Use of bacillus subtilis in degradation of deoxynivalenol

A technology of deoxynivalenol and Bacillus subtilis, which is applied in the directions of application, medical preparations containing active ingredients, bacteria, etc., to achieve the effect of avoiding incomplete degradation

Inactive Publication Date: 2009-04-22
NANCHANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, there are few research reports on the degradati

Method used

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  • Use of bacillus subtilis in degradation of deoxynivalenol
  • Use of bacillus subtilis in degradation of deoxynivalenol
  • Use of bacillus subtilis in degradation of deoxynivalenol

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Experimental program
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Embodiment 1

[0020] 1. Connect the strains to be tested from the preserved freeze-dried tubes to the plates for activation twice, incubate at 37°C for 18 hours and take them out for use;

[0021] 2. Insert the activated strain to be tested into MRS or nutrient broth (NB) liquid medium, and simulate it into a food contamination model of 1 μg / mL according to the literature. Cultivate at 37°C in an anaerobic or 180rpm shaker for 12 hours, centrifuge the culture solution at 3000rpm for 15 minutes, and use the supernatant for later use;

[0022] 3. Dilute 8 times with ultra-pure water adjusted to be slightly alkaline in advance, and confirm that its pH value is neutral;

[0023] 4. Use the indirect competitive ELISA kit to detect the culture fluid of all strains to be tested;

[0024] 5. Experimental results such as figure 1 Shown: the abscissa is the test strain number (No. 1 is Bacillus subtilis ZZ; No. 2 is Bacillus subtilis KM; No. 3 is Bacillus licheniformis; No. 4 is Lactobacillus rhamn...

Embodiment 2

[0026] 1. Bacillus was cultured at 37°C with shaking at 180rpm for 12h, and centrifuged at 4000rpm for 10min to separate the supernatant and bacteria.

[0027] 2. The supernatant was filtered with a 0.22 μm microfiltration membrane, and the filtrate was collected;

[0028] 3. The cells were washed twice with normal saline to remove the remaining supernatant, and finally suspended with normal saline so that the cell concentration was 10 10 CFU / mL;

[0029] 4. Take 980 μL of the filter-sterilized supernatant and bacterial suspension, and add 20 μL of 50 μg / mL deoxynivalenol aqueous solution, so that the final concentration of deoxynivalenol is 1 μg / mL, 37 Incubate at ℃ for 3h and 14h for sampling;

[0030] 5. Use an indirect competitive ELISA kit to detect the residual concentration of deoxynivalenol in the sample;

[0031] 6. The experimental results are as follows: figure 2 Shown: the abscissa represents the test strain number (No. 1 is Bacillus subtilis ZZ; No. 2 is Baci...

Embodiment 3

[0033] 1. collect the supernatant of Bacillus subtilis by the method in embodiment 2 for subsequent use;

[0034] 2. Heat the supernatant at 65, 75, 85, 95, and 100°C for 10 minutes, and then add deoxynivalenol (final concentration: 1000ng / mL) for co-cultivation for 3 hours;

[0035] 3. All samples were tested for the residual concentration of deoxynivalenol by indirect competitive ELISA;

[0036] 4. The experimental results are as follows: image 3 Shown: the abscissa represents the heat treatment temperature (° C.), and the ordinate represents the residual concentration of deoxynivalenol in the test sample (ng / mL). ND means that the residual concentration of deoxynivalenol in the sample was not detected. from image 3 It can be seen that the effective degradation components in the supernatant gradually lose their activity with the increase of heat treatment temperature, and the activity loss ratio is larger at 65-75°C.

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Abstract

The invention relates to application of a bacillus subtilis in the degradation of mycotoxins deoxynivalenol enol. The application method is as follows: 1. a method for eliminating the interference of a liquid medium to an indirect competitive ELISA which detects the deoxynivalenol enol; 2. a method for preparing an enzyme preparation from a strain cultured product; 3. a method for preparing a living bacteria preparation; and 4. a method for preparing an oral preparation or a drug for human body or animal body detoxification. The invention has the advantages that the bacillus subtilis can completely degrade the polluted mycotoxin deoxynivalenol enol in food or feed through biosynthesis and zymological approach under a moderate condition, and can avoid the disadvantages of incomplete degradation and destruction of nutrient caused by using a physical or a chemical method to process mycotoxin.

Description

technical field [0001] The invention relates to an application of a bacillus subtilis, in particular to an application of a bacillus subtilis in degrading the mycotoxin deoxynivalenol. Background technique [0002] As people pay more attention to food safety, many countries and regions have adopted a variety of production quality management practices, such as Good Agricultural Practices (Good Agricultural Practices, GAP), Good Manufacturing Practices (Good Manufacturing Practice, GMP) and hazard analysis and Hazard Analysis and Critical Control Point (HACCP), etc., to ensure the food safety supply chain system from "field to table". In 2002 and 2006, my country's Ministry of Science and Technology launched the "Tenth Five-Year Plan" and "Eleventh Five-Year Plan" national science and technology major projects "Key Technologies for Food Safety", emphasizing the construction and strengthening of my country's food safety support system. As a major hidden danger affecting food s...

Claims

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Application Information

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IPC IPC(8): C12N1/20A62D3/02A23K1/16A61K35/74A61P39/02C12R1/125A62D101/28A23K10/16A61K35/742
Inventor 魏华程波财万翠香曾明徐锋许恒毅郭亮扬史良
Owner NANCHANG UNIV
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