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Quantitative detection method of intracellular active thiol compounds in white rot fungi under heavy metal stress

A quantitative detection method, the technology of mercapto compounds, applied in the field of environmental microbiology, can solve the problems of low crushing efficiency, easy frostbite, troublesome operation, etc., and achieve the effect of improving crushing efficiency, promoting application, and simple and convenient detection method

Inactive Publication Date: 2015-09-30
HUNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] The existing detection methods for active thiol compounds are mainly used for the detection of active thiol compounds in higher organisms such as animals and plants. The crushing methods used are basically liquid nitrogen grinding and crushing, and the crushing efficiency is low, the work intensity is high, and the operation is difficult. Troublesome, not suitable for batch crushing, and there is a safety hazard that is prone to frostbite

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  • Quantitative detection method of intracellular active thiol compounds in white rot fungi under heavy metal stress
  • Quantitative detection method of intracellular active thiol compounds in white rot fungi under heavy metal stress

Examples

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Embodiment 1

[0023] In this example, the bacterial species used for white-rot fungi is Phanerochaete chrysosporium, a model strain of white-rot fungi ( Phanerochaete chrysosporium ) BKMF-1767 (the deposit number in the American Type Culture Collection is ATCC 24725, this strain is preferred, but not limited to).

[0024] A method for quantitative detection of intracellular active sulfhydryl compounds of white rot fungi under heavy metal stress of the present invention, comprising the following steps:

[0025] (1) Scrape the Phanerochaete chrysosporium spores on the plate into sterile water to form a Phanerochaete chrysosporium spore suspension with a turbidity of 60 (the amount of spores contained is about 2.5×10 6 pcs / L); then add 4ml of Phanerochaete chrysosporium spore suspension into the liquid medium (that is, the culture medium) (the liquid medium has been sterilized at 105°C for 30min) for shaking culture, shaking culture The temperature is 37°C, the rotation speed is 150r / min, and...

Embodiment 2

[0035] In this example, the strains used by white rot fungi are Phanerochaete chrysosporium, a model strain of white rot fungi. Phanerochaete chrysosporium BKMF-1767.

[0036] A method for quantitative detection of intracellular active sulfhydryl compounds of white rot fungi under heavy metal stress of the present invention, comprising the following steps:

[0037] (1) Scrape the Phanerochaete chrysosporium spores on the plate into sterile water to form a Phanerochaete chrysosporium spore suspension with a turbidity of 60 (the amount of spores contained is about 2.5×10 6 pcs / L); then add 4ml of Phanerochaete chrysosporium spore suspension into the liquid medium for shaking culture, the temperature of shaking culture is 37°C, the rotation speed is 150r / min, and the shaking culture time is 60h;

[0038] (2) Add a concentration of 15mg L -1 The cadmium-containing solution, when cultured for 0h, 4h, 8h, 16h, and 24h, respectively take 3 cultures (that is, 3 parallel samples) to...

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Abstract

The invention discloses a quantitative detection method for intracellular active sulfhydryl compound of white-rot fungi under heavy metal stress. The method comprises the following steps of: adding white-rot fungi spore into a culture solution, and performing shaking culture on the fungi spore till the logarithmic phase of the white-rot fungi; subsequently, adding a heavy metal solution into the white-rot fungi culture solution, performing shaking culture on the solution continually till a set sampling time, filtering and separating the culture, and washing the culture by buffer liquid at 0-4 DEG C; furthermore, mixing obtained mycelium pellets with the buffer liquid at 0-4 DEG C, carrying out ice-water bath ultrasonication on the mixture, and carrying out refrigerated centrifugation on obtained homogenate to obtain supernate which is taken as a sample; and measuring the absorbance of the sample by a method with a 5.5'-disulfenyl-2-nitrobenzene acid solution, and calculating the content of the active sulfhydryl compound in the sample. The detection method is simple, convenient, high in crushing efficiency and capable of rapidly and reliably acquiring and quantitatively detecting the content of the intracellular active sulfhydryl compound of the white-rot fungi.

Description

technical field [0001] The invention belongs to the field of environmental microbiology, and relates to a method for measuring the content of active sulfhydryl compounds in white-rot fungal cells, in particular to a quantitative detection method for active sulfhydryl compounds in white-rot fungal cells under heavy metal stress. Background technique [0002] With the development of society, the content of heavy metals and refractory organic substances in the environment is increasing day by day. These bioxenobiotics can be enriched and persist in organisms through the food chain, seriously threatening the ecological environment and human health. In recent years, due to the advantages of low cost, low impact on the environment and high treatment efficiency, microbial treatment technology has attracted more and more attention from environmental protection workers and scientific and technological personnel. [0003] White rot fungi have a unique ability to degrade and process x...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/31
Inventor 黄健陈桂秋曾光明晏铭陈安伟张企华易斌杜坚坚王璐尚翠周颖李欢可
Owner HUNAN UNIV