Aspergillus niger capable of rapidly degrading zearalenone and application of aspergillus niger
A technology of zearalenone and Aspergillus niger, applied in the field of microorganisms, can solve the problems of long action time, unfavorable preservation of raw materials, unsatisfactory degradation effect, etc., and achieve the effects of high food safety, fast binding speed and high degradation efficiency
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Embodiment 1
[0025] Embodiment 1: the acquisition of mutant Aspergillus niger
[0026] Mutagenesis steps:
[0027] Mutagenesis method: use high-concentration zearalenone (ZEN) to stimulate Aspergillus niger FS-Z1 for 24 hours, and cycle 5 times, compare the degradation rate of each generation for screening, and select the one with the highest degradation rate as FS- 10-YJ.
[0028] 1. Prepare a sterile petri dish, pour PDA medium into the aseptic operating table (light the alcohol lamp in the table), the amount of the medium exceeds half of the capacity of the petri dish, and prepare 5 blank plates. After it solidifies, invert it.
[0029] 2. Take 0.25mL of the bacterial liquid (Aspergillus niger FS-Z1) in the freezing tube and put it on the PDA plate, spread it evenly with a spreader, and cultivate it in a 28°C incubator for 2-3 days.
[0030] 3. When Aspergillus niger grows on the plate, scrape the spores with a cotton swab, rinse them in sterilized normal saline, and make a suspensio...
Embodiment 2
[0039] Example 2: Degradation of ZEN by Aspergillus niger FS-10-YJ
[0040] Degrade 500ppb of ZEN toxin in PDB medium.
[0041] 1. Prepare 20mL of PDB medium in a fungal culture bottle, add ZEN toxin, and control the toxin content in the medium to 500ppb, and co-cultivate for 60h;
[0042] 2. Add the mutagenic Aspergillus niger spore suspension, and control the spore concentration in the medium to 10 3 、10 4 、10 5 、10 6 CFU / mL;
[0043] 3. Put the culture bottle into a 28°C, 180rpm fungal shaker for cultivation, take out the corresponding samples every 4 hours, extract with chloroform, and use high performance liquid chromatography to detect the toxin content.
[0044] High performance liquid chromatography conditions:
[0045] Type of HPLC: Agilent Technologies 1260infinity;
[0046] Chromatographic column: C18 column, column length: 150mm, inner diameter: 4.6mm, viscosity: 4μm;
[0047] Mobile phase: acetonitrile: water: methanol = 46:46:8;
[0048] Flow rate: 1.0mL...
Embodiment 3
[0057] Embodiment 3: the passage stability of Aspergillus niger FS-10-YJ
[0058]This mutagenesis Aspergillus niger is carried out PDA culture medium plate coating plate, subculture 5 generations, according to method in embodiment 2 (bacteria concentration is 10 6 CFU / mL, ZEN concentration is 1ppm), use Aspergillus niger FS-10-YJ to treat the ZEN solution for 36h, and measure the degradation rate after the reaction. It can be known from Table 2 that the degradation rate of the mutagenized Aspergillus niger can be stabilized above 95% after 5 generations, and the genetic stability is better.
[0059] The passage stability of table 2 Aspergillus niger FS-10-YJ
[0060] 1st generation 2nd generation 3rd generation 4th generation 5th generation ZEN degradation rate% 95.24% 96.84% 95.16% 96.84% 96.84%
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