Detection of environmental pollutants through photobacterium leiognathi
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A technology of environmental pollutants and luminescent bacteria, applied in the field of microorganisms, can solve the problems of detection of environmental pollutants without luminescent bacteria
Inactive Publication Date: 2011-05-25
OCEAN UNIV OF CHINA
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Except for Luminescent bacteria, there is no report on the use of other species of luminescent bacteria to detect environmental pollutants.
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Embodiment 1
[0015] Common seafood caught from the coastal waters of Qingdao, including squid, prawns, yellow croaker, blackhead, monkfish, etc. Divide various aquatic products into small pieces and place them on solid medium plates, culture them in a water-proof constant temperature incubator at 25°C for 10 hours, and observe the luminescence in a dark room. Luminescent colonies were marked and streaked on plates. After several times of separation and purification, the luminescent bacteria YL was obtained.
Embodiment 2
[0017] Take a single colony of luminescent bacteria YL and inoculate it in 100ml liquid medium (medium composition: peptone 5g, yeast extract 1g, FePO 4 0.1g, NaCl 20g, glycerin 3ml, distilled water 1000ml), cultured on a shaking table at 27°C for 8hrs to make the bacteria reach the terminal logarithmic growth phase.
[0018] Take 5ml of heavy metal zinc solutions of different concentrations and add them to 5ml of bacterial solution, and incubate and react on a shaking table at 27°C for 20min. After the reaction is completed, measure the luminescence value in a weak luminescence measuring instrument for 50 s, and take the average value of the results. It can be seen from the measurement results that the inhibition rate and Zn 2+ Concentration is proportional to ( Figure 4 ), so when measuring this kind of pollutant in food or environment, the content of the pollutant can be inferred according to the inhibition rate.
Embodiment 3
[0020] Take a single colony of luminescent bacteria YL and inoculate it in 100ml liquid medium (medium composition: peptone 5g, yeast extract 1g, FePO 4 0.1g, NaCl 20g, glycerin 3ml, distilled water 1000ml), cultured on a shaking table at 27°C for 8hrs to make the bacteria reach the terminal logarithmic growth phase.
[0021] Take 5ml of heavy metal copper solutions of different concentrations and add them to 5ml of bacterial solution, and incubate and react on a shaking table at 27°C for 20min. After the reaction is completed, measure the luminescence value in a weak luminescence measuring instrument for 50 s, and take the average value of the results. From the measurement results, it can be seen that the inhibition rate and Cu 2+ The concentration is proportional to ( Figure 5 ), so when measuring this kind of pollutant in food or environment, the content of the pollutant can be inferred according to the inhibition rate.
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Abstract
The invention provides photobacterium leiognathi (Photobacterium leiognathi YL) from a marine environment and quantitative detection of environmental pollutants by using the strain. A strain of luminous bacterium YL from the marine environment is separated and purified, and the strain is Gram-negative bacterium and has a rod shape. The luminous bacterium YL is identified as the Photobacterium leiognathi according to a phylogenesis tree constructed according to a 16S rDNA sequencing result and physiological and biochemical results. The strain was preserved in China Center for Type Culture Collection (CCTCC) on 27, December, 2006, and the collection number is M 206139. The 16S rDNA gene sequence of the luminous bacterium has been submitted to a GenBank nucleotide sequence database, and the access number is EF017227. The strain has the characteristics of fast growth, stable luminous performance, low nutrition requirement, sensitivity to environmental pollutants, and the like, and the luminous intensity of the luminous bacterium is determined to form the maximum peak at 474nm through fluorescent scanning. The luminous intensity inhibition rate is in a direct proportion to the concentration of various environmental pollutants, and the quantitative detection of the environmental pollutants can be realized. The method for detecting the environmental pollutants by the strain has the characteristics that: the cost is low and the method is easy and convenient to operate, high in sensitivity and the like, and can meet the requirement on rapid detection of the pollutants.
Description
technical field [0001] The invention relates to a microorganism, in particular to a marine luminescent bacterium Photobacterium leiognathi YL strain which can be used for the detection of environmental pollutants. Background technique [0002] According to the inventor's information and literature search, there are only three known genera of marine luminescent bacteria, which are Vibrio, Photobacterium, and Shewanella. On land, there are different short bacillus (Xenorhabdus). In an aerobic natural environment, luminescent bacteria can produce luciferase (LE), catalyze flavin mononucleotide (FMNH 2 ) and long-chain aliphatic aldehydes undergo oxidation reactions to emit blue-green fluorescence. [0003] At present, Photobacterium phosphoreum is widely used in our country. There are some reports on the use of Photobacterium phosphoreum to detect environmental pollutants. Except for Luminescent bacteria, there is no report on using other species of luminescent bacteria to d...
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