Microbial flora capable of rapidly degrading estrogen and analogs in chicken manure
A microbial flora and rapid degradation technology, applied in the field of microbial engineering, can solve problems such as unclear and complete metabolic pathways, and achieve the effect of improving environmental adaptability and reducing risks to ecology and human health.
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Embodiment 1
[0041] Embodiment 1, the cultivation of bacterial classification and the preparation of flora
[0042] Comamonas testosteroni ATCC11996, nine strains of KF1-Fe, Ca3, Cb7, M6, M2, A3, H5 and S19-1, respectively take 200 μL bacterial liquid from the mother liquid of each strain, add 2 mL LB medium. Among them, 40 μg / mL kanamycin sulfate was added to the LB medium of Ca3, Cb7, M6, M2 and A3; 2% NaCl was added to the LB medium of H5 and S19-1. The above-mentioned nine bacterial solutions were cultivated overnight at 27° C. on a shaker at 180 r / min. Use LB medium to adjust the OD of the above nine cultured bacterial solutions 600nm =1. Take the same volume of bacteria solution and mix them to get the composite bacteria group.
Embodiment 2
[0043] Embodiment 2, test of the response characteristics of estrogen in chicken manure degraded by compound flora:
[0044] (1) Degradation of chicken manure in the flora:
[0045] The composite bacterial solution and LB medium were mixed at a volume ratio of 0:1; 1:99; 1:9; 1:0 to prepare a composite bacterial solution with OD values of 0; 0.01; 0.1; 1.
[0046] Take 1 g of chicken manure and add it to the composite bacterial solution with OD values of 0; 0.01; 0.1; 1, and artificially add 0.5 mM estradiol solution, and culture at 27 °C and 180 r / min for 10 days.
[0047] (2) Extraction of estrogen and analogues in chicken manure:
[0048] Since estrogen and its analogues are easily soluble in chloroform, we use chloroform extraction to extract estrogen and its analogues from chicken manure or chicken manure degraded by microbial flora. That is, add 4 mL of sterile water and 1 mL of chloroform respectively, transfer to a centrifuge tube, and shake for 10 minutes. Cent...
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