Structural and Biochemical Characterization of the Curcumin-Reducing Activity of CurA from Vibrio vulnificus,Journal of Agricultural and Food Chemistry

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Structural and Biochemical Characterization of the Curcumin-Reducing Activity of CurA from Vibrio vulnificus
Journal of Agricultural and Food Chemistry ( IF 5.7 ) Pub Date : 2018-09-25 00:00:00 , DOI: 10.1021/acs.jafc.8b03647
Soo-Bong Park ₁ , Da-Woon Bae ₁ , Nina Abigail B. Clavio ₂ , Lei Zhao ₃ , Chang-Sook Jeong _{4,

5} , Bo Mee Choi ₁ , Stephani Joy Y. Macalino ₂ , Hee-Jeong Cha ₆ , Jin-Byung Park ₆ , Jun Hyuck Lee _{4,

5} , Sang-Jip Nam ₁ , Sun Choi ₂ , Min-Kyu Kim _{3,

4} , Sun-Shin Cha ₁

Affiliation

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.

中文翻译：

创伤弧菌CurA降低姜黄素活性的结构和生化特征

姜黄素是膳食香料姜黄（一种黄色的成分姜黄属）。这种无毒的多酚具有抗肿瘤，消炎，凋亡和抗氧化的活性。通过共生大肠杆菌中的姜黄素还原酶（CurA）将摄入的姜黄素还原为具有更强治疗潜力的多水合形式。在这项研究中，我们证明了与大肠杆菌CurA具有87％序列相似性的弧菌弧菌CurA（Vv CurA）通过分光光度法检测NADPH氧化和高效液相色谱分析姜黄素的消耗量和产物产生了姜黄素的还原活性。之后，我们确定了Vv CurA和Vv CurA / NADPH复合物，并通过诱导拟合对接制作了Vv CurA / NADPH /姜黄素三元复合物的计算机模型。根据结构信息，通过突变和动力学研究鉴定并表征了在催化中起关键作用的活性位点残基，使我们提出了CurA的反应机理。

更新日期：2018-09-25

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