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A method for detecting biological toxicity based on bacteria possessing β-galactosidase

A galactosidase, biotoxicity technology, applied in the measurement of color/spectral properties, analysis by chemical reaction of materials, and material analysis by observing the effect on chemical indicators, which can solve problems such as cumbersome induction process , to achieve the effect of low operator requirements, simple operation and broad application prospects

Active Publication Date: 2019-02-15
TECHNICAL INST OF PHYSICS & CHEMISTRY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Toxi-Chromo The induced Escherichia coli that can produce β-galactosidase is used, and the induction process is relatively cumbersome

Method used

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  • A method for detecting biological toxicity based on bacteria possessing β-galactosidase
  • A method for detecting biological toxicity based on bacteria possessing β-galactosidase
  • A method for detecting biological toxicity based on bacteria possessing β-galactosidase

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] A method for detecting biological toxicity based on bacteria possessing β-galactosidase, comprising the following steps:

[0044]1. Prepare LB medium: Dissolve 0.5g of beef extract, 1g of peptone and 0.5g of sodium chloride in 100mL of deionized water, adjust the pH to 7.0-7.4, and sterilize in high-pressure steam sterilization. After the LB medium is cooled, inoculate the medium with Escherichia coli (ATCC 25922). Then the inoculated culture medium was placed in a constant temperature water bath shaker at 37°C and cultured for 16 hours. At this time, the Escherichia coli was just in the logarithmic growth phase. Escherichia coli was separated by centrifugation, washed twice with 0.8% NaCl solution, and then Escherichia coli was dispersed in 0.8% NaCl solution to obtain a certain concentration of Escherichia coli bacteria liquid.

[0045] 2. Take five 1.5mL test tubes, numbered a, b, c, d, e. Add 400 μL of Escherichia coli bacteria solution to each test tube respectiv...

Embodiment 2

[0055] With embodiment 1, difference is: add the Cu(NO 3 ) 2 solution instead of Cd(NO 3 ) 2 solution.

[0056] Figure 4 It is the detection of different concentrations of Cu in Example 2 2+ UV-Vis absorption spectrum of biological toxicity to Escherichia coli (Cu in a2-f2 2+ The concentrations are 0, 10, 30, 40, 50, 70 mg / L in turn). The larger the absorbance value, the larger the amount of ONP, the concentration of Cu 2+ The toxic effect on Escherichia coli is low; the smaller the absorbance value, the smaller the amount of ONP, the concentration of Cu 2+ The toxic effect on Escherichia coli is greater. It is easy to see that Cu 2+ The biological toxicity to Escherichia coli is much greater than that of Cd 2+ Biotoxicity to Escherichia coli.

Embodiment 3

[0058] 1. Prepare the medium required for the growth of Bacillus subtilis (bacteria number: 1.1086). Dissolve 1 g of peptone, 0.3 g of beef extract, and 0.5 g of sodium chloride in 100 mL of deionized water, adjust the pH to 7.0, and sterilize in a high-pressure steam sterilizer. After the medium is cooled, inoculate the medium with Bacillus subtilis. Subsequently, the inoculated culture medium was placed in a constant temperature water-bath shaker at 37° C. for 24 hours, and Bacillus subtilis was just in the logarithmic growth phase at this time. Bacillus subtilis was centrifuged, washed twice with 0.8% NaCl solution, and then the bacteria were dispersed in 0.8% NaCl solution to obtain a certain concentration of Bacillus subtilis bacteria liquid.

[0059] 2,3,4,5 are basically the same as embodiment 1.

[0060] Figure 5 It is the detection of different concentrations of Cd in Example 3 2+ UV-Vis absorption spectrum of biological toxicity to Bacillus subtilis (Cd in a3-d3...

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Abstract

The invention discloses a method for detecting biological toxicity based on bacteria having beta-galactosidase, wherein the method comprises the steps: mixing a bacteria liquid with a to-be-detected substance, to obtain a mixed liquid; adding ONPG (2-nitrophenyl-beta-D-galactoside) into the mixed liquid, to obtain a mixed reaction liquid; adding sodium carbonate into the mixed reaction liquid, to obtain a final liquid; testing the absorbancy of the final liquid, and evaluating the biological toxicity according to the absorbancy, wherein the bacteria can decompose ONPG to produce beta-galactosidase. The method has no need of induction, rapidly detects the biological toxicity in about 1 h, is simple to operate, has no need of special large instruments, and has low requirements on operators.

Description

technical field [0001] The invention relates to the field of biotoxicity detection, in particular to a method for detecting biotoxicity by using bacteria possessing β-galactosidase. Background technique [0002] With the increasingly prominent problem of environmental pollution, the detection of biological toxicity, especially the rapid detection of biological toxicity, has become more and more important. Traditionally, fish and daphnia are used to detect biological toxicity, but it often has problems such as slow response, long time-consuming, and high cost. The rapid reproduction and low price of microorganisms are ideal materials for detecting biological toxicity. [0003] It is currently the most mature product in the field of biological toxicity detection, which uses Vibrio fischeri. This bacterium is a luminescent bacterium from the ocean. It is rare and expensive. It emits visible fluorescence during normal metabolism; when toxic substances exist, the normal metab...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/78G01N21/31
Inventor 只金芳方德煜李久铭钱俊
Owner TECHNICAL INST OF PHYSICS & CHEMISTRY - CHINESE ACAD OF SCI