Compound bacteria capable of rapidly degrading 17-beta-estradiol as well as preparation method and application of compound bacteria

A technology of rapid degradation and compound bacteria, applied in the field of biomedicine, can solve the problems of late start of research, less research on microbial degradation characteristics and degradation mechanism of environmental estrogens, etc., and achieve the effect of high efficiency degradation, good synergistic effect, and high efficiency degradation performance.

Inactive Publication Date: 2017-12-29
NORTHEAST NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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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 re

Method used

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  • Compound bacteria capable of rapidly degrading 17-beta-estradiol as well as preparation method and application of compound bacteria
  • Compound bacteria capable of rapidly degrading 17-beta-estradiol as well as preparation method and application of compound bacteria
  • Compound bacteria capable of rapidly degrading 17-beta-estradiol as well as preparation method and application of compound bacteria

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 compound bacteria capable of rapidly degrading 17-beta-estradiol as well as a preparation method and application of the compound bacteria. The compound bacteria contains rhodococcus rhodococcus DSH with a preservation number of CGMCC No.12392 and comamonas testosterone QYY with a preservation number of CGMCC No.15223, wherein the sequences of rhodococcus rhodococcus DSH and comamonas testosterone QYY are respectively presented by SEQ ID NO:1 and SEQ ID NO:2. The preparation method comprises the steps of respectively inoculating the strain DSH and the strain QYY into an inorganic salt culture medium, carrying out culturing so as to prepare DSH bacteria liquid and QYY bacteria liquid, and mixing the DSH bacteria liquid with the QYY bacteria liquid in a volume ratio of 1.5 to 1, so as to obtain the compound bacteria capable of rapidly degrading 17-beta-estradiol. The provided compound bacteria can rapidly degrade 17-beta-estradiol and has strong adaptive capacity to external environments. The compound bacteria is continuously cultured in the inorganic salt culture medium utilizing 17-beta-estradiol as a unique carbon source for 72 hours, so that 17-beta-estradiol with a concentration lower than 50mg/L can be rapidly and efficiently degraded, and the degradation rate reaches above 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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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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