Method for purifying xylanase

A technology of xylanase and polycanase, applied in the field of biological enzymes, can solve the problems of inconvenient long-term storage and transportation, low resource utilization rate, and high operating cost of dextran gel filtration chromatography

Inactive Publication Date: 2021-03-19
CHONGQING TECH & BUSINESS UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The main disadvantages of this method are: ① Dextran gel filtration chromatography not only has high operating costs, but also has limited production capacity, which is not convenient for industrial scale production; ② The xylan in the xylanase enzyme liquid product obtained by elution The enzyme concentration is low, which is not convenient for subsequent industrial application
The main disadvantages of this method are: 1. the raw material of xylanase stock solution is not pretreated, the resource utilization rate is low, and the product produced has low purity, which cannot meet the requirements of preparing high value-added products; 2. the obtained product is liquid, Not convenient for long-term storage and transportation

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] A method for purifying xylanase, the specific method steps are as follows:

[0027] (1) Preparation of crude xylanase solution

[0028] Using commercially available crude xylanase powder as raw material, the crude xylanase powder was dispersed in the purified In water, use 0.1mol / L sodium hydroxide solution to adjust the pH to 8.0, stir evenly to obtain a mixed solution; then pump the mixed solution into a hollow fiber membrane filter for filtration, control the pore size of the filter membrane to 0.2µm, and the pumping speed At 500mL / min, the membrane filtrate and membrane retentate were collected separately. The collected membrane retentate mainly contains protein, which can be used as a feed additive after being dried and pulverized; the collected membrane filtrate is crude xylanase liquid, which is used for the next step of treatment.

[0029] (2) Preparation of xylanase ultrafiltrate

[0030] After step (1) is completed, pump the crude xylan enzyme solution coll...

Embodiment 2

[0036] A method for purifying xylanase, the specific method steps are as follows:

[0037] (1) Preparation of crude xylanase solution

[0038] Using commercially available crude xylanase powder as raw material, the crude xylanase powder was dispersed in the purified In water, use 0.2mol / L sodium hydroxide solution to adjust the pH to 9.0, stir evenly to obtain a mixed solution; then pump the mixed solution into a hollow fiber membrane filter for filtration, control the pore size of the filter membrane to 0.2µm, and the pumping speed At 800mL / min, the membrane filtrate and membrane retentate were collected separately. The collected membrane retentate mainly contains protein, which can be used as a feed additive after being dried and pulverized; the collected membrane filtrate is crude xylanase liquid, which is used for the next step of treatment.

[0039] (2) Preparation of xylanase ultrafiltrate

[0040] After step (1) is completed, pump the crude xylanase solution collecte...

Embodiment 3

[0046] A method for purifying xylanase, the specific method steps are as follows:

[0047] (1) Preparation of crude xylanase solution

[0048] Using commercially available crude xylanase powder as raw material, the crude xylanase powder was dispersed in the purified In water, use 0.4mol / L sodium hydroxide solution to adjust the pH to 10.0, stir evenly to obtain a mixed solution; then pump the mixed solution into a hollow fiber membrane filter for filtration, control the pore size of the filter membrane to 0.2µm, and the pumping speed At 1000mL / min, the membrane filtrate and membrane retentate were collected separately. The collected membrane retentate mainly contains protein, which can be used as a feed additive after being dried and pulverized; the collected membrane filtrate is crude xylanase liquid, which is used for the next step of treatment.

[0049] (2) Preparation of xylanase ultrafiltrate

[0050] After step (1) is completed, pump the crude xylan enzyme liquid coll...

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Abstract

The invention belongs to the technical field of biological enzymes, and particularly relates to a method for purifying xylanase. The method disclosed by the invention takes coarse xylanase powder which is sold in the market as a raw material, and the raw materials can be produced through steps, including pH regulation, hollow fibrous membrane filtering, ultrafiltration, freeze drying and the like,to obtain purified xylanase. By use of the method disclosed by the invention, ultrafiltration stage treatment is adopted, a purification multiple and efficiency can be effectively improved, production cost is lowered, in addition, liquid nitrogen can be adopted to realize quick cryopreserving of enzyme liquid, enzyme crystal structure change and enzyme activity loss caused by slow pre-freezing can be reduced, and product performance is improved. The method disclosed by the invention has the characteristics of a simple production technology, convenience in production, a high product yield, a low equipment investment and operation cost, a high resource comprehensive utilization rate and the like. The xylanase product purified by the method disclosed by the invention has high activity, can meet the requirement of the medicine industry for high-purity enzyme preparations, and can be widely applied to industries, including medicines, food, chemical engineering and the like.

Description

technical field [0001] The invention belongs to the technical field of biological enzymes, in particular to a method for purifying xylanase. Background technique [0002] Xylanase belongs to the class of hydrolytic enzymes and is an enzyme system that degrades xylan into xylooligosaccharides. The enzyme system mainly includes β-1,4-endoxylanase (EC 3.2.1.8), β-D - Xylosidase (EC 3.2.1.37), α-D-glucuronidase (EC 3.2.1.139), α-L-arabinofuranosidase (EC 3.2.1.55) and acetylxylanase ( EC 3.3.1.72), etc. Among them, β-1,4-endoxylanase is the most critical enzyme to completely hydrolyze xylan. It hydrolyzes xylan into small Xylooligosaccharides such as oligosaccharides and xylobiose, and small amounts of xylose and arabinose. Xylanase is widely used in the fields of medicine, food, energy, paper and feed, and is one of the fastest-growing industrial enzyme preparations in recent years, with high commercial value and market potential. However, the commercially available xylanas...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/42
CPCC12N9/2482C12Y302/01008
Inventor 张杰李小琴王星敏蔡琼
Owner CHONGQING TECH & BUSINESS UNIV
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