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Method for increasing yield of bacillus amyloliquefaciens antimicrobial lipopeptide fengycin
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A technology of amylolytic spores and antibacterial lipopeptides, which is applied in the field of increasing the output of Bacillus amyloliquefaciens antibacterial lipopeptide fengycin, can solve the problems that there are no relevant reports on the interaction of montmorillonite powder, and achieve the improvement of Fengycin production, wide application range, and highly reproducible effect
Pending Publication Date: 2022-03-22
HUAIYIN INSTITUTE OF TECHNOLOGY
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Problems solved by technology
However, there is no relevant report on the interaction of montmorillonite powder in the process of microbial metabolism.
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Embodiment 1
[0035] Streak the preserved Bacillus amyloliquefaciens CGMCC NO.18719 on the solid activation medium, pick out a single colony and connect it to the liquid activation medium. The activation culture conditions are 35° C., 180 rpm, and culture for 12 hours. When the OD600 is about 1.0 to be detected, the seed solution is ready for use.
[0038] (1) Weighing and UV sterilization of montmorillonite powder
[0039] Accurately weigh 0.05g, 0.1g, 0.5g and 1g of montmorillonite powder, and expose to ultraviolet light for 60min;
[0040] (2) Preparation of fermentation medium and high temperature sterilization
[0041] Accurately weigh 1g of beef extract, 14g of sodium glutamate, 0.5g of KCl, 2.5g of NaCl, KH 2 PO 4 1g, MgSO 4 0.5g, MnSO 4 5mg, CuSO 4 0.16mg, FeSO 4 ·7H 2 O 0.15mg, ZnSO4 0.12g, fructose 40g, glucose 25g, distilled water to 1L, pH=7.0, sterilized at 115°C for 30min;
[0042] In Example 1, the seed liquid was inoculated in the fermentation medium (100 mL) with an inoculum size of 5% by volume, and then accurately weighed 0.1 g of montmorillonite powder and added to the above-mentioned fermentation liquid. The culture bottle of montmorillonite powder was used as blank control. After the addition was completed, the culture flask was incubat...
Embodiment 3
[0044] Separation and Content Detection of Fengycin
[0046] Divide the fermented liquid of Example 2 into 500ml centrifuge cups, centrifuge in a low-temperature refrigerated centrifuge at 4°C, 5,000×g for 20min; discard the precipitated mixture of bacteria and montmorillonite powder to obtain a cell-free supernatant;
[0047] (2) Preparation of Fengycin Crude Extract
[0048] Adjust the pH value of the cell-free supernatant obtained in (1) to 2.0 with 6 mol / L HCl, and place it in the refrigerator overnight. Centrifuge the acid-precipitated system again, centrifuge at 5,000×g for 20 minutes at 4°C, discard the supernatant, add distilled water and stir at low temperature to adjust the pH value to 7.0, lyophilize into powder and extract with methanol, and extract the methanol mixture at 45°C Prepare the lipopeptide crude extract dry powder in a rotary vacuum evaporator, and store it in the refrigerator for subsequent use;
[004...
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Abstract
The invention discloses a method for increasing the yield of antibacterial lipopeptide fengmycin of bacillus amyloliquefaciens, which is characterized in that the bacillus amyloliquefaciens is inoculated into a fermentation culture medium containing montmorillonitepowder for fermentation culture, so that the yield of the antibacterial lipopeptide fengmycin is increased. According to the method, bacillus amyloliquefaciens is taken as a strain, and montmorillonitepowder with a certain concentration is added into a fermentationsystem to improve medium transfer of the fermentation system, so that the yield of Fengycin can be remarkably increased. A powdery crude extract of the Fengycin is obtained through fermentation, concentration, extraction and freeze-drying, a weighing method and HPLC (High Performance Liquid Chromatography) detection result show that the yield of the Fengycin can be remarkably increased by montmorillonitepowder with a proper concentration, the maximum yield can reach 5.89 g / L, and the yield of the Fengycin is increased by 2.65 times compared with that of a control group without the montmorillonite powder. According to the method, the yield of fengycin can be remarkably increased, and the production cost is reduced.
Description
technical field [0001] The invention belongs to the technical field of industrial microorganisms, and in particular relates to a method for increasing the output of the antibacterial lipopeptide fengycin of Bacillus amyloliquefaciens. Background technique [0002] With the promotion of the concept of green health, environmental protection and sustainable development, and the increase of resistance of pathogenic bacteria caused by the abuse of antibiotics, antibiotic substitutes have become a research hotspot. Fengycin is a lipopeptide synthesized by Bacillus amyloliquefaciens through the non-ribosomal pathway and can strongly inhibit filamentous fungi. Because of its advantages of good stability, high safety and resistance to drugs, it is considered to be a new type of preservative, Antimicrobial drugs are also regarded as one of the future alternatives to antibiotics, and are expected to make up for the prevalent problem of antibiotic resistance today. The latest research ...
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