Inhibitory kinetics and bioactivities of Nuciferine and Methyl Ganoderate on Mucor miehei lipase and 3T3-L1 preadipocytes

• Published in: International journal of biological macromolecules 2020-11, Vol.163, p.1719-1728
• Main Authors: Liu, Xiao-Tian, Liu, Tian-Tian, Xu, Hui-Long, Chen, Qing-Xi, Wang, Qin
• Format: Article
• Language: English
• Subjects: 3T3-L1 Cells; 3 T3-L1; Adipocytes - drug effects; Adipogenesis - drug effects; Animals; Anti-Obesity Agents - pharmacology; Aporphines - pharmacology; Catalytic Domain - drug effects; Cell Differentiation - drug effects; Cell Line; Cell Proliferation - drug effects; Cholesterol - metabolism; Kinetics; Lipase; Lipase - metabolism; Methyl Ganoderate; Mice; Molecular Docking Simulation - methods; Nuciferine; Obesity; Obesity - drug therapy; Obesity - metabolism; Protein Conformation, alpha-Helical - drug effects; Protein Conformation, beta-Strand - drug effects; Reishi - chemistry; Rhizomucor - drug effects; Triglycerides - metabolism; Triterpenes - pharmacology
• ISSN: 0141-8130; 1879-0003
• DOI: 10.1016/j.ijbiomac.2020.09.127

In this study, inhibitory kinetics of Nuciferine and Methyl Ganoderate extrated from Lotus Leaves and Ganoderma lucidum on Mucor miehei Lipase were studied first. The molecular structure of Nuciferine and Methyl Ganoderate were determined. The inhibitory effects of two extracts on lipase were reversible, with the IC50 values of 0.194 and 0.332 mg/mL, respectively. The inhibition kinetic analysis by Lineweaver-Burk plots showed that they were a mixed-type inhibitor of lipase, with inhibition constants KI of 0.16 and 0.29 mg/mL, and KIS of 0.36 and 0.49 mg/mL, respectively. Results of spectral analysis showed that the UV absorption and the molecule fluorescence spectrum of the lipase hydrolyzate were significantly decreased after the inhibitor was added. The molecular docking further suggested that the interaction site between the active substance and inhibitor was located in an α-helix and a β-sheet of the lipase, and the lipase active site was interfered by the inhibitor near the cap structure. In addition, the proliferation and differentiation of 3 T3-L1 preadipocytes were inhibited by two extracts. Total triglycerides and cholesterol were significantly reduced in the cells. The results confirmed that Nuciferine and Methyl Ganoderate can be used as potential obesity treatment drugs.