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Enzyme immobilization method

A technology of immobilizing enzyme and calcium chloride solution, applied in the field of bioengineering, can solve the problems of loss of enzyme activity, limitation of three-dimensional conformation flexibility, increase of cell immobilization cost, etc., and achieve the effect of improving enzyme activity

Active Publication Date: 2016-09-21
QINGDAO INST OF BIOENERGY & BIOPROCESS TECH CHINESE ACADEMY OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Taking the carrier as an example, the carrier materials used in domestic research on immobilization are almost all concentrated on organic materials such as alginate, carrageenan or some macroporous resins. Although the immobilization effect is good, the carrier material has poor corrosion resistance and pressure exists. Deformable over time and unable to regenerate
In addition, the equipment required for continuous operation is relatively complicated. These factors greatly increase the cost of cell immobilization and limit the industrial application of this technology in China.
[0007] (2) Immobilization has a greater impact on activity
Other immobilization methods, such as covalent methods, are limited by the three-dimensional conformation and flexibility of the enzyme, and the immobilization carrier used may touch the active sensitive area of ​​the enzyme, often accompanied by a large loss of enzyme activity

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] (1) Add 0.9ml of CaCl with a concentration of 0.2mol / L to 45ml of ALDC with a concentration of 1.2mg / ml in PBS (4mmol / L, pH=7.4) buffer solution 2 In the solution, react at 25°C for 12 hours, centrifuge at 12000rpm for 10min, and wash three times with distilled water to obtain Ca 3 (PO 4 ) 2 – ALDC nanoflowers.

[0045] (2) Calculating the supernatant obtained in step (1) and the enzyme content in the washing liquid with a UV-visible spectrophotometer can obtain Ca 3 (PO 4 ) 2 – Enzyme loading on ALDC nanoflowers is 1.9 mg / Ca 3 (PO 4 ) 2 (1 mg). The nanoflowers loaded with 17 mg of ALDC enzyme were dispersed in 10 ml of 0.03 mol / L sodium alginate solution, and stirred for 1 hour to obtain a uniform dispersion.

[0046] (3) The concentration of 100ml of the dispersion obtained in step (2) is 2.0mol / L CaCl dripped into 100ml with a syringe at a speed of 3ml / min. 2 solution, stirred and solidified for 15 min, centrifuged and washed three times with distilled wate...

Embodiment 2

[0050] Except that the concentration of ALDC in the PBS (4 mmol / L, pH=7.4) buffer solution of ALDC in step (1) is set to 0 mg / ml, that is, ALDC is not included, the calcium alginate coating is prepared according to the same method as in Example 1 Ca 3 (PO 4 ) 2 Nanoflower microspheres.

Embodiment 3

[0052] Except that the concentration of ALDC in the PBS (4mmol / L, pH=7.4) buffer of ALDC in step (1) was set to 0.05mg / ml, the calcium alginate-coated Ca 3 (PO 4 ) 2 – ALDC nanoflower microspheres.

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Abstract

The invention provides an enzyme immobilization method, and particularly relates to a method for immobilization of enzyme for beer production. According to the method, alpha-acetolactate decarboxylase buffer solution is added to calcium chloride solution for reaction to obtain a nanoflower structure provided with preliminary immobilized enzyme, then obtained nanoflower is dispersed in sodium alga acid solution, obtained dispersion liquid is added to the calcium chloride solution dropwise at a certain speed, and stirring and curing are conducted to obtain a final microsphere product. According to microspheres prepared with the enzyme immobilization method, immobilized enzyme activity is higher than free enzyme activity by 98% or more, and immobilized enzyme is stable and recyclable and has broad prospects in industrial application.

Description

technical field [0001] The invention relates to the field of bioengineering, in particular to an enzyme immobilization method, in particular to an enzyme immobilization method for beer production. Background technique [0002] As one of the fruits of bioengineering research, enzyme preparation has been popularized and improved in the beer industry. In recent years, in order to improve the quality, reduce the cost and increase the benefit, the beer enterprises use more and more enzyme preparations. Through the use of enzyme preparations, it can make up for the lack of enzyme activity in malt, increase the amount of auxiliary materials, improve the composition of wort, increase the degree of fermentation of beer, shorten the fermentation cycle, and increase production. Supplementing external enzymes in the production process runs through the whole process of beer production, which can make the operation easy and the benefit improved. For example, when high-temperature-resist...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N11/14C12N11/10C12N11/04C12C5/00
CPCC12C5/004C12N9/88C12N11/04C12N11/10C12N11/14C12Y401/01005
Inventor 董建军侯同刚余俊红王建勋尹花王兰常宗明黄淑霞咸漠
Owner QINGDAO INST OF BIOENERGY & BIOPROCESS TECH CHINESE ACADEMY OF SCI
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