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Biomimetic affinity purification method of p-aminobenzamidine biomimetic affinity ligand

A p-aminobenzamidine, amination technology, applied in chemical instruments and methods, other chemical processes, ion exchange regeneration, etc., can solve the problems of increasing steps and costs, affecting the utilization of metalloproteases, etc., to achieve simple process and activity recovery high rate effect

Active Publication Date: 2020-07-17
YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Moreover, in the process of metal chelation chromatography, imidazole with certain toxicity needs to be used in the elution process, which will affect the utilization of metalloproteases as pharmaceuticals, and the use of desalting columns to process imidazole will increase additional steps and costs.

Method used

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Experimental program
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Embodiment 1

[0035] 1. Synthesis of biomimetic affinity materials

[0036] 1), epichlorohydrin activated agarose gel

[0037] The agarose gel (Sepharose 6B) was thoroughly washed with double distilled water at a ratio of 1:10 V / V until the pH of the solution reached 7.0, and the washed agarose gel was air-dried and then dissolved in the activation solution (1M Sodium hydroxide, 2.5g dimethyl sulfoxide and 10ml epichlorohydrin), shake at 100rpm in a shaker at 40°C for 2.5h for activation to obtain epichlorohydrin-activated sepharose 6B (see process a).

[0038] 2), amination of epichlorohydrin activated agarose gel

[0039] Add 35% saturated ammonia to the activated agarose gel 6B, then add a certain amount of double distilled water, and incubate overnight at 100 rpm in a shaker at 30°C, so that the activated agarose gel 6B is added with amino groups to obtain aminated activated agarose gel. Glue 6B (see procedure b).

[0040] 3), connecting the cyanuric chloride linking arm

[0041] Ad...

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Abstract

The invention relates to a bionic affinity purification method of p-aminobenzamidine bionic affinity ligand, including the preparation of bionic affinity materials and the bionic affinity of p-aminobenzamidine agarose gel column to metalloprotease MP purification. The purification method has the advantages of simple flow, short period, high efficiency and high purity of 98.7%.

Description

technical field [0001] The invention relates to the technical field of marine microorganisms, in particular to a biomimetic affinity purification method of p-aminobenzamidine biomimetic affinity ligand. Background technique [0002] The biomimetic affinity ligands of marine microbial enzymes are usually derived from specific inhibitors of the enzymes. In previous studies, based on the crystal structure information of marine low-temperature alkaline protease MP, it was prepared by fermentation of marine bacteria YS-80 (marine bacteria YS-80 was isolated from deep sea mud marine bacteria (YS-80), and was preserved in CCTCC, Wuhan, China , the preservation number is CCTCC NO:M209177, see patent application number 200910177301.0, a small molecule inhibitor library of marine low-temperature alkaline protease MP was constructed through virtual screening. Among the 15 most effective small molecule reversible inhibitors predicted by fine screening Five of them are benzamidine deriv...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): B01J20/22B01J20/30B01D15/08C12N9/50
CPCB01D15/08B01J20/22C12N9/50
Inventor 李尚勇孙谧郝建华刘均忠王跃军马子宾
Owner YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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