Solid phase support crosslinking enzyme aggregation and method for preparing the same

A technology of aggregates and cross-linked enzymes, applied to biochemical equipment and methods, fixed on or in inorganic carriers, fixed on/in organic carriers, etc., can solve problems such as inability to recycle and reuse, and achieve The effect of easy optimization of reaction conditions, strong antitrypsin ability, and high liquid flow rate

Inactive Publication Date: 2009-07-29
THE INST OF BIOTECHNOLOGY OF THE CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The first technical problem to be solved in the present invention is to overcome the disadvantage that aggregate cross-linking enzymes cannot be recycled and reused, and provide a cross-linking enzyme aggregate immobilized on a carrier. The cross-linking enzyme aggregate has thermal stability In addition to the advantages of high anti-trypsin ability, etc., it also has a certain physical form and stable structure, can be recycled, and can be continuously used in production

Method used

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  • Solid phase support crosslinking enzyme aggregation and method for preparing the same
  • Solid phase support crosslinking enzyme aggregation and method for preparing the same
  • Solid phase support crosslinking enzyme aggregation and method for preparing the same

Examples

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Embodiment 1

[0027] Example 1 Preparation of immobilized cross-linked organophosphate degrading enzyme aggregates

[0028] Weigh the organophosphorus degrading enzyme and dissolve it in phosphate buffer solution to prepare an enzyme solution with a protein concentration of 0.4 mg / ml, and add porous microspheres (polystyrene type non-polar adsorption resin, model ADS-5; average pore diameter The particle size is 30nm and the particle size is greater than 100μm; performance: it does not adsorb proteins, sugars, inorganic acids, alkalis, salts, and small-molecular hydrophilic organic substances; it has high resistance to heat, acid, alkali, organic solvents, etc. Stability. Tianjin Nankai Hecheng Science and Technology Co., Ltd. formula), stirred slowly at 4°C for 3 hours; then added ammonium sulfate to a saturation of 67.5%, stirred slowly at 4°C for 5 hours; added glutaraldehyde (25%) The solution was stirred slowly at 4°C for 18 hours to a final concentration of 2.2%; the porous microspher...

Embodiment 2

[0030] Weigh lactase and dissolve it in phosphate buffered saline, prepare an enzyme solution with a protein concentration of 0.5 mg / ml, add porous microspheres (polystyrene type non-polar adsorption resin, model is ADS-5. The average pore size is 30nm , the particle size is greater than 100 μm; performance: it does not adsorb proteins, sugars, inorganic acids, alkalis, salts, and small-molecular hydrophilic organic compounds; it has high stability under conditions such as heat, acid, alkali, and organic solvents (produced by Tianjin Nankai Hecheng Technology Co., Ltd.), and stirred slowly at 4°C for 3 hours. Add ammonium sulfate to a saturation of 67.5%, and stir slowly at 4°C for 5 hours. Add glutaraldehyde (25%) solution to a final concentration of 2.2%, and stir slowly at 4°C for 18 hours. The porous microspheres were taken out by filtration, and rinsed repeatedly until the solution was clear.

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Abstract

The invention discloses immobilized crosslinked enzyme aggregates (CLEAs) and a preparation method thereof. The crosslinked enzyme aggregates are fixed on a porous microsphere; the preparation method thereof comprises: dispersing detached enzyme needing to be immobilized into the porous microsphere; carrying out depositing and crosslinking on zymoprotein in the porous microsphere in sequence, and finally rinsing the porous microsphere, thus preparing the immobilized crosslinked enzyme aggregates. The method of the immobilized enzyme can be used for immobilizing the enzyme of various types or sources; the immobilized method is simple and easy to be carried out, is easy to be operated, does not need special devices and has low production cost; the prepared crosslinked enzyme aggregates have high stability and strong capacity for resisting trypsinase, has a certain physical form and a stable structure, can be recycled and repeatedly used and can be used for continuous industrial production.

Description

technical field [0001] The invention relates to immobilized enzymes, in particular to a cross-linked enzyme aggregate fixed on a solid phase carrier and a preparation method thereof, belonging to the field of enzyme immobilization. Background technique [0002] Since the start of enzyme immobilization research in the 1960s, people have discussed and studied enzyme immobilization technologies from various aspects, and have successfully used physical and chemical methods to immobilize enzymes, many of which are And products have been applied in industrial production. But the fixed objects are basically free enzymes, because free enzymes are relatively fragile soluble proteins that have high requirements on the environment, and are generally unstable to heat, strong acids, and strong alkalis. In the commonly used immobilization methods, due to the strong chemical reaction, the properties of the protein are usually greatly affected, and because the enzyme has a certain advanced...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N11/04C12N11/14C12N9/24C12N9/20C12N9/04C12N9/00C12N9/14
Inventor 郭军伍宁丰张伟范云六田健
Owner THE INST OF BIOTECHNOLOGY OF THE CHINESE ACAD OF AGRI SCI
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