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High-flux fast screening method for antibiotics generated bacterium and device thereof

A technology for antibiotics and bacteria production, applied in the direction of microorganism-based methods, biochemical cleaning devices, biochemical equipment and methods, etc., can solve the problems of cumbersome operations, difficult conditions to control, and long time consumption, so as to improve screening efficiency and reduce labor costs. Effects of cost and workload reduction

Inactive Publication Date: 2008-03-26
SOUTH CHINA SEA INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Generally speaking, the above methods all need to purify the bacteria to be screened first, and it is impossible to realize the simultaneous isolation and screening of the strains
In addition to the agar block method, other methods require liquid fermentation of the bacteria to be screened. Not only is the operation cumbersome, it takes a long time, the conditions of liquid fermentation are difficult to control, and the liquid fermentation conditions of some strains to be screened are not suitable for the production of antibiotics.
Although the steps of the agar block method are relatively simple, when screening antagonistic bacteria, it is necessary to transfer the agar block of the bacteria to be screened from the culture plate to the indicator plate, and only a few bacteria to be screened can be screened at a time. During operation, it is inconvenient to transfer the agar block containing the bacteria to be screened, and requires a lot of materials, which is time-consuming and laborious

Method used

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  • High-flux fast screening method for antibiotics generated bacterium and device thereof
  • High-flux fast screening method for antibiotics generated bacterium and device thereof
  • High-flux fast screening method for antibiotics generated bacterium and device thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1: From the sponge leachate, screen the bacterial strains that have antagonistic effect with Bacillus subtilis while isolating bacterial strains

[0029] The devices shown in Figures 2 to 4 are used. The upper and lower layers of each device have an area of ​​120 mm × 80 mm. The bottom of each petri dish on the upper layer has a small hole with a diameter of about 4 mm that communicates with the lower petri dish. microporous membrane.

[0030] Open the upper cover of the device, add sterile modified 2216E liquid medium containing agar 15g / L to the 96 petri dishes on the upper layer, and store it at 4°C for later use after solidification;

[0031] Break 1g of fresh sponge into small fragments in 10mL of sterilized seawater, mix well and let it stand for 5min to take the supernatant, dilute the supernatant 10 times to form 4 gradients in sequence, and then take 100μL of each gradient dilution and apply them evenly on On the above-mentioned upper medium, culture i...

Embodiment 2

[0034] Example 2: Separation and screening of bacterial strains with antagonistic effects on Staphylococcus aureus from sponges

[0035] More than 300 strains were isolated and purified from South China Sea sponge samples with 2216E medium.

[0036] Four devices identical to those in Example 1 were used. Open the upper cover, add sterile agar 10g / L 2216E solid medium into the upper culture dish of the device, cover the upper cover after solidification, and turn it upside down; open the lower cover, and pour in sterilized LB liquid at about 50°C Culture medium (agar 15g / L, other conditions are the same as in Example 1), cover the upper and lower covers after it solidifies, and put the device upright; in each petri dish on the upper layer, inoculate different isolated bacterial strains, and about 5 μL of bacterial strains can be inserted. or use an inoculation loop to pick a single colony; then place it in a 28°C incubator with the upper cover facing up and culture it upright f...

Embodiment 3

[0038] Example 3: Isolation and screening of strains with antagonistic effects on yeast from soil

[0039] More than 80 strains were isolated and purified from soil samples collected from Shennongjia Nature Reserve with ISP4 medium;

[0040] A device as shown in accompanying drawings 5-7 is adopted, the upper and lower layers of the device are each 120 mm × 80 mm in area, the upper layer is composed of 96 petri dishes, the lower layer is a flat petri dish with edges, and the bottom of the upper layer is 96 petri dishes. There are small holes with a diameter of about 6mm communicating with the lower layer, and a semi-permeable microporous membrane is fixed close to each hole. The lower layer cover is made of alloy material, and the size is 130m×90mm×10mm.

[0041] Open the loam cake, inoculate after adding the aseptic improved ISP4 culture medium (agar 10g / L) in 96 petri dishes on the upper layer of the device. Each petri dish is inoculated with different isolated strains, and...

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Abstract

The present invention relates to process and apparatus for screening antibiotic producing strain. The apparatus has one upper layer of culture dishes and one lower layer of culture dishes separated with one semi-permeable microporous film. After abacterial liquid culture medium connecting agar in 10-15 g / l for the bacterial strain to be screened is fed to the culture dishes in one layer and solidified and the bacterial strain to be screened is inoculated and cultured for sufficient time, indicator bacterium is inoculated to the culture dishes in the other layer, the bacterial strain to be screened and the indicator bacterium are cultured simultaneously and the indicator bacterium culturing medium is observed for whether to have inhibition zone. The process of the present invention is simple, fast, high in screening efficiency and low in cost.

Description

technical field [0001] The invention relates to a new method for high-throughput rapid screening of antibiotic-producing bacteria, and also relates to a new device used in the method. Background technique [0002] The discovery of antibiotics such as penicillin and their clinical application have played a huge role in the control of human infectious diseases. However, with the widespread use of antibiotics, especially the abuse of antibiotics, various drug-resistant pathogens have emerged one after another, and the speed of emergence is getting faster and faster. For example, Methicillin-resistant Staphylococcus (MRSA), a common infectious bacterium in hospitals, can greatly increase the mortality rate of infectious diseases. Another example is the emergence of superbugs that can resist various antibiotics such as methicillin, tetracycline, and vancomycin. The emergence of these bacteria greatly threatens human health and quality of life. [0003] Although rational use of ...

Claims

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Application Information

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IPC IPC(8): C12M1/34C12R1/01C12R1/04C12R1/125C12R1/19C12R1/38C12R1/385C12R1/44C12R1/645C12R1/80C12R1/85C12R1/865
Inventor 欧阳永长李翔谢练武姜淑梅戴世鲲秦岭
Owner SOUTH CHINA SEA INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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