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High-yield laccase fungus-trametes hirsute La-7 and application thereof

A technology of Trametes trachees and Trametess, applied in the field of microorganisms, can solve the problems of environmental secondary pollution, inability to remove steroid estrogen, etc., and achieve the effects of low production cost, mild conditions and simple operation

Active Publication Date: 2020-02-28
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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  • High-yield laccase fungus-trametes hirsute La-7 and application thereof
  • High-yield laccase fungus-trametes hirsute La-7 and application thereof
  • High-yield laccase fungus-trametes hirsute La-7 and application thereof

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 (0-10cm) of the sewage outfall of Qianshan County, Qianshan County, Anqing City, Anhui Province. 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 d...

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 of 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 an ultraviolet-visible spectrometer (UV-2550, Shimadzu, Japan), by oxidation 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 μL cru...

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 high-yield laccase fungus named as trametes hirsute La-7 which is collected at the China General Microbiological Culture Collection Center at 3# of Yard 1, Beichenxi Road, Chaoyang District, Beijing on Sept. 27th, 2019 under the number of CGMCC No 5.2199. Extracellular crude laccase produced by fermenting trametes hirsute La-7 can effectively remove 17 beta-estradiol in natural water and livestock-poultry manure.

Description

technical field [0001] The invention belongs to the field of microbes, and in particular relates to a high-yielding laccase-producing fungus Trametes trachees 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 estro...

Claims

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Application Information

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Patent Type & Authority Applications(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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