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Method for increasing diethylstilbestrol degradation rate of laccase

A technology of diethylstilbestrol and degradation rate, applied in chemical instruments and methods, biochemical equipment and methods, enzymes, etc., can solve problems such as the decline of laccase processing ability

Active Publication Date: 2020-02-28
GUILIN UNIVERSITY OF TECHNOLOGY
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  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, when it is used for DES to degrade sewage, it generally takes about 6 hours to complete the degradation. However, in sewage with DES concentration higher than 50mg / L, the treatment capacity of laccase decreases, and the highest degradation rate is only 70%. about
Therefore, the degradation efficiency and degradation rate of laccase for DES sewage need to be further improved.

Method used

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  • Method for increasing diethylstilbestrol degradation rate of laccase
  • Method for increasing diethylstilbestrol degradation rate of laccase

Examples

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Effect test

Embodiment 1

[0031] (1) Preparation of standard solution: dissolve diethylstilbestrol in methanol to form a 2.5g / L diethylstilbestrol mother liquor, dilute the diethylstilbestrol mother liquor into standard solutions with concentrations of 1, 5, 10, 20, 30, 50mg / L, and 100mg / L respectively, The standard curve of diethylstilbestrol was prepared by detecting the peak area by HPLC.

[0032] HPLC detection conditions are as follows:

[0033] Chromatographic column: ZORBAX SB-C18 (150mm×4.6mm×5μm); detection conditions are: injection volume 10μL, equal volume of acetonitrile and water as flow equal gradient elution for 10min, pump flow rate 1.0mL / min, column temperature : 30°C, the ultraviolet detector detects the peak at 240nm wavelength, and records the peak area value.

Embodiment 2

[0035] (1) use the 0.01mol / L acetic acid-sodium acetate buffer solution that the pH that acetic acid and sodium acetate are formulated is 5.5;

[0036] (2) Use the acetic acid-sodium acetate buffer solution prepared in step (1) to prepare a laccase mother solution with a concentration of 1 mg / mL and an ABTS solution with a concentration of 0.01mol / L, wherein the laccase is produced by Coriolus versicolor, The enzyme activity is 0.94U / mg;

[0037] (3) Add 0.5mL laccase mother liquor, 0mL ABTS solution and 0.4mL DES mother liquor to 10mL reaction system, and make up the rest with buffer solution, put them in a shaking incubator at 55°C for reaction, set the speed at 120r / min, and react After 0.5 h, the solution was extracted twice with ethyl acetate, and the organic phase was extracted and then rotary evaporated to dryness. Methanol was used to make up the volume, and a vacuum filter was used to take samples, and the concentration of the reacted samples was measured by HPLC.

Embodiment 3

[0039] Same as Example 2, the difference is that in step (3), 0.5mL of laccase mother liquor, 0.1mL of ABTS and 0.4mL of DES mother liquor are added to 10mL of the reaction system, and the rest is made up by buffer solution.

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Abstract

The invention relates to a method for increasing the diethylstilbestrol degradation rate of laccase. The method specifically comprises: combining ABTS and a laccase solution to form a laccase-ABTS medium solution, mixing the laccase-ABTS medium solution and a diethylstilbestrol solution in a reaction system, placing the mixture into a shaking incubator, carrying out a reaction for a certain time,and extracting diethylstilbestrol by utilizing ethyl acetate to terminate the reaction, so that the degradation of diethylstilbestrol is realized. According to the invention, the laccase-ABTS medium solution is prepared and used for degrading diethylstilbestrol, so that the diethylstilbestrol degradation rate of laccase is increased to 99.9% from original 44.5% so as to easily achieve the treatment of diethylstilbestrol sewage, and the method has practical application value in the field of sewage treatment.

Description

technical field [0001] The invention relates to the technical field of sewage treatment, in particular to a method for improving the degradation rate of diethylstilbestrol by laccase. Background technique [0002] Diethylstilbestrol (DES) is a synthetic non-steroidal estrogen or xenoestrogen, which has stronger estrogenic activity than estradiol. It has been used in animal husbandry as an oral contraceptive and growth promoter since 1938. Later studies found that it is an endocrine disrupting substance, DES exposure has a strong negative impact on humans and animals, and even exposure to this synthetic estrogen has brought cancer risks to the world. Zeng Jin, a National Cancer Institute study reported that the use of DES during pregnancy was associated with an increased relative risk of reproductive tract abnormalities, infertility, pregnancy complications, cervical intraepithelial neoplasia, and definite cell adenocarcinoma. For safety reasons, therefore, some countries p...

Claims

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Application Information

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IPC IPC(8): C02F3/34C12N9/96C12N9/02C02F101/34
CPCC02F3/342C02F2101/345C12N9/0061C12N9/96C12Y110/03002
Inventor 刘红艳谢世伟林晓莲李海云李子院张会香单杨
Owner GUILIN UNIVERSITY OF TECHNOLOGY
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