The Cloning and Prokaryotic Expression of Polyphenol Oxidase Gene in Pear (Pyrus Pyrifolia Nakai)

CHEN Dongsheng; WANG Kunbo; LI Qin; LI Juan; HUANG Jian&#x02032;an; LIU Zhonghua

doi:10.13305/j.cnki.jts.2015.01.004

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Journal of Tea Science ›› 2015, Vol. 35 ›› Issue (1) : 17-23. DOI: 10.13305/j.cnki.jts.2015.01.004

The Cloning and Prokaryotic Expression of Polyphenol Oxidase Gene in Pear (Pyrus Pyrifolia Nakai)

CHEN Dongsheng, WANG Kunbo^*, LI Qin, LI Juan, HUANG Jian′an, LIU Zhonghua^*

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Abstract

：The polyphenol oxidase (PPO, GenBank accession No. JQ861265) genes were cloned by PCR from Pyrus Pyrifolia Nakai. The full length of PPO gene was 1β782βbp without introns, coding a precursor peptide. PPO precursor consists of 593 amino acids with a molecular weight of about 65.8βkDa. It has a theoretical PI of 8.4 and has a transit peptide consisting of 47 amino acids. The mature PPO without transit peptide consists of 547 amino acids with a molecular weight of about 60.8βkDa and a theoretical PI of 6.69. There are 2 Cu-binding domain in the α-helix of PPOs. The protein accumulation got a peak in 3-6 hours. The induced proteins (precursor PPO and mature PPO) can oxidize catechins into theaflavins in vitro.

Key words

enzymatic synthesis / gene clone / polyphenol oxidase / prokaryotic expression / theaflavins

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CHEN Dongsheng, WANG Kunbo, LI Qin, LI Juan, HUANG Jian′an, LIU Zhonghua. The Cloning and Prokaryotic Expression of Polyphenol Oxidase Gene in Pear (Pyrus Pyrifolia Nakai)[J]. Journal of Tea Science. 2015, 35(1): 17-23 https://doi.org/10.13305/j.cnki.jts.2015.01.004

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