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Lyophilized powder of Photobacter lumina and preparation method thereof

A technology of bright photobacteria and freeze-dried powder, applied in biochemical equipment and methods, methods based on microorganisms, bacteria, etc., can solve problems such as differences in cryoprotective effects, and achieve good stability

Active Publication Date: 2017-12-29
EAST CHINA UNIV OF SCI & TECH +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, because the concentration of each protective agent, the ability of cell penetration, etc. are not the same, the effect of cryoprotection will be different.

Method used

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  • Lyophilized powder of Photobacter lumina and preparation method thereof
  • Lyophilized powder of Photobacter lumina and preparation method thereof
  • Lyophilized powder of Photobacter lumina and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] formula:

[0053] Skim milk 30.0%

[0054] Trehalose 22.1%

[0055] Arginine 0.9%

[0056] Sodium Chloride 1.3%

[0057] Bacteria balance

[0058] The bacteria are collected after cultivation, that is, the bacteria are centrifuged to remove the supernatant, and the preparation method is as follows:

[0059] Take a Photobacterium phosphoreum EP tube and inoculate the bacterial solution into a 250ml Erlenmeyer flask containing 50ml of culture medium. According to the previously optimized culture conditions, the inoculation amount is 2%, 200r / min, pH 7.0, Shake culture at 18°C ​​for 16-20h.

[0060] The formula of described culture medium:

[0061]

[0062]

[0063] Place the cultured bacterial solution in a centrifuge tube, centrifuge at 7000r / min for 30min, remove the supernatant, wash with 1ml of 3% NaCl solution, centrifuge, remove the supernatant, and collect the precipitate, which is the obtained said bacteria;

[0064] The preparation method of the lyo...

Embodiment 2

[0090] formula:

[0091] Skim Milk 29%

[0092] Trehalose 23%

[0093] Arginine 0.8%

[0094] Sodium Chloride 1.1%

[0095] Bacteria balance

[0096] The thalline is the material collected after cultivation, that is, the thalline after the bacterial liquid is centrifuged to remove the supernatant, and the preparation method is the same as in Example 1.

[0097] Place the cultured bacterial solution in a centrifuge tube, centrifuge at 9000r / min for 20min, remove the supernatant, wash with 1ml of 3% NaCl solution, centrifuge, remove the supernatant, and collect the precipitate, which is the obtained said bacteria;

[0098] The preparation method of the lyophilized powder of Photobacterium lumina is as follows:

[0099] (1) The bacteria are added to the mixture of skim milk, trehalose, arginine and sodium chloride, dispersed, then placed in an ampoule bottle, pre-cooled at -20°C for 1 hour, and then pre-frozen at -70°C 3 hours;

[0100] (2) Put the pre-frozen ampoules int...

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Abstract

The invention discloses photobacterium phosphoreum freeze-dried powder and a preparation method thereof. The photobacterium phosphoreum freeze-dried powder comprises the following components in percentage by weight: 25-35 percent of skimmed milk, 20-24 percent of mycose, 0.7-1.0 percent of arginine, 1-1.5 percent of sodium chloride and the balance of thalli. The photobacterium phosphoreum freeze-dried powder adopts a compound cryoprotectant; and experiments prove that the compound cryoprotectant is greatly improved in freezing resistance, has a higher protective property than a simplex protective agent and has good stability. The average value of luminous intensity of the photobacterium phosphoreum freeze-dried powder provided by the invention is about 1,843mv and approximate to a model expected value 1,820. When the toxicity of ZnSO4.7H2O, K2Cr2O7 and C6H5OH is detected with photobacterium phosphoreum freeze-dried powder prepared in different batches, the results show that the EC50 value variation coefficients detected by the photobacterium phosphoreum freeze-dried powder prepared in different batches are less than 6%.

Description

technical field [0001] The invention relates to a preparation of luminescent bacteria and a preparation method thereof. Background technique [0002] Based on the current situation of environmental pollution in my country, the detection method of luminescent bacteria toxicity is an efficient and convenient means to determine the biological toxicity of pollutants. The biggest advantage of the luminescent bacteria toxicity detection method is that it can be detected on-site, the method is fast, accurate, and easy to operate. There is a significant negative correlation between the relative luminosity of luminescent bacteria and the comprehensive toxicity of water. Using photoelectric detection technology to use luminescent bacteria that are sensitive to the environment for the detection of water toxicity has the advantages of simplicity and speed. On-site toxicity testing requires fast and simple operation, but fresh bacterial solution is inconvenient to carry and difficult to...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/04C12N1/20C12R1/01
CPCC12N1/04C12N1/20
Inventor 冯耀宇李佩苏嘉缘孟芹胡越蔡敏姚红利吴晓袁旭华
Owner EAST CHINA UNIV OF SCI & TECH
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