Method for synergistically immobilizing L-arabinose isomerase and ionic liquid

A technology of ionic liquid and arabinose, applied in the direction of isomerase, immobilized on/in organic carrier, biochemical equipment and method, etc., can solve the problem of low enzyme production level, separation and purification of unfavorable products, large amount of addition, etc. problems, to achieve the effect of increasing the amount of enzyme protein immobilization, broad commercial application prospects, and improving activity

Inactive Publication Date: 2018-05-15
TIANJIN MODERN VOCATIONAL TECH COLLEGE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In recent years, research at home and abroad has reported some microorganisms that can be used to ferment and produce L-arabinose isomerase, but the enzyme production levels of these microorganisms are all low. If L-ribulose is produced in the ...

Method used

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  • Method for synergistically immobilizing L-arabinose isomerase and ionic liquid
  • Method for synergistically immobilizing L-arabinose isomerase and ionic liquid

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] The preparation of step 1 PANF-I

[0026] Add 2.0g of dry acrylic fiber (PANF), ethylenediamine (0.12mol) and 30mL of deionized water into a 100mL three-neck flask, reflux for 4h under electromagnetic stirring, then take out the fiber sample, filter it with suction, and use a 60-70℃ Wash with water repeatedly until the lotion is neutral. After drying in the air, then vacuum-dry at 60°C for 12 hours to obtain PANF-I.

[0027] The preparation of step 2 PANF-II

[0028] Dried PANF-I was stirred in 1M, 2eq nitric acid solution for 3h at room temperature, and then the formed fiber-immobilized ionic liquid was taken out, filtered with suction, and rinsed with deionized water until neutral. After drying in the air, it was dried in vacuum (60°C) for 12h.

[0029] Step 3 Preparation of PANF-III

[0030] Add PANF-II and 5 mL of free enzyme solution to 28 mL of 0.5% glutaraldehyde aqueous solution, shake and cross-link at room temperature (25°C) for 1 h, pour off the solution,...

Embodiment 2

[0037] The preparation of step 1 PANF-I

[0038] Add 2.0g of dry acrylic fiber (PANF), 0.12mol of butanediamine and 30mL of deionized water into a 100mL three-neck flask, reflux for 4h under electromagnetic stirring, then take out the fiber sample, filter it with suction, and use 60-70℃ water Wash repeatedly until the lotion is neutral. After drying in the air, then vacuum-dry at 60°C for 12 hours to obtain PANF-I.

[0039] The preparation of step 2 PANF-II

[0040] Dried PANF-I was stirred in 1M, 2eq sulfuric acid solution at room temperature for 3h, and then the formed fiber-immobilized ionic liquid was taken out, filtered with suction, and rinsed with deionized water until neutral. After drying in the air, it was dried in vacuum (60°C) for 12h.

[0041] Step 3 Preparation of PANF-III

[0042] Add PANF-II and 5 mL of free enzyme solution to 28 mL of 0.5% glutaraldehyde aqueous solution, shake and cross-link at room temperature (25°C) for 1 h, pour off the solution, and w...

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Abstract

The invention provides a method for synergistically immobilizing L-arabinose isomerase and ionic liquid. Acrylic fibers are used as a carrier to synergistically immobilize ion liquid and L-arabinose isomerase. The method comprises the following steps: adding acrylic fibers, a compound I and 30mL of deionized water into a three-opening bottle, refluxing, taking out a fiber test sample, suction filtering, washing by using water, air drying, vacuum drying, and obtaining PANF-I, wherein a molecular formula of the compound I is H2N(CH2)nCH2NH2; stirring the dry PANF-I in an HY acid solution, takingout the formed fiber immobilized ion liquid, suction filtering, sprinkling, air drying, and drying; and adding PANF-II and 5mL of freeze enzyme solution into a cross-linking agent, oscillating cross-linking, pouring out the solution, washing the immobilized enzyme particles separately by using 0.9 percent sodium chloride and deionized water, suction filtering, and drying, thus obtaining the immobilized enzyme, and storing at a low temperature. According to the method, the acrylic fibers are adopted as the carrier to synergistically immobilize the ethylamine nitrate ion liquid and L-arabinoseisomerase. The method is mild in condition, high in yield, simple in method, and easy in mass preparation.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular to a method for synergistically immobilizing L-arabinose isomerase and ionic liquid. Background technique [0002] L-arabinose isomerase is an intracellular enzyme that can catalyze the synthesis of rare functional sugars such as D-tagatose and L-ribulose, and has important applications in the fields of food and medicine. Among them, L-ribulose can be converted into L-ribose through one-step biocatalysis. L-ribose has huge potential biological activity in food and medicine, and L-ribulose is the most effective biosynthetic pathway for preparing L-ribose. Among them, L-arabinose isomerase is one of the most effective enzymes for biological production of L-ribulose. In recent years, domestic and foreign studies have reported some microorganisms that can be used to ferment and produce L-arabinose isomerase, but the enzyme production levels of these microorganisms are all low. If L-ribulos...

Claims

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Application Information

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IPC IPC(8): C12N11/08C12N9/90
CPCC12N9/90C12N11/08C12Y503/01004
Inventor 袁文蛟王静刘鹏齐菲刘佳刘美玲
Owner TIANJIN MODERN VOCATIONAL TECH COLLEGE
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