Open AccessPurification and biochemical characterization of polyphenol oxidase from seeds of melon (Citrullus colocynthis)
Author(s) -
David Morakinyo Sanni,
Catherine Joke Adeseko,
Samuel Olufemi Bamidele
Publication year - 2020
Publication title -
nova biotechnologica et chimica
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.212
H-Index - 9
eISSN - 1339-004X
pISSN - 1338-6905
DOI - 10.36547/nbc.v19i2.777
Subject(s) - chemistry , gallic acid , polyphenol oxidase , catechol , citrullus colocynthis , ascorbic acid , catechol oxidase , substrate (aquarium) , tyrosinase , browning , enzyme , enzyme assay , food science , nuclear chemistry , biochemistry , chromatography , botany , antioxidant , peroxidase , oceanography , biology , geology
Polyphenol oxidase (PPO) is an enzyme that is responsible for the enzymatic browning of fruits and vegetables. This is generally undesired process and need to be prevented in food technology. PPO from seeds of Citrullus colocynthis was purified, the physicochemical properties such as effects of pH and temperature, substrate specificity, effects of inhibitors and cations on PPO activity and the kinetic parameters for four substrates namely, catechol, L-DOPA, gallic acid and tyrosine, were determined. The purification steps resulted in 41-fold with 10 % yield, and the optima pH and temperature values for PPO from C. colocynthis were found to be pH 7.0 and 60 °C, respectively using catechol as substrate. About 9 % enzyme initial activity was retained after 60 min of incubation at 80 °C, and the apparent molecular weight was determined as 42 kDa by partially denaturing SDS-PAGE. PPO activity was inhibited by ascorbic acid, SDS and certain divalent (Ca2+, Zn2+, Mg2+ and, Fe2+) and monovalent (Na+) metal. Moreover, purified enzyme solution showed diphenolase activity toward catechol, gallic acid, L-DOPA and monophenolase activity toward tyrosine, therefore, tyrosinase was identified as the only one PPO in C. colocynthis seeds. This study revealed the use of temperature above 80 °C to inhibit PPO activity during processing and storage of melon seeds.