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Bacillus velezensis AM6, microbial inoculum, and preparation method and application of microbial inoculum

A technology of bacillus and bacterial agent, applied in the field of Bacillus velei AM6, bacterial agent and its preparation, to achieve excellent emulsification performance, good resistance to salt and alkali and drought, and to assist and promote digestion and absorption

Active Publication Date: 2021-07-09
RES CENT FOR ECO ENVIRONMENTAL SCI THE CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are still few studies on Bacillus with broader antibacterial properties, and it is of great significance to screen out Bacillus with strong environmental adaptability and broad antibacterial activity.

Method used

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  • Bacillus velezensis AM6, microbial inoculum, and preparation method and application of microbial inoculum
  • Bacillus velezensis AM6, microbial inoculum, and preparation method and application of microbial inoculum
  • Bacillus velezensis AM6, microbial inoculum, and preparation method and application of microbial inoculum

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preparation example Construction

[0026] According to some embodiments of the present invention, there is also provided a method for preparing the above bacterial agent, comprising the following steps: inoculating Bacillus velei AM6 on the LB1 culture plate, and culturing at 37° C. for 12 hours; The colonies were transferred to LB1 liquid medium, and cultured at 37°C for 12 hours to obtain seed liquid; the seed liquid was inoculated into LB1 liquid medium at a volume ratio of 1%, and fermented and cultivated at 37°C for 30 hours to obtain a fermented liquid as a liquid bacterial agent, and fermented and cultivated The ventilation volume is 500~1000m 3 / h, the tank pressure is 0.04Mpa, and the rotation speed is 80rpm.

[0027] In some embodiments, the preparation method further includes centrifuging the fermentation broth to obtain the cells, adding skim milk with a mass concentration of 5% to the cells as a protective agent, and spray-drying at an air inlet temperature of 90-100°C , to obtain solid microbial ...

Embodiment 1

[0035] Screening and Identification of Bacillus Velez AM6 Strain

[0036] 1.1 Separation and purification

[0037] (1) Inoculate 5 g of the saline-alkali environment sample collected in a sterilized inorganic salt medium with crude oil as the only carbon source, and culture in a shake flask at 37°C and 180 rpm for 7 days;

[0038] (2) Transfer to fresh aseptic inorganic salt medium with crude oil as the only carbon source at an inoculation rate of 10%, repeat 3 times;

[0039] (3) Dilute the enriched culture in a gradient, and make the gradient 10 5 、10 6 、10 7 Spread the diluted solution on the LB plate, and select single bacteria to streak and purify on the LB plate;

[0040] (4) Continue to purify the single bacteria isolated by streaking until a colony consisting of a single bacterial strain is formed on the plate.

[0041] 1.2 Molecular biological identification

[0042] (1) The Gram staining of the Bacillus veleisi AM6 strain was identified as Gram-positive; the mo...

Embodiment 2

[0046] Preparation of bacterial agent of Bacillus Velez AM6 strain

[0047] (1) Take a tube of preserved strains of Bacillus Velez AM6, inoculate it on an LB1 culture plate, and incubate in a biochemical incubator at 37°C for 12 hours. Aseptically transfer the bacterial colonies to LB1 liquid medium, shake and culture at 37°C for 12 hours to obtain the seed liquid, and the bacterial cells are short rod-shaped ( figure 2 ).

[0048] (2) The Bacillus Velez AM6 seed solution obtained in step (1) is inoculated in a large amount of LB1 liquid medium by 1% (volume ratio), and fermented and cultivated at 37°C, and the ventilation rate is 500~1000m 3 / h, tank pressure 0.04Mpa, rotation speed 80rpm, culture time 30h.

[0049] (3) after the fermentation is completed, the Bacillus velei AM6 fermented liquid is obtained, wherein the number of viable bacteria in the fermented liquid is measured by the plate counting method, and the result shows that the number of viable bacteria can be ...

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Abstract

The invention discloses bacillus velezensis AM6, a microbial inoculum, and a preparation method and application of the microbial inoculum. The bacillus velezensis AM6 is preserved in the China General Microbiological Culture Collection Center, and the preservation number is CGMCC No. 21320. The strain has low requirements on growth conditions, has wide environmental adaptability, and can be used as food with antibacterial activity, animal and aquatic product feed with antibacterial activity, and an antibacterial agent for plants and crops.

Description

technical field [0001] The invention belongs to the field of biological technology, and in particular relates to a bacillus veleisi AM6, a bacterial agent, a preparation method and an application thereof. Background technique [0002] Since the last century, the large-scale use of chemicals such as pesticides and antibiotics in agricultural production and livestock and poultry breeding has solved the problem of food sources for billions of people. However, the environmental impact of the application of these chemicals has been highlighted. In response to these current situations, various countries have also formulated a series of regulations to get rid of the dependence on these chemicals. For example, the European Union has recently set a series of targets including a 50% reduction in chemical agents and toxic pesticides to drastically reduce the use of agrochemicals in Europe in the next decade. Scientists have begun to develop biological solutions such as biotechnology ...

Claims

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Application Information

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IPC IPC(8): C12N1/20A01G7/06A23L3/3571C12R1/07
CPCC12N1/20A01G7/06A23L3/3571
Inventor 马安周尹珺庄国强
Owner RES CENT FOR ECO ENVIRONMENTAL SCI THE CHINESE ACAD OF SCI
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