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Immobilization method of xylanase

A technology of xylanase and immobilized carrier, applied in the direction of immobilization on/in organic carrier, can solve the problems of immobilized enzyme mechanical properties, poor stability, no involvement, etc. The effect of stability and high application value

Inactive Publication Date: 2015-12-16
TIANJIN MODERN VOCATIONAL TECH COLLEGE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

As a part of the immobilized enzyme, the carrier material of the immobilized enzyme has a great influence on the various properties of the immobilized enzyme. Chitosan has unique properties because of its rich hydroxyl and amino groups in the molecule, which makes it easy to carry out chemical reactions. , especially the rich amino group has a very high affinity with various proteins, it is a good carrier for enzyme immobilization, and there is no residual monomer, which makes it more advantageous as a carrier than other high polymers, but now In the prior art, chitosan is used as a carrier in combination with a cross-linking agent and a suitable immobilized enzyme preparation process is selected. Although the conversion rate and activity recovery rate of immobilized xylanase have been improved, the mechanical properties of the immobilized xylanase , poor stability, and there is no technology involving bacterial cellulose / chitosan composite microspheres as a carrier to realize the immobilization of enzymes in the prior art

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] A kind of immobilization method of xylanase of the present embodiment uses bacterial cellulose / chitosan composite microsphere as immobilized carrier, comprises the preparation of immobilized carrier and the preparation of immobilized xylanase, and the steps are as follows:

[0026] (1) Preparation of immobilized carrier:

[0027] a. Seed culture: insert Acetobacter xylinum CGMCC No.1.1812 slant seeds purchased from China Industrial Microbiology Culture Collection and Management Center into 100ml HS seed culture solution, and vibrate at 30°C for 24 hours at 160rpm to obtain seed culture liquid;

[0028] b. Preparation of bacterial cellulose: Inoculate an Erlenmeyer flask equipped with HS fermentation medium with an inoculum of 2% (v / v), and culture it statically in an incubator at 30°C for 6 days to form bacterial cellulose gel. glue;

[0029] c. Beating of bacterial cellulose: soak the bacterial cellulose gel prepared in step b in 0.1mol / l NaOH solution, boil at 100°C...

Embodiment 2

[0036] A kind of immobilization method of xylanase of the present embodiment uses bacterial cellulose / chitosan composite microsphere as immobilized carrier, comprises the preparation of immobilized carrier and the preparation of immobilized xylanase, and the steps are as follows:

[0037] (1) Preparation of immobilized carrier:

[0038] a. Seed culture: insert Acetobacter xylinum CGMCC No.1.1812 slant seeds purchased from China Industrial Microbiology Culture Collection and Management Center into 100ml HS seed culture solution, and vibrate at 30°C for 24 hours at 160rpm to obtain seed culture liquid;

[0039] b. Preparation of bacterial cellulose: Inoculate an Erlenmeyer flask equipped with HS fermentation medium with a 10% (v / v) inoculation amount, and culture it statically in an incubator at 30° C. for 2 days to form bacterial cellulose gel. glue;

[0040] c. Bacterial cellulose beating: soak the bacterial cellulose gel prepared in step b in 0.1mol / l NaOH solution, boil at...

Embodiment 3

[0047] A kind of immobilization method of xylanase of the present embodiment uses bacterial cellulose / chitosan composite microsphere as immobilized carrier, comprises the preparation of immobilized carrier and the preparation of immobilized xylanase, and the steps are as follows:

[0048] (1) Preparation of immobilized carrier:

[0049] a. Seed culture: insert Acetobacter xylinum CGMCC No.1.1812 slant seeds purchased from China Industrial Microbiology Culture Collection and Management Center into 100ml HS seed culture solution, and vibrate at 30°C for 24 hours at 160rpm to obtain seed culture liquid;

[0050] b. Preparation of bacterial cellulose: Inoculate an Erlenmeyer flask equipped with HS fermentation medium with an inoculum of 5% (v / v), and culture it statically in an incubator for 4 days at 30° C. to form bacterial cellulose gel. glue;

[0051] c. Bacterial cellulose beating: soak the bacterial cellulose gel prepared in step b in 0.1mol / l NaOH solution, boil at 100°C ...

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Abstract

The invention provides an immobilization method of xylanase, which comprises the processes of immobilization carrier preparation and immobilized xylanase preparation. The immobilization carrier preparation process mainly comprises the following steps: bacteria cellulose seed culture, bacteria cellulose preparation, bacteria cellulose pulping, and bacteria cellulose / chitosan composite microsphere preparation. The immobilized xylanase preparation process comprises the following steps: crosslinking the bacteria cellulose / chitosan microsphere and glutaric dialdehyde, carrying out crosslinking reaction on the crosslinked-formyl-containing bacteria cellulose / chitosan microsphere and a xylanase solution at 4 DEG C, centrifuging, precipitating and carrying out vacuum drying to obtain the immobilized xylanase. The activity recovery rate of the immobilized enzyme is up to 75.3%, and the relative enzyme activity reaches 80.5%. After the immobilized enzyme is continuously used 5 times, the enzyme activity is kept at 83.6%, and thus, the immobilized enzyme has favorable stability and higher application value.

Description

technical field [0001] The invention relates to an enzyme immobilization method, in particular to a xylanase immobilization method. Background technique [0002] Xylanase is a complex enzyme system that degrades xylan molecules. Its composition is relatively complex. It is the most critical hydrolase in the xylan hydrolase system. Xylan is hydrolyzed into low xylans such as xylooligosaccharides and xylobiose and a small amount of xylose and arabinose. Xylanases include endo-β-1,4-xylanase, β-D-xylosidase, α-L-furano-arabinosidase, acetylxylan esterase, phenol esterase, and the like. Xylanase is widely distributed in nature and exists in marine and terrestrial bacteria, marine algae, fungi, etc. The reported microorganisms capable of producing xylanase include bacteria, Streptomyces, Aspergillus and Penicillium, etc. The high-quality xylanase is mainly processed by extracting, concentrating and other processes after fermenting with Aspergillus oryzae, Aspergillus niger, Bac...

Claims

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Application Information

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IPC IPC(8): C12N11/12C12N11/08
Inventor 汤卫华殷海松魏纪平陈珊张乐龙尾程秀玮苑鹏黄艳玲高芦宝闫雪冰
Owner TIANJIN MODERN VOCATIONAL TECH COLLEGE
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