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Mechanism and Nature of Inhibition of Trypsin by Ligupurpuroside A, a Ku-Ding Tea Extract, Studied by Spectroscopic and Docking Methods

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Abstract

Ligupurpuroside A is a glycoside extracted from Ku-Ding tea. As extracts from Ku-Ding tea exhibit anti-inflammatory property, we hypothesize that Ligupurpuroside A may be an active compound which inhibits trypsin activity during the anti-inflammatory process. The mechanism and nature of inhibition of trypsin by Ligupurpuroside A have been studied by multi-spectroscopic method, enzyme-activity assay and molecular docking. Enzyme activity assay reveals that Ligupurpuroside A significantly inhibits the activity of trypsin through a competitive manner with an IC50 value of 3.08 × 10−3 mol L−1. Fluorescence titration together with thermodynamic analysis indicate that a Ligupurpuroside A-trypsin complex is formed, and that hydrophobic force and hydrogen bonding are the main forces stabilizing the complex. UV-vis absorption, synchronous fluorescence and circular dichroism spectra show that the interaction between Ligupurpuroside A and trypsin induces conformational changes of trypsin with a decrease in the contents of α-helix and β-sheet. Finally, molecular docking further suggests that Ligupurpuroside A molecule binds within the active pocket of trypsin via hydrophobic force and hydrogen bond. Results from this study of the interaction of trypsin with its natural inhibitor should be useful to minimize the antinutritional effects and make full use of tea extracts in the food industry, and be also helpful to the design of the drugs for the diseases related to overexpression of trypsin.

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Acknowledgements

This work was supported by the National Natural Science Foundation of China (Grant 31540012, 31470431, 30570421, 81501213), Guangdong Natural Science Foundation for Major cultivation project (2014A030308017), Shenzhen Science and Technology Innovation Committee Grants (JSGG20160229120821300, JCYJ20150625103526744, JCYJ20150324140036823, JCYJ20120613112512654, JCYJ20140414090541801, JSGG20130411160539208, KQCX20140522111508785, CXZZ20150601110000604, ZDSYS201506031617582), Shenzhen special funds for Bio-industry development (NYSW20140327010012) and Shenzhen Medical Scientific Research Project (201401077).

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Authors and Affiliations

  1. College of Life Sciences and Oceanography, Shenzhen Key Laboratory of Marine Bioresources and Ecology/Shenzhen Key Laboratory of Microbial Genetic Engineering, Shenzhen University, Nanhai Ave 3688, Shenzhen, Guangdong, 518060, People’s Republic of China

    Zhibing Wu, Liangliang Shen, Qingguo Han, Jun Lu, Xu Xu, Hong Xu, Fengwen Huang, Jiangfeng Xie & Zhangli Hu

  2. School of Science, School of Interprofessional Health Studies, Faculty of Health and Environmental Sciences, Institute of Biomedical Technology, Auckland University of Technology, Auckland, New Zealand

    Jun Lu

  3. The University of Hong Kong-Shenzhen Hospital, Shenzhen, 518053, China

    Haifeng Tang

  4. School of Medicine, Shenzhen University, Shenzhen, 518060, China

    Zhendan He

  5. State Key Laboratory of Food Science and Technology/School of Resources, Environmental and Chemical Engineering, Nanchang University, Nanchang, 330031, China

    Zheling Zeng

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  1. Zhibing Wu
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  2. Liangliang Shen
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  3. Qingguo Han
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  4. Jun Lu
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  6. Xu Xu
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  7. Hong Xu
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  9. Jiangfeng Xie
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  10. Zhendan He
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  11. Zheling Zeng
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Corresponding authors

Correspondence to Hong Xu or Zhangli Hu.

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Zhibing Wu, Liangliang Shen and Qingguo Han contributed equally to this work.

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Wu, Z., Shen, L., Han, Q. et al. Mechanism and Nature of Inhibition of Trypsin by Ligupurpuroside A, a Ku-Ding Tea Extract, Studied by Spectroscopic and Docking Methods. Food Biophysics 12, 78–87 (2017). https://doi.org/10.1007/s11483-016-9465-0

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  • DOI: https://doi.org/10.1007/s11483-016-9465-0

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