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Application of antibacterial lipopeptide fengycin in inhibiting the growth and toxin synthesis of Aspergillus flavus

An antimicrobial lipopeptide and Aspergillus flavus technology, applied in the field of bioengineering, can solve the problem that there are no marine microorganisms to inhibit Aspergillus flavus, and achieve the effect of inhibiting the biosynthesis of aflatoxin and inhibiting spore germination

Active Publication Date: 2021-12-28
OCEAN UNIV OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are many studies on antibacterial marine microorganisms and antibacterial marine active substances at home and abroad, mainly distributed in Alteromonas sp., Arthrobacter sp., Bacillus sp., Flavobacterium sp. Flavobacterium sp.), Micrococcus sp., Pseudomonas sp., Pseudoalteromonas sp., Staphylococcus sp., these marine microorganisms or their metabolism The product has a good inhibitory effect on some pathogenic bacteria and some plant pathogenic fungi, but there are few reports on marine microorganisms as the source of anti-Aspergillus flavus activity, and there are no reports on marine microorganisms inhibiting the growth of Aspergillus flavus and the synthesis of toxins

Method used

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  • Application of antibacterial lipopeptide fengycin in inhibiting the growth and toxin synthesis of Aspergillus flavus
  • Application of antibacterial lipopeptide fengycin in inhibiting the growth and toxin synthesis of Aspergillus flavus
  • Application of antibacterial lipopeptide fengycin in inhibiting the growth and toxin synthesis of Aspergillus flavus

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Isolation of marine microorganisms

[0031] The retrieved marine samples were treated as follows: seaweed, seaweed and other marine organisms were fully washed with sterile normal saline, then cut into pieces with sterile equipment and then diluted; marine fish were first fully washed with sterile normal saline, and then washed with sterile Dissect the sterilized utensils, take their intestinal tract and body surface respectively, add 50mL of sterile seawater to each, vortex shaker for 1min, let it stand until clarified, take 3mL of the supernatant and dilute it in 30mL of sterile normal saline ; Sediment was diluted with sterile normal saline at a ratio of 1:10 (V / V) to make a suspension; seawater was directly diluted with sterile normal saline at a ratio of 1:10 (V / V). Pipette 100 μL of the diluted fish intestine, fish body, sediment, seawater, and seaweed 5 groups of sample solutions and spread them on Gaoshi No. 1, beef extract peptone, MRS, and PDA medium plates. ...

Embodiment 2

[0046] Culture of marine Bacillus megaterium

[0047] Pick a ring from the fresh Bacillus megaterium slant and inoculate it into the seed medium (100mL / 250mL), culture it on a shaker at 160r / min at 36°C for 18-22h, and then insert the inoculation amount into the foundation Fermentation medium (100mL / 250mL) was shaken for 72 hours under the same culture conditions as above.

[0048] Among them, regarding the fermentation medium: the Plackett-Burman design method was used to screen the main influencing factors affecting the target product, and it was found that glucose (P=0.0004) had a positive effect on the production of antimicrobial lipopeptides, with a large degree of influence, and it was the most important factor in the fermentation medium. sole carbon source. However, due to the excessive addition of glucose, it is easy to cause the fermented product to be too viscous, which is not conducive to the separation and purification of bacterial cells and antibacterial substanc...

Embodiment 3

[0053] Purification of lipopeptides (two-step ultrafiltration method)

[0054] The lipopeptide aqueous solution was passed through a 0.22 μm cellulose acetate filter membrane (PALL Gelman Laboratory, New York, USA) to remove tiny impurities, and a polyethersulfone ultrafiltration membrane separation device with a molecular weight cut-off of 30 kDa was selected to preliminarily purify the lipopeptide by a two-step ultrafiltration method antibiotic. Specifically:

[0055] In the first step, the ultrafiltration membrane separation device is used to intercept macromolecular substances such as lipopeptide micelles and macromolecular miscellaneous proteins with a molecular weight exceeding 30 kDa and collect the intercepted products. Gently remove the retentate with a 200 μL pipette, dissolve it with Milli-Q water and add a certain amount of chromatographic grade methanol to disperse the lipopeptide micelles into monomers, adjust the pH of the solution to 7.0 with 1.0M NaOH and coo...

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Abstract

The invention proposes the application of an antibacterial lipopeptide Fengycin in inhibiting the growth of Aspergillus flavus and the synthesis of toxin, which belongs to the technical field of bioengineering. The antibacterial lipopeptide Fengycin provided by the present invention is a mixture of antibacterial lipopeptide Fengycin A and antibacterial lipopeptide Fengycin B. The antibacterial lipopeptide Fengycin is derived from the fermentation extract of marine Bacillus megaterium. It has been proved by experiments that the antibacterial lipopeptide Fengycin It has obvious inhibitory effect on Aspergillus flavus hyphae elongation, spore germination and aflatoxin biosynthesis. In addition, the present invention also optimizes the fermentation medium of Bacillus megaterium, and the antimicrobial lipopeptide content produced by the optimized fermentation medium is (571.26±9.72) mg / L, which is 1.82 times of the output before optimization , laying the foundation for the large-scale production of antibacterial lipopeptides that inhibit Aspergillus flavus.

Description

technical field [0001] The invention belongs to the technical field of bioengineering, and in particular relates to the application of an antibacterial lipopeptide Fengycin in inhibiting the growth of Aspergillus flavus and the synthesis of toxins. Background technique [0002] Aflatoxin is a kind of furanoxinone derived from polyketide, which belongs to the secondary metabolite of Aspergillus. It has been confirmed that the strains that can produce aflatoxin are mainly Aspergillus flavus and Aspergillus parasiticus. A large number of experimental data show that aflatoxin can induce experimental liver cancer in humans and various animals, and is the strongest chemical carcinogen discovered so far. Aflatoxin removal methods can be divided into conventional methods and biological detoxification methods. Conventional methods include alkali treatment, oxidation treatment, adsorbent treatment, etc., but conventional methods are prone to nutrient loss and high cost; biological det...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K7/06C07K1/34C07K1/14C12P21/02A01N37/46A01P3/00A01P21/00C12R1/11
Inventor 孔青牟海津张芳马云晓
Owner OCEAN UNIV OF CHINA
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