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A screening method for heavy metal-resistant microorganisms

A screening method and microbial technology, applied in the field of environmental microorganisms, can solve the problems of substandard concentration of culture medium, false positives of strains, and easy destruction of culture medium components.

Active Publication Date: 2021-09-10
SUZHOU ACADEMY OF AGRI SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, in complex culture medium, especially a variety of carbon sources or medium containing reducing substances, there are many problems, for example, it is easy to cause the concentration of Cr(VI) in the medium to be substandard, and the actual concentration of Cr(VI) is small; The form of Cr(VI) in the medium may change, and Cr(VI) may be reduced by the medium components; the medium components are easily destroyed
The above problems will lead to false positives or false negatives in the screened Cr(VI) resistant strains

Method used

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Embodiment 1

[0030] Embodiment 1: A kind of screening method of resistance to Cr (VI) microorganism

[0031] 1. The method of screening Cr(VI)-resistant microorganisms by soaking in phosphate-Cr(VI) buffer solution (hereinafter referred to as PCB method)

[0032] Prepare pH 7.4 disodium hydrogen phosphate-potassium dihydrogen phosphate buffer solution, weigh 8 g NaCl, 0.2 g KCl, 1.42 g NaCl into a 1 L beaker 2 HPO 4 and 0.27 g KH 2 PO 4 , then add about 800 mL of deionized water into the beaker, stir well to dissolve, adjust the pH to 7.4 with concentrated hydrochloric acid, and dilute the solution to 1 L with deionized water. Configuration 20 g K 2 Cr 2 o 7 L -1 Mother liquor, weighed 20 g and dried to constant weight K 2 Cr 2 o 7 To a 1 L beaker, add about 800 mL of deionized water, fully dissolve and dilute to 1 L. pH 7.4 phosphate buffer with 20 g K 2 Cr 2 o 7 L -1The mother liquor was mixed according to a certain volume, and filtered and sterilized with a 0.22 μm filt...

Embodiment 2

[0045] Embodiment 2: a kind of screening method of resistant Fe(Ⅲ) microorganism

[0046] Ferric ions are important trace elements for microorganisms, but high concentrations of ferric iron can also be toxic to microorganisms. At the same time, ferric iron is oxidizing, and it is easy to produce redox reactions with reducing sugars in the medium, such as glucose. The method can be implemented through the following steps, so as to avoid the reduction reaction and improve the microorganism screening efficiency.

[0047] Step (1). Configure an inorganic buffer solution system containing ferric iron. In this example, disodium hydrogen phosphate-potassium dihydrogen phosphate buffer solution is used to weigh 8 g of NaCl, 0.2 g of KCl, and 0.2 g of NaCl into the beaker. 2 HPO 4 1.42 g, KH 2 PO 4 0.27 g, and about 800 ml of deionized water was added to the beaker, and stirred thoroughly to dissolve. Concentrated hydrochloric acid was added dropwise to adjust the pH to 7.4, an...

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Abstract

A screening method for heavy metal-resistant microorganisms, characterized in that: sequentially comprising the following steps: preparing a buffer solution, adding a heavy metal salt solution to the buffer solution, and then preparing buffer solutions containing heavy metal ions with different concentration gradients, and filtering to sterilize For later use, the buffer solution is a neutral inorganic salt buffer solution; the buffer solution containing heavy metal ions is mixed with the environmental sample to prepare an environmental sample suspension, and after mixing, incubate at a constant temperature of 20-40°C for 0.5-5 Hours, let the microorganisms in the environmental samples accept heavy metal ion immersion screening; after the immersion, carry out gradient dilution to the environmental sample suspension to obtain a dilution, transfer the dilution to a medium without heavy metal ions, and then cultivate to obtain a stable Resistant to heavy metal microorganisms. The method of the invention has the characteristics of quickness, simplicity, no interference to the culture medium, no selectivity, high recognition rate for heavy metal-resistant microorganisms, and sensitivity to non-heavy metal-resistant microorganisms.

Description

technical field [0001] The invention belongs to the field of environmental microorganisms, and in particular relates to a screening method for heavy metal-resistant microorganisms. Background technique [0002] Chromium is an extremely important heavy metal raw material in modern industry. It is widely used in electroplating, tanning, dyeing industry, anti-corrosion and fungicide and other fields. It is estimated that more than 170,000 tons of chromium-containing products enter the environment every year worldwide. [1] . The common chromium in the environment is Cr(III) and Cr(VI), among which Cr(III) has low toxicity and weak mobility in a neutral pH environment, and is also an important trace element for organisms; while Cr(VI) is highly toxic , easily soluble in water, has strong migration ability in the environment, can quickly penetrate into the cell membrane of organisms, destroy intracellular proteins and nucleic acids and affect metabolic processes, short-term and l...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12R1/085C12R1/125C12R1/01
CPCC12N1/20
Inventor 梅丽娟施林林殷士学
Owner SUZHOU ACADEMY OF AGRI SCI
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