Immobilized cutinase, preparation method and application of removing phthalates in water

A technology of phthalic acid and cutinase, applied in the direction of immobilization on or in inorganic carriers, chemical instruments and methods, water pollutants, etc., can solve the problems affecting the application of immobilized enzymes, general biocompatibility, General problems such as adsorption stability, to achieve the effect of eliminating tissue defects, good adsorption stability, and high activity recovery rate

Inactive Publication Date: 2016-08-24
HUNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, using general carriers to immobilize enzymes by physical adsorption method has the problems of relatively small adsorption capacity, general adsorption stability, and general biocompatibility, which affects the application of immobilized enzymes.

Method used

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  • Immobilized cutinase, preparation method and application of removing phthalates in water
  • Immobilized cutinase, preparation method and application of removing phthalates in water
  • Immobilized cutinase, preparation method and application of removing phthalates in water

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] The immobilized cutinase uses a porous gold nanometer material as a carrier, and the cutinase is fixed on the porous gold nanometer material through physical adsorption to obtain the immobilized cutinase. The steps are as follows: 10 mg of porous gold nanomaterials are added to the cutinase solution, the cutinase solution is prepared from cutinase and Tris-HCl buffer (concentration is 20 mM, pH value is 8.0), the cutinase in the cutinase solution The concentration is 0.8 mg / mL, the mass of cutinase in the cutinase solution is 4 mg, shake and adsorb at 120 rpm at 50 ° C for 6 h, then wash several times with Tris-HCl buffer solution with a pH value of 8.0, and take out the sheet-shaped porous gold The nanometer material is freeze-dried at -40° C. to obtain immobilized cutinase. The cutinase adsorption capacity of the porous gold nanomaterial is 39.2mg / g, and the pore diameter of the porous gold nanomaterial is 40-50nm. The recovery rate of the enzyme activity of the immo...

Embodiment 2

[0040]The immobilized cutinase uses a porous gold nanometer material as a carrier, and the cutinase is fixed on the porous gold nanometer material through physical adsorption to obtain the immobilized cutinase. The steps are: 10 mg of porous gold nanomaterials are added to the cutinase solution, the cutinase solution is prepared from cutinase and phosphate buffer, the concentration of cutinase in the cutinase solution is 0.2 mg / mL, and the cutinase solution The mass of cutinase in the medium is 6 mg, the pH value of the cutinase solution is 9.0, oscillating and adsorbing at 150 rpm at 45 ° C for 4 h, and then washed several times with Tris-HCl buffer solution with a pH value of 8.0, and the sheet-shaped porous gold nanomaterial is taken out, Freeze and vacuum-dry at -40°C to obtain immobilized cutinase. The cutinase adsorption capacity of the porous gold nanomaterial is 36.5mg / g, the pore diameter of the porous gold nanomaterial is 40-50nm, the enzyme activity recovery rate of...

Embodiment 3

[0045] Preparation of immobilized cutinase under different oscillation adsorption time conditions

[0046] Take 10 mg of the porous gold nanomaterial prepared in Example 1 as a carrier, add 10 mL of Tris-HCl buffer containing 4 mg of cutinase (concentration is 20 mM, pH value is 8.0), and absorb at 50 ° C at a speed of 150 rpm for 30 ~360min, after the immobilization is completed, wash several times with Tris-HCl buffer solution with a pH value of 8.0, take out the flake-shaped porous gold nanomaterial, and freeze and vacuum-dry it at -40°C to obtain the immobilized cutinase.

[0047] The amount of cutinase in the buffer solution before fixation and the remaining cutinase amount in the buffer solution after fixation were respectively measured by the Lowry method, and the cutinase adsorbed by the unit carrier in different time was calculated, and the results were shown in Table 1; from the data analysis in Table 1, it can be seen that the appropriate The range of vibration adso...

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Abstract

The invention relates to the field of enzyme immobilization, and in particular relates to an immobilized cutinase and a preparation method and an application thereof in removal of phthalic acid esters in water. The immobilized cutinase is fixed on a porous gold nanomaterial through a physical absorption function by taking the porous gold nanomaterial as a carrier, wherein the adsorption quantity of the cutinase on the porous gold nanomaterial is more than 30mg / g; the bore diameter of the porous gold nanomaterial is between 40nm and 50nm. The preparation method comprises the following steps: adding the porous gold nanomaterial into a cutinase solution; adsorbing in an oscillation manner at 45 DEG C to 55 DEG C; and performing washing, freezing and vacuum drying, thereby obtaining the immobilized cutinase. The immobilized cutinase can be used for degrading the phthalic acid esters in the water, and is good in biocompatibility, high in adsorption quantity and good in adsorption stability.

Description

technical field [0001] The invention relates to the field of enzyme immobilization, in particular to an immobilized cutinase, a preparation method and its application in removing phthalates in water bodies. Background technique [0002] Cutinase (cutinase, EC3.1.1.74) is a hydrolase that can degrade cutin and produce a large amount of fatty acid monomers. It is a member of the α / β hydrolase family with a small molecular weight and belongs to serine esterase. Cutinase can not only catalyze the hydrolysis of the ester bonds of insoluble polymer plant cutin, but also act on other long-chain and short-chain fatty acid esters, emulsified triglycerides and soluble synthetic esters. Cutinase can realize biorefining of cotton fiber and biomodification of synthetic fiber, and is a key enzyme preparation to promote clean production and green processing in the textile industry; Professor Richard Grosse of New York Institute of Technology has developed plastics produced by processing bi...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N11/14C02F3/00C02F101/34
Inventor 贺迅张辰曾光明黄丹莲刘亮赖萃赵美花黄超李宁杰危臻许飘程敏
Owner HUNAN UNIV
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