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J Agric Food Chem. 2018 Oct 10;66(40):10608-10616. doi: 10.1021/acs.jafc.8b03647. Epub 2018 Oct 2.

Structural and Biochemical Characterization of the Curcumin-Reducing Activity of CurA from Vibrio vulnificus.

Author information

1
Department of Chemistry & Nanoscience , Ewha Womans University , Seoul 03760 , Republic of Korea.
2
College of Pharmacy and Graduate School of Pharmaceutical Sciences , Ewha Womans University , Seoul 03760 , Republic of Korea.
3
Biotechnology Research Division , Korea Atomic Energy Research Institute (KAERI) , Jeongeup 56212 , Republic of Korea.
4
University of Science and Technology , Daejeon , Republic of Korea.
5
Unit of Polar Genomics , Korea Polar Research Institute , Incheon 21990 , Republic of Korea.
6
Department of Food Science and Engineering , Ewha Womans University , Seoul 03760 , Republic of Korea.

Abstract

Curcumin is a yellow-colored ingredient in dietary spice turmeric ( Curcuma longa Linn). This nontoxic polyphenol has antitumor, anti-inflammatory, apoptotic, and antioxidant activities. The ingested curcumin is reduced to multihydrated forms with more potent therapeutic potentials by the curcumin reductase (CurA) from commensal Escherichia coli. In this study, we demonstrated that Vibrio vulnificus CurA ( VvCurA) with 87% sequence similarity to the E. coli CurA exhibits the curcumin-reducing activity through spectrophotometric detection of NADPH oxidation and high performance liquid chromatographic analysis of curcumin consumption and product generation. Afterward, we determined the crystal structures of VvCurA and the VvCurA/NADPH complex, and made the in silico model of the VvCurA/NADPH/curcumin ternary complex through induced fit docking. Based on structural information, active site residues that play critical roles in catalysis have been identified and characterized by mutational and kinetic studies, leading us to propose the reaction mechanism of CurA.

KEYWORDS:

crystal structure of apo VvCurA; crystal structure of the VvCurA/NADPH complex; curcumin-reducing enzyme; enzyme mechanism; in silico model of the VvCurA/NADPH/curcumin complex

PMID:
30251539
DOI:
10.1021/acs.jafc.8b03647
[Indexed for MEDLINE]

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