Biochemical and Functional Characterization of Anthocyanidin Reductase (ANR) from Mangifera indica L

Biochemical and Functional Characterization of Anthocyanidin Reductase (ANR) from Mangifera indica L

Mango ( L.) is abundant in proanthocyanidins (PAs) that are important for human health and plant response to abiotic stresses. However, the molecular mechanisms involved in PA biosynthesis still need to be elucidated. Anthocyanidin reductase (ANR) catalyzes a key step in PA biosynthesis. In this stu...

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Bibliographic Details
Published in:Molecules (Basel, Switzerland) Vol. 23; no. 11; p. 2876
Main Authors: Tan, Lin, Wang, Mei, Kang, Youfa, Azeem, Farrukh, Zhou, Zhaoxi, Tuo, Decai, María Preciado Rojo, Lina, Khan, Ikhlas A, Pan, Zhiqiang
Format: Journal Article
Language:English
Published: Switzerland MDPI AG 05-11-2018
MDPI
Subjects:
(−)-catechin
(−)-epicatechin
Amino acids
Anthocyanidin reductase
Biosynthesis
Catechin
Citric acid
Cloning
Cobalt
E coli
Enzymes
Epicatechin
Flavonoids
Fruits
Investigations
Mangifera indica
Mangifera indica L
Mangoes
Metabolites
Molecular modelling
NADH
Phylogenetics
proanthocyanidins
Proteins
Reductases
Substrates
(−)-catechin
Mangifera indica L
anthocyanidin reductase
proanthocyanidins
(−)-epicatechin
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Summary:Mango ( L.) is abundant in proanthocyanidins (PAs) that are important for human health and plant response to abiotic stresses. However, the molecular mechanisms involved in PA biosynthesis still need to be elucidated. Anthocyanidin reductase (ANR) catalyzes a key step in PA biosynthesis. In this study, three ANR cDNAs ( , , ) were isolated from mango, and expressed in . In vitro enzyme assay showed MiANR proteins convert cyanidin to their corresponding flavan-3-ols, such as (-)-catechin and (-)-epicatechin. Despite high amino acid similarity, the recombinant ANR proteins exhibited differences in enzyme kinetics and cosubstrate preference. MiANR1-2 and MiANR1-3 have the same optimum pH of 4.0 in citrate buffer, while the optimum pH for MiANR1-1 is pH 3.0 in phosphate buffer. MiANR1-1 does not use either NADPH or NADH as co-substrate while MiANR1-2/1-3 use only NADPH as co-substrate. MiANR1-2 has the highest Km and Vmax for cyanidin, followed by MiANR1-3 and MiANR1-1. The overexpression of in ban mutant reconstructed the biosynthetic pathway of PAs in the seed coat. These data demonstrate MiANRs can form the ANR pathway, leading to the formation of two types of isomeric flavan-3-ols and PAs in mango.
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ISSN:1420-3049
1420-3049
DOI:10.3390/molecules23112876