Separating, culturing and application methods for Rhodococcus erythropolis strain used for degrading aflatoxin B1

A technology of Rhodococcus rhodochrous and aflatoxin is applied in the field of culturing Rhodococcus erythropolis, which can solve the problems of low toxin removal efficiency, low toxin degradation efficiency, complicated degradation operation process, etc., and achieves high detoxification activity. , the effect of solving the source of raw materials and simple process

Inactive Publication Date: 2015-07-01
COFCO NUTRITION & HEALTH RES INST +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Studies on lactic acid bacteria, bifidobacteria and yeast have shown that most bacteria remove toxins less than 50% efficiently
The research on detoxification of some other bacteria and fungi also found that the toxin degradation efficiency of most bacteria is not high, and there are many factors affecting the degradation efficiency, including action time, solution pH value, solution composition, number of bacteria, toxin concentration and Metal ions, etc., while the degradation operation process is complex and difficult to implement on a large scale

Method used

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  • Separating, culturing and application methods for Rhodococcus erythropolis strain used for degrading aflatoxin B1
  • Separating, culturing and application methods for Rhodococcus erythropolis strain used for degrading aflatoxin B1
  • Separating, culturing and application methods for Rhodococcus erythropolis strain used for degrading aflatoxin B1

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0060] Example 1 Preliminary screening and preliminary identification of Rhodococcus strains that degrade AFB1

[0061] Using moldy grains (a mixture of moldy peanuts, corn, rice and sorghum) as materials, the PBS gradient dilution separation method was used for primary screening using the primary screening medium with AFB1 as the only carbon source. The specific operation is:

[0062] Use PBS (0.01mol / L) to prepare AFB1 solution with a concentration of 3.00% as component B of the primary screening medium, filter and sterilize; Sodium dihydrogen 0.25%, dipotassium hydrogen phosphate 0.20%, magnesium sulfate 0.05%, sodium nitrate 0.30%, agar 1.50%; pH7.2; 121 ℃ autoclave for 20min) uniformly mixed according to the volume ratio of 1:9 to prepare the primary screening The medium was poured onto the plate, and the final concentration of AFB1 in the primary screening medium was 0.3%.

[0063] Use PBS (0.01mol / L) to prepare PBS moldy grain sample dilutions with different concentra...

Embodiment 2

[0066] Example 2 Re-screening and identification of Rhodococcus strains that degrade AFB1

[0067] Use PBS (0.01mol / L) to prepare AFB1 solution with a concentration of 3.00% as component B of the re-screening medium, filter and sterilize; , sodium chloride 0.50%; pH7.2; 121°C autoclave for 20 minutes) were uniformly mixed according to the volume ratio of 1:9 to prepare a re-screening liquid medium, and the final concentration of AFB1 in the medium was 0.3%.

[0068] The refrigerated colonies of 7 strains of Rhodococcus erythropolis strains NHRI-1, NHRI-2, NHRI-3, NHRI-4, NHRI-5, NHRI-6 and NHRI-7 obtained in Example 1 were respectively inoculated in 5.0 In mL of re-screened liquid medium, the re-screened liquid medium without adding bacteria was used as a blank control, placed in a constant temperature shaker, and cultured at 30°C and 100r / min for 72h.

[0069] After the cultivation, the concentration of AFB1 in the re-screening medium was detected by HPLC-MS, and the degrada...

Embodiment 3

[0081] Example 3 Preservation and Preservation of Rhodococcus Rhodococcus Strains Degrading AFB1

[0082] Inoculate the pure colonies of seven identified strains of Rhodococcus erythropolis into solid medium (composition: 1.00% peptone, 0.50% beef extract, 0.50% glucose, 0.50% sodium chloride, 1.50% agar; pH7.2- 7.4) Incubate on a plate for 72 hours in a 30°C incubator, and verify its purity by microscopy. Pick a single colony with a purity ≥ 99.5% and transfer it to a test tube for slant culture, then refrigerate for use, and store a backup copy of the strain in a glycerol tube, and store it at -80°C for low temperature storage.

[0083] In addition, the seven identified Rhodococcus strains NHRI-1, NHRI-2, NHRI-3, NHRI-4, NHRI-5, NHRI-6 and NHRI-7 were deposited on December 17, 2013 at General Microbiology Center (CGMCC), China Committee for Culture Collection of Microorganisms, the preservation numbers are CGMCC No.8590, CGMCC No.8591, CGMCC No.8592, CGMCC No.8593, CGMCC No...

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Abstract

The invention relates to separating, culturing and application methods for a Rhodococcus erythropolis strain used for degrading aflatoxin B1 (AFB1), specifically to a separating method for the Rhodococcus erythropolis strain used for degrading AFB1, a culturing method for the Rhodococcus erythropolis strain obtained by using the separating method and a method for applying the Rhodococcus erythropolis strain in degradation of AFB1. The separating method for the strain has high specificity and high efficiency and is simple to operate; and the obtained Rhodococcus erythropolis strain has the advantages of a wide source, simple preparation, mild reaction conditions, high detoxification activity, etc.

Description

technical field [0001] The invention relates to a method for isolating, cultivating and using strains of Rhodococcus erythropolis. Specifically, the present invention relates to a method for isolating a strain of Rhodococcus erythropolis that degrades aflatoxin B1 (AFB1), a method for cultivating the strain of Rhodococcus erythropolis obtained by the separation method, and using the strain of Rhodococcus erythropolis to degrade The method of AFB1. Background technique [0002] Mycotoxins are secondary metabolites produced during the growth of molds. Mycotoxin contamination has always been a major threat to the global food, feed and agricultural products industries, bringing huge economic losses to production every year. At present, the mycotoxins that pose the greatest threat to related industries include aflatoxin (Aflatoxin, AFT), trichothecenes, and zearalenone. Among them, as the most toxic of all mycotoxins, aflatoxin is a secondary metabolite produced by fungi such ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A23L1/015C12R1/01A23L5/20
CPCC12N1/20C12N1/205C12R2001/01
Inventor 黄蔚霞蔡军倪媛媛刘佳佳李慧刘洋欧静堃
Owner COFCO NUTRITION & HEALTH RES INST
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