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Method of enzymatic degradation of 4-aminosalicylic acid

A technology for enzymatic degradation of aminosalicylic acid, which is applied in the field of enzymatic degradation of 4-aminosalicylic acid, can solve the problems of low decomposition efficiency and high energy consumption, and achieve reverse reaction prevention, low energy consumption, and improved enzyme reaction activity Effect

Inactive Publication Date: 2019-09-06
JINING UNIV
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Problems solved by technology

In the prior art, the decomposition of 4-aminosalicylic acid mainly relies on heating means, and industrial application will consume a lot of heat energy, and the decomposition efficiency will be low as the reverse reaction proceeds

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  • Method of enzymatic degradation of 4-aminosalicylic acid
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  • Method of enzymatic degradation of 4-aminosalicylic acid

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Embodiment Construction

[0023] The present invention will be described in further detail below in combination with specific embodiments.

[0024] figure 1 A method for enzymatic degradation of 4-aminosalicylic acid shown, comprising the following steps:

[0025] 1. Extract and amplify salicylic acid decarboxylase Sdc gene; activate Trichosporon moniliiforme WU-0401 in LB liquid medium and extract Sdc gene from Trichosporon yeast strain; Trichosporon moniliiforme WU-0401, the species Trichosporium yeast can metabolize salicylic acid decarboxylase, and salicylic acid decarboxylase has decarboxylation to 4-aminosalicylic acid to generate meta-aminophenol; the Sdc gene of salicylic acid decarboxylase Sdc is amplified by PCR polymerase chain reaction, To prepare a large amount of salicylate decarboxylase.

[0026] Ⅱ. Make a recombinant plasmid containing the Sdc gene; carry out double digestion of the Sdc gene and the carrier plasmid after PCR amplification, and culture overnight at 16°C under the actio...

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Abstract

The invention discloses a method of enzymatic degradation of 4-aminosalicylic acid. The method includes: extracting and amplifying salicylic acid decarboxylase Sdc genes; preparing recombinant plasmids containing the Sdc genes; constructing Escherichia coli Sdc expression strains for inducible expression; performing ultrasonic fragmentation of Escherichia coli Sdc and purification of salicylic acid decarboxylase; arranging a reaction box and a reaction system. In the method, technical conditions that high temperature is needed for degradation of 4-aminosalicylic acid are replaced, so that thedecomposition reaction is easier to carry out, and the degradation efficiency is higher; a large number of salicylic acid decarboxylase can be prepared by genetic engineering, so that the applicationof the salicylic acid decarboxylase can be industrialized, and the metabolic yield per unit time can be increased; the reaction system with the highest metabolic efficiency is prepared, ferrous chloride is added in the reaction system, a pumping pump is arranged on the reaction box, the enzyme reaction activity can be improved, the occurrence of reverse reaction is prevented, and the decompositionefficiency is improved; mass spectrometry is more accurate than colorimetric method in the prior art in measuring the enzyme activity.

Description

technical field [0001] The invention relates to a method, in particular to a method for enzymatically degrading 4-aminosalicylic acid. Background technique [0002] Salicylic acid and its derivatives are widely used in medicine, chemical industry and other fields, but due to their toxicity and long natural degradation time, they will affect the natural ecological balance and human health. Therefore, it is important and necessary to study their degradation methods for realizing green production. Sex [1-3]. Moreover, the decarboxylation products of salicylic acid and its derivatives - phenol and its derivatives are also important organic chemical raw materials, widely used in the chemical industry and pharmaceutical industry, and can be used as intermediates in chemical synthesis and important Chemical raw material. [0003] 4-Aminosalicylic acid is unstable in aqueous solution above 40°C, decomposes and releases carbon dioxide to become m-aminophenol. In the prior art, the...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/88C12N15/60C12N15/70C12Q1/527
CPCC12N9/88C12N15/70C12Q1/527C12Y401/01091
Inventor 胡彦营张爱民李薇韩小龙杨永涛
Owner JINING UNIV
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