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A culture medium for luminescent bacteria

A culture medium and technology of luminescent bacteria, applied in the direction of bacteria, microorganism-based methods, microorganisms, etc., can solve the problems of unstable bacterial content, high cost, unfavorable experimental research and popularization of luminescent bacteria in liquid culture of luminescent bacteria, and achieve Conducive to promotion and application, low cost, and the effect of reducing research costs

Active Publication Date: 2016-03-02
北京尚洋东方环境科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, the cost of liquid medium for luminescent bacteria in the market is relatively high, which is not conducive to the experimental research and popularization and application of luminescent bacteria
And there is no mature plan for the cultivation conditions of luminescent bacteria, which makes the content of liquid culture bacteria of luminescent bacteria unstable

Method used

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  • A culture medium for luminescent bacteria
  • A culture medium for luminescent bacteria
  • A culture medium for luminescent bacteria

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] Embodiment 1 Contrast test of luminescent bacteria content and production cost

[0017] Prepare luminescent bacteria culture medium with different ratios for comparative experiments. The specific components of each experimental group are shown in Table 1:

[0018] Table 1: Components of the medium

[0019]

[0020]

[0021] The components in each of the above experimental groups were configured into 100ml of culture medium, and then used after high pressure. And 0.5g of tryptone, 0.5g of yeast extract powder, 0.3mL of glycerin, 0.1g of potassium dihydrogen phosphate, 1.26g of disodium hydrogen phosphate dodecahydrate, and 3g of sodium chloride were dissolved in 100mL of water to make a culture medium, as Control group, the components of the control group are currently used medium for the cultivation of luminescent bacteria.

[0022] Add samples to the culture plate, add 3 wells for each group of experiments, add 350uL of medium to each well, inoculate 7uL of sha...

Embodiment 2

[0026] The determination of embodiment 2 optimal culture conditions

[0027] Dissolve the freeze-dried powder of Vibrio fischeri with 3% NaCl solution, recover for 10 minutes, inoculate into 150mL optimized liquid medium (experimental group 2 in Example 1) in a 500mL Erlenmeyer flask, and culture at 180rpm 18h for enrichment. Inoculate the 1%, 1.5%, 2%, 3%, and 4% of the culture medium into 150mL culture medium in a 500mL Erlenmeyer flask, and incubate at 180rpm for 18h. The bacterial solution was diluted with 3% NaCl, and the colonies were counted, and the relative fluorescence intensity was measured.

[0028] Inoculate the recovered bacterium solution into the 200mL liquid medium (control group in Example 1) contained in the 500mL Erlenmeyer flask, the inoculum amounts were 1%, 1.5%, 2%, 3%, 4%, 25% respectively. Cultivate at 180rpm for 18h. The colonies were counted after serially diluting the bacterial solution with 3% NaCl solution, and their relative fluorescence inte...

Embodiment 3

[0032] The comparison of the luminous intensity of embodiment 3 luminescent bacteria

[0033] Add the inoculated two culture media into the luminometer, and dynamically measure the luminosity of the luminescent bacteria. The results are as follows: figure 1 shown. The results showed that the luminous intensity of the culture medium provided by the present invention continued to increase in the luminescence measuring instrument, while the luminous intensity of the control group was distributed in the range of 48-245. It can be seen that the culture medium of the present invention can ensure that the duration of luminous intensity of luminescent bacteria is extended.

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Abstract

The invention provides a culture medium for a light-emitting bacterium. Each litre of the culture medium contains 5-10 g of tryptone, 5-10 g of beef extract, 2-4 mL of glycerol, 0.3-1.5 g of potassium dihydrogen phosphate, 11-13 g of disodium hydrogen phosphate dodecahydrate and 21-35 g of sodium chloride. The invention further provides a method for culturing the light-emitting bacterium by using the culture medium. When the method is used for culturing the light-emitting bacterium, the volume of inoculated bacterium liquid is 2% of that of the culture medium. The bacterium content of the light-emitting bacterium in the liquid culture medium can stably surpass 10 to the power of 8 CFU / mL, so that the research and application of the light-emitting bacterium and the production of freeze-dried powder thereof are facilitated; and the culture medium for the light-emitting bacterium is low in cost, so that popularization and application of the light-emitting bacterium are facilitated.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to the preparation of a medium for luminescent bacteria and a method for cultivating the luminescent bacteria using the medium. Background technique [0002] Luminescent bacteria are a class of bacteria that perform bioluminescence. Most of them are marine, and they are the same cause of the glow of the sea surface as the luminescent plankton. Although the luminescent bacteria have various forms, their physiological characteristics are very similar. Generally, it does not liquefy gelatin, and does not form poison after decomposing proteins. It often parasitizes various animals and causes "luminescence disease", that is, parasitic luminescence. At present, three kinds of luminescent bacteria commonly used in China are: Photobacterium luminosus, Vibrio fischeri, and Vibrio qinghai. [0003] Among them, Luminescent Bacillus is a Gram-negative bacterium, which can be used in the determin...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12R1/63
Inventor 袁恒
Owner 北京尚洋东方环境科技有限公司
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