title: Fabrication of enzyme-immobilized halloysite nanotubes for affinity enrichment of lipase inhibitors from complex mixtures.
abstract: Lipase is the key enzyme for catalyzing triglyceride hydrolysis in vivo, and lipase inhibitors have been used in the management of obesity. We present the first report on the use of lipase-adsorbed halloysite nanotubes as an efficient medium for the selective enrichment of lipase inhibitors from natural products. A simple and rapid approach was proposed to fabricate lipase-adsorbed nanotubes through electrostatic interaction. Results showed that more than 85% lipase was adsorbed into nanotubes in 90 min, and approximately 80% of the catalytic activity was maintained compared with free lipase. The specificity and reproducibility of the proposed approach were validated by screening a known lipase inhibitor (i.e., orlistat) from a mixture that contains active and inactive compounds. Moreover, we applied this approach with high performance liquid chromatography-mass spectrometry technique to screen lipase inhibitors from the Magnoliae cortex extract, a medicinal plant used for treating obesity. Two novel biphenyl-type natural lipase inhibitors magnotriol A and magnaldehyde B were identified, and their IC50 values were determined as 213.03 and 96.96 μM, respectively. The ligand-enzyme interactions of magnaldehyde B were further investigated by molecular docking. Our findings proved that enzyme-adsorbed nanotube could be used as a feasible and selective affinity medium for the rapid screening of enzyme inhibitors from complex mixtures.
authors: Pharmaceutical Informatics Institute, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou 310058, China.
authors: Haibo
authors: H
authors: Wang
authors: College of Preclinical Medicine, Zhejiang Chinese Medical University, Hangzhou 310053, China.
authors: Xiaoping
authors: X
authors: Zhao
authors: Pharmaceutical Informatics Institute, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou 310058, China.
authors: Shufang
authors: S
authors: Wang
authors: Pharmaceutical Informatics Institute, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou 310058, China.
authors: Shan
authors: S
authors: Tao
authors: Pharmaceutical Informatics Institute, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou 310058, China.
authors: Ni
authors: N
authors: Ai
authors: Pharmaceutical Informatics Institute, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou 310058, China. Electronic address: mysky@zju.edu.cn.
authors: Yi
authors: Y
authors: Wang
mesh_terms: D000538:Aluminum Silicates; D001688:Biological Products / Q000737:chemistry*; D001713:Biphenyl Compounds / Q000737:chemistry; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D000077215:Clay; D045424:Complex Mixtures; D004791:Enzyme Inhibitors / Q000737:chemistry*; D004800:Enzymes, Immobilized; D017705:Lignans / Q000737:chemistry; D008049:Lipase / Q000037:antagonists & inhibitors*; D031566:Magnolia / Q000737:chemistry; D013058:Mass Spectrometry / Q000379:methods; D062105:Molecular Docking Simulation; D043942:Nanotubes; D010936:Plant Extracts / Q000737:chemistry; D015203:Reproducibility of Results; D055672:Static Electricity
chemical_list: D000538:Aluminum Silicates; D001688:Biological Products; D001713:Biphenyl Compounds; D045424:Complex Mixtures; D004791:Enzyme Inhibitors; D004800:Enzymes, Immobilized; D017705:Lignans; D010936:Plant Extracts; C587664:magnaldehyde B; D008049:Lipase; D000077215:Clay
keywords: Affinity selection mass spectrometry; Enzyme inhibitor; Microextraction; Rapid screening; Traditional Chinese medicine