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Bacillus amyloliquefaciens MBRC1 strain and application thereof

A technology of bacterial strains and antibacterial activity, applied in the field of microorganisms, can solve the same or similar problems, and achieve the effects of enhanced screening and cultivation, strong antibacterial activity, and improved accuracy and selectivity

Active Publication Date: 2015-05-13
绿川(浙江)环境科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

For example, Chinese patent application (publication number: CN103695360A) discloses a method and application of a marine Bacillus amyloliquefaciens 1A, which is a strain screened from marine microorganisms, but its main It is an antibacterial and disease-preventing application to fruit postpartum pathogenic fungi; said fruit postpartum pathogenic fungi are: apple anthracnose (Colletotrichum gloeos porioides), grape white rot (Coniella diplodiella), pear alternaria (Alternaria kikuchiana), Citrus Penicillium italicum (Penicllium italicum), but not related to the application in the preparation of antibacterial drugs, and there are many kinds of Bacillus amyloliquefaciens, the role of different Bacillus amyloliquefaciens is not necessarily the same or similar

Method used

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  • Bacillus amyloliquefaciens MBRC1 strain and application thereof
  • Bacillus amyloliquefaciens MBRC1 strain and application thereof
  • Bacillus amyloliquefaciens MBRC1 strain and application thereof

Examples

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Effect test

Embodiment 1

[0020] Obtaining of the Bacillus amyloliquefaciens MBRC1 bacterial strain of the present invention

[0021] The sample seabed mud used in this example was collected from the seabed mud of Jianmen Port, Taizhou City, Zhejiang Province (28°1′57″N, 121°36′38″E).

[0022] The LB solid medium, LB liquid medium, fermentation medium I, fermentation medium II, fermentation medium III, fermentation medium IV and agar double-layer solid medium used in this example are all prepared from seawater;

[0023] The LB solid medium described in this embodiment specifically includes the weight ratio of the following components:

[0024] Yeast extract: 5.0g / L; Tryptone: 10.0g / L; Sodium chloride: 10.0g / L; Agar: 15.0g / L; PH 7.2~7.4.

[0025] The liquid seed culture medium (LB liquid culture medium) described in the present embodiment specifically comprises the weight ratio of following composition:

[0026] Yeast extract: 5.0g / L; Tryptone: 10.0g / L; Sodium chloride: 10.0g / L; PH 7.2~7.4.

[0027] Th...

Embodiment 2

[0042] Morphological, physiological and biochemical identification and phylogenetic analysis of the Bacillus amyloliquefaciens MBRC1 bacterial strain of the present invention:

[0043] The LB solid medium used in this example is prepared from seawater, which includes the weight ratio of the following components:

[0044] Yeast extract: 5.0g / L; Tryptone: 10.0g / L; Sodium chloride: 10.0g / L; Agar: 15.0g / L; PH 7.2~7.4.

[0045] Inoculate the Bacillus amyloliquefaciens MBRC1 strain obtained in the present invention on an LB solid medium plate, culture it upside down at 30°C for 24 hours, and observe its colony growth state.

[0046] Strain morphology and physiological and biochemical experiments can refer to industrial microbiological experimental biology (Zhuge Jian, Wang Zhengxiang, industrial microbiological experiment technology manual [M], China Light Industry Press, 1994) and Berger's bacterial identification manual (R.E. Buchanan, N.E. Gibbons, Berger Bacteria Identification...

Embodiment 3

[0055] Molecular biological research of Bacillus amyloliquefaciens MBRC1 strain of the present invention

[0056] The PCR amplification of the 16S rRNA of the Bacillus amyloliquefaciens MBRC1 strain of the present invention can be performed by conventional methods in the art. More specifically, the DNA extraction in the PCR amplification of the present invention adopts the bacterial genome extraction kit of Shanghai Saibaisheng Gene Technology Co., Ltd., and the specific extraction method is carried out according to the method of the kit, and the primers used in the amplification are universal primers. Synthesized by Shanghai Saibaisheng Gene Technology Co., Ltd., the primers are:

[0057] 5'-ACAAGCCCTGGAAACGGGGT-3';

[0058] 5'-CACCAGGAATTCCGATCT-3';

[0059] The conditions of the PCR amplification reaction were: pre-denaturation at 94°C for 4 min, cycle, denaturation at 94°C for 50 s, annealing at 55°C for 50 s, extension at 72°C for 2 min, 35 cycles, extension at 72°C for...

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Abstract

The invention relates to a bacillus amyloliquefaciens MBRC1 strain and application thereof and belongs to the technical field of micro-organisms. The strain is used for solving the difficult problem that a novel strain with antibacterial activity is screened from terrigenous microorganisms. The invention provides the bacillus amyloliquefaciens MBRC1 strain and application thereof. The strain is collected in China Center for Type Culture Collection (CCTCC), has a collection number of CCTCC NO: M2014015, has antibacterial activity and can be used for preparing antibacterial drugs and food preservatives. The strain is a novel strain screened from the seabed slurry, belongs to bacillus amyloliquefaciens, has stronger antibacterial activity and especially has stronger antibacterial effect on bacillus subtilis, escherichia coli and staphylococcus aureus, the difficult problem of screening from the terrigenous microorganisms is solved and a new way for the research on the antibacterial drugs is opened.

Description

technical field [0001] The invention relates to a Bacillus amyloliquefaciens MBRC1 bacterial strain and an application thereof, belonging to the technical field of microorganisms. Background technique [0002] Antibacterial drugs are commonly used and important drugs in clinical practice. With the long-term use of antibacterial drugs, environmental bacteria are constantly developing resistance to them. Therefore, it is required to continuously develop new antibacterial drugs to deal with this situation. Most of the antibacterial compounds currently used clinically are primary and secondary metabolites produced by terrestrial soil microorganisms. They are of various types and have different functions, and have made considerable contributions to the development of the health of human society. After long-term research and development, it is difficult to obtain new terrestrial antibacterial active substances at this stage, which is also a major problem in the screening of terres...

Claims

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Application Information

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IPC IPC(8): C12N1/20A01N63/00A01P5/00A01P1/00A01P3/00A61K35/742A61P31/04A23L3/3571C12R1/07
CPCA01N63/00A23L3/3571C12N1/205C12R2001/07Y02A50/30
Inventor 陈建军郝之奎吴翰桂廖祥儒项彬彬王思思
Owner 绿川(浙江)环境科技有限公司
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