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Application of synergy of mannosylerythritol lipid and ultrasound in inhibition of growth of drug-resistant bacteria

The technology of sugar erythritol and sugar alcohol lipid is applied in the application field of mannose erythritol lipid and ultrasonic synergy in inhibiting the growth of drug-resistant bacteria, which can solve the problems of difficult food safety control and achieve inhibition of biofilm formation. , Expand the scope of application, inhibit the effect of growth

Active Publication Date: 2021-02-12
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Recently, MRSA has been continuously detected in beef, pork, milk and other foods, which has become a difficult problem for food safety control

Method used

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  • Application of synergy of mannosylerythritol lipid and ultrasound in inhibition of growth of drug-resistant bacteria
  • Application of synergy of mannosylerythritol lipid and ultrasound in inhibition of growth of drug-resistant bacteria
  • Application of synergy of mannosylerythritol lipid and ultrasound in inhibition of growth of drug-resistant bacteria

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Preparation of Mannose Erythritol Lipid

[0034] Mannose erythritol lipid is produced by Pseudozyma aphidis DSM70725. The production and purification method of mannose erythritol lipid: first, inoculate Torulopsis aphids into the activation medium containing 3% yeast extract, 3% malt extract, 10% glucose and 5% peptone, and inoculate at 28°C and 180rpm for 36 hours. After activation, inoculate 1mL of bacterial culture into NaNO 3 3g / L, MgSO 4 ·7H 2 O 0.3g / L, KH 2 PO 4 Seed medium composed of 0.3g / L, yeast extract 1g / L, glucose 40g / L and distilled water. After 2 days of cultivation at 28°C and 180 rpm, the seed culture was centrifuged and washed twice with saline. The resulting cells were inoculated in 80 mL / L soybean oil containing MgSO 4 ·7H 2 O 0.3g / L, KH 2 PO 4 0.3g / L, NaNO 3 3g / L, yeast extract 1g / L in the fermentation medium. The fermentation process was carried out at 28° C. and 180 rpm for 7 days. After the fermentation, the same volume of ethyl...

Embodiment 2

[0037] Synergistic antibacterial effect of MEL-A and ultrasonic treatment

[0038] (1) Preparation of bacterial suspension

[0039] For MRSA: under sterile conditions, use an inoculation loop to pick 1-2 rings of bacteria preserved on the slant, inoculate them into 10 mL of LB medium, and culture them in a full-temperature shaker at a temperature of 37°C and a rotation speed of 180r / min After 12 hours, take out 1.8mL of bacterial liquid into two 2mL centrifuge tubes, centrifuge at 8000rpm and 4°C for 3min to collect the bacterial cells, wash and resuspend with sterilized normal saline, repeat the above centrifugation and washing operation 3 times, and finally Prepare bacterial suspension with sterile saline.

[0040] (2) Determination of synergistic antibacterial activity of mannose erythritol lipid and ultrasound

[0041] Prepare physiological saline containing 256 μg / mL MEL-A and without MEL-A, and sterilize. Take the bacterial suspension and inoculate it into a 50mL centri...

Embodiment 3

[0048] Synergistic Antibacterial Mechanism Detection of Mannose Erythritol Lipid and Ultrasound

[0049] Carry out according to the experimental operation of embodiment 2, experimental condition is ultrasonic power 200W, processing time 15min, get four groups of samples (control group; ultrasonic group; MEL-A group; MEL-A and ultrasonic group) in 2mL centrifuge tube.

[0050] (1) Intracellular protein and nucleic acid leakage detection

[0051] The samples were centrifuged for 3 min at 8000 rpm. The supernatant was passed through a 0.22 μm membrane, and the absorbance at 260 nm and 280 nm of the filtrate was measured by an ultraviolet spectrophotometer.

[0052] (2) Detection of cell membrane integrity

[0053] Wash the sample twice with pre-cooled phosphate buffer, resuspend the pellet in 100 μL of buffer, then add 10 μL of PI dye, mix well, react at room temperature in the dark for 10-15 minutes, finally add 400 μL of buffer, and place on ice On, detected by flow cytometr...

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Abstract

The invention discloses an application of synergy of mannosylerythritol lipid and ultrasound in inhibition of growth of drug-resistant bacteria, and belongs to the technical field of food safety. Theapplication comprises the steps: mixing a sample to be sterilized with mannosylerythritol lipid to prepare an ultrasonic solution system, and carrying out ultrasonic treatment. Quantitative characterization shows that MEL-A and ultrasound have a synergistic effect which is remarkably superior to that of a single effect, and the method can effectively inhibit the growth of drug-resistant bacteria and planktonic bacteria and can remarkably inhibit the formation of a biological film of the drug-resistant bacteria. In combination with good emulsibility, surface activity, moisture retention and degradability of MEL-A, the MEL-A can be developed into antibacterial products such as natural food preservatives, food antibacterial films and the like, and has potential market value.

Description

technical field [0001] The invention relates to the technical field of food safety, in particular to the application of synergy of mannose erythritol lipid and ultrasound in inhibiting the growth of drug-resistant bacteria. Background technique [0002] Surfactants are a class of amphiphilic molecules, usually composed of lipophilic groups (such as hydrocarbon chains or their substitutes, etc.) and hydrophilic groups (such as sugar alcohols, etc.). Biosurfactants can be divided into five categories according to their structure: glycolipids, lipopeptides and lipoproteins, phospholipids and fatty acids, polymeric surfactants and particulate surfactants. Glycolipids are the most important class of biosurfactants, and there are many varieties, such as sophorolipids, trehalolipids, rhamnolipids, and cellobiolipids. MannosylerythritoI lipids (MannosylerythritoI lipids, MELs) is a glycolipid biosurfactant. MELs not only have good emulsification, biodegradability, surface activity...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A23L3/3562A01N43/16A01P1/00A61L2/025
CPCA01N43/16A23L3/3562A61L2/025A61L2/0011A61L2202/21
Inventor 陈启和舒琴魏天予娄行行
Owner ZHEJIANG UNIV
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