Biochemical Analysis of Recombinant Pea Seed Coat-Specific Polyphenol Oxidase (PeaPPO) in Relation to Various Phenolic Substrates

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F61989592%3A15640%2F25%3A73634527" target="_blank" >RIV/61989592:15640/25:73634527 - isvavai.cz</a>

Nalezeny alternativní kódy

RIV/61989592:15310/25:73634527 RIV/00027006:_____/25:10179196

Výsledek na webu

<a href="https://pubs.acs.org/doi/pdf/10.1021/acs.jafc.5c01839" target="_blank" >https://pubs.acs.org/doi/pdf/10.1021/acs.jafc.5c01839</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1021/acs.jafc.5c01839" target="_blank" >10.1021/acs.jafc.5c01839</a>

Jazyk výsledku

angličtina

Název v původním jazyce

Biochemical Analysis of Recombinant Pea Seed Coat-Specific Polyphenol Oxidase (PeaPPO) in Relation to Various Phenolic Substrates

Popis výsledku v původním jazyce

The seed coat serves as the primary protective barrier, offering mechanical and chemical defense for the embryo. It contains various metabolites, including phenolic compounds, which can be oxidized by polyphenol oxidase (PPO) to form oligomers. In this study, we heterologously expressed a 515 amino acid protein derived from wild pea (Pisum elatius), omitting its Nterminal signal sequence, and analyzed its biochemical properties. The recombinant PeaPPO required sodium dodecyl sulfate (SDS) for activation and exhibited activity between pHs 5.2 and 7.0, peaking at pH 6.0 with 0.25 mM SDS. Tropolone and its isomer thujaplicin were the most effective inhibitors. PeaPPO catalyzed reactions with seed coat-derived substrates, displaying activity toward phenols, catechols, and pyrogallols, with the highest affinity for catechols. Principal component analysis of LC-MS/MSderived phenolic profiles demonstrated that PPO+ and ppo− genotypes differ significantly in their accumulation of PPO substrates and inhibitors. These findings confirm that PeaPPO possesses both monophenolase and catechol oxidase activities, identifying it as a tyrosinase.

Název v anglickém jazyce

Biochemical Analysis of Recombinant Pea Seed Coat-Specific Polyphenol Oxidase (PeaPPO) in Relation to Various Phenolic Substrates

Popis výsledku anglicky

The seed coat serves as the primary protective barrier, offering mechanical and chemical defense for the embryo. It contains various metabolites, including phenolic compounds, which can be oxidized by polyphenol oxidase (PPO) to form oligomers. In this study, we heterologously expressed a 515 amino acid protein derived from wild pea (Pisum elatius), omitting its Nterminal signal sequence, and analyzed its biochemical properties. The recombinant PeaPPO required sodium dodecyl sulfate (SDS) for activation and exhibited activity between pHs 5.2 and 7.0, peaking at pH 6.0 with 0.25 mM SDS. Tropolone and its isomer thujaplicin were the most effective inhibitors. PeaPPO catalyzed reactions with seed coat-derived substrates, displaying activity toward phenols, catechols, and pyrogallols, with the highest affinity for catechols. Principal component analysis of LC-MS/MSderived phenolic profiles demonstrated that PPO+ and ppo− genotypes differ significantly in their accumulation of PPO substrates and inhibitors. These findings confirm that PeaPPO possesses both monophenolase and catechol oxidase activities, identifying it as a tyrosinase.

Druh

J_imp - Článek v periodiku v databázi Web of Science

CEP obor

—

OECD FORD obor

10611 - Plant sciences, botany

Projekt

Výsledek vznikl pri realizaci vícero projektů. Více informací v záložce Projekty.

Návaznosti

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)

Rok uplatnění

2025

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Název periodika

JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY

ISSN

0021-8561

e-ISSN

1520-5118

Svazek periodika

73

Číslo periodika v rámci svazku

35

Stát vydavatele periodika

US - Spojené státy americké

Počet stran výsledku

15

Strana od-do

21754-21768

Kód UT WoS článku

001554775100001

EID výsledku v databázi Scopus

2-s2.0-105015387526