Wu Qian, Zhang Yang, Tang Hu, Chen Yashu, Xie Bijun, Wang Chao, Sun Zhida
Hubei Collaborative Innovation Center for Industrial Fermentation, Research Center of Food Fermentation Engineering and Technology of Hubei, Hubei University of Technology , Wuhan, Hubei 430068, China.
Natural Product Laboratory, Department of Food Science and Technology, Huazhong AgriculturalUniversity , Wuhan, Hubei 430070, People's Republic of China.
J Agric Food Chem. 2017 Jan 11;65(1):216-223. doi: 10.1021/acs.jafc.6b04244. Epub 2016 Dec 23.
Anthocyanins were isolated from blueberry wine lees using Sephadex LH-20 column chromatography and semipreparative high-performance liquid chromatography (semipreparative HPLC) and then identified by HPLC-DAD-ESI-MS/MS. Our results show that malvidin-3-hexose (Mv-3-hex) and malvidin-3-(6'acetyl)-hexose (Mv-3-ace-hex) are the major components in the anthocyanin extracts of blueberry wine lees (>90%). The binding characteristics of Mv-3-hex and Mv-3-ace-hex with β-glucosidase were investigated by fluorescence spectroscopy, circular dichroism (CD) spectroscopy, and molecular docking. Spectroscopic analysis revealed that β-glucosidase fluorescence quenched by Mv-3-hex and Mv-3-ace-hex follows a static mode. Binding of Mv-3-hex and Mv-3-ace-hex to β-glucosidase mainly depends on electrostatic force. The result from CD spectra shows that adaptive structure rearrangement and increase of β-sheet structure occur only in the presence of Mv-3-ace-hex. A molecular docking study suggests that Mv-3-ace-hex has stronger binding with β-glucosidase than Mv-3-hex.
采用葡聚糖凝胶LH - 20柱色谱法和半制备高效液相色谱法（半制备HPLC）从蓝莓酒糟中分离出花色苷，然后通过HPLC - DAD - ESI - MS/MS进行鉴定。我们的结果表明，矢车菊素 - 3 - 己糖苷（Mv - 3 - hex）和矢车菊素 - 3 -（6'- 乙酰基）- 己糖苷（Mv - 3 - ace - hex）是蓝莓酒糟花色苷提取物中的主要成分（>90%）。通过荧光光谱法、圆二色性（CD）光谱法和分子对接研究了Mv - 3 - hex和Mv - 3 - ace - hex与β - 葡萄糖苷酶的结合特性。光谱分析表明，Mv - 3 - hex和Mv - 3 - ace - hex对β - 葡萄糖苷酶荧光的猝灭遵循静态模式。Mv - 3 - hex和Mv - 3 - ace - hex与β - 葡萄糖苷酶的结合主要取决于静电力。CD光谱结果表明，仅在Mv - 3 - ace - hex存在的情况下才会发生适应性结构重排和β - 折叠结构增加。分子对接研究表明，Mv - 3 - ace - hex与β - 葡萄糖苷酶的结合比Mv - 3 - hex更强。
