Laccase-Mediated Flavonoid Polymerization Leads to the Pericarp Browning of Litchi Fruit.

Litchi pericarp turns brown rapidly after fruit harvest, while the mechanism remains obscure. The contents of (-)-epicatechin (EC) and procyanidins (PCs) A2/B1/B2/C1 decreased during the pericarp browning, and a previously identified laccase (ADE/LAC) showed activity to these compounds, with brown products observed in the reactions. By UPLC-DAD-QTOF-MS/MS, isomers of dimeric, trimeric, and tetrameric PCs were detected in the EC-ADE/LAC reaction. In the presence of cyanidin-3-O-rutiside and rutin, anthocyanin-EC and rutin-EC adducts were, respectively, produced, and darker brown precipitation was observed in these reactions relatively to the EC-ADE/LAC reaction alone. ADE/LAC catalyzed the conversion of PC B2 to A-type PC dimers and B-type PC tetramers. ADE/LAC complemented the transparent testa of Arabidopsis LAC15-loss-of-function mutant (tt10) to wild-type dark brown seed coat. The results demonstrated that ADE/LAC-mediated flavonoid polymerization played an important role in the browning of pericarp.