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Reduction of Dihydrokaempferol by Vitis vinfera Dihydroflavonol 4-Reductase to Produce Orange Pelargonidin-Type Anthocyanins
- Xie, Sha, Zhao, Ting, Zhang, Zhenwen, Meng, Jiangfei
- Journal of agricultural and food chemistry 2018 v.66 no.13 pp. 3524-3532
- Escherichia coli, Vitis vinifera, anthocyanins, biosynthesis, cultivars, enzymatic treatment, enzyme activity, grapes, pelargonidin, taxifolin
- Vitis vinifera has been thought to be unable to produce pelargonidin-type anthocyanins because its dihydroflavonol 4-reductase (DFR) does not efficiently reduce dihydrokaempferol. However, in this study, pelargonidin 3-O-glucoside was detected in the skins of V. vinifera ‘Pinot Noir’, ‘Cabernet Sauvignon’, and ‘Yan73’, as well as in the flesh of ‘Yan73’ by HPLC–ESI-MS/MS. Additionally, pelargonidin 3-O-(6-acetyl)-glucoside was detected in ‘Yan73’ skin and flesh for the first time. To further confirm the presence of pelargonidin-type anthocyanins in these grape cultivars, their DFRs were cloned, expressed in Escherichia coli, and purified. An enzyme-activity analysis revealed that V. vinifera DFR can reduce dihydrokaempferol to produce leucopelargonidin, although it prefers dihydroquercetin and dihydromyricetin as substrates. Thus, the existence of a pelargonidin-based anthocyanin-biosynthetic pathway was confirmed in V. vinifera via mass-spectrometric and enzymatic methods and redirected anthocyanin biosynthesis in V. vinifera L. cultivars.