Immobilization method of xylanase and immobilized xylanase

A xylanase and modification technology, applied in the field of enzyme engineering, can solve the problems of high price of enzymes, limited application, difficult recovery, etc. Effect

Inactive Publication Date: 2017-02-22
FUYANG NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, xylanase is mostly hydrolyzed in the form of free enzymes, but due to the high price of enzymes, difficulty in recycling, and easy inactivation by heat, acid-base or metal ions during the reaction process, it is largely limited. Its application in industrial production

Method used

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  • Immobilization method of xylanase and immobilized xylanase
  • Immobilization method of xylanase and immobilized xylanase
  • Immobilization method of xylanase and immobilized xylanase

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Weigh APTES modified Fe 3 o 4 @SiO 2 Nanoparticles 1.0057g was placed in 100mL (pH=5.5) of acetic acid-sodium acetate buffer solution, added 4% (v / v) 19.05mL concentration of 25wt% glutaraldehyde solution, mechanically stirred in a water bath at 25°C, 180rpm After stirring for 4 hours, APTES modified Fe 3 o 4 / SiO 2 Nanoparticles loaded with glutaraldehyde;

[0054] Add 0.1295 g of xylanase dissolved in acetic acid-sodium acetate buffer solution, and mechanically stir at 180 rpm for 3 hours to obtain immobilized xylanase;

[0055] Magnetically separate, wash with acetic acid-sodium acetate buffer solution 3 to 4 times, and finally use acetic acid-sodium acetate buffer solution to make up to 100mL; put it in the refrigerator for later use.

[0056]Test the impact of different influencing factors on the enzymatic activity of the immobilized xylanase obtained in the present embodiment and the free state xylanase:

[0057] 1. Preparation of solutions and samples

[...

Embodiment 2

[0107] Weigh APTES modified Fe 3 o 4 @SiO 2 Nanoparticle 5g is placed in the acetic acid-sodium acetate buffer solution of 150mL (pH=5.5), add 4% (v / v) 95mL concentration and be 25wt% glutaraldehyde solution, carry out mechanical stirring in 25 ℃ of water baths, 180rpm stirs 4 hours, get APTES modified Fe 3 o 4 / SiO 2 Nanoparticles loaded with glutaraldehyde;

[0108] Add 0.65 g of xylanase dissolved in acetic acid-sodium acetate buffer solution, and mechanically stir at 180 rpm for 3 hours to obtain immobilized xylanase;

[0109] Magnetically separate, wash with acetic acid-sodium acetate buffer solution 3 to 4 times, and finally use acetic acid-sodium acetate buffer solution to make up to 100mL; put it in the refrigerator for later use.

[0110] The immobilized xylanase obtained in the present embodiment is tested for performance using the experimental means in Example 1, and the results are as follows:

[0111] 60°C is the best pretreatment temperature for immobilize...

Embodiment 3

[0118] Weigh APTES modified Fe 3 o 4 @SiO 2 Nanoparticle 15g is placed in the acetic acid-sodium acetate buffer solution of 500mL (pH=5.5), add 4% (v / v) 285mL concentration and be 25wt% glutaraldehyde solution, carry out mechanical stirring in 25 ℃ of water baths, 180rpm stirs 4 hours, get APTES modified Fe 3 o 4 / SiO 2 Nanoparticles loaded with glutaraldehyde;

[0119] Add 1.95 g of xylanase dissolved in acetic acid-sodium acetate buffer solution, and mechanically stir at 180 rpm for 3 hours to obtain immobilized xylanase;

[0120] Magnetically separate, wash with acetic acid-sodium acetate buffer solution 3 to 4 times, and finally use acetic acid-sodium acetate buffer solution to make up to 300mL; put it in the refrigerator for later use.

[0121] The immobilized xylanase obtained in the present embodiment is tested for performance using the experimental means in Example 1, and the results are as follows:

[0122] 60°C is the best pretreatment temperature for immobili...

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Abstract

The invention provides an immobilization method of xylanase and immobilized xylanase. In the invention, xylanase is immobilized on the surface of a 3-aminopropyltriethoxysilane-modified Fe3O4@SiO2 nanoparticle carrier by a covalent bond (peptide bond) through the binding function of a bifunctional reagent to obtain immobilized xylanase, which limits the spatial configuration transformation of xylanase, so as to realize the heat tolerance, acid and alkali resistance, and resistance to interference of metal ions and inhibitors of xylanase; and the immobilized xylanase has relatively high tolerance to heat (40-70 DEG C), relatively wide adaptability to extreme pH values (3.0-8.0), relatively high tolerance to metal ions (Ba<2+>, Fe<2+>, Mn<2+>, Ca<2+>) and interfering agents (blue moon hand sanitizer, Triton X-100, Tween 20, EDTA solution, ethanol), and excellent operational stability. The activity of the immobilized xylanase can still be maintained at 73% or higher after being repeatedly used for 9 times.

Description

technical field [0001] The invention relates to the technical field of enzyme engineering, in particular to a method for immobilizing xylanase and the immobilized xylanase. Background technique [0002] Since the term "hemicellulose" was first used in 1891 to describe components extracted and isolated from plant materials with dilute lye, xylan is the most abundant renewable resource in nature after cellulose in terms of carbohydrate content One has received widespread attention. Due to the complexity of xylan structure, its complete degradation requires the participation of various hydrolytic enzymes to complete. Xylanase (Xylanase) is a kind of hydrolytic enzyme, which can degrade hemicellulose. It mainly contains three enzymes: the first is β-xylosidase, and the second is exo-β-1,4-xylanase. Carbohydrase, the third is endo-β-1,4-xylanase. [0003] Xylanase is widely used in the food industry, paper industry, animal husbandry, etc., and has health care functions for the...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N11/14C12N9/42
CPCC12N9/2482C12N11/14C12Y302/01008
Inventor 宋崇富盛良全戴亚
Owner FUYANG NORMAL UNIVERSITY
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