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Prokaryotic expression and purification of Camellia sinensis polyphenol oxidase
Author(s) -
Liu Jingwei,
Huang Youyi,
Ding Jian,
Liu Cong,
Xiao Xiudan,
Ni Dejiang
Publication year - 2010
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.4111
Subject(s) - polyphenol oxidase , camellia sinensis , catechol , escherichia coli , chemistry , catechol oxidase , polyphenol , biochemistry , enzyme , oxidase test , food science , biology , botany , gene , antioxidant , peroxidase
BACKGROUND: Polyphenol oxidase (PPO) causes the postharvest loss of fruits and vegetables but is also a key factor in the quality development of tea. However, there are no reports on engineered active plant PPO purified from prokaryotic cells. RESULTS: In this study the ppo gene of about 1800 bp from Camellia sinensis cv. Yihongzao was successfully cloned and expressed in Escherichia coli . The PPOs purified from both the soluble fraction and the inclusion bodies showed activity. In addition, 1.0 × 10 −7 mol L −1 Cu 2+ and acidic conditions were found to be favourable for the engineered PPO catalysis of catechol oxidation. CONCLUSION: This paper represents the first report on C. sinensis ppo expression in E. coli and engineered active PPO purification. The results of the study provide a basis for the large‐scale preparation and application of PPO. Copyright © 2010 Society of Chemical Industry