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A Laccase-Producing Fungus Trametes la-7 and Its Application

A technology of Trametes trachees and Trametess, which is applied in the field of microorganisms, can solve the problems of secondary pollution of the environment and the inability to remove steroid estrogen, etc., and achieve the effects of low production cost, mild conditions and high catalytic efficiency

Active Publication Date: 2020-11-27
ANHUI AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These technologies can effectively reduce high-concentration steroid estrogen pollution in the environment, but cannot remove low-concentration steroid estrogen pollution, and may form toxic transformation products, causing secondary pollution to the environment

Method used

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  • A Laccase-Producing Fungus Trametes la-7 and Its Application
  • A Laccase-Producing Fungus Trametes la-7 and Its Application
  • A Laccase-Producing Fungus Trametes la-7 and Its Application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1 1

[0026] Example 1 Isolation, screening and identification of a strain of laccase-producing fungus Trametes trachees La-7

[0027] The soil samples contaminated by livestock and poultry feces were collected from the surface soil (0-10 cm) of the sewage outlet of Qianshan County, Anqing City, Anhui Province, and the average concentration of 17β-estradiol in the soil was 134.7 μg kg -1 . Weigh 2 g of fresh soil samples and place them in sterile water containing 0.9% NaCl for 1-2 minutes to vigorously shake to promote the dissociation of fungi from the soil. The cultivable fungi in the soil were isolated and purified by the method of streaking on a plate. Select the hyphae with better growth and inoculate it on a solid medium containing 0.05% guaiacol, and incubate it upside down at 30°C for 4-8 days. If reddish brown appears on the edge of the colony, it indicates that laccase is produced. According to the degree of color development and color development The ring diameter size ...

Embodiment 2

[0029] The optimization of the liquid fermentation culture condition that embodiment 2 Trametes trachees La-7 produces laccase

[0030] Use 150mL Erlenmeyer flask to pack 50mL laccase-producing liquid fermentation medium (pH 3-7), inoculate bacteria cake with a diameter of 0.6cm, 30°C, 150r min -1 Shaking culture 7d. Within the set incubation time interval, take 5mL fermentation broth and place it at 4°C, 12,000r min -1 Centrifuge at high speed for 5 minutes, and the obtained supernatant is the crude laccase solution. Study pH, temperature, carbon source, nitrogen source and Cu 2+ Effects of ion concentration and other single factors on the laccase-producing ability of Tracherichtes tracherichae La-7. Using ultraviolet-visible spectrometer (UV-2550, Shimadzu, Japan), through the oxidation of 1mmol L -1 2,6-dimethoxyphenol (2,6-DMP) was used to determine the change of laccase activity in the fermentation broth before and after the reaction. The reaction system contained 20 ...

Embodiment 3

[0032] 17β-estradiol in the removal buffer of embodiment 3 Trametes trachees La-7 laccase

[0033] Put the laccase fermentation broth produced by Trametes trachees La-7 at 4°C, 12,000r·min -1 Centrifuge at high speed for 5 minutes to obtain crude extracellular laccase supernatant. Preliminarily separate and purify the crude laccase liquid by salting out method, add 70% saturated ammonium sulfate to the crude laccase supernatant, 4°C, 150r min -1 Shake for 24h to precipitate the crude laccase, then adjust the enzyme activity of the collected precipitate to 200U·mL with ultrapure water -1 As a concentrated crude laccase stock solution, store in a 4°C refrigerator for later use.

[0034] The effect of pH on the removal of 17β-estradiol in the experimental system of La-7 laccase from Tracherichtes tracherichtes was studied by batch equilibrium test. The reaction system was constructed in 25mL glass reactors, each containing 5mL 10mmol L -1 C-PBS buffer and 5 μmol L -1 17β-est...

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Abstract

The invention discloses a strain of laccase-producing fungus, which is named as trachephrine ( Trametes hirsuta ) La‑7, was deposited in the General Microbiology Center of the China Microbiological Culture Collection Management Committee on September 27, 2019. The culture preservation number is CGMCC No 5.2199, and the preservation address is No. 3, No. 1, Beichen West Road, Chaoyang District, Beijing. Utilize the tracheomyces of the present invention ( Trametes hirsuta ) La‑7 fermented extracellular crude laccase can effectively remove 17 β ‑Estradiol.

Description

technical field [0001] The invention belongs to the field of microorganisms, and in particular relates to a strain of laccase-producing fungus trachenotus La-7 and its application. Background technique [0002] Steroid estrogens in the environment mainly come from industrial wastewater, livestock and poultry waste, and domestic garbage. They can be released into natural water bodies and farmland soil, thereby causing health risks to wild animals and humans. Steroid estrogen enters the body of animals and humans, and by simulating, interfering or antagonizing the synthesis, transport, metabolism and release of biological endocrine substances, it leads to endocrine disorders and disorders, thereby destroying its normal regulation on the organism. In recent years, researchers have frequently detected steroid estrogens such as estrone, 17β-estradiol and 17α-ethinyl estradiol in surface water, groundwater and farmland soil environment in my country. Exposure of steroid estrogens...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/14C12N9/02C02F3/34C02F11/02C02F103/20C02F101/34C12R1/645
CPCC02F3/342C02F11/02C02F2101/34C02F2103/20C12N9/0061C12Y110/03002C12N1/145C12R2001/645
Inventor 孙凯司友斌
Owner ANHUI AGRICULTURAL UNIVERSITY
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