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Literature summary extracted from

  • Wu, Y.; Pan, L.; Yu, S.; Li, H.
    Cloning, microbial expression and structure-activity relationship of polyphenol oxidases from Camellia sinensis (2010), J. Biotechnol., 145, 66-72.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.10.3.1 expressed in Escherichia coli BL21(DE3) cells using pET30c expression vector (high expression) and in Pichia pastoris GS115 using both the secretory and non-secretory vectors pPICZ-A and pPICZA (low expression) Camellia sinensis

Organism

EC Number Organism UniProt Comment Textmining
1.10.3.1 Camellia sinensis A6N8J4 cultivar Tieguanyin
-
1.10.3.1 Camellia sinensis A6NAA0 cultivar Longjing
-
1.10.3.1 Camellia sinensis A6YS04 cultivar Qihong
-
1.10.3.1 Camellia sinensis A6YS05 cultivar Xiangbolv
-
1.10.3.1 Camellia sinensis C5MLZ1 cultivar Puer
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.10.3.1 ammonium sulfate precipitation, column chromatography, gel filtration Camellia sinensis

Renatured (Commentary)

EC Number Renatured (Comment) Organism
1.10.3.1 method one is to resuspend the proteins twice in 50 mM Tris-HCl buffer (pH 7.5) with 5 mM dithiothreitol, 2% (v/v) Triton X-100 and 5 mM NaCl, followed by centrifugation at 14000 x g for 5 min. Method two is to resuspend the proteins in one-tenth volume of its original culture of inclusion body isolation buffer containing 20 mM Tris-HCl and 1% (v/v) Triton X-100, pH 7.5, and then in 50 mM Tris-HCl, pH 8.0, each followed by centrifugation at 14000 x g for 5 min Camellia sinensis

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.10.3.1 2 catechol + O2
-
 Camellia sinensis 2 1,2-benzoquinone + 2 H2O
-
 ?

Synonyms

EC Number Synonyms Comment Organism
1.10.3.1 polyphenol oxidase
-
 Camellia sinensis
1.10.3.1 PPO
-
 Camellia sinensis

Expression

EC Number Organism Comment Expression
1.10.3.1 Camellia sinensis a 3.1fold increase in PPO activity over non-recombinant controls is obtained by expressing the PPO fragment without signal sequences and the CuC domain in Escherichia coli BL21 (DE3) using the pET30c vector up