Ultrasound-assisted three-phase partitioning of polyphenol oxidase from potato peel (Solanum tuberosum)

AbstractPolyphenol oxidase (PPO) from the wild edible mushroom Lepiota procera is partially purified and biochemically characterized using three-phase partitioning (TPP), which is an easily applied and effective method. This method includes ammonium sulfate saturation at different concentrations, t-butanol addition (1:1; 1:5), and adjustment of pH. Optimum purification parameters with 20% ammonium sulfate saturation and 1:1 t-butanol conditions led to the highest activity at bottom phase with 8.4 fold purification. Sodium dodecyl sulfate- polyacrylamide gel electrophoresis (SDS-PAGE) analysis showed the molecular weight of the purified enzyme to be 35 kDa. The partially purified PPO presented a high level of activity with L-DOPA (Michaelis-Menten constant, Km – 0.12 mM), followed by caffeic acid (0.27 mM) and 4-methylcatechol (0.46 mM) and it is classified as a catecholase type of PPO. The enzyme had peak activity at a temperature of 40 °C and a pH value of 7.0. These results demonstrate a new enzyme source and easy purification method, useful for various industrial applications.

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Comparative study of ultrasonic pretreatment and ultrasound assisted three phase partitioning for extraction of custard apple seed oil

Ultrasonics Sonochemistry ◽

10.1016/j.ultsonch.2019.104821 ◽

2020 ◽

Vol 61 ◽

pp. 104821 ◽

Cited By ~ 2

Author(s):

Dhanashree C. Panadare ◽

Akash Gondaliya ◽

Virendra K. Rathod

Keyword(s):

Comparative Study ◽

Seed Oil ◽

Ultrasonic Pretreatment ◽

Phase Partitioning ◽

Apple Seed ◽

Three Phase ◽

Custard Apple ◽

Ultrasound Assisted

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Application of three-phase partitioning to the purification and characterization of polyphenol oxidase from antioxidant rosemary (Rosmarinus officinalis L.)

International Journal of Food Engineering ◽

10.1515/ijfe-2020-0118 ◽

2020 ◽

Vol 0 (0) ◽

Author(s):

Yonca Yuzugullu Karakus ◽

Busra Kahveci ◽

Arda Acemi ◽

Gulden Kocak

Keyword(s):

Enzyme Activity ◽

Polyphenol Oxidase ◽

Cetyltrimethylammonium Bromide ◽

Rosmarinus Officinalis ◽

Phase Partitioning ◽

Three Phase ◽

Diammonium Salt ◽

The Common ◽

Rosmarinus Officinalis L

AbstractPolyphenol oxidase (PPO) has been purified from the rosemary plant (Rosmarinus officinalis L.) through three-phase partitioning (TPP) and has been biochemically characterized. The optimized TPP consisted of 50% (w/v) ammonium sulfate and equal volumes of crude extract and tert-butanol prepared at pH 6.5 and room temperature. Using this system, PPO was purified 14-fold, with 230% recovery of activity from the middle phase. The partitioned enzyme had a molecular mass of 53 kDa. The highest enzyme activity was detected at 30 °C and pH 7.0 against catechol. In substrate specificity tests, the enzyme displayed activity towards catechol, 4-methylcatechol, caffeic acid, hydroquinone, 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS), pyrogallol, syringaldezine, and 3,4-dihydroxy-L-phenylalanine but no activity towards L-tyrosine. The enzyme was inhibited by the common PPO inhibitors; salicylhydroxamic acid (SHAM), cetyltrimethylammonium bromide (CTAB), polyvinylpyrrolidone (PVP), and the organic solvent dimethyl sulfoxide (DMSO). Enzyme activity increased in the presence of the organic solvents acetone, ethanol, and methanol.

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