Literature summary for 1.3.1.77 extracted from

  • Zhu, Y.; Peng, Q.Z.; Li, K.G.; Xie, D.Y.
    Molecular cloning and functional characterization of the anthocyanidin reductase gene from Vitis bellula (2014), Planta, 240, 381-398 .
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
gene ANR, DNA and amino acid sequence determination and analysis, sequence comparisons and phylogenetic analysis, recombinant expression of His-Trx-tagged enzyme in Escherichia coli strain BL21(DE3), recombinant expression of GFP-tagged enzyme in Arabidopsis thaliana ban mutant Vitis bellula

Engineering

Protein Variants Comment Organism
additional information generation of an ANR T-DNA insertion that lack flavan-3-ols and proanthocyanidins Vitis bellula

Inhibitors

Inhibitors Comment Organism Structure
NaCl purified recombinant VbANR almost loses its catalytic activity at 500 mM Vitis bellula

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
additional information kinetics of Trx-VbANR with NADPH and NADH, overview Vitis bellula
0.1308
-
delphinidin pH 4.0, 40°C, recombinant enzyme Vitis bellula
0.1929
-
NADPH pH 4.0, 40°C, recombinant enzyme Vitis bellula
0.8169
-
NADH pH 4.0, 40°C, recombinant enzyme Vitis bellula
0.1757
-
cyanidin pH 4.0, 40°C, recombinant enzyme Vitis bellula
0.1345
-
pelargonidin pH 4.0, 40°C, recombinant enzyme Vitis bellula

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
cyanidin + 2 NADPH + H+ Vitis bellula
-
(2R,3R)-epicatechin + 2 NADP+ ? ?
delphinidin + 2 NADPH + H+ Vitis bellula
-
(2R,3R)-epigallocatechin + 2 NADP+ ? ?
pelargonidin + 2 NADPH + H+ Vitis bellula
-
(2R,3R)-epiafzelechin + 2 NADP+ ? ?

Organism

Organism UniProt Comment Textmining
Vitis bellula H9TZS7
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His-Trx-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography Vitis bellula

Source Tissue

Source Tissue Comment Organism Textmining
leaf
-
Vitis bellula
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
additional information the recombinant protein in vitro converts anthocyanidins to 2R,3R-cis-flavan-3-ols and 2S,3R-trans-flavan-3-ols, two stereo configurations at the C2 and C3 positions, such as (-)-epicatechin and (-)-catechin. flavan-3-ols produced from the catalysis of Trx-VbANR result from the reduction activity of VbANR but not Trx. Stereochemistry of the enzyme reaction, overview Vitis bellula ?
-
?
cyanidin + 2 NADPH + H+
-
Vitis bellula (2R,3R)-epicatechin + 2 NADP+ ? ?
delphinidin + 2 NADPH + H+
-
Vitis bellula (2R,3R)-epigallocatechin + 2 NADP+ ? ?
pelargonidin + 2 NADPH + H+
-
Vitis bellula (2R,3R)-epiafzelechin + 2 NADP+ ? ?
anthocyanidin + 2 NADPH + H+
-
Vitis bellula 2,3-trans-(2S,3R)-flavan-3-ol + 2 NADP+ ? ?
cyanidin + 2 NADH + H+
-
Vitis bellula (2R,3R)-epicatechin + 2 NAD+ ? ?
delphinidin + 2 NADH + H+
-
Vitis bellula (2R,3R)-epigallocatechin + 2 NAD+ ? ?
pelargonidin + 2 NADH + H+
-
Vitis bellula (2R,3R)-epiafzelechin + 2 NAD+ ? ?

Subunits

Subunits Comment Organism
? x * 56000, recombinant His-Trx-tagged enzyme, SDS-PAGE Vitis bellula

Synonyms

Synonyms Comment Organism
ANR
-
Vitis bellula
anthocyanidin reductase
-
Vitis bellula
VbANR
-
Vitis bellula

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
40
-
recombinant enzyme Vitis bellula

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
30 50 activity profile, overview Vitis bellula

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2
-
pelargonidin pH 4.0, 40°C, recombinant enzyme Vitis bellula
1.665
-
cyanidin pH 4.0, 40°C, recombinant enzyme Vitis bellula
2.012
-
delphinidin pH 4.0, 40°C, recombinant enzyme Vitis bellula

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
4
-
recombinant enzyme Vitis bellula

pH Range

pH Minimum pH Maximum Comment Organism
3 8 activity range, profile overview Vitis bellula

Cofactor

Cofactor Comment Organism Structure
NADPH
-
Vitis bellula
NADH
-
Vitis bellula
additional information enzyme VbANR uses both NADPH and NADH but prefers to employ NADPH Vitis bellula

General Information

General Information Comment Organism
physiological function anthocyanidin reductase (ANR) is an NADPH-/NADH-dependent enzyme that transfers two hydrides to anthocyanidins to produce three types of isomeric flavan-3-ols Vitis bellula
additional information the putative enzymatic activity triad is composed of Ser136, Tyr173, and Lys177 Vitis bellula

KCat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
14.87
-
pelargonidin pH 4.0, 40°C, recombinant enzyme Vitis bellula
9.48
-
cyanidin pH 4.0, 40°C, recombinant enzyme Vitis bellula
15.38
-
delphinidin pH 4.0, 40°C, recombinant enzyme Vitis bellula