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Double-enzyme co-immobilizing method and immobilized enzyme prepared through method

A technology of immobilizing enzymes and setting temperature, applied in the field of bioengineering, can solve the problem of high cost of immobilizing multi-enzymes, and achieve the effects of improving catalytic efficiency, broadening tolerance and satisfying operating conditions

Inactive Publication Date: 2017-03-15
TSINGTAO BREWERY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the high cost of immobilized multi-enzymes, there are few industrial applications. Therefore, how to provide a method for immobilizing multi-enzymes to obtain high catalytic efficiency and low-cost immobilized multi-enzymes will be an important issue in this field. important topics studied

Method used

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  • Double-enzyme co-immobilizing method and immobilized enzyme prepared through method
  • Double-enzyme co-immobilizing method and immobilized enzyme prepared through method
  • Double-enzyme co-immobilizing method and immobilized enzyme prepared through method

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Effect test

Embodiment 1

[0035] Add 0.9ml, 0.2 mol / L calcium chloride solution, reacted for 12 hours at 25°C, centrifuged at 12000rpm for 10min, and washed three times with distilled water centrifugally to obtain α-acetolactate decarboxylase-calcium phosphate-proline endonuclease (PSEP-Ca 3 (PO 4 ) 2 -ALDC) immobilized carrier, its electron micrograph is as figure 1 shown. It can be seen from the figure that when the initial enzyme is not contained, the formed carrier is mainly a stacked lamellar structure, the structure is relatively dense, and the specific surface area is not high.

Embodiment 2

[0037] Add 0.9ml, 0.2 mol / L calcium chloride solution, reacted for 12 hours at 25°C, centrifuged at 12000rpm for 10min, and washed three times with distilled water to obtain α-acetolactate decarboxylase-calcium phosphate-proline endonuclease (PSEP-Ca 3 (PO 4 ) 2 -ALDC) immobilized enzyme, in the prepared immobilized enzyme, the content of immobilized enzyme per milligram of calcium phosphate loaded is 13.38mg, and its electron micrograph is as follows figure 2 shown. It can be seen from the figure that at this initial enzyme concentration, compared with the structure without enzyme, the lamellar structure is loose and the specific surface area increases. At this time, the activities of the two enzymes that have been immobilized are about 30% of the activities of the free enzymes.

Embodiment 3

[0039] Add 0.9ml, 0.2 mol / L calcium chloride solution, reacted for 12 hours at 25°C, centrifuged at 12000rpm for 10min, and washed three times with distilled water to obtain α-acetolactate decarboxylase-calcium phosphate-proline endonuclease (PSEP-Ca 3 (PO 4 ) 2 -ALDC) immobilized enzyme, in the prepared immobilized enzyme, the content of immobilized enzyme per milligram of calcium phosphate loaded is 28.84mg, and its electron micrograph is as follows image 3 shown. It can be seen from the figure that at this initial enzyme concentration, the sheet structure is further loosened, forming a honeycomb shape, with a better nanoflower structure, and the specific surface area is further increased. At this time, the activities of the two enzymes that have been immobilized are basically the same as the activity of the free enzyme, and even higher than the activity of the free enzyme.

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Abstract

The invention provides a double-enzyme co-immobilizing method and an immobilized enzyme prepared through the method and belongs to the field of bioengineering, and multiple immobilized enzymes with high catalysis efficiency and low cost can be obtained. The double-enzyme co-immobilizing method comprises the following steps: adding a calcium chloride solution into a buffer solution containing alpha-acetolacetate decearboxylase and proline endoprotease which are to be immobilized, enabling full reaction at set temperature, and centrifugally separating and washing to obtain the immobilized enzymes of nanoflower structures. The double-enzyme co-immobilizing method can be applied to beer production, in particular to the final filter stage of clear wine in a beer production process.

Description

technical field [0001] The invention belongs to the field of bioengineering, and in particular relates to an enzyme immobilization method, in particular to a double-enzyme co-immobilization method and the immobilized enzyme prepared by the method. Background technique [0002] As one of the fruits of bioengineering research, enzyme preparations have been popularized in the beer industry. By using enzyme preparations, it can make up for the lack of enzyme activity in malt, improve wort components, increase beer fermentation degree, shorten fermentation cycle, increase production, etc. 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, adding proline endonuclease to the fermentation broth can enhance the abiotic stability of beer; adding α-acetolactate decarboxylase to the fermentation broth can reduce the diacetyl content conveniently and quickly....

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N11/14C12C5/00
CPCC12N11/14C12C5/004C12N9/50C12N9/88C12Y304/21026C12Y401/01005
Inventor 董建军侯同刚余俊红王建勋尹花王乔常宗明黄淑霞咸漠胡淑敏
Owner TSINGTAO BREWERY
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