Trehalase inhibitor and application thereof
A trehalase and inhibitor technology, applied in the field of trehalase inhibitors, can solve problems such as research reports that have not yet seen the effect of trehalase activity
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Embodiment 1
[0037] Example 1 Theoretical calculation of inhibitors isomalum and human trehalose
[0038] 1. Human seaweed sugar enzyme homologous modeling and verification
[0039] Since human and other mammal sources have not been parsed, this study was homologous to HSTRE in order to obtain HSTRE three-dimensional structural information. Submitting the HSTRE sequence to SWSS-Model, search the Swiss-Model Template library with BLAST and HHBLITS to find the related structure that matches the target sequence. Search for the trendy saccharin structure PDB ID from the insect intestinal citariopaus troine space: 5Z66, select the template to model and evaluate and verify.
[0040] 2. Treatment of receptor proteins
[0041] The QUICKPREP module of the Molecular Simulation Software MoE Dock 2019 performs the correction, protonation and energy of the HSTRE's three-dimensional structure, the protonization, and energy of the protein HSTRE side chain, and hydrogen is added, and the residue protonation, ...
Embodiment 2
[0049] Example 2 List of inhibitory kinetics of algae sugar inhibitors
[0050] 1. Influence of inhibitor concentration on inhibitory effect
[0051] The reaction system assay was determined by: 20 enzyme active trehalose; 15 enzyme; the reaction trehalose solution, 100 mM pH 5.5 citrate buffer, the total reaction volume of 200 punch, the above solutions After 2 minutes of preheating at 50 ° C, it was added to the trehalose, and 10 min was reacted at 50 ° C constant temperature conditions, and then placed in a boiling water bath for 20 min, and then added 200 μl of DNS solution to boiling water bath for 5 minutes, and after the cooling was measured at 540 nm. The absorbent value; if the reaction liquid contains a reducing component, the determination of glucose content is performed using a glucose oxidase method (GOPOD oxidase method), and the measurement method is carried out in accordance with the kit operand provided by Suzhou Geni Biotechnology Co., Ltd.. Set three parallel pe...
Embodiment 3
[0059] Example 3 is a isomaliosteal simulated digestion experiment, which is used to illustrate the feasibility of the application of isomalt sugar in the preparation of the use of human algae hydrolyzase to regulate the disease.
[0060] 1, in vitro simulation oral digestion
[0061] Configure soluble starch solutions (1%, w / w), isomorphoid 0.1 mol / L and saliva amylase solution (20 U / mL), 50 mmol / L pH 7.4 sodium phosphate buffer. According to the following Table 2, the orally simulated digestive system is formulated. The total volume of the reaction system is 200 μl, placed in a reactive system in a boiling water bath after 37 ° C, 120 rpm, and 30 min, and then uses a glucose oxidase method (GOPOD oxidase method). The determination of glucose was carried out, and each group was repeatedly measured three times.
[0062] Table 2 In vitro simulation oral digestion system
[0063]
[0064]
[0065] 2, in vitro simulated gastrointestinal digestion
[0066] Gastrointestinal...
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