Bacillus mangrove-2008, bacterial agent and its use and method in inhibiting the growth of Aspergillus niger
A technology for Bacillus and Aspergillus niger, applied in the field of inhibiting the growth of Aspergillus niger, Bacillus Mangrove-2008, can solve the problems of food safety threats, inconvenient use, drug residues, etc., to avoid adverse effects, improve antibacterial effect and Effects of antimicrobial efficiency, good ecological benefit and prospect of economic benefit
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Embodiment 1
[0040] The separation and acquisition of embodiment 1 bacillus Mangrove-2008
[0041] Select Shenzhen Baguang mangrove forest (N22°39′, E114°30′) as the collection point, take a small amount of collected sea mud homogenate, and perform conventional 10x gradient dilution in a sterile test tube to obtain a dilution of 10- 2 -10-7 6 gradient samples. from 10- 5 -10- 7 Take 200 μL sample dilutions from the three concentration gradient samples and spread them on the solid separation medium of LB, NA, potato medium PDA, and beef extract peptone medium ND, and observe the colonies after culturing at 37°C for 2-3 days. According to the difference in edge shape, color, and shape, pick typical colonies from the plate and inoculate them on the corresponding isolation medium using the three-section line method to obtain pure single strains. Finally, the isolated pure culture was inoculated into the corresponding slant medium, and stored in a refrigerator at 4°C for future use.
[0042...
Embodiment 2
[0043] Observation and identification of embodiment 2 bacillus Mangrove-2008
[0044] Inoculate the strain Mangrove-2008 on the LB medium, observe and record the state of the colony, and its colony morphology is as follows: figure 1 As shown, the streak culture results are as follows figure 2 shown. Measure the diameter of the colony, the presence of a sterile film, etc.; and perform spore staining and Gram staining on the Mangrove-2008 strain with chemical dyes, and then observe and record the results under an optical microscope. The colony characteristics are shown in Table 1, and the spore staining is as follows image 3 shown.
[0045] The 16S DNA was extracted by conventional methods and amplified by PCR, and the products were sequenced by Hunan Qingke Biotechnology Co., Ltd. The sequencing results were used to construct a phylogenetic tree with MEGA 6.0, such as Figure 4 shown.
[0046] Combining the physiological, biochemical and microbiological detection result...
Embodiment 3
[0049] The culture condition optimization of embodiment 3 bacillus Mangrove-2008
[0050] 1. Screening of strain basal medium
[0051] Five culture media were selected for screening: LB medium, PDA medium, YEB medium, NA medium and broth agar medium. Pick mature strains, inoculate them in the basic liquid medium, and cultivate them at 200r / min, 37°C for 12 hours to make seed liquids, and take 3% of the seed liquids and inoculate them in different mediums, 200r / min, 37°C Constant temperature and shaking culture for 12 hours, observe and compare the results, the concentration of the bacterial solution is measured by the spectrophotometer OD 600 Determination, such as Figure 5 As shown, the results showed that Bacillus Mangrove-2008 had the best growth effect on NA medium, so NA medium was selected as the basic medium for subsequent research.
[0052] 2. Determination of strain growth curve
[0053] Pick mature strains, inoculate them in the basic liquid medium, culture at 2...
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