Screening method of reversible inhibition protective agent of chitosan hydrolase

A hydrolase and chitosan technology, applied in biochemical equipment and methods, microbial determination/inspection, instruments, etc., can solve the problem that enzyme reversible inhibitors do not have a specific screening model structure information database, the research field is difficult, and there is no effective methods, etc., to achieve the effect of rapid screening methods, strong pertinence, and reliable methods

Inactive Publication Date: 2021-03-30
枣庄市杰诺生物酶有限公司
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Problems solved by technology

[0004] There is no ready-made specific screening model and structural information database for industrial enzyme reversible inhibitors, which has always been a difficult problem in the research field, and no breakthrough has been made in China
And there is no effective method in the technical field of chitosanase reversible inhibition protective agent screening

Method used

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  • Screening method of reversible inhibition protective agent of chitosan hydrolase
  • Screening method of reversible inhibition protective agent of chitosan hydrolase
  • Screening method of reversible inhibition protective agent of chitosan hydrolase

Examples

Experimental program
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Effect test

Embodiment 1

[0028] The screening method of the reversible inhibition protective agent of embodiment 1 chitosan hydrolase

[0029] The screening method of the reversible inhibition protective agent of chitosan hydrolase comprises the following steps:

[0030] (1) According to the sequence structure of known chitosan hydrolase, the structural data of inhibitory protective agent, and the experimental data of the reaction of enzyme and small molecular compound (Table 1), the inhibitor contains sodium acetate (Sodium acetate), benzalkonium benzalkonium chloride, ethyl acetimidate, EDTA, glucosamine, guanidinium hydrochloride, cysteine , iodoacetic acid (monoiodoacetate);

[0031] Table 1: Table of Inhibition Constants for Chitosan Hydrolase Inhibitor Reaction

[0032]

[0033]

[0034] The inhibition constant (Ki) can reflect the ability of the compound to bind to the enzyme. Among these 18 compounds, the ability to bind to the chitosan hydrolase is too strong (Ki<0.3), which will easi...

Embodiment 2

[0050] Example 2 Docking Simulation of Compound ZINC01006243 and Chitosan Hydrolase

[0051] Select the compound 5,7-Diphenyl-1,7-dihydro-pyrrolo[2,3-d]pyrimidine-2,4-dithione with the first scoring value for research, dissolve it in ethanol to prepare a 1mM solution, and follow the 25μM The amount that joins in the concentrated chitosan hydrolase solution (8000u / ml), enzymatic activity is suppressed, retains activity 18.76%. After being stored at 40°C for 30 days, it was diluted according to 1:10 times and tested. The result was that the activity was 652u / ml, and the activity was recovered, and the relative enzyme activity exceeded 80%.

[0052] The above results show that the theoretical and experimental results of the screening method established by the present invention complement each other and have high reliability.

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Abstract

The technical problem to be solved by the invention is to provide a screening method of a reversible inhibition protective agent of chitosan hydrolase so as to overcome the defects in the prior art, thereby being suitable for application of the chitosan hydrolase to an industrial enzyme preparation and improving the stability of the chitosan hydrolase. The design thought of the method comprises the steps of firstly, establishing a known reversible inhibition protective agent training set, performing molecular docking and sorting according to existing chitosan hydrolase structure data, and performing verification in combination with a test result to determine the reliability of the method; and then establishing a screening rule and a screening model, respectively screening, sorting and scoring the small molecule compound database by using the rule and the model, gradually narrowing the range, and finally determining the compound which can be used as the reversible inhibition protectiveagent of the chitosan hydrolase according to the sorting results of several times from top to bottom.

Description

technical field [0001] The invention belongs to the technical field of active substance screening, and in particular relates to a screening method for a protective agent for reversible inhibition of chitosan hydrolase. Background technique [0002] Enzymes are characterized by less dosage, high catalytic efficiency and strong specificity. After more than half a century of rapid development, the enzyme preparation industry has now formed a dynamic high-tech industry, which is widely used in food, textile, washing, papermaking, feed and bio-energy industries. At present, compared with similar products, liquid enzyme preparation technology saves water by 30% to 50% and saves energy by more than 50%, which is more in line with the development needs of low-carbon economy; there is no doubt that high-activity, high-purity, liquid enzymes will become The development direction of enzyme preparation in the future. [0003] The activity and function of an enzyme depend on the integr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G16B5/00G16B35/20G16B50/00C12Q1/34
CPCG16B5/00G16B35/20G16B50/00C12Q1/34
Inventor 许向阳宋在伟殷允超种法国宋均营黄家超叶王立王鹏
Owner 枣庄市杰诺生物酶有限公司
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