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Process for fixing amino acylatease by large pore anion resin

An anion resin, aminoacylase technology, applied in the direction of being fixed on/in an organic carrier, can solve the problems of high price, poor hydrodynamic performance and mechanical strength, and high use cost, and achieves low price and is not easy to microorganisms. Mild effects of invasion, immobilization conditions

Inactive Publication Date: 2006-06-28
TIANJIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in China, due to the high price of the carrier DEAE-SephadexA-25, the domestic use cost is relatively high, and the hydrodynamic properties and mechanical strength of the carrier are relatively poor

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] 1. Transformation of macroporous anion resin

[0032] Put macroporous anion resin DEAE-E / H in 0.1mol.l -1 In the hydrochloric acid solution, it is converted to Cl after treatment at room temperature - After transformation, the carrier was fully washed with deionized water, and filtered to dryness.

[0033] 2. Preparation of free aminoacylase solution

[0034] Dissolve the aminoacylase with deionized water and dilute to a specific enzyme activity of 120U.ml -1 , with a concentration of 0.05mol L -1 NaOH solution to adjust the pH to 6.5.

[0035] 3. Immobilized aminoacylase

[0036] Take 1g step 1 to process transformation to obtain Cl - The type DEAE-E / H carrier was put into a 100ml Erlenmeyer flask, added to 16ml of the free aminoacylase solution prepared in step 2, shaken at 25°C at a speed of 150rpm for 5h, then stood still for 1h, and filtered with suction.

[0037] 4. Washing of immobilized enzymes

[0038] Take the immobilized enzyme prepared in step 3, put...

Embodiment 4

[0040] 1. Transformation of macroporous anion resin

[0041] According to the method of embodiment two, macroporous anion resin D380 is processed and transformed into Ac - type resin.

[0042] 2. Preparation of free aminoacylase solution

[0043] The free aminoacylase solution was prepared according to the method in Example 1.

[0044] According to the method of Example 1, the immobilized enzyme was cleaned and the enzyme activity was measured, and the specific enzyme activity of the prepared immobilized enzyme was 1050U·g -1 , The recovery rate of enzyme activity was 54.7%.

Embodiment 5

[0046] The operating conditions are the same as in Example 1, changing the concentration of the free enzyme solution to 180U·ml -1 , the specific activity of the immobilized aminoacylase prepared was 1030U·g -1 , The recovery rate of enzyme activity is 35.7%.

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PUM

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Abstract

The invention discloses macropore anion resin immobilization amino group acylation enzyme. It belongs to immobilization amino group acylation enzyme technique. The method includes the following steps: dissolving macropore anion resin in different acid solution with 0.05mol.l-1-0.15mol.1-1 density; transferring to Ac-, Cl-, SO42- or NO3- type vector; diluting to ratio enzyme active 80-180U.ml-1; adjusting pH value to 5-10 by NaOH solution; adding into free enzyme solution at 20-45 centigrade degree; vibrating; pumping filtering; discharging ion water; vibrating; pumping filtering; dipping in sodium acetate solution; pumping filtering; washing by ion water until without enzyme activity; pumping and filtering to dry. The operation is simple. Immobilization enzyme has good mechanical property and chemical stability. Ration enzyme active can reach 1800U.ml-1. And the reclaiming of enzyme active is more than 60%. Thus the immobilization amino group acylation enzyme can be extensively applied to amino acid resolution.

Description

technical field [0001] The invention relates to a method for immobilizing aminoacylase with macroporous anion resin. It belongs to the technique of immobilizing aminoacylase. Background technique [0002] Enzymes can efficiently catalyze reactions at normal temperature and pressure, and have high stereoselectivity and specificity. However, enzymes, as a kind of protein, have poor stability and are unstable to heat, strong acid, strong alkali, etc. Even under the optimal reaction conditions of enzymes, they tend to be inactivated quickly. Moreover, it is difficult to separate the free enzyme from the product after the reaction, and the product is likely to cause protein contamination. In order to make full use of the activity of the enzyme and overcome the various shortcomings of the above-mentioned free enzyme, the enzyme is usually immobilized and then used in biological reactions. [0003] Although immobilized enzymes are superior to free enzymes in many aspects, only a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N11/08
Inventor 王燕朱怀工王淑兰张凤宝吴世梅
Owner TIANJIN UNIV
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