Literature summary for 1.10.3.1 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.

Cloned(Commentary)

Cloned (Comment)	Organism
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

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

Purification (Commentary)

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

Renatured (Commentary)

Renatured (Comment)	Organism
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

Renatured (Comment)

Organism

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)

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

Synonyms

Synonyms	Comment	Organism
polyphenol oxidase	-	Camellia sinensis
PPO	-	Camellia sinensis

Expression

Organism	Comment	Expression
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