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Method for fixing key enzymes and cofactors in formaldehyde decomposition path and application thereof

A formaldehyde decomposition, key enzyme technology, applied in the field of enzyme engineering

Inactive Publication Date: 2012-12-19
KUNMING UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Formate dehydrogenase (formate dehydrogenase, EC 1.2.1.2, FDH) can oxidize formic acid and finally generate CO 2 (Labrou and Rigden, Biochem.J.2001, 354:455-463), but so far, there is no research and development at home and abroad to use enzymes to control the method of formaldehyde pollution

Method used

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  • Method for fixing key enzymes and cofactors in formaldehyde decomposition path and application thereof
  • Method for fixing key enzymes and cofactors in formaldehyde decomposition path and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Immobilization of recombinant protein of PADH and recombinant protein of FDH and its cofactor NAD:

[0023] Add the following solutions to the 30ml reaction system: 12ml of 30% acrylamide, 7.8ml of 1.5mol / L (pH7.5) potassium phosphate buffer, 7.9ml of sterile water, 1.5mg of PADH recombinant protein, 2.25mg of FDH recombinant protein, 100mmol / L cofactor NAD + 2ml, stirred slowly at room temperature for 20min to mix the enzyme protein, cofactor and acrylamide carrier matrix evenly, then added 0.3ml of 10% ammonium persulfate (APS), 0.012ml of TEMED, stirred slowly for 2min to completely cross-link the acrylamide monomer into After polyacrylamide, quickly pour the mixture into the gel maker that has been installed, and place it at room temperature for 3-4 hours to make the polyacrylamide completely solidify, carefully peel off the gel, and cut it into a length of 5cm with a clean scalpel , 1cm wide and 0.5cm thick strips, put them into a 6ml empty chromatographic column....

Embodiment 2

[0025] Analysis of the absorption effect of immobilized enzyme on formaldehyde:

[0026] Use a syringe to simultaneously inject 6ml of gaseous formaldehyde into the upper end of the chromatography tube and the control tube (blank tube) equipped with immobilized enzyme strips, and the outlet of the tube is connected to a hand-held formaldehyde analyzer ( figure 1 B), the measured initial concentration of formaldehyde in the tube is 0.47ppm, and the remaining formaldehyde concentration in the tube is measured every 1 hour. Take the remaining formaldehyde concentration in the tube as the ordinate, and take the measurement time as the abscissa to draw the formaldehyde absorption curve as follows: figure 2 shown. The results show that compared with the blank tube, the immobilized enzyme does have an absorption effect on formaldehyde, especially in the first 3 hours, the formaldehyde decline rate is faster, from the initial 0.47ppm to 0.32ppm, and 32% of the injected gas formaldeh...

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Abstract

The invention discloses a method for fixing key enzymes and cofactors in a formaldehyde decomposition path. The two key enzymes and the cofactors NAD in the formaldehyde decomposition path are fixed in polyacrylamide gel to prepare an immobilized enzyme which can be used for decomposing and absorbing formaldehyde gas.

Description

Technical field: [0001] The invention belongs to the technical field of enzyme engineering, and in particular relates to a method for controlling formaldehyde pollution by enzymatic methods. Two enzyme proteins with catalytic activity and thermal stability—GSH-independent formaldehyde dehydrogenase (PADH) and formic acid Dehydrogenase (FDH) is immobilized inside the polyacrylamide gel and prepared as an immobilized enzyme, which can be used to decompose and absorb gaseous formaldehyde. Background technique: [0002] Many building decoration materials such as wood-based panels, adhesives, paints, wallpapers, furniture, etc. can release formaldehyde, making formaldehyde one of the main gases of indoor air pollution. Due to the presence of formaldehyde inside the board, the volatilization period can be as long as 3 to 15 years, and it is difficult to completely volatilize within one or two years after decoration. High concentrations of formaldehyde are toxic to the nervous sys...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N11/08C12N11/04B01D53/84B01D53/72
CPCY02A50/20
Inventor 陈丽梅张婧
Owner KUNMING UNIV OF SCI & TECH