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Method for fixing pullulanase with chitosan magnetic nanoparticles

A magnetic nanoparticle, pullulanase technology, applied in the field of enzyme engineering, can solve the problems of complicated process, limited application, harsh conditions, etc., and achieve the effects of high enzyme activity, increased use value, and simple steps.

Inactive Publication Date: 2014-10-15
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] At present, the synthesis of magnetic chitosan / ferric oxide nanoparticles mainly adopts microemulsion and emulsion polymerization, but these synthesis methods still have complex processes and harsh conditions (higher temperature and pressure, protective atmosphere, organic solvent and organic surfactants, etc.); and most of these synthetic methods are carried out in water / oil systems, however, since most of the actual production uses water systems, this may also limit its application in actual production

Method used

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  • Method for fixing pullulanase with chitosan magnetic nanoparticles
  • Method for fixing pullulanase with chitosan magnetic nanoparticles
  • Method for fixing pullulanase with chitosan magnetic nanoparticles

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Add 3g of chitosan powder to 100ml of 2% acetic acid solution, stir until the chitosan is completely dissolved; slowly add 0.4ml of 25% glutaraldehyde solution, continue to stir at a constant speed for 15min, and then let stand for 24h to obtain chitosan hydrogel Gel; soak the hydrogel in 0.15mol / L Fe 3+ Aqueous solution, water, 0.075mol / L Fe 2+ In the aqueous solution, it was circulated four times, at this time, the chitosan amino group chelated with iron and ferrous ions; the chitosan hydrogel containing iron ions was soaked in 1.25 mol / L NaOH solution for alkalization treatment for 12 hours, and obtained Black hydrogel, the hydrogel is hydrolyzed with 3% acetic acid and 5% hydrogen peroxide solution to obtain black nanoparticles with magnetic response.

[0036]The magnetically responsive substances were collected under an external magnetic field, and the chitosan magnetic particles aggregated within 10 s. The supernatant was discarded, and then rinsed repeatedly wit...

Embodiment 2

[0041] Add 4 g of chitosan powder into 100 ml of 2% acetic acid solution, stir until the chitosan is completely dissolved; slowly add 0.6 ml of glutaraldehyde solution, continue to stir at a constant speed for 20 minutes, and then let it stand for 12 hours to obtain a chitosan hydrogel; Soak the hydrogel sequentially in 0.2mol / L Fe 3+ Aqueous solution, water, 0.1mol / L Fe 2+ In the aqueous solution, cycle four times; then soak the chitosan hydrogel containing iron ions in 1.5 mol / L NaOH solution for alkaline treatment for 12 hours to obtain a black hydrogel, which is treated with 2% acetic acid and 4 % hydrogen peroxide solution was hydrolyzed to obtain black magnetically responsive nanoparticles, and the magnetically responsive substances were collected under an external magnetic field. The chitosan magnetic particles aggregated within 10 s, and the supernatant was discarded, and then rinsed repeatedly with distilled water to neutral; the obtained magnetic nanoparticles have ...

Embodiment 3

[0046] Add 5 g of chitosan powder to 100 ml of 3% acetic acid solution, stir until the chitosan is completely dissolved; slowly add 0.8 ml of glutaraldehyde solution, continue to stir at a constant speed for 30 min, and then let stand for 24 h to obtain chitosan hydrogel ; Soak the hydrogel sequentially in 0.2mol / L Fe 3+ Aqueous solution, water, 0.1mol / L Fe 2+ In the aqueous solution, cycle four times; soak the chitosan hydrogel containing iron ions in 1.25 mol / L NaOH solution for alkalization treatment for 12 hours, and obtain a black hydrogel, which is treated with 4% acetic acid and 8% The hydrogen peroxide solution was hydrolyzed to obtain black magnetically responsive nanoparticles. The magnetically responsive substances were collected under an external magnetic field. The chitosan magnetic particles aggregated within 10 s. The supernatant was discarded, and then rinsed repeatedly with distilled water until Neutral; the obtained magnetic nanoparticles have good dispersio...

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Abstract

A method for fixing pullulanase with chitosan magnetic nanoparticles comprises the steps as follows: (1) the chitosan magnetic nanoparticles are prepared; (2) amination treatment is performed on surfaces of the chitosan magnetic nanoparticles; (3) the chitosan magnetic nanoparticles after amination treatment and the pullulanase form covalent bonds in the presence of a cross-linking agent to fix the pullulanase. Compared with a traditional pullulanase fixing method, the method has the advantages that a carrier prepared with a chitosan hydrogel induction method is firstly used for fixing the pullulanase, and the method is simple in step, mild in condition and environment-friendly.

Description

technical field [0001] The invention relates to the technical field of enzyme engineering, in particular to a new method for immobilizing pullulanase with chitosan magnetic nanoparticles prepared by a hydrogel induction method. Background technique [0002] Pullulanase (Pullulanase, EC 3.2.1.41) is a debranching enzyme that has important uses in the food processing industry that uses starch as raw material. It can decompose the smallest unit of branched chain and maximize the use of starch raw materials. . At present, it is mainly used in the industry to increase the output and purity of high glucose syrup and ultra-high maltose syrup; to increase the maltose content of beer syrup; to increase the content of resistant starch in potato starch, corn starch, and sweet potato starch and to produce high amylose. Wait. [0003] Compared with free enzymes, immobilized enzymes have many advantages, so research on various immobilized enzyme technologies and carrier materials has em...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N11/14C12N11/10B01J13/04
Inventor 金征宇龙杰焦爱权徐学明谢正军田耀旗周星赵建伟杨哪王金鹏
Owner JIANGNAN UNIV
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