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Effects of L-Malic Acid on Alpha-Glucosidase: Inhibition Kinetics and Computational Molecular Dynamics Simulations

Abstract

The inhibitory effect of L-malic acid (MA) on alpha-glucosidase (EC 3.2.1.20) was investigated by combination study between inhibition kinetics and computational simulations. The results from the serial kinetics demonstrated that MA could directly inactivate the enzyme activity in a dose-dependent manner and a typical non-competitive type, as well as in a fast inactivate process without detectable time course. The tertiary conformation study with an application of spectrofluorimetry showed that MA modulated the tertiary structural conformation of alpha-glucosidase both on the overall and on regional active site pocket, which monitored by red-shift intrinsic fluorescence peak with decreases intensities, and the significant intensity increasing of 1-anilinonaphthalene-8-sulfonate (ANS)-binding fluorescence, respectively. To have more insight, we also adapted the computational molecular dynamics (MD) simulations. The results showed that MA was located in the entrance of active pocket for the catalytic reaction and blocked the passage of substrate. It confirmed that MA inhibits as a non-competitive type, not direct docking to the glucose binding site. Our study provides important molecular mechanisms to figure out alpha-glucosidase inhibition that might associate to development of type 2 diabetes mellitus drug.

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Abbreviations

MA:

L-Malic acid

PNPG:

p-Nitrophenyl α-D-glucopyranoside

PNP:

4-Nitrophenol

ANS:

1-Anilinonaphthalene-8-sulfonate

MD:

Molecular dynamics

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Acknowledgments

This study was supported by the National Natural Science Foundation of China (No. 31071433). Dr. Hai-Meng Zhou was supported by the fund from Science and Technology of Xiaoshan District (No. 2012121) and the China Natural Science Foundation (No. 31170732 and No. 31270854). Dr. Jun-Mo Yang was supported by a grant from the Korea Health Technology R&D Project, Ministry of Health & Welfare, Republic of Korea (No. HI12C1299) and by grants from the Samsung Biomedical Research Institute (SMX1131301; GL1-B2-181-1; CA6-216-3). Dr. Jinhyuk Lee was supported by grants from the Korea Research Institute of Bioscience and Biotechnology (KRIBB) Research Initiative Program, the Korean Ministry of Education, Science and Technology (MEST) (2012R1A1A2002676), and the Pioneer Research Center Program through the National Research Foundation of Korea funded by the Ministry of Science, ICT & Future Planning (2013M3C1A3064780). Dr. Zhi-Rong Lü was supported by a grant from the Science and Technology Planning Project of Hangzhou (No. 20120232B39). Dr. Yong-Doo Park was supported by a grant from Zhejiang Provincial Natural Science Foundation of China, “Towards studying the function of C3dg protein and elucidating its role in the pathogenesis of atopic dermatitis” (Grant No. LY14H110001). This subject is supported by Korea Ministry of Environment as “The Environmental Health Action Program.”

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Correspondence to Lin Gou or Jun-Mo Yang.

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Lin Gou and Yi Zhan equally contributed to this study.

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Gou, L., Zhan, Y., Lee, J. et al. Effects of L-Malic Acid on Alpha-Glucosidase: Inhibition Kinetics and Computational Molecular Dynamics Simulations. Appl Biochem Biotechnol 175, 2232–2245 (2015). https://doi.org/10.1007/s12010-014-1429-6

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Keywords

  • Alpha-glucosidase
  • Malic acid
  • Inhibition