pISSN - 0022-5142
DOI - 10.1002/jsfa.6503
Subject(s) - persea , peroxidase , ascorbic acid , chemistry , polyphenol oxidase , abts , pleurotus , isoelectric point , chromatography , substrate (aquarium) , enzyme , biochemistry , food science , mushroom , botany , antioxidant , biology , dpph , ecology
BACKGROUND Avocado ( Persea americana Mill, cv. Hass) fruit ranks tenth in terms of the most important products for Mexico. Avocado products are quite unstable due to the presence of oxidative enzymes such as polyphenol oxidase and peroxidase. The present study is to characterize the activity of purified avocado peroxidase from avocado in order to ascertain the biochemical and kinetic properties and their inhibition conditions . RESULTS Purification was performed by Sephacryl S 200 HR gel filtration chromatography and its estimated molecular weight was 40 kDa . The zymogram showed an isoelectric point of 4.7. Six substrates were tested in order to ascertain the affinity of the enzyme for these substrates. The purified peroxidase was found to have low K m (0.296 mM ) and high catalytic efficiency (2688 mM −1 s −1 ) using 2,2′‐azino‐bis(3‐ethylbenzthiazoline‐6‐sulfonic acid), optimum activity being reached at 51°C, pH 3.8. The addition of dithiothreitol, β ‐mercaptoethanol, ascorbic acid, sodium azide, l‐cysteine and Tween‐20 had high inhibitory effects, while metals ions such as Cu + , Fe 2+ and M n 2+ had weak inhibitory activity on purified avocado peroxidase . CONCLUSION The purified avocado peroxidase exhibits high inhibition ( K i = 0.37 µM ) with 1.97 µM n ‐propyl gallate using ABTS as substrate at 51°C, pH 3.8 for 10 min. © 2013 Society of Chemical Industry