Shinella oryzae Z-25 for producing mycotoxin zearalenone degrading enzyme, and application thereof
A technology of zearalenone and Shinella, applied in the field of microorganisms, can solve the problems of residual toxic substances, high cost, loss of nutrients, etc.
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[0026] In another aspect, the present disclosure also provides a method for preparing zearalenone-degrading enzyme, wherein the preparation method includes: inoculating the Shenbacillus oryzae Z-25 described in the first aspect into the medium for culturing , to obtain cultures.
[0027] Among them, the medium and culture conditions can be selected from various known suitable ones. The cultivated material contains the protein of zearalenone degrading enzyme, so it has the activity of zearalenone degrading enzyme, and can be directly used as zearalenone degrading enzyme composition according to needs, and can also be purified according to needs. The erythralenone degrades the protein with enzymatic activity before use.
[0028] Preferably, the medium is at least one of LB liquid medium and MSM medium; the culture conditions of the culture include: the culture temperature is 25-30°C, the culture time is 2-4 days, and the shaking speed is 140 -160rpm, transfer 1-3 rounds.
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Embodiment 1
[0035] This example is used to illustrate the preparation and identification process of Shinbacillus oryzae Z-25.
[0036]Weigh 1.0g sample (rice rhizosphere soil sample), add it to 100ml normal saline, resuspend and shake for 1h, then use 5% inoculum size, draw the sample suspension to 50mL basal salt culture containing 1μg / mL ZEN toxin culture medium (MineralSalt Medium, MSM) at 30°C and 180rpm for 7 days. Subsequently, the bacterial suspension was transferred to 50 mL MSM medium containing 10 μg / mL ZEN toxin at an inoculum amount of 5%. The transfer was repeated four times and the concentration of ZEN was continuously increased, so that the final concentrations of ZEN in the medium were 1 μg / mL, 10 μg / mL, 20 μg / mL and 40 μg / mL. Place it upside down in an incubator at 28°C for 3 days to obtain isolated strains.
[0037] The components of MSM medium include: K 2 HPO 4 2.0g, KH 2 PO 4 1g, MgSO 4 ·7H 2 O 0.5g, NaNO 3 0.5g, (NH 4 ) 2 SO 4 0.5 g, 1 mL filter-steri...
Embodiment 2
[0042] This example is used to illustrate that Shinella oryzae Z-25 has zearalenone degradation activity.
[0043] Inoculate the Shennella Z-25 isolated in Example 1 into LB liquid medium, culture and activate it on a shaker at 220 rpm at 28°C for 1-2 days, and inoculate the bacteria liquid on corn red On the plate of inorganic salt medium with mycocetone as the sole carbon source. The concentration gradient of ZEN was set so that the final concentrations of ZEN in the inorganic salt medium were 5 μg / mL, 10 μg / mL, and 20 μg / mL, respectively. After culturing at 28°C for 72-96 hours, observe the morphology and growth of each strain, and record the ZEN tolerance of the strains.
[0044] The degradative activity of ZEN by the strain of Shinibacterium oryzae Z-25 was detected by HPLC, and the seed solution of Shenuntella Z-25 was inoculated into the MSM medium containing 40 μg / mL ZEN as the only carbon source according to the inoculum amount of 5%, 28 Cultivate in a shaker at 220...
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