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A kind of compound bacteria capable of rapidly degrading 17β-estradiol and its preparation method and application

A rapid degradation and estradiol technology, applied in the field of biomedicine, can solve the problems of late start of research, less research on environmental estrogen microbial degradation characteristics and degradation mechanism, etc., to achieve high-efficiency degradation performance, good synergistic effect, and adapt to the external environment powerful effect

Inactive Publication Date: 2021-04-30
NORTHEAST NORMAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Regarding the topic of using biological methods to remove estrogen from the environment, the domestic research started relatively late, and the relevant researches also mainly focused on the toxicology research of estrogen, the exploration of detection methods and the risk assessment of estrogen pollution. There are few studies on the microbial degradation characteristics and degradation mechanism of the

Method used

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  • A kind of compound bacteria capable of rapidly degrading 17β-estradiol and its preparation method and application
  • A kind of compound bacteria capable of rapidly degrading 17β-estradiol and its preparation method and application
  • A kind of compound bacteria capable of rapidly degrading 17β-estradiol and its preparation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1 Acquisition and preservation of bacteria degrading 17β-estradiol

[0032] Sludge samples from pharmaceutical factories and sewage treatment plants were collected as microbial sources, refrigerated and transported back to the laboratory. And the samples were cultured with only carbon source. Dissolve 15 mg of 17β-estradiol in 30 mL of methanol to prepare a 17β-estradiol mother solution with a concentration of 500 mg / L. Take 1mL of 17β-estradiol mother liquor and add it to the Erlenmeyer flask containing 100mL of inorganic salt medium, so that the final concentration of 17β-estradiol in the inorganic salt medium is 5mg / L (the composition of inorganic salt medium is g / L) : Na 2 HPO 4 4.260; KH 2 PO 4 2.650; MgSO 4 ·7H 2 O 0.200; (NH 4 ) 2 SO 4 1.500; CaCl 2 0.020. Adjust the pH to 7.0 with 0.1mol / L NaOH and 0.1mol / L HCl, and add 1mL of trace elements. Trace element composition (g / L): NiCl 2 ·6H 2 O 0.024; CoCl 2 ·6H 2 O 0.190; H 3 BO 3 0.006; Z...

Embodiment 2

[0035] Example 2 Identification of bacteria degrading 17β-estradiol

[0036] The DNA was amplified by PCR amplification technique using universal primers for bacteria. The sequencing results were searched for homology comparison with the 16S rDNA gene sequence in the GenBank database using the BLAST tool in NCBI. The comparison results show that the 16S rDNA gene sequence of Rhodococcus DSH has high homology with the 16S rDNA gene sequences of multiple strains of Rhodococcus and Rhodococcus equi in the genus Rhodococcus (Rhodococcus), combined with the morphology of Rhodococcus DSH Characteristics and physiological and biochemical indicators, strain DSH was preliminarily identified as Rhodococcus, named Rhodococcus (Rhodococcus sp.) DSH. The 16S rDNA gene sequence of strain QYY is close to the genetic distance of Comamonas testosteroni, combined with the morphological characteristics and physiological and biochemical indicators of strain QYY, it was identified as Comamonas te...

Embodiment 3

[0037] Embodiment 3 Composite bacteria are to the degradation ability of 17β-estradiol

[0038] Dilute the prepared composite bacterial solution to OD with phosphate buffer solution 600 When the value is 1.0, the bacterial suspension is prepared for use. Take out 200 μL of composite bacteria liquid and inoculate it into 17β-estradiol as the only carbon source, and inoculate it into inorganic salt medium with a concentration of 50 mg / L, and culture it continuously for 7 days in a constant temperature shaking incubator at 30°C and 120 rpm, and take samples once a day . After the inorganic salt medium was pretreated, the ability of the complex bacteria to degrade 17β-estradiol was detected by high-performance liquid chromatography (HPLC). The detector is UV (DualλAbsorbance Detector, Water2487), and the chromatographic column is Zorbax Eclipse Plus C18 column (150×4.6mm, 3.5mm). The mobile phase volume ratio is acetonitrile: water = 1:1, the detector wavelength is 275 nm, the ...

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Abstract

The invention discloses a compound bacterium capable of rapidly degrading 17β-estradiol and its preparation method and application. The compound bacterium consists of Rhodococcus DSH with a preservation number of CGMCC No. 12392 and testosterone with a preservation number of CGMCC No. 15223. The composition of Comamonas QYY, the 16S rDNA sequences of Rhodococcus DSH and Comamonas testosteroni QYY are respectively shown in SEQ ID NO: 1 and SEQ ID NO: 2 in the sequence listing. The strain DSH and the strain QYY were respectively inoculated into the inorganic salt medium to obtain the DSH bacterial liquid and the QYY bacterial liquid, and the volume ratio of the two bacterial liquids of DSH and QYY was 1.5:1, and the efficient degradation of 17β-estradiol was obtained. Complex bacteria. The composite bacterium provided by the invention has high-efficiency degradation performance on 17β-estradiol, and has strong ability to adapt to the external environment. Cultivate the complex bacteria in the inorganic salt medium with 17β-estradiol as the only carbon source for continuous culture for 72 hours, and can quickly and efficiently degrade 17β-estradiol with a concentration lower than 50mg / L, and the degradation rate can reach more than 98%. .

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to a compound bacterium capable of rapidly degrading 17β-estradiol and its preparation method and application. Background technique [0002] As a common endocrine disruptor, environmental hormones are similar in structure and function to estrogen secreted by the body under normal conditions. Once it enters the body, it can not only disrupt the normal synthesis and metabolism of endocrine hormones, affect growth and sex differentiation, but also cause developmental abnormalities and reproductive disorders in multiple systems such as nerves, endocrine, and immunity, and even cause teratogenicity and carcinogenesis. [0003] Environmental hormones can take advantage of ocean currents and atmospheric circulation to transfer from low-latitude regions to high-latitude regions, and even reach polar ecosystems. In addition, because environmental hormones can be enriched and amplified i...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C02F3/34B09C1/10A62D3/02C12R1/01A62D101/28
CPCA62D3/02A62D2101/28B09C1/10C02F3/34C12N1/20
Inventor 霍洪亮于清淼邵缓缓王平朱遂一杨武马喆
Owner NORTHEAST NORMAL UNIVERSITY
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