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Immobilized lysine decarboxylase, preparation thereof, 1, 5-pentane diamine preparation method and product

A technology of lysine decarboxylase and amine group, which is applied in the direction of biochemical equipment and methods, fixed on/in organic carriers, enzymes, etc., can solve the problem of less immobilized lysine decarboxylase, etc., and achieve a solution The effects of poor stability in use, high vitality, and convenient recycling

Pending Publication Date: 2018-03-09
CATHAY R&D CENT CO LTD +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] At present, there are few reports on the preparation and stability of immobilized lysine decarboxylase, and there are no reports on the chemical modification of cheap carriers to make it suitable for the preparation of immobilized lysine decarboxylase

Method used

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Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0100] The preparation process of the immobilized lysine decarboxylase of the present invention can be obtained by reacting the aldehyde-modified polymer carrier and the lysine decarboxylase in a buffer solution.

[0101] In some embodiments, the amount of lysine decarboxylase relative to the aldehyde-modified polymer carrier can be adjusted, and the resulting immobilized lysine decarboxylase can maintain high enzyme activity. In a preferred embodiment, the amount of lysine decarboxylase is 1500-4000 U, more preferably 2000-3500 U, most preferably 2500-3000 U per gram of the polymer carrier.

[0102] In some embodiments, the activity concentration of the lysine decarboxylase solution can be controlled at 100-1000 U / mL.

[0103] The aldehyde-modified polymer carrier and the lysine decarboxylase generally react in a buffer solution. The buffer solution can be selected from acetate buffer, phosphate buffer, citrate buffer, etc., preferably phosphate buffer.

[0104] The pH valu...

Embodiment 1

[0124] Modification of Cellulose Acetate Hydrolyzate Carrier and Preparation of Immobilized Lysine Decarboxylase

[0125] (1) Add 100 g of cellulose acetate to 1 L of NaOH solution with a concentration of 1 mol / L, add it to a boiling water bath for hydrolysis for 1-2 hours, pour out the hydrolyzate, and wash the hydrolyzed cellulose acetate with water for 3-5 times, and then stored in water for later use.

[0126] (2) 50g of hydrolyzed cellulose acetate is added to a concentration of 10% of 500g of β-sulfone ethylsulfone aniline solution, and under the condition of heating in a water bath at 60°C, etherification reaction is carried out, and sodium hydroxide solution is added dropwise at the same time. , manual stirring, until the pH is about 11, stop the reaction, wash the cellulose acetate with hot water to neutral.

[0127] (3) Add 30 g of etherified cellulose acetate to 200 g of 10% glutaraldehyde solution, stand at 20° C. for 2 hours, wash the fibers with water, and colle...

Embodiment 2

[0134] Modification of Polyvinyl Alcohol Carrier and Preparation of Immobilized Lysine Decarboxylase

[0135] (1) Take 30g of polyvinyl alcohol, put it into 500g of β-sulphate isosulfone-aniline solution with a concentration of 10%, and carry out etherification reaction under 60°C water bath heating condition, add dropwise sodium hydroxide solution at the same time, manually Stir until the pH is about 11, stop the reaction, and wash the polyvinyl alcohol fiber with hot water to neutrality.

[0136] (2) Add 30 g of etherified PVA to 200 g of 10% glutaraldehyde solution, react at room temperature for 2 hours, wash the fibers with water, and collect them for later use.

[0137] (3) Put the above-mentioned PVA fiber activated by glutaraldehyde into 90g of lysine decarboxylase solution with a pH value of 7.0, the amount of lysine decarboxylase is 3000U based on per gram of the polymer carrier, and the solution is ionic concentration Prepare immobilized lysine decarboxylase with 0....

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PUM

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Abstract

The invention relates to immobilized lysine decarboxylase, a preparation thereof, a 1, 5-pentane diamine preparation method and the prepared 1, 5-pentane diamine. The invention specifically relates toimmobilized lysine decarboxylase comprising an aldehyde group-modified polymer carrier and lysine decarboxylase, and a method for preparing immobilized lysine decarboxylase by the aldehyde group-modified polymer carrier; a method for preparing 1, 5-pentane diamine by the immobilized lysine decarboxylase, and the prepared 1, 5-pentane diamine thereof. The immobilization method has high enzyme immobilization efficiency and good stability in use, improves the use efficiency of enzyme, solves the problem of poor use stability of free lysine decarboxylase (cells) in traditional processes, greatlyreduces the use cost of the carrier and the biological production cost of 1, 5-pentanediamine, simplifies the separation step of 1, 5-pentane diamine solution and enzyme, increases the degree of automation of the production of 1, 5-pentane diamine, and promotes the process of industrialization of biological production of 1, 5-pentanediamine.

Description

technical field [0001] The present invention relates to the field of biochemical engineering, in particular to an immobilized lysine decarboxylase, a preparation method thereof, a method for preparing 1,5-pentanediamine using the immobilized lysine decarboxylase, and the prepared 1,5-pentanediamine 5-pentanediamine. Background technique [0002] Biocatalytic reactions have the characteristics of mild reaction conditions, high selectivity, and high reaction efficiency, so they are increasingly used in the field of chemical synthesis. However, due to the poor stability of biological enzymes in in vitro industrial environments and the high cost of enzyme preparations, the practical application of biocatalysis in the industrial field is greatly limited. [0003] Immobilized enzyme technology not only effectively solves the problem of repeated use of catalysts, but also significantly improves the stability and catalytic properties of biological enzymes (substrate affinity, stere...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N11/12C12N11/08C12N11/02C12P13/00
CPCC12N9/88C12N11/02C12N11/08C12N11/12C12P13/001C12Y401/01018
Inventor 徐冠珠于丽珺刘修才
Owner CATHAY R&D CENT CO LTD
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