Citation

Jiang S, Penner MH. Foods 2021; 10(10).

Copyright

(Copyright © 2021, MDPI: Multidisciplinary Digital Publications Institute)

DOI

10.3390/foods10102486

PMID

unavailable

Abstract

The ability to accurately measure the activity of polyphenol oxidase (PPO) in complex matrices is essential. A problem encountered when using spectrophotometric methods is interference due to ascorbic acid (AA), often used as an enzyme "protecting agent" during PPO extraction. This study focuses on the nature of AA's effect on spectrophotometric determinations of PPO activity as well as enzyme extraction. Potato extracts and semi-purified PPO were used as enzyme sources. The inactivation of PPO attributed to AA is substrate-mediated. The extent of AA-dependent inactivation of PPO in model systems varied between substrates. AA only slows mechanism-based inactivation of PPO induced by catechol, possibly owing to the prevention of quinone formation. AA minimally protects PPO activity during enzyme extraction. The problem associated with AA in PPO assay could be circumvented by using ascorbate oxidase to remove AA when catechol is the primary substrate or by using chlorogenic acid as the primary substrate. © 2021 by the authors. Licensee MDPI, Basel, Switzerland.

Language: en

Keywords

Ascorbic acid; Extraction; Activity quantification; Mechanism-based (suicide) inactivation; Polyphenol oxidase