Sectional oxygen supply fermentation technology of insect pathogenic nematode symbiotic bacteria and use of its fermented material

A technology of entomopathogenic nematodes and fermentation technology, applied in the direction of bacteria, application, biocide, etc., to achieve the effect of improving utilization rate, reducing production cost and increasing yield

Inactive Publication Date: 2008-02-20
馥稷生物科技发展(上海)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There are no reports or patents on the related technologies and methods of the cultivation and fermentation of symbiotic bacteria and the production of secondary metabolites

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] 1. Streak the pathogenic bacteria Xenorhabdus nematophilus YL001 stored in the seed tube on the NA medium plate, culture at 28°C for 24 hours, pick a single colony, then streak on the NBTA medium plate, and culture at 28°C for 48h;

[0032] 2. Use an inoculation loop to pick up the blue colony on the NBTA medium plate, inoculate it in a 250ml Erlenmeyer flask filled with 50ml NB medium, and culture it with shaking at 28°C and 180r / min for 16-24h to become a first-grade seed solution;

[0033] 3. Put the first-grade seed liquid into a 250ml Erlenmeyer flask containing seed medium at an inoculation amount of 9.0%, the liquid volume is 25ml, pH7.25, shake and cultivate at 26°C and 220rpm for 16-24h, and become a second-grade seed liquid;

[0034] 4. Add 4.5L fermentation medium, 0.01% defoamer, pH 7.2, 0.1-0.15MPa off-site steam sterilization for 30min to 28°C in 7L stirred fermenter, add 9.0% secondary seed liquid, Under the conditions of 26° C., ventilation rate of 2.5 ...

Embodiment 2

[0039] 1. Streak the symbiotic bacteria stored in the seed tube on the NA medium plate, incubate at 28°C for 24-48h, pick a single colony, and then streak on the NBTA medium plate, incubate at 28°C for 24-48h;

[0040] 2. Use an inoculation loop to pick up the blue colony on the NBTA medium plate, inoculate it in a 250ml Erlenmeyer flask filled with 50ml NB medium, and culture it with shaking at 28°C and 180r / min for 16-24h to become a first-grade seed solution;

[0041] 3. Put the first-grade seed liquid into a 250ml Erlenmeyer flask with seed medium at an inoculation amount of 9.0%, the liquid volume is 25ml, pH 7.2, and vibrate at 26°C and 220rpm for 16-24h to become a second-grade seed liquid;

[0042] 4. Add 4.5L fermentation medium, 0.01% defoamer, pH 7.2, 0.1-0.15MPa off-site steam sterilization for 30min to 26°C, add 9.0% secondary seed liquid into the 7L stirred fermenter, Cultivate at 26°C for 72 hours to obtain a fermented product. During the fermentation process,...

Embodiment 3

[0053] The application of Xenorhabdus nematophilus YL001 fermented liquid of Xenorhabdus nematophilus YL001 carries out on the basis of embodiment 1,2:

[0054] 1. The fermentation broth obtained in the process is further centrifuged at 14000r / min, 4°C for 10min, and the supernatant is collected;

[0055] 2. Dilute the supernatant and fermented product 10 times with the melted PDA medium, put 10ml of the medium into a sterilized petri dish with a diameter of 9.0cm to make a plate, and use the PDA with sterile medium as a control . From the edge of the phytopathogenic fungal colonies cultivated for 3 to 6 days, use a puncher to cut into 7.5mm-diameter bacterial blocks, place them in the center of the plate, and cultivate them in the dark at 20°C for 3 days to measure the diameter of each pathogenic fungal colony;

[0056] 3. The supernatant of Xenorhabdus nematophilus YL001 fermentation, at 100-300ml (fermentation liquid) / L, can grow on the hyphae of Alternaria tabacum, Tomato...

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PUM

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Abstract

The invention relates to a segmented oxygen supply fermenting technology of entomiasis initial line insect symbiosis fungus and the application of fermenting liquid. It uses Xenorhabdus nematophilus YL001 as raw material and adopts controlling airflow quantity and whisking rotating speed to realize segmented oxygen supply during fermenting process. The cell growth and antibacterial activity is higher than single oxygen supply fermenting method. The fermenting liquid could be used to manufacture antiseptic.

Description

technical field [0001] The invention relates to a bacterial fermentation process, in particular to a staged oxygen-supply fermentation process for improving the antibacterial activity of entomopathogenic nematode symbiotic bacteria and the use of the fermented product. Background technique [0002] Entomopathogenic nematode symbionts are a type of bacteria parasitic in the intestinal tract of entomopathogenic nematodes, belonging to the family Enterobacteriaceae, facultatively anaerobic, chemoheterotrophic, and Gram stain negative. Including Xenorhabdus and Photorhabdus, they live in symbiosis with Steinernema and Heterorhabditis nematodes respectively. In nature, such bacteria exist in the intestines of nematodes at the third instar infection stage. As the nematodes infect insects, the symbiotic bacteria are carried into the insect and released into the host blood cavity, and the symbiotic bacteria reproduce in large numbers, producing toxins and Bacteriostatic substances,...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A01N63/02
Inventor 王永宏何军刘霞李广泽冯俊涛马志卿张兴
Owner 馥稷生物科技发展(上海)有限公司
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