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Method for preparing immobilized enzymes and immobilized strains

A technology for immobilizing enzymes and strains, which is applied in the direction of immobilizing on or in inorganic carriers, or in or on organic carriers, and can solve the problems of severe cross-linking conditions, severe coupling conditions, and enzyme transfer capacity. Small problems, to achieve the effect of heavy recycling utilization rate and enzyme activity improvement, good membrane regeneration performance, and low production cost

Inactive Publication Date: 2014-02-26
LIUZHOU JINGYUAN BIOLOGICAL SCI & TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But each method has disadvantages
The adsorption method is sensitive to factors such as ionic strength, pH, temperature, etc., so the enzyme is easily damaged and leaked, and the enzyme transfer capacity is small; the coupling reagents in the covalent coupling method are often toxic to living tissues, and the coupling conditions are intense. Causes enzyme failure and high cost; cross-linking method has severe conditions and often has diffusion limitations, making it difficult to use
The embedding method can only be used for low molecular weight substrates, not suitable for column systems, the reaction is often limited by diffusion, not all monomeric materials and solvents are suitable for various enzymes

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] An elastase aqueous solution was prepared, and after testing, the enzyme activity unit was 3500 U / mL, and the pH of the solution was adjusted to 6.5 with sodium bicarbonate. Prepare a bifidobacterium aqueous solution, control the live bacteria content in the strain solution to 1 billion / mL; place a ceramic membrane with a membrane pore size of 0.1 μm in a 4% sodium dodecyl sulfate solution for activation by soaking for 3 Hours, the solution needs to submerge the ceramic membrane. Mix 1L elastase aqueous solution, 1L acetic acid bacteria aqueous solution, 1cm 2 The ceramic membrane was mixed in a reactor, stirred for 1 hour, soaked at 25° C. for 58 hours, and filtered to remove the solution to obtain a ceramic membrane adsorbed with enzymes and strains. Soak the ceramic membrane with enzymes and strains adsorbed in a polyethyleneimine solution with a pH of 8 and a mass fraction of 1.2% for 15 hours. Bacteria. Example 2

Embodiment 2

[0028] An aqueous carboxypeptidase solution was prepared. After testing, the enzyme activity unit was 4000 U / mL, and the pH of the solution was adjusted to 7.0 with sodium bicarbonate. Prepare an aqueous solution of actinomycetes, and control the live bacteria content in the strain solution to 1.2 billion / mL; place a ceramic membrane with a membrane pore size of 0.3 μm in a 6% sodium dodecyl sulfate solution for activation by soaking for 4 Hours, the solution needs to submerge the ceramic membrane. Mix 1L carboxypeptidase aqueous solution, 1L actinomycete aqueous solution, 1cm 2 The ceramic membrane was mixed in a reactor, stirred for 1.5 hours, soaked at 28° C. for 60 hours, and filtered to remove the solution to obtain a ceramic membrane adsorbed with enzymes and bacteria. Soak the ceramic membrane with enzymes and strains adsorbed in a polyethyleneimine solution with a pH of 8.5 and a mass fraction of 1.3% for 18 hours. Bacteria.

Embodiment 3

[0030] Prepare a mixed aqueous solution of pepsin, chymotrypsin, trypsin, carboxypeptidase, and elastase. After testing, the enzyme activity unit is 4500 U / mL, and the pH of the solution is adjusted to 7.5 with sodium bicarbonate. Prepare a mixed aqueous solution of bifidobacteria, lactic acid bacteria, aerobic bacillus, photosynthetic bacteria, saccharomyces, actinomycetes, and acetic acid bacteria, and control the live bacteria content in the strain solution to 1.5 billion / mL; The ceramic membrane was soaked and activated for 5 hours in an 8% sodium dodecyl sulfate solution, and the solution had to submerge the ceramic membrane. Mix 100L enzyme solution, 100L strain solution, 100cm 2 The ceramic membrane was mixed in a reactor, stirred for 2 hours, soaked at 30° C. for 65 hours, and filtered to remove the solution to obtain a ceramic membrane adsorbed with enzymes and strains. Put the ceramic membrane adsorbed with enzymes and strains at a pH of 9 and soak in a polyethylene...

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Abstract

The invention relates to a method for preparing immobilized enzymes and immobilized strains, belonging to the field of biotechnology. The method for preparing immobilized enzymes and immobilized strains comprises the steps of preparing an enzyme solution and a strain solution, adsorbing enzymes and strains by using a ceramic membrane, inoculating the ceramic membrane on which enzymes and strains are adsorbed to a polyethyleneimine crosslinking agent solution to soak, and filtering and drying the obtained product so as to obtain immobilized enzymes and immobilized strains. The method disclosed by the invention is high in activity of prepared enzymes and strains, simple in extraction process, low in production cost and less in equipment investment, and easily expands the scale of production.

Description

technical field [0001] The invention relates to a method for preparing immobilized enzymes and immobilized strains, belonging to the field of biotechnology. Background technique [0002] In alcohol production, high-depth organic wastewater has become a huge pollution treatment problem. The discharge of these pollutants into the water will consume a large amount of dissolved oxygen in the water, destroy the balance of oxygen, lead to the deterioration of water quality, and bring serious pollution chains to the environment. reaction. Alcohol waste liquid has a high suspended content, with an average suspended solid content of up to 40,000 mg / L; high pollution concentration, and the COD of wastewater is as high as 20,000 to 30,000, including suspended solids, dissolved COD and colloids, organic matter accounts for 93% to 94%, and inorganic matter accounts for 93% to 94%. 6% to 7%, the organic matter is composed of carbohydrates, followed by nitrogenous compounds, biological ba...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N11/14C12N11/08
Inventor 张宝龙
Owner LIUZHOU JINGYUAN BIOLOGICAL SCI & TECH CO LTD
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