Aspergillus niger strain and application thereof in ochratoxin A degradation

A technology for ochratoxin and mycotoxins, applied in the field of genetic engineering, can solve the problems of lack of biodegradation effect, poor degradation effect, long time, etc., and achieve the effect of good passage stability and good degradation effect.

Active Publication Date: 2021-12-03
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the Aspergillus niger strain currently used to degrade OTA has a poor degradation effect and takes a long time
For example, the Aspergillus niger W35 disclosed in the patent CN 107312719 B, when the pH value of its fermented liquid is 6.5, the 48h degradation rate is 56.0%, but its fermented liquid takes a long time to prepare
And the existing Aspergillus niger can only have a single effect, that is, it can only act on the mycotoxin OTA alone, but in the actual application environment, it may be a mixed environment of various mycotoxins, such as: ochratoxin A, yellow Aspergillus toxin B1 (AFB1), zearalenone (ZEN) and deoxynivalenol (DON)
In addition, many studies only stop at the degradation in the medium, lacking the biodegradation effect in the actual sample

Method used

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  • Aspergillus niger strain and application thereof in ochratoxin A degradation
  • Aspergillus niger strain and application thereof in ochratoxin A degradation
  • Aspergillus niger strain and application thereof in ochratoxin A degradation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0061] Embodiment 1: the preparation of Aspergillus niger (Aspergillus niger) FS-UV-21

[0062] Specific steps are as follows:

[0063] Aspergillus niger FS-Z1 was mutated by ultraviolet irradiation and repeated 5 times. 40 irradiated strains were selected, and the OTA degradation rate of each strain was compared. The strain with the highest degradation rate was selected for preservation and identification.

[0064] The specific mutagenesis steps are:

[0065] 1. Inoculate the parent Aspergillus niger screened from the fermented soy sauce into PDA solid medium, culture at 28°C for 120 hours, take 50mL of normal saline to wash the spores, oscillate evenly, form a single spore suspension, and make 1×10 6 CFU / mL bacterial suspension.

[0066] 2. Take 5 mL of the above-mentioned spore suspension and irradiate it with 200W ultraviolet light for 30 minutes, repeating 5 times.

[0067] 3. Pick 40 strains after mutagenesis, numbered 1-40 respectively, inoculate these 40 strains on ...

Embodiment 2

[0071] Embodiment 2: the dynamic change of Aspergillus niger (Aspergillus niger) FS-UV-21 degradation OTA

[0072] Specific steps are as follows:

[0073] 1. Inoculate Aspergillus niger FS-UV-21 and Aspergillus niger FS-Z1 into PDA solid medium respectively, after culturing at 28°C for 120h, take 50mL of normal saline containing 0.02% (v / v) Tween 80 and wash the spores , oscillate evenly to form a single spore suspension, the concentration of the spore suspension at this time is: 1×10 6 CFU / mL.

[0074] 2. Take the above-mentioned spore suspension respectively, inoculate it into 50mL PDB medium containing 1ppm OTA according to the inoculum amount of 2% (v / v), cultivate it on a shaking table at 28°C and 180rpm for 72h, and extract corresponding samples to detect OTA every 6h Concentration changes, the results are shown in Table 1 and image 3 shown.

[0075] Table 1: Degradation rate (%) of OTA with different reaction times

[0076]

[0077] Depend on image 3 It can b...

Embodiment 3

[0078] Embodiment 3: the passage stability of Aspergillus niger FS-UV-21

[0079] Specific steps are as follows:

[0080] 1. Inoculate Aspergillus niger FS-UV-21 into PDA solid medium and pass for 5 generations. Method: After culturing at 28°C for 7 days, it becomes the first generation, and transfers the first-generation strain to a new PDA solid medium, 28 After culturing at ℃ for 7 days, it was the second generation, and the operation was repeated to the fifth generation, and the degradation effects were detected respectively.

[0081] 2, according to the method in embodiment 2 (bacterial concentration is 10 6CFU / mL, OTA concentration is 1ppm), utilizes Aspergillus niger FS-UV-21 to process the solution containing OTA 48h, detects the degradation rate of Aspergillus niger FS-UV-21 after the determination reaction of OTA after each passage respectively.

[0082] Depend on Figure 4 It can be seen that the degradation rate of the mutagenized Aspergillus niger can be stabil...

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Abstract

The invention discloses an aspergillus niger strain and application thereof in ochratoxin A degradation, and belongs to the field of gene engineering. The invention provides the Aspergillus niger FS-UV-21 strain which is preserved in the China General Microbiological Culture Collection Center on November 9, 2020, and the preservation number of the Aspergillus niger FS-UV-21 strain is CGMCC No. 20751. The aspergillus niger FS-UV-21 has a good degradation effect on ochratoxin A, the degradation rate reaches 74.49%, and the degradation rate is improved by 18.23% compared with that of an original strain; the time for achieving the optimal degradation effect is advanced and is shortened by 18 hours; aspergillus niger belongs to food-grade microorganisms, has relatively high food safety, and can realize efficient biological degradation of fungaltoxin.

Description

technical field [0001] The invention relates to an Aspergillus niger strain and its application in the degradation of ochratoxin A, belonging to the field of genetic engineering. Background technique [0002] Mycotoxins are secondary metabolites produced by fungi and widely exist in agricultural products and their products. Due to their serious harm to humans and animals, mycotoxin contamination has become one of the major global food-borne risks to human health. There are more than 400 kinds of mycotoxins that have been discovered by scientists. Although they all exhibit different physiological and biochemical properties, they are all toxic, carcinogenic, and mutagenic. There is a certain relationship. At present, the most common mycotoxins include ochratoxin, aflatoxin, zearalenol, and vomitoxin, etc. These mycotoxins are the most harmful mycotoxins found so far to grain, feed, animals and humans. Ochratoxins, especially ochratoxin A (OTA), are considered to be among the...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/14C12N15/01C12N13/00A62D3/02A23K10/12A23L5/20C12R1/685A62D101/28
CPCC12N15/01C12N13/00C12N1/14A62D3/02A23K10/12A23L5/28A62D2101/28Y02P60/87
Inventor 孙秀兰邹东纪剑孙嘉笛叶永丽张银志
Owner JIANGNAN UNIV
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