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.
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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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