Recombinant L-PaPPO Expression and Purification

The expression and the purification of L-PaPPO were performed as described previously for tomato PPO (Kampatsikas et al., 2019 (link)) with some modifications. L-PaPPO was expressed in E. coli SHuffle® T7 cells (NEB), in modified 2xYT medium. The culture was grown at 18 °C under shaking (185 rpm) for 72 h. Enzyme purification was performed using an ÄKTA fast protein liquid chromatography system (FPLC). The detailed methods are presented in the SM.

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Derardja A.E., Pretzler M., Kampatsikas I., Radovic M., Fabisikova A., Zehl M., Barkat M, & Rompel A. (2022). Polyphenol oxidase and enzymatic browning in apricot (Prunus armeniaca L.): Effect on phenolic composition and deduction of main substrates. Current Research in Food Science, 5, 196-206.

Publication 2022

E. coli SHuffle® T7 cells

Cells E coli Enzyme Liquid chromatography Protein Tomato

Corresponding Organization : University of Vienna

Other organizations : Constantine 1 University

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Variable analysis

independent variables

Expression of L-PaPPO in E. coli SHuffle® T7 cells
Culture conditions (18 °C, 185 rpm, 72 h)

dependent variables

Purification of L-PaPPO using ÄKTA fast protein liquid chromatography system (FPLC)

control variables

Modified 2xYT medium for L-PaPPO expression

controls

No positive or negative controls were explicitly mentioned in the provided information.

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