Application of three-phase partitioning to the purification and characterization of polyphenol oxidase from antioxidant rosemary (Rosmarinus officinalis L.)
-
Yonca Yuzugullu Karakus
,
Busra Kahveci
Abstract
Polyphenol 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.
Funding source: Kocaeli Üniversitesi
Award Identifier / Grant number: 2017-057
Acknowledgments
The present study was granted by Kocaeli University (BAP Project number: 2017/057).
Author contribution: All the authors have accepted responsibility for the entire content of this submitted manuscript and approved submission.
Research funding: The present study was granted by Kocaeli University (BAP Project number: 2017/057).
Conflict of interest statement: The authors declare no conflicts of interest regarding this article.
Ethical approval: This article does not contain any studies with human participants or animals performed by any of the authors.
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© 2020 Walter de Gruyter GmbH, Berlin/Boston
Articles in the same Issue
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- Application of three-phase partitioning to the purification and characterization of polyphenol oxidase from antioxidant rosemary (Rosmarinus officinalis L.)
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Articles in the same Issue
- Articles
- The effect of slight milling on nutritional composition and morphology of quinoa (Chenopodium) grain
- Application of three-phase partitioning to the purification and characterization of polyphenol oxidase from antioxidant rosemary (Rosmarinus officinalis L.)
- Frozen kinetics models for sensory, chemical, and microbial spoilage of preserved razor clam (Sinonovacula constricta) at different temperatures
- Low-molecular-weight peptides with potential cardiovascular regulatory functions from Atlantic salmon skin
- The effect of the stale bread flour addition on flour and bread quality
- Control of spoilage fungi in yogurt using MicroGARD 200™, Lyofast-FPR2™ and HOLDBAC-YMC™ as bioprotectants
- A study on correlations between antimicrobial effects and diffusion coefficient, zeta potential and droplet size of essential oils
- Optimization of ultrasonic extraction of Lycium barbarum polysaccharides using response surface methodology
