Purification and Characterization of Latent Polyphenol Oxidase from Apricot (Prunus armeniaca L.)

被引：79

作者：

Derardja, Ala Eddine ^{[1
,2
]}

Pretzler, Matthias ^{[1
]}

Kampatsikas, Ioannis ^{[1
]}

Barkat, Malika ^{[2
]}

Rompel, Annette ^{[1
]}

机构：

[1] Univ Vienna, Fak Chem, Inst Biophys Chem, Althanstr 14, A-1090 Vienna, Austria

[2] Univ Freres Mentouri Constantine 1, INATAA, Lab Bioqual, Route Ain El Bey, Constantine 25000, Algeria

来源：

JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY | 2017年 / 65卷 / 37期

基金：

奥地利科学基金会;

关键词：

apricot; tyrosinase; polyphenol oxidase; protein purification; characterization; activation; MUSHROOM TYROSINASE; BIOCHEMICAL-PROPERTIES; FRUIT; PLANT; MATURITY; PERICARP; VARIETY; FLOWER; ABPPO4; ACID;

D O I：

10.1021/acs.jafc.7b03210

中图分类号：

S [农业科学];

学科分类号：

09 ;

摘要：

Polyphenol oxidase from apricot (Prunus armeniaca) (PaPPO) was purified in its latent form (L-PaPPO), and the molecular weight was determined to be 63 kDa by SDS-PAGE. L-PaPPO was activated in the presence of substrate at low pH. The activity was enhanced by CuSO4 and low concentrations (<= 2 mM) of SDS. PaPPO has its pH and temperature optimum at pH 4.5 and 45 degrees C for catechol as substrate. It showed diphenolase activity and highest affinity toward 4-methylcatechol (K-M = 2.0 mM) and chlorogenic acid (K-M = 2.7 mM). L-PaPPO was found to be spontaneously activated during storage at 4 degrees C, creating a new band at 38 kDa representing the activated form (A-PaPPO). The mass of A-PaPPO was determined by mass spectrometry as 37455.6 Da (Asp102 Leu429). Both L-PaPPO and A-PaPPO were identified as polyphenol oxidase corresponding to the known PaPPO sequence (UniProt 081103) by means of peptide mass fingerprinting.

引用

页码：8203 / 8212

页数：10

共 59 条

[1] Purification of polyphenol oxidase from borage (Trachystemon orientalis L.) by using three-phase partitioning and investigation of kinetic properties [J].