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High stability immobilized enzyme preparation method

An immobilized enzyme, high stability technology, applied in the direction of immobilization on/in the organic carrier, can solve the problems of poor mechanical strength of the immobilized enzyme, and achieve a wide range of applicable environments, good stability, and high enzyme activity. high rate effect

Inactive Publication Date: 2006-05-03
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The embedding method includes grid embedding, microcapsule embedding and liposome embedding, etc. In the embedding method, because the enzyme itself does not participate in the chemical binding reaction, it can obtain higher enzyme activity recovery, but the prepared Immobilized enzymes are usually mechanically weak

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] First, take lipase powder (or any one of the above enzyme powders, the same below) and dissolve it in a buffer solution to prepare a 0.1% to 2% enzyme solution for later use. Weigh sodium dodecylsulfonate (SDS) and Deionized water was prepared into a 20% aqueous solution for later use. Then, measure 10ml of 20% SDS aqueous solution, add 5ml of n-heptane, 10g of polyethylene glycol diacrylate, 10g of trimethylolpropane trimethacrylate, 23g of methyl methacrylate and 4ml of enzyme solution and stir After uniformity, let stand. When the solution becomes transparent, add 1% to 3% of the reaction substrate lauric acid, and finally add 1 to 2 g of azobisisobutyronitrile, polymerize at 40 to 60°C until the system is solidified, and continue to react for 2 hours. After the polymerization is completed, the polymer is taken out, and the granular polymer is obtained after crushing, grinding, grading and screening, and placed in a Soxhlet extractor to extract water and lauric acid...

Embodiment 2

[0025] Firstly, lipase powder is weighed and dissolved in a buffer solution to prepare a 1%-4% enzyme solution for later use. Then, measure 10ml of n-heptane, add 8-10g of bis(2-ethylhexyl)sodium succinate (AOT), 2-4ml of enzyme solution, 20g of polyethylene glycol dimethacrylate, trimethylol Stir 10g of propane triacrylate until transparent, add 1% to 3% of the reaction substrate octanoic acid, add 0.10 to 0.30g of photoinitiator benzoin methyl ether, stir well, pour it into a mold, and carry out photoinitiated polymerization under ultraviolet light Until the system solidifies and forms, continue to illuminate for 3-8 seconds. After the polymerization is completed, the polymer is taken out, and the granular polymer is obtained after crushing, grinding, grading and screening, and placed in a Soxhlet extractor to extract water and substrate with acetone or petroleum ether organic solvent, and then vacuum-dried to obtain the immobilized enzyme product. The activity yield of th...

Embodiment 3

[0027] Firstly, lipase powder is weighed and dissolved in a buffer solution to prepare a 0.05%-1% enzyme solution for later use. Then, measure 5ml of n-heptane, add Tween802~6g, enzyme solution 2~4ml, polyethylene glycol dimethacrylate 20g, pentaerythritol tetraacrylate (PETA) 20g, stir until transparent, add 2% oleic acid, After stirring evenly, pour it into an ampere irradiation bottle, vacuumize or seal it with high-purity nitrogen, and place it in a 60Co γ-ray irradiation field to irradiate until the system solidifies and forms. After the polymerization is completed, the polymer is taken out, and the granular polymer is obtained after crushing, grinding, grading and screening, placed in a Soxhlet extractor to extract water and substrate with acetone or petroleum ether organic solvent, and then vacuum-dried to obtain the immobilized enzyme product . The activity yield of the enzyme was measured to be 133%. Under the condition of 40-50°C, the immobilized enzyme is used con...

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Abstract

The invention discloses a preparation method for high-stable immobilized enzyme, which comprises: using enzyme, water, surfactant, organic solvent, monomer or prepolymer, and crosslinking agent to construct microemulsion system; adding a little reaction substrate to induce enzyme active conformation; heating, ultraviolet irradiating or direct radiating to initiate free radical polymerization and realize the embedding fix in high-crosslinking carrier. This invention is simple, has high yield and well stability, and fits to wide range.

Description

technical field [0001] The invention relates to a preparation method of highly stable immobilized enzyme. Background technique [0002] It is well known that enzymes are efficient and specific biocatalysts. Many difficult organic chemical reactions can be carried out smoothly under mild conditions of normal temperature and pressure when enzymes are used as catalysts, and side reactions can be avoided or reduced. Therefore, enzymes have been widely used in brewing, food, medicine and other fields since ancient times. Especially in modern times, due to the development of biochemistry, the mechanism of action of enzymes has become increasingly clear, new enzyme sources have been continuously developed, and the application of enzymes has become more active. However, enzymes are composed of proteins. In most cases, they are not stable enough to heat, strong acids, strong alkalis, organic solvents, etc. Even under the optimal conditions of enzyme reactions, they will often be in...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N11/08C12N11/04
Inventor 宋锡瑾王杰
Owner ZHEJIANG UNIV
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