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Method for screening trimethylamine lyase inhibitor by utilizing microorganisms

A technology of trimethylamine and lyase, applied in the field of microorganisms, can solve the problems of accelerated drug screening, time-consuming and labor-intensive

Inactive Publication Date: 2020-12-04
CHINA PHARM UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Analyzing the matching degree between ligands and receptors can greatly speed up drug screening by using computer-aided drug design (CADD). The isotope labeling method requires specific isotope detection instruments; while animal experiments need to raise animals, which is time-consuming and laborious

Method used

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  • Method for screening trimethylamine lyase inhibitor by utilizing microorganisms
  • Method for screening trimethylamine lyase inhibitor by utilizing microorganisms

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Experimental program
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Embodiment 1

[0022] 1) Selection and cultivation of indicator bacteria based on choline conversion to trimethylamine

[0023] After purchasing the indicator bacteria Klebsiella pneumoniae from ATCC (( Klebsiella Pneumoniae )) ATCC 13883, Klebsiella pneumoniae ATCC 13883 was inoculated onto Nutrient Broth (NB) solid plates from a slant and incubated anaerobically at 37°C for 24 hours. Inoculate the activated bacterial species into the test tube containing the NB liquid medium, and incubate at 37 ° C for 24 hours to prepare the indicator bacteria liquid.

[0024] Among them: NB solid medium: each liter of medium contains: peptone 10g, beef extract 3g, sodium chloride 5g, agar 20g, distilled water 1000mL, adjust the pH to 7.2; 121 ℃ autoclave for 15min.

[0025] NB liquid medium: Each liter of medium contains: peptone 10g, beef extract 3g, sodium chloride 5g, distilled water 1000mL, pH adjusted to 7.2; autoclaved at 121°C for 15min.

[0026] 2) Determination of minimum inhibitory concentra...

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Abstract

The invention relates to microorganisms, in particular to a method for screening a trimethylamine lyase inhibitor by utilizing microorganisms. The method for screening the trimethylamine lyase inhibitor by utilizing microorganisms is characterized by comprising the following steps: inoculating a specific culture medium only containing cholineinoculator with Klebsiella pneumoniae ATCC 13883 servingas an indicator; adding a substance to be screened into the culture medium at a concentration lower than the minimum inhibitory concentration, and carrying out anaerobic culture for 12-48 hours at 37DEG C; and taking a culture solution, determining the content of trimethylamine, taking a culture solution without the to-be-detected substance as a contrast, and indicating that the substance can inhibit microorganisms from converting choline to generate trimethylamine and is a trimethylamine lyase inhibitor when the content of trimethylamine is lower than that of a control group; The method hasthe advantages of quickness, simplicity, convenience, easiness in operation, low reagent demand, capability of performing high-throughput screening and accurate result.

Description

technical field [0001] The invention relates to microorganisms, in particular to a method for screening trimethylamine lyase inhibitors using microorganisms. Background technique [0002] Cardiovascular disease (CVD) is a group of diseases that affect the heart and blood vessels, including coronary heart disease, angina pectoris, myocardial infarction, congenital heart disease, hypertension, stroke, valvular heart disease, cardiomyopathy and other atherosclerosis An important pathological factor in atherosclerosis (AS)-induced cardiovascular disease, characterized by slowly progressive inflammation in conductive and resistance arteries with accumulation of cholesterol-containing low-density lipoprotein (LDL) particles in the subendothelial layer. Finding effective strategies to inhibit atherosclerosis is of great scientific significance for reducing the morbidity and mortality of cardiovascular disease. [0003] The development of atherosclerosis is based on a combination o...

Claims

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Application Information

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IPC IPC(8): C12Q1/527C12Q1/02C12R1/22
CPCC12Q1/527C12Q1/025G01N2333/26
Inventor 储卫华衡星
Owner CHINA PHARM UNIV
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